From Hurdles to Breakthroughs: Building and Validating a PSMA-positive Triple-Negative Breast Cancer Mouse Model for Preclinical Targeted Radionuclide Therapy

From Hurdles to Breakthroughs: Building and Validating a PSMA-positive Triple-Negative Breast Cancer Mouse Model for Preclinical Targeted Radionuclide Therapy | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article From Hurdles to Breakthroughs: Building and Validating a PSMA-positive Triple-Negative Breast Cancer Mouse Model for Preclinical Targeted Radionuclide Therapy Benjamin Chaussin, Lucie Sanchez, Sophie Levesque, Alban Revy, and 15 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7904335/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 16 Feb, 2026 Read the published version in Scientific Reports → Version 1 posted 12 You are reading this latest preprint version Abstract Background Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer, for which targeted therapies are emerging. Prostate-specific membrane antigen (PSMA), expressed mainly by the endothelial cells of the neovascularization of TNBC, or tumor cells, is an interesting target in TNBC that needs to be studied. This study aims to validate a reproductible, PSMA-positive murine model for Targeted Radionuclide Therapy (TRT) assessment. Methods Twenty-five syngeneic or xenograft murine models were produced by modifying the TNBC cell line, the implantation site (ectopic and orthotopic), cell numbers and matrix (e.g Matrigel®) use. Tumor growth and engraftment were recorded. PSMA expression was evaluated histologically using immunohistochemistry (IHC) and PSMA-targeted positron emission tomography (PET)- computed tomography (CT) with [ 18 F]-DCFPyL. The tumor-to-liver ratio (TLR) was used to quantify the uptake of the radiopharmaceutical. To address the absence of PSMA expression in native models, MDA-MB-231 cells were genetically modified via lentiviral transduction to overexpress PSMA for in vivo evaluation. Results Despite high cellular proliferation and extensive tumor neovascularization, immunohistochemical analyses revealed an absence of PSMA expression in all non-transfected models tested. This finding was further confirmed by [¹⁸F]-DCFPyL PET/CT imaging, which showed a TLR below 1, indicating negligible radiotracer uptake within tumoral tissues. In contrast, the MDA-MB-231 psma model achieved 100% tumor engraftment and a tumor volume of 208 ± 61 mm 3 at 28 days post-injection (not significantly different from the MDA-MB-231 wt model). This model showed a TLR of 10.3 ± 4.3 one hour after intravenous injection of [ 18 F]-DCFPyL. Conclusions This study emphasizes the challenge of creating a reproducible murine model with PSMA expression. Nevertheless, the initial transfection of MDA-MB-231 cells to express PSMA resulted in the development of an innovative TNBC murine model that expresses PSMA. This model is reproducible and exhibits heterogeneous PSMA expression, and is representative of clinical observations. It is also suitable for evaluating PSMA-targeted TRT in TNBC. Biological sciences/Cancer Health sciences/Oncology Targeted Radionuclide Therapy Triple-Negative Breast Cancer Prostate-Specific Membrane Antigen Syngeneic model Xenograft model Orthotopic model Lentiviral transduction Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 BACKGROUND Triple-negative breast cancer (TNBC) accounts for 15% of breast cancers and is considered as the most aggressive subtype, with the poorest prognosis. Approximatively 46% of TNBC patients are expected to develop metastases which are associated with high microvascular densities [ 1 , 2 ]. The median overall survival for metastatic TNBC (mTNBC) is 14,8 months [ 3 ]. Despite the development of antibody-drug conjugates which provide more therapeutic options beyond checkpoint and PARP inhibitors in the metastatic stage, the prognosis for metastatic triple-negative breast cancer (mTNBC) remains poor [ 4 , 5 ]. Consequently, research in this field is very active, seeking new therapeutic approaches that can either provide alternative options or be combined with existing treatments to improve outcomes [ 6 ]. The ongoing development of Targeted Radionuclide Therapy (TRT) is opening new avenues in oncology by providing additional therapeutics options. Notably, it represents an attractive strategy for addressing metastasis. In 2022, the European Medicines Agency approved the radiopharmaceutical lutetium ( 177 Lu) vipivotide tetraxetan which targets the Prostate Specific Membrane Antigen (PSMA) for therapy in metastatic castration-resistant prostate cancer (mCRPC) [ 7 ]. This approval has prompted the investigation of additional applications of PSMA-TRT in cancer beyond the prostate. Indeed, although PSMA was initially discovered and characterized in the prostate, it is also expressed in other solid cancers [ 8 , 9 ]. Therefore, expanding PSMA-targeted TRT to non-prostate malignancies that overexpress PSMA is of interest, particularly glioblastoma, renal cell carcinoma and breast cancer (BC) [ 10 – 15 ]. However, PSMA expression is generally weaker and more heterogeneous in non-prostate solid tumors than in prostate cancer. In particular, PSMA has been shown to be expressed in 66%-74% of primary BC and 75%-100% of distant metastases with the highest rates observed in TNBC [ 10 , 16 , 17 ]. Furthermore, immunohistochemistry (IHC) studies have demonstrated that PSMA is predominantly expressed by endothelial cells of tumor neovessels (46%-83%) and also by tumor cells (3%-72%), in BC [ 10 , 16 – 18 ]. PSMA overexpression has also been shown to be higher in neovessels from metastatic sites than in primary tumor neovascularization [ 17 ], supporting the use of PSMA-targeted TRT for mTNBC. Nevertheless, predominant localization in tumor neovascularization raises questions about therapeutic efficacy. Therefore, it is essential to evaluate the efficacy and relevance of PSMA-targeted TRT in mouse models representative of these tumors to demonstrate proof of concept for clinical trials. To the best of our knowledge, only two teams have successfully developed TNBC murine models with PSMA expression in preclinical studies. However, these models exhibited necrosis cores and heterogeneous PSMA expression [ 19 – 21 ]. Shirke et al. described the level of PSMA expression as insufficient for therapy efficacy. In contrast, Heesch et al. successfully performed [ 177 Lu]Lu-PSMA-I&T assessments in mice, highlighting the heterogeneity between models and even among mice within the same model. This study aimed to develop a preclinical TNBC murine model that can express PSMA and is highly reproducible and suitable for future PSMA-targeted TRT research. To achieve this goal, we evaluated 25 syngeneic and xenograft models (4T1, MDA-MB-231, MDA-MB-468, MDA-MB-436, BT-20), including co-inoculations with endothelial cells to enhance tumor neovascularization. We also evaluated a novel PSMA-positive model generated via lentiviral transduction of MDA-MB-231 cells. We assessed engraftment, tumor growth, PSMA expression and PSMA-targeted radiopharmaceuticals, such as [ 18 F]-DCFPyL uptake. METHODS Cell lines The murine TNBC cell line 4T1 (CRL-2539), and Human TNBC cell lines MDA-MB-231 (HTB-26), MDA-MB-468 (HTB-132), MDA-MB-436 (HTB-130) and BT-20 (HTB-19) cell lines were purchased from the American Type Culture Collection (ATCC, Table 1 ). The MDA-MB-436 and 4T1 cell lines were cultured in RPMI-1640 medium supplemented with 10% Fetal Bovine Serum (FBS, Dutscher, Bernolsheim, France) and 4 µg / mL gentamicin. The MDA-MB-231 wt , MDA-MB-468 and BT-20 cell lines were maintained with MEM-Glutamax supplemented with 10% FBS, 1mM of sodium pyruvate, 1X of Non-Essential Amino Acids, 1X of vitamins and 4 µg / mL of gentamicin (50 mg / mL). Except for FBS, all culture reagents were purchased from Gibco® (ThermoFisher Scientific, Strasbourg, France). The human primary endothelial cell line, HUVEC cells (c-12208, PromoCell GmbH, Heidelberg, Germany) was maintained in the Endothelial Cell Growth Medium 2 (ready-to-use) (Promocell, GmbH, Heidelberg, Germany). All cells were maintained at 37°C with 5% of CO 2 and were routinely tested for mycoplasma infection. PSMA overexpression and lentiviral transduction The PSMA and LacZ-V5 tags were synthesized by Vector Builder and cloned into a third-generation self-inactivating lentiviral expression vector (Vector Builder) under the eukaryotic translation elongation factor 1 α (EF1A) promoter: PSMA plasmid pLV[Exp]-Bsd-EF1A > hFOLH1[NM_004476.3]; LacZV5 plasmid pLV[Exp]-Bsd-EF1A>{LacZV5}. Lentiviruses were produced by triply transfecting HEK-293T cells with the lentiviral transfer vector and the packaging plasmids psPAX2 (a gift from Didier Trono; Addgene plasmid #12260; http://n2t.net/addgene:12260 ; RRID: Addgene_12260) and pMD2.G (a gift from Didier Trono; Addgene plasmid #12259; http://n2t.net/addgene:12259 ; RRID: Addgene_12259) at a ratio of 0.3:0.3:0.1. Transfection was performed using jetPEI (101000053, Polyplus). Viral supernatants were collected 48 hours post-transfection, filtered through a 0.22 µm filter, and used for transduction. MDA-MB-231 wt cells were transduced with lentiviral particles at a multiplicity of infection (MOI) of 1 for MDA-MB-231 wt in the presence of hexadimethrine bromide (8 µg/mL). MDA-MB-231 psma cells were cultured with the same culture medium as MDA-MB-231 wt , but with the addition of blasticidin (9 µg/mL, InvivoGen, ant-bl-05) for antibiotic selection. In vivo studies All experiments were approved by the Auvergne local Ethic committee C2E2A and the French Ministry of Education and Research (approval Nos. 49893 &. 23955) and were conducted in accordance with ARRIVE guidelines ( https://arriveguidelines.org ). Mice were housed under environmentally controlled conditions (4–5 animals per cage, ventilated racks, 60% relative humidity, temperature maintained at 21–24°C, and a 12 h/12 h light/dark cycle). Animals had free access to standard food and water. Tumor xenograft and sygeneic model were established by injecting tumor cells into anesthetized mice. Anesthesia was induced and maintained with gaseous isoflurane (Iso-Vet, 1.5%, 1 L/min, air/O₂ 70/30, v/v) using an animal anesthesia system (Minerve). For syngeneic models, 7–9 week old female BALB/cAnNRj mice from Janvier Labs (Le Genest-Saint-Isle, France) were inoculated with 4T1 cells at different concentrations (Table 2 ; Additional File. Table S1 ). For the xenograft models, 7–9 week old female NMRI-Foxn1nu/Fox1nu and female Balb/cAnNRj-Foxn1 nu/nu from Janviers Labs (Le Genest-Saint-Isle, France) were inoculated with human TNBC cell lines with or without the co-injection of different amount of HUVEC cells (Table 2 ). For both the syngeneic and xenograft models, subcutaneous injections were performed into the right flank with 100 µL of cells suspension. For orthotopic injection, the cells were injected into the third thoracic mammary fat pad (MFP) with a 100 µL cell suspension (Table 2 ) (see Additional File Table S1 ). The cells were resuspended in 100 µL of one of the following solutions: i) PBS (ThermoFisher Scientific, Strasbourg, France), ii) 1:1 volume ratio of PBS and Geltrex® (ThermoFisher Scientific, Strasbourg, France), iii) 1:1 volume ratio of PBS and Matrigel® (Corning®, New York, USA). Tumor volume and body weight were measured once a week for the xenograft model and twice a week for the syngeneic model by the same operator. At different time points (at least 2 mice) between 19 days and 81 days post inoculation (p.i.), we re-anesthetized the mice with isoflurane (5%), prior to confirmation of anesthesia with a tail pinch, and performance of cervical dislocation to sacrifice them. Metastases were examined in the liver and lungs. The tumors were split into two parts, one half oriented toward the periphery and the other oriented toward the center, before paraffin embedding and hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) for Ki-67, CD31, PSMA, and HIF-1α. Tumor volume (TV) was calculated using caliper measurements as TV (mm³) = [length × (width)²]/2 and doubling time (DT) as DT = Ln2/a , where “a” is the slope of the exponential growth phase. The maximum tumor size permitted by the local ethics committee, (1500 mm 3 ), was never reached by any mouse in this study. Nuclear PET imaging Positron emission tomography / computed tomography (PET/CT) scans were performed using the IRIS PET/CT (Inviscan) scanner. Acquisitions were performed for 15 minutes, one hour after intravenous (i.v.) injection of 7.50 ± 1.42 MBq of [ 18 F]-DCFPyL (Curium France, Saint-Beauzire, France). Then, the mice were anesthetized in an induction chamber containing 2.5% isoflurane in air and followed by 1.5% isoflurane in air to maintain anesthesia using an animal anesthesia machine (Minerve). The energy window used was 250–750 keV. Images were reconstructed according to the OSEM 3D algorithm including corrections for dead time, decay, and randoms. For in vivo biodistribution, images were analyzed using InterView™ Fusion Software (ver. 3.11.011). The volume of interest (VOI) of tumor, liver and muscle was delineated to calculate the tumor-to-liver ratio (TLR) and the tumor-to-muscle ratio (TMR) for each mouse. For ex vivo biodistribution, mice were anesthetized with isoflurane (5%) 1.5 hours p.i. Anesthesia was confirmed by a tail pinch, followed by cervical dislocation to euthanize the animals. The organs and tumors were then collected and weighed. Radioactivity was measured using a γ-counter (Wizard 1480, Perkin Elmer) and the gamma counting data were corrected for physical decay (109.8 minutes) and background. All uptake data were expressed as percent injected activity per gram (%IA/g). The experimental part of this study was performed on IVIA platform, Clermont-Ferrand, France ( https://doi.org/10.18145/ivia ). Immunohistochemistry and H&E staining protocol Immunohistochemistry and H&E staining protocol Five µm sections were cut and adhered to poly-L-Lysine-coated glass microscope slides. The paraffin was then removed with two 15 minutes baths of SafeSolv (#00699464, VWR, France). The tissues were then rehydrated with successive ethanol baths at decreasing concentrations (100%, 100%, 95%, 70%, H 2 O; five minutes each). For the H&E protocol, nuclei were stained by a four-minute bath of hematoxylin (#C0303, Diapath, France), then rinsed with a four-minute bath of tap water. The sections were stained by eosin Y bath for four minutes and rinsed in the same manner before dehydration and mounting (described below). For IHC, heating-mediated antigen retrieval was performed using either a citrate buffer (10 mM sodium citrate tribasic (Sigma #066K0042); 0.05% Tween 20; pH 6) for Ki-67, cleaved-caspase3 and HIF-1α staining or a Tris-EDTA buffer (10 mM Base Trizma® (Sigma #T1503); 0.25 mM EDTA (Invitrogen, #15575-038); 0.05% Tween 20; pH 9) for CD31 staining. This was performed for 30 minutes at 95°C for both. The slides were cooled for 20 minutes, then rinsed in PBS 1X for three five-minute intervals. The tissues were blocked for one hour at room temperature (RT) in PBS 1X-BSA (1%)-Avidin (Vector laboratories, SP-2001). After rinsing, endogenous peroxidases were quenched using 0.3% H 2 O 2 in PBS 1X for 30 minutes at RT and rinsed again. Tumor sections were then incubated overnight at 4°C with the primary antibody in the staining solution (PBS 1X-BSA (1%)-Biotin (Vector laboratories, SP-2001)). We used the following antibodies i) anti-CD31 antibody ( 1/2000 , Abcam # ab182981) ii) anti-Ki-67 antibody ( 1/100 , Abcam # ab15580) iii) anti-PSMA antibody ( 1/100 , Cell Signaling Technology #12815) iv) anti-HIF-1α ( 1/100 , Cell Signaling Technology #48085). After rinsing, the tissue sections were stained with a biotinylated goat anti-rabbit IgG antibody ( 1/500 , Invitrogen # 656140) for one hour at RT. After rinsing, amplification was performed using Vectastain ABC Reagent (Vector laboratories, PK-4000) with a 30-minute incubation at RT. Revelation was performed after rinsing with a DAB kit (Vector laboratories, SK-4105). Tissue sections were rinsed and counterstained with a two-minute bath of hematoxylin and washed for two minutes in tap water. For H&E and IHC protocols, the tissues were dehydrated using successive ethanol baths at increasing concentrations (70%, 95%, 100%, SafeSolv, two minutes each prior to mounting with Shandon-Mount (Epredia, #1900333). Histological images were acquired using an Axio Scan.Z1 slide scanner and the ZEISS Colibri 7 camera. Antibody controls for PSMA, CD31 and Ki-67 stainings are shown in Fig S6 (Additional File 1). Immunohistochemical staining was evaluated using the H-score method, which integrates both the intensity of staining and the percentage of stained cells. Staining intensity was categorized as follows: 0, no staining; 1, weak staining; 2, moderate staining and 3, strong staining. For PSMA, the H-score was calculated as a continuous variable (range 0–300) using the following formula: H-score = 1×(% cells with weak staining) + 2×(% cells with moderate staining) + 3×(% cells with strong staining) . Statistical Analysis Statistical analyses and graphical representations were performed using GraphPad Prism (version 10.3.1, GraphPad Software, San Diego, CA, USA). Variables were presented as mean with standard deviation. A Kruskal-Wallis test was performed to compare the doubling time of the MDA-MB-231 wt , MDA-MB-231 wt/ MDA-MB-231 psma and MDA-MB- 231psma mouse models. The difference between the groups was tested using the Dunnett's multiple comparison test, and a p value of 0.05 or less was considered statistically significant. RESULTS Site of inoculation, matrix and number of cells modulate tumor engraftment however PSMA remains unexpressed Different cell lines were tested and are summarized in Table 1 . Table 1 List of Triple-negative breast cancer (TNBC) and endothelial cell line for preclinical in vivo studies Type Cell line Disease Organism Source Mutation Endogenous PSMA expression (by Flow Cytometry) Triple-Negative Breast Cancer MDA-MB-231 Adenocarcinoma (pleural effusion) / Homo sapiens ATCC (HTB-26) CDKN2A/B, BRAF, KRAS, TERT, TP53 No MDA-MB-468 Adenocarcinoma (pleural effusion) Homo sapiens ATCC (HTB-132) TP53, PTEN, RB1 No MDA-MB-436 Adenocarcinoma (pleural effusion) Homo sapiens ATCC (HTB-130) BRCA1, FHIT, TP53, RB1 No BT-20 Carcinoma (primary tumor) Homo sapiens ATCC (HTB-19) CDKN2A, PIK3CA, RB1, TP53 No 4T1 Mimics human breast cancer, Stage IV Mus musculus ATCC (CRL-2539) TP53, PIK3G No Endothelial cells HUVEC Primary Human Umbilical Vein Endothelial Cells Homo sapiens Promocell (C-12208) / No The murine 4T1 cell line was tested for the syngeneic model, and the human TNBC cell lines for xenograft models. Primary human endothelial cell line HUVEC was tested for co-inoculation. Figure 1, summarizes the different strategies that were explored to improve tumor engraftment and PSMA expression in order to develop a preclinical TNBC murine model that expresses PSMA (Fig. 1). Figure 1: Flow Chart of Strategies Enhancing Engraftment, Growth, Vascularization and PSMA in TNBC Models. Murine models were developed by modifying the TNBC cell line, cell numbers, site of implantation, use of matrices and the co-inoculation with endothelial cells (HUVEC). For each model, PSMA expression was analyzed by immunohistochemistry. For few models, [ 18 F]-DCFPyL PET images was harvested to study radiotracer accumulation and confirmed PSMA expression. TNBC: Triple-negative breast cancer Prior to in vivo experiments, all cell lines were tested negative for PSMA expression by flow cytometry (Additional file 1: Fig. S1 ). Table 2 presents a curated selection of the most relevant murine models, highlighting key characteristics such as mouse strain, cell line, inoculation parameters, and tumor growth. The remaining 19 models are detailed in Supplementary Table S1 (Additional File 1). Table 2 Characterization of syngeneic and xenograft murine models for triple-negative breast cancer development *MFP: Mammary fat pad. TNBC: Triple-negative breast cancer. PBS: Phosphate buffer saline Female Balb/c mice inoculated with 10,000 4T1 cells in the MFP with PBS exhibited a tumor volume of 1,132 ± 89 mm 3 at 25–30 days post-inoculation (n = 12) (Fig. 2 A). Hematoxylin-eosin staining has revealed necrosis for each syngeneic model (5%-35% of the tumor surface between 13–25 days p.i.) (Fig. 2 B). Ki-67 IHC showed a high proliferation. However, despite strong CD31 staining indicating neovascularization in the syngeneic model, no PSMA expression was detected at the same location as CD31 staining (Fig. 2 B). Considering the tumor uptake of [¹⁸F]-DCFPyL, whole-body and axial, coronal and sagittal PET views at one hour post-injection (Fig. 2 C, D) revealed physiological radiotracer accumulation in the salivary and lacrimal glands, as well as in the liver. There was a rapid elimination of activity from the blood pool, as well as significant renal excretion, with radiotracer accumulation in the bladder. No radiotracer uptake was observed in the tumor. The TLR was 0.38 ± 0.03 (Table 3 ). Table 3 Ex vivo and in vivo biodistribution of [ 18 F]-DCFPyL on TNBC murine models Models 4T1, 1.10 4 , PBS, MFP (n = 4) MDA-MB-468, 1.10 6 , Matrigel®, MFP (n = 5) MDA-MB-231 wt , 1.10 6 , Matrigel®, MFP (n = 5) MDA-MB-231 wt /MDA-MB-231 psma , 5.10 5 /5.10 5 , Matrigel®, MFP (n = 6) MDA-MB-231 psma , 1.10 6 , Matrigel®, MFP (n = 6) Ex vivo biodistribution (IA%/g) Tumor 0.40 ± 0.06 0.43 ± 0.18 0.22 ± 0.05 15.05 ± 3.77 17.84 ± 5.15 Blood 0.45 ± 0.07 0.13 ± 0.04 0.13 ± 0.03 0.17 ± 0.06 0.16 ± 0.05 Mammary Fat Pad 0.52 ± 0.09 0.27 ± 0.07 0.19 ± 0.05 0.24 ± 0.04* 0.24 ± 0.04* Salivary gland 0.73 ± 0.20 0.27 ± 0.61 0.24 ± 0.04 0.28 ± 0.05* 0.25 ± 0.05* Liver 1.79 ± 0.34 1.036 ± 0.12 1.01 ± 0.22 1.04 ± 0.12 1.10 ± 0.21 Tail 2.79 ± 0.34 0.74 ± 0.15 0.75 ± 0.10 0.74 ± 0.07* 0.74 ± 0.15* Muscle 0.38 ± 0.30 0.48 ± 0.61 0.07 ± 0.01 0.34 ± 0.60 0.09 ± 0.2 Kidney 143.70 ± 23.59 59.21 ± 23.99 61.98 ± 13.88 50.37 ± 12.59* 57.59 ± 18.09* PET imaging TLR 0.38 ± 0.03 0.46 ± 0.18 0.40 ± 0.11 9.04 ± 2.51 10.26 ± 4.28 TMR 1.76 ± 0.73 1.48 ± 0.54 1.50 ± 0.87 22.51 ± 0.03 32.75 ± 0.03 The groups are described as follows: Cell line, number of cells, solution of inoculation, site of inoculation. Values are mean ± SD. %IA/g: percentage of injected activity per gram. * n = 4. TNBC: Triple-negative breast cancer. TLR: Tumor-to-liver ratio. TMR: Tumor-to-muscle ratio. PET: Positron emission tomography. Variations in the inoculation site (ectopic/orthotopic) and the addition of a matrix (Geltrex®/Matrigel®) affected tumor morphology and neovascularization (see Fig. 3 and Additional file 1: Fig. S2), yet PSMA expression was universally absent. This sets the stage for a detailed examination of the most informative TNBC models. For the xenograft models, MDA-MB-231 wild-type (called MDA-MB-231 wt ) and MDA-MB-468 cells were inoculated subcutaneously into the right flank in PBS (Additional file 1: Fig. S2) or in the MFP using Matrigel® (Fig. 3 ). At 23–26 days p.i., tumor volumes were 246 ± 49 mm 3 and 144 ± 65 mm 3 for MDA-MB-231 wt and MDA-MB-468 inoculated with Matrigel®, respectively (Fig. 3 A, B.). IHC staining of MFP models revealed higher levels of neovascularization and necrosis than subcutaneous models when examined using H&E staining (30%-70% for MDA-MB-468 models and 50%-80% for MDA-MB-231 wt models, respectively, at 26-81- and 44-78-days post-inoculation (p.i.)). However, no PSMA expression was detected by IHC in any of the MDA-MB-231 wt or MDA-MB-468 models (Fig. 3 C, D.). In these TNBC models inoculated with Matrigel®, whole-body and axial, coronal and sagittal [¹⁸F]-DCFPyL PET views showed a similar distribution one hour post-injection of the radiopharmaceuticals compared to the 4T1 model, with lower uptake in the salivary and lacrimal glands (Fig. 3 E-H.) (Table 3 ). These results were confirmed by an ex vivo biodistribution study showing an uptake in the salivary glands of 0.24 ± 0.04 IA%/g and 0.27 ± 0.61 IA%/g compared to 0.73 ± 0.20 IA%/g for 4T1, MDA-MB-231 wt and MDA-MB-468 models respectively. The calculated TLRs were 0.40 ± 0.11 and 0.46 ± 0.18 for MDA-MB-231 wt and MDA-MB-468 models respectively (Table 3 ). Co-inoculation of HUVEC and TNBC cells failed to induce PSMA expression in the neovasculature. Primary HUVEC cells were co-inoculated with TNBC cells to promote human neovascularization and potentially induce PSMA expression. MDA-MB-231 wt and MDA-MB-231 wt /HUVEC cells inoculated with Matrigel® exhibited tumor volumes of 303 ± 64 mm 3 (n = 6, 100% tumor engraftment) and 242 ± 103 mm 3 (n = 10, 100% tumor engraftment), respectively, at 27–30 days p.i. (Fig. 3 A, Fig. 4 A). It should be mentioned that tumoral tissue evidenced proliferation (Ki-67 staining) and neovascularization (CD31 staining), but no PSMA expression. Additionally, no qualitative differences in neovascularization were observed between TNBC-only and TNBC/HUVEC models (Fig. 3 C and 4 B). Validation and characterization of the PSMA-positive TNBC murine model Due to the lack of syngeneic or xenograft TNBC models that express PSMA, MDA-MB-231 cells were modified by lentiviral transduction to overexpressed PSMA (MDA-MB-231 psma ). To evaluate the effects of tumor PSMA expression and mimic intratumoral heterogeneity, female mice were inoculated with 1 million MDA-MB-231 psma or a 1:1 mixture of 500,000 MDA-MB-231 wt and 500,000 MDA-MB-231 psma cells both of which were suspended in Matrigel®. These groups were compared to mice inoculated with 1 million MDA-MB-231 wt with Matrigel®. At 28–30 days post-inoculation, tumor volume were 208 ± 61 mm 3 for MDA-MB-231 psma , 212 ± 97 mm 3 for the MDA-MB-231 wt /MDA-MB-231 psma models and 305 ± 65 mm 3 for MDA-MB-231 wt . Tumor doubling times (DT) of MDA-MB-231 psma , MDA-MB-231 wt /MDA-MB-231 psma and MDA-MB-231 wt were 20.2 ± 8.3 days; 14.7 ± 12.5 days; 24.9 ± 6.6 days, respectively with no significant differences ( Kruskal-Wallis test ) (Fig. 5 A.). PET imaging showed physiological accumulation of [ 18 F]-DCFPyL in excretory organs, including the bladder and kidneys for all models (Fig. 5 B-E). Unlike MDA-MB-231 wt , both PSMA-positive models demonstrated strong tracer uptake, with TLRs of 15.1 ± 3.8 for the MDA-MB-231 wt /MDA-MB-231 psma model and 17.8 ± 5.2 for MDA-MB-231 psma (Table 3 ). No radiopharmaceutical uptake was observed in central tumor regions, which is consistent with necrosis that was confirmed by dissection and H&E staining (Fig. 5 F). Necrotic areas increased as tumor developed in both PSMA-positive models: 5–40% (mean = 20% ± 16%) at 21–34 days p.i. to 25–60% (mean = 51% ± 14%) at 42–55 days p.i. Higher PSMA expression was observed near hypoxic and apoptotic tumor cores. MDA-MB-231 wt /MDA-MB-231 psma and MDA-MB-231 psma models were characterized by a H-score: 180 ± 28 and 192 ± 10 respectively. No-PSMA-positive endothelial cells were detected in any models. DISCUSSION TNBC is the most aggressive breast cancer subtype. It has recently benefited from advances in antibody-drug conjugated and immunotherapy, as well as combination [ 6 ]. Nevertheless, the prognosis remains poor. The oncology therapeutic arsenal has recently expanded to include TRT. Syngeneic and xenograft models are essential tools for assessing therapeutic efficacy and tumor–host interactions in a reproducible, controlled setting. Out of 23 murine models characterized as PSMA-negative, some were limited by poor tumor growth (BT-20 line), indicating low tumorigenicity [ 22 ]. These observations have led to the establishment of the first transfected model of TNBC with PSMA expression. One observation from this experimental work is the difficulty of reproducing existing models from the literature. Shirke et al. reported three models inoculated in MFP: i) 4T1 cells in BALB/c mice (10 000 cells) ii-iii) MDA-MB-231 and MDA-MB-468 in BALB/c Nude mice (1.10 6 cells). Interestingly, in our study, female BALB/c mice inoculated with 10 000 4T1 cells in the mammary fat pads with PBS exhibited a tumor volume of 618 ± 122 mm 3 versus 250 mm 3 for Shirke et al. model at the same stage [ 21 ]. The tumor volume of MDA-MB-468 cells inoculated in the MFP with PBS was not indicated in this study. In 2024, Heesch et al. have published a model of Rj:NMRI- Foxn1 nu/nu inoculated with 5.10 6 MDA-MB-231 in the MFP that underwent [ 177 ]Lu-PSMA-I&T therapy. Unfortunately, the tumor volume was relative (normalized to the first day of treatment) so we could not compare the tumor development of our model. Despite four reproduced models described in the literature characterized by neovascular expression of PSMA, we were unable to find PSMA expression in our model as evidenced by IHC as well as [ 18 F]-DCFPyL PET images (Fig. 2 C; Fig. 3 E-H; Fig. 5 B-E). Heesch el al. demonstrated that hypoxia can induce PSMA expression in both tumoral and endothelial cell [ 23 ]. Hypoxic models were harvested (subcutaneously [ 24 – 26 ]) by Morgenroth et al. in 2019. MDA-MB-231 cells inoculated in the right flank have resulted in tumors with [ 68 Ga]Ga-PSMA-11 uptake by PET imaging (target mean to background mean ratio of 43.3 ± 0.9) [ 19 ], while our subcutaneous models showed no PSMA expression . This lack of reproducibility is increasingly recognized as a major issue in biomedical research, with both scientific and ethical implications notably concerning the Reduction aspect of the 3Rs (Replacement, Reduction and Refinement) [ 27 – 30 ]. Since we were unable to reproduce these models, we investigated how to induce PSMA expression in tumor neovascularization. In the non-prostatic cancers, PSMA is thought to increase the neovascularization and invasion via nitric oxide production or the FAK-PAK pathway from laminin derivatives mediated by integrin β1 activation [ 31 – 34 ]. We hypothesize that limited vascularization in the models may impair PSMA expression. Thus, orthotopic injection appeared as the first option to increase neovascularization, followed by the addition of matrix and human endothelial cells. Moreover, previous studies have demonstrated higher radiopharmaceutical uptake in orthotopic models than in ectopic models of the same volume, which may be due to a higher vascular perfusion [ 25 , 26 ]. Unfortunately, Matrigel®, Geltrex® and co-inoculation of TNBC cell lines with endothelial cells did not result in PSMA expression by IHC in any of the 23 models tested. Considering the heterogeneous expression of PSMA, analyzing a single 5 µm tissue section in IHC study is limiting because the analyzed section may not contain regions where the signal is present, which could lead to false-negative results. Since PSMA expression is heterogeneous within tumoral tissue [ 35 ], the absence of PSMA expression in our study could be a consequence of the tumor intra-heterogeneity. Thus, nuclear imaging could be a relevant alternative method. The main limitations of nuclear imaging are the access of the radiopharmaceuticals, unlike IHC which is widely accessible and the limited resolution of the TEP camera which can miss low levels of radiotracer accumulation in cases of low PSMA expression. As the main aim of this study was to evaluate PSMA expression ex vivo , only a few mice in the selected groups were injected with PSMA-targeting radiopharmaceuticals. In our case, Tumor-to-Liver ratio was consistently under one, which would be considered a PSMA-negative tumor for therapeutic clinical selection [ 7 ]. As the specificity of [ 18 F]-DCPyL to murine PSMA has to be investigated, our results showed an [ 18 F]-DCPyL uptake of 0.73 ± 0.20 IA%/g in the salivary gland of BALB/c mice. These results suggest that the [ 18 F]-DCPyL may target murine PSMA, a topic that has not yet been studied to the best of our knowledge. Given the inability to induce PSMA in tumor neovessels and its lack of expression in human TNBC cells, MDA-MB-231 wt cells were transfected via lentiviral transduction to induce PSMA expression. Transfecting cancer cells to express PSMA has already been explored for Renal Cell Carcinoma (RCC) [ 36 ]. To mimic the intra-tumoral heterogeneity observed clinically, MDA-MB-231 psma cells were implanted alone or together with MDA-MB-231 wt cells. Interestingly, we observed a higher PSMA expression near the hypoxic and apoptotic core. Further investigation is needed to understand the modulation of the transgene or PSMA in these zones. Despite PSMA expression in cancer cells, we were unable to demonstrate PSMA induction in tumor neovessel endothelial cells in this model. It would be interesting to transfect endothelial cells to create a model with neovascular PSMA expression [ 37 , 38 ]. If validated, this model could provide a complementary approach to address questions related to the residence time of the radiopharmaceutical in PSMA-positive tumor neovascularization and its potential impact on therapeutic efficacy. PET imaging with [ 18 F]-DCFPyL showed comparable radiotracer uptake in tumors composed solely of MDA-MB-231 psma cells and in tumors containing a mixture of MDA-MB-231 wt and MDA-MB-231 psma cells. These results are consistent with IHC studies in which PSMA expression heterogeneity was observed on both models. For comparison, PC3-PIP (PSMA+, generated by lentiviral transduction of the PC-3 cell line [ 39 ]), prostate cancer xenografts exhibit [¹⁸F]-DCFPyL uptake of 44.2 ± 9.7 and 84.3 ± 12.3%IA/g at 1 h p.i. [ 40 , 41 ] whereas our MDA-MB-231 wt / MDA-MB-231 psma and MDA-MB-231 psma tumors showed an uptake of 15.05 ± 3.77 and 17.84 ± 5.15 respectively. This is consistent with clinical observations showing lower PSMA expression in TNBC than in mCRPC. Finally, ex vivo biodistribution and dosimetric studies with the therapeutic radiopharmaceutical, [ 177 Lu]Lu-PSMA- should be conducted before initiating TRT studies. Our data suggest that early-stage treatment (< tumors less than 250 mm³) may help avoid necrotic cores and improve therapeutic outcomes. CONCLUSIONS Based on our experience and in accordance with literature reports, the TNBC tumor model and the endothelial cell and TNBC tumor co-inoculated models display faint or no PSMA expression. The reproducibility of these models is questionable. Here, to our knowledge, we present the first PSMA-transfected TNBC model exhibiting high PSMA expression via IHC and high PSMA tracer accumulation via PET in TNBC tumor cells. This new model guarantees high reproducibility and is suitable for preclinical evaluation of TRT. Abbreviations %IA/g Percentage of injected activity per gram ATCC American type culture collection BC Breast cancer CT Computed tomography DT Doubling time FBS Fetal bovine serum H&E Hematoxylin-eosin HU Hounsfield units IHC Immunohistochemistry i.v. intra-venous IVIA In vivo imaging Auvergne MOI Multiplicity of infection mCRPC metastatic castration-resistant prostate cancer MDA-MB-231 wt MDA-MB-231 wild-type mTNBC metastatic triple-negative breast cancer MFP Mammary fat pad OCT Optimal cutting temperature PBS Phosphate buffer saline PET Positron emission tomography p.i. post inoculation PSMA Prostate specific membrane antigen RT Room temperature TLM Tumor-to-muscle ratio TLR Tumor-to-liver ratio TNBC Triple-negative breast cancer TRT Targeted radionuclide therapy TV Tumor volume Declarations ETHICS APPROVAL AND CONSENT TO PARTICIPATE The study was conducted in accordance with ARRIVE guidelines (https://arriveguidelines.org) and approved protocols outlined by the Auvergne Local Ethics Committee C2E2A and the French Ministry of Education and Research (approval numbers 49893 and 23955). No human studies were conducted. The maximum tumor size permitted by the local ethics committee, (1500 mm 3 ), was never reached by any mouse in this study. CONSENT FOR PUBLICATION Not applicable AVAILABILITY OF DATA AND MATERIALS All data generated or analyzed during this study are included in this published article [and its supplementary information files]. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. COMPETING INTERESTS The authors declare that they have no competing interests related to this work FUNDING This research was supported by grants from the Ligue contre le cancer comité de l’Allier et du Cantal. AUTHORS' CONTRIBUTIONS BC and EJ performed in vivo experiments, analyzed data, and interpreted results. LS, EB, SB, MA and AV participated in vivo experiments. MK performed histological and immunohistochemical analysis. CMo and JD performed lentiviral transduction of TNBC cell lines. SL, AR, MT and SS performed (radio)chemical analysis. BC, EJ, FC drafted the first draft of the manuscript. EC, EMN, FPL, CMe, FC and EJ supervised and contributed intellectually to the work and carefully revised the manuscript. All authors read, revised and approved the final manuscript. ACKNOWLEDGEMENTS We would like to thank Christelle Soubeyrand-Damon and Anipath histopathology facility (Institute of Genetics, Reproduction and Development, UMR 6293 CNRS/U1103 INSERM/Clermont Auvergne University) for their technical assistance with the histochemical studies. We would also like to thank Christelle Blavignac and Clémence Chapuis, Centre Imagerie Cellulaire Sante _UCA PARTNER, for their technical support and expertise in flow cytometry. We would like to thank CURIUM France (Saint-Beauzire, France) for supplying the [ 18 F]-DCFPyL, that made it possible to perform PET/CT scans on mouse models. 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16:38:29","extension":"html","order_by":17,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":144661,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7904335/v1/b484c93f293201819f3c44d5.html"},{"id":95663205,"identity":"4a1a8772-f21f-4950-8216-6a825f53b87c","added_by":"auto","created_at":"2025-11-11 16:38:31","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":123441,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFlow Chart of Strategies Enhancing Engraftment, Growth, Vascularization and PSMA in TNBC Models. \u003c/strong\u003eMurine models were developed by modifying the TNBC cell line, cell numbers, site of implantation, use of matrices and the co-inoculation with endothelial cells (HUVEC). For each model, PSMA expression was analyzed by immunohistochemistry. For few models, [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL PET images was harvested to study radiotracer accumulation and confirmed PSMA expression. TNBC: Triple-negative breast cancer\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7904335/v1/fa7f19c110a67e33b17dc1b3.jpg"},{"id":95663185,"identity":"ee731b2c-af27-4d67-be3c-2ee0b3078cdd","added_by":"auto","created_at":"2025-11-11 16:38:29","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":123445,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCharacterization of syngeneic model and absence of PSMA expression. (A) \u003c/strong\u003eTumors volume after inoculation in syngeneic models (n=6 per group). \u003cstrong\u003e(B) \u003c/strong\u003eHistological analyses of the TNBC syngeneic model. Tumor sections were stained with hematoxylin-eosin (H\u0026amp;E) or immunostained for Ki-67, PSMA and CD31. A zoom was performed on a central and peripheral area of each tumor for Ki-67, PSMA and CD31 staining. \u003cstrong\u003e(C) \u003c/strong\u003eMaximal Intensity Projection (MIP) and \u003cstrong\u003e(D) \u003c/strong\u003eaxial, coronal and sagittal views and of [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL PET imaging in syngeneic TNBC murine model. The groups are described as follows: Cell line, number of cells, solution of inoculation, site of inoculation. Mice were sacrificed at different time points. Each curve represents one individual tumor. Black arrows correspond to the different stages of sacrifice. PET imaging was performed 1-hour post-injection with a 15 minutes acquisition time. Blue circles represent the anatomical tumor location. White arrow represents organs. Bl: Bladder, Kd: Kidney, Sg: salivary gland, Lg: Lacrimal gland, L: Liver. For MIP, PET color scale range is 0 - 626 kBq/mL, CT color scale range is 645 - 3055 HU. For axial, coronal and sagittal views, PET color scale range is 0 – 626 kBq/mL, CT color scale range is -643 – 555 HU. PBS: Phosphate buffer saline. MFP: Mammary fat pad. TNBC: Triple-negative breast cancer. PET/CT : Positron emission tomography / Computed tomography.\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7904335/v1/b06678cea2c49bbe97b819ac.jpg"},{"id":95663183,"identity":"fc017d60-362a-401b-a0ef-f6adfa76ccf1","added_by":"auto","created_at":"2025-11-11 16:38:29","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1494329,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCharacterization of MDA-MB-231\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ewt\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e and MDA-MB-468 xenograft mouse models and absence of PSMA expression. (A-B) \u003c/strong\u003eTumors volume after inoculation in xenograft models (n=6 per group). \u003cstrong\u003e(C-D) \u003c/strong\u003eHistological analyses of the TNBC xenograft model. Tumor sections were stained with hematoxylin-eosin (H\u0026amp;E) or immunostained for Ki-67, PSMA and CD31. A zoom was performed on a central and peripheral area of each tumor for Ki-67, PSMA and CD31 staining. \u003cstrong\u003e(E,G) \u003c/strong\u003eMaximal Intensity Projection and \u003cstrong\u003e(F,H)\u003c/strong\u003e axial, coronal and sagittal views of [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL PET imaging in xenograft TNBC murine model. The groups are described as follows: Cell line, number of cells, solution of inoculation, site of inoculation. Mice were sacrificed at different time points. Black arrows correspond to the different stages of sacrifice. PET imaging was performed 1-hour post-injection with a 15 minutes acquisition time. Blue circles represent the anatomical tumor location. White arrow represents organs. Bl: Bladder, Kd: Kidney, Sg: salivary gland, Lg: Lacrimal gland, L: Liver. For MIP, PET color scale range is 0 - 626 kBq/mL, CT color scale range is 645 - 3055 HU. For axial, coronal and sagittal views, PET color scale range is 0 – 626 kBq/mL, CT color scale range is -643 – 555 HU. PBS: Phosphate buffer saline. MFP: Mammary fat pad. TNBC: Triple-negative breast cancer. PET/CT : Positron emission tomography / Computed tomography.\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7904335/v1/add88b0c72d493bc282ed80f.jpg"},{"id":95663180,"identity":"a3ab02a2-f3b3-4858-87bb-7832457a045e","added_by":"auto","created_at":"2025-11-11 16:38:28","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":96426,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCharacterization of co-inoculated TNBC/endothelial cells xenograft mouse model and absence of PSMA expression. (A) \u003c/strong\u003eTumors volume of co-inoculated TNBC/HUVEC models (n=12 per group). \u003cstrong\u003e(B) \u003c/strong\u003eHistological analyses of the TNBC/HUVEC xenograft model. Tumor sections were stained with hematoxylin-eosin (H\u0026amp;E) or immunostained for Ki-67, PSMA and CD31. A zoom was performed on a central and peripheral area of each tumor for Ki-67, PSMA and CD31 staining. The groups are described as follows: Cell line, number of cells, solution of inoculation, site of inoculation. Mice were sacrificed at different time points. Each curve represents one individual tumor. Black arrows correspond to the different stages of sacrifice. PBS: Phosphate buffer saline. MFP: Mammary fat pad. TNBC: Triple-negative breast cancer.\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7904335/v1/81d147fa0c0f7bd73f344839.jpg"},{"id":95663274,"identity":"361cb4ca-7d80-43bc-b72d-344f14e3916a","added_by":"auto","created_at":"2025-11-11 16:38:39","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1791124,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eValidation of PSMA positive TNBC murine model. (A)\u003c/strong\u003e Tumor doubling time of MDA-MB-231\u003cem\u003ewt\u003c/em\u003e; MDA-MB-231\u003cem\u003ewt\u003c/em\u003e/MDA-MB-231\u003cem\u003epsma\u003c/em\u003e and MDA-MB-231\u003cem\u003epsma\u003c/em\u003e models were characterized and compared. \u003cstrong\u003e(B, C) \u003c/strong\u003eMaximal Intensity Projection (MIP) and \u003cstrong\u003e(D, E) \u003c/strong\u003eaxial, coronal and sagittal views of [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL PET/CT imaging. \u003cstrong\u003e(F)\u003c/strong\u003e Hematoxylin-eosin; ki-67; PSMA and CD31 immunostaining of tumor sections. Experimental groups are described as follows: cell line, number of cells injected, solution of inoculation, site of inoculation. Black arrows correspond to the different stages of sacrifice. PET imaging was performed 1-hour post-injection with a 15 minutes acquisition time. Blue circles represent the anatomical tumor location. White arrow represents organs. Bl: Bladder, Kd: Kidney, L: Liver. For MIP, PET color scale range is 0 - 626 kBq/mL, CT color scale range is 645 - 3055 HU. For axial, coronal and sagittal views, PET color scale range is 0 – 626 kBq/mL, CT color scale range is -643 – 555 HU. PBS: Phosphate Buffer Saline. MFP: Mammary Fat Pad. TNBC: Triple-negative breast cancer. PET/CT : Positron emission tomography / Computed tomography. Kruskal-Wallis test; ns: not significant. Values are mean ± SD.\u003c/p\u003e","description":"","filename":"5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7904335/v1/0384c470b2ea7d0ea8b0a62f.jpg"},{"id":103251066,"identity":"bed26f74-8da3-4289-baca-cfac671540b3","added_by":"auto","created_at":"2026-02-23 16:03:17","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":5015749,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7904335/v1/550401d7-5cc6-4fa0-a506-034a5bf372d3.pdf"},{"id":95663207,"identity":"99f46865-dfa7-4719-9516-5c40a86dd034","added_by":"auto","created_at":"2025-11-11 16:38:31","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":52089379,"visible":true,"origin":"","legend":"","description":"","filename":"Additionalfile1final.docx","url":"https://assets-eu.researchsquare.com/files/rs-7904335/v1/7e96a5d94f2a6e6e9496f386.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"From Hurdles to Breakthroughs: Building and Validating a PSMA-positive Triple-Negative Breast Cancer Mouse Model for Preclinical Targeted Radionuclide Therapy","fulltext":[{"header":"BACKGROUND","content":"\u003cp\u003eTriple-negative breast cancer (TNBC) accounts for 15% of breast cancers and is considered as the most aggressive subtype, with the poorest prognosis. Approximatively 46% of TNBC patients are expected to develop metastases which are associated with high microvascular densities [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. The median overall survival for metastatic TNBC (mTNBC) is 14,8 months [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Despite the development of antibody-drug conjugates which provide more therapeutic options beyond checkpoint and PARP inhibitors in the metastatic stage, the prognosis for metastatic triple-negative breast cancer (mTNBC) remains poor [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Consequently, research in this field is very active, seeking new therapeutic approaches that can either provide alternative options or be combined with existing treatments to improve outcomes [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe ongoing development of Targeted Radionuclide Therapy (TRT) is opening new avenues in oncology by providing additional therapeutics options. Notably, it represents an attractive strategy for addressing metastasis. In 2022, the European Medicines Agency approved the radiopharmaceutical lutetium (\u003csup\u003e177\u003c/sup\u003eLu) vipivotide tetraxetan which targets the Prostate Specific Membrane Antigen (PSMA) for therapy in metastatic castration-resistant prostate cancer (mCRPC) [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. This approval has prompted the investigation of additional applications of PSMA-TRT in cancer beyond the prostate. Indeed, although PSMA was initially discovered and characterized in the prostate, it is also expressed in other solid cancers [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Therefore, expanding PSMA-targeted TRT to non-prostate malignancies that overexpress PSMA is of interest, particularly glioblastoma, renal cell carcinoma and breast cancer (BC) [\u003cspan additionalcitationids=\"CR11 CR12 CR13 CR14\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. However, PSMA expression is generally weaker and more heterogeneous in non-prostate solid tumors than in prostate cancer. In particular, PSMA has been shown to be expressed in 66%-74% of primary BC and 75%-100% of distant metastases with the highest rates observed in TNBC [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Furthermore, immunohistochemistry (IHC) studies have demonstrated that PSMA is predominantly expressed by endothelial cells of tumor neovessels (46%-83%) and also by tumor cells (3%-72%), in BC [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan additionalcitationids=\"CR17\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. PSMA overexpression has also been shown to be higher in neovessels from metastatic sites than in primary tumor neovascularization [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e], supporting the use of PSMA-targeted TRT for mTNBC. Nevertheless, predominant localization in tumor neovascularization raises questions about therapeutic efficacy. Therefore, it is essential to evaluate the efficacy and relevance of PSMA-targeted TRT in mouse models representative of these tumors to demonstrate proof of concept for clinical trials.\u003c/p\u003e\u003cp\u003eTo the best of our knowledge, only two teams have successfully developed TNBC murine models with PSMA expression in preclinical studies. However, these models exhibited necrosis cores and heterogeneous PSMA expression [\u003cspan additionalcitationids=\"CR20\" citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Shirke et al. described the level of PSMA expression as insufficient for therapy efficacy. In contrast, Heesch et al. successfully performed [\u003csup\u003e177\u003c/sup\u003eLu]Lu-PSMA-I\u0026amp;T assessments in mice, highlighting the heterogeneity between models and even among mice within the same model.\u003c/p\u003e\u003cp\u003eThis study aimed to develop a preclinical TNBC murine model that can express PSMA and is highly reproducible and suitable for future PSMA-targeted TRT research. To achieve this goal, we evaluated 25 syngeneic and xenograft models (4T1, MDA-MB-231, MDA-MB-468, MDA-MB-436, BT-20), including co-inoculations with endothelial cells to enhance tumor neovascularization. We also evaluated a novel PSMA-positive model generated \u003cem\u003evia\u003c/em\u003e lentiviral transduction of MDA-MB-231 cells. We assessed engraftment, tumor growth, PSMA expression and PSMA-targeted radiopharmaceuticals, such as [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL uptake.\u003c/p\u003e"},{"header":"METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eCell lines\u003c/h2\u003e\u003cp\u003eThe murine TNBC cell line 4T1 (CRL-2539), and Human TNBC cell lines MDA-MB-231 (HTB-26), MDA-MB-468 (HTB-132), MDA-MB-436 (HTB-130) and BT-20 (HTB-19) cell lines were purchased from the American Type Culture Collection (ATCC, Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The MDA-MB-436 and 4T1 cell lines were cultured in RPMI-1640 medium supplemented with 10% Fetal Bovine Serum (FBS, Dutscher, Bernolsheim, France) and 4 \u0026micro;g / mL gentamicin. The MDA-MB-231\u003cem\u003ewt\u003c/em\u003e, MDA-MB-468 and BT-20 cell lines were maintained with MEM-Glutamax supplemented with 10% FBS, 1mM of sodium pyruvate, 1X of Non-Essential Amino Acids, 1X of vitamins and 4 \u0026micro;g / mL of gentamicin (50 mg / mL). Except for FBS, all culture reagents were purchased from Gibco\u0026reg; (ThermoFisher Scientific, Strasbourg, France). The human primary endothelial cell line, HUVEC cells (c-12208, PromoCell GmbH, Heidelberg, Germany) was maintained in the Endothelial Cell Growth Medium 2 (ready-to-use) (Promocell, GmbH, Heidelberg, Germany). All cells were maintained at 37\u0026deg;C with 5% of CO\u003csub\u003e2\u003c/sub\u003e and were routinely tested for mycoplasma infection.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003ePSMA overexpression and lentiviral transduction\u003c/h3\u003e\n\u003cp\u003eThe PSMA and LacZ-V5 tags were synthesized by Vector Builder and cloned into a third-generation self-inactivating lentiviral expression vector (Vector Builder) under the eukaryotic translation elongation factor 1 α (EF1A) promoter: PSMA plasmid pLV[Exp]-Bsd-EF1A\u0026thinsp;\u0026gt;\u0026thinsp;hFOLH1[NM_004476.3]; LacZV5 plasmid pLV[Exp]-Bsd-EF1A\u0026gt;{LacZV5}. Lentiviruses were produced by triply transfecting HEK-293T cells with the lentiviral transfer vector and the packaging plasmids psPAX2 (a gift from Didier Trono; Addgene plasmid #12260; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://n2t.net/addgene:12260\u003c/span\u003e\u003cspan address=\"http://n2t.net/addgene:12260\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e; RRID: Addgene_12260) and pMD2.G (a gift from Didier Trono; Addgene plasmid #12259; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://n2t.net/addgene:12259\u003c/span\u003e\u003cspan address=\"http://n2t.net/addgene:12259\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e; RRID: Addgene_12259) at a ratio of 0.3:0.3:0.1. Transfection was performed using jetPEI (101000053, Polyplus). Viral supernatants were collected 48 hours post-transfection, filtered through a 0.22 \u0026micro;m filter, and used for transduction. MDA-MB-231\u003cem\u003ewt\u003c/em\u003e cells were transduced with lentiviral particles at a multiplicity of infection (MOI) of 1 for MDA-MB-231\u003cem\u003ewt\u003c/em\u003e in the presence of hexadimethrine bromide (8 \u0026micro;g/mL). MDA-MB-231\u003cem\u003epsma\u003c/em\u003e cells were cultured with the same culture medium as MDA-MB-231\u003cem\u003ewt\u003c/em\u003e, but with the addition of blasticidin (9 \u0026micro;g/mL, InvivoGen, ant-bl-05) for antibiotic selection.\u003c/p\u003e\u003cp\u003e\u003cspan type=\"BoldItalicUnderline\" class=\"BoldItalicUnderline\" name=\"Emphasis\"\u003eIn vivo\u003c/span\u003e \u003cspan type=\"BoldUnderline\" class=\"BoldUnderline\" name=\"Emphasis\"\u003estudies\u003c/span\u003e\u003c/p\u003e\u003cp\u003eAll experiments were approved by the Auvergne local Ethic committee C2E2A and the French Ministry of Education and Research (approval Nos. 49893 \u0026amp;. 23955) and were conducted in accordance with ARRIVE guidelines (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://arriveguidelines.org\u003c/span\u003e\u003cspan address=\"https://arriveguidelines.org\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Mice were housed under environmentally controlled conditions (4\u0026ndash;5 animals per cage, ventilated racks, 60% relative humidity, temperature maintained at 21\u0026ndash;24\u0026deg;C, and a 12 h/12 h light/dark cycle). Animals had free access to standard food and water. Tumor xenograft and sygeneic model were established by injecting tumor cells into anesthetized mice. Anesthesia was induced and maintained with gaseous isoflurane (Iso-Vet, 1.5%, 1 L/min, air/O₂ 70/30, v/v) using an animal anesthesia system (Minerve). For syngeneic models, 7\u0026ndash;9 week old female BALB/cAnNRj mice from Janvier Labs (Le Genest-Saint-Isle, France) were inoculated with 4T1 cells at different concentrations (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e; Additional File. Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). For the xenograft models, 7\u0026ndash;9 week old female NMRI-Foxn1nu/Fox1nu and female Balb/cAnNRj-Foxn1 nu/nu from Janviers Labs (Le Genest-Saint-Isle, France) were inoculated with human TNBC cell lines with or without the co-injection of different amount of HUVEC cells (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). For both the syngeneic and xenograft models, subcutaneous injections were performed into the right flank with 100 \u0026micro;L of cells suspension. For orthotopic injection, the cells were injected into the third thoracic mammary fat pad (MFP) with a 100 \u0026micro;L cell suspension (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) (see Additional File Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). The cells were resuspended in 100 \u0026micro;L of one of the following solutions: i) PBS (ThermoFisher Scientific, Strasbourg, France), ii) 1:1 volume ratio of PBS and Geltrex\u0026reg; (ThermoFisher Scientific, Strasbourg, France), iii) 1:1 volume ratio of PBS and Matrigel\u0026reg; (Corning\u0026reg;, New York, USA). Tumor volume and body weight were measured once a week for the xenograft model and twice a week for the syngeneic model by the same operator.\u003c/p\u003e\u003cp\u003eAt different time points (at least 2 mice) between 19 days and 81 days post inoculation (p.i.), we re-anesthetized the mice with isoflurane (5%), prior to confirmation of anesthesia with a tail pinch, and performance of cervical dislocation to sacrifice them. Metastases were examined in the liver and lungs. The tumors were split into two parts, one half oriented toward the periphery and the other oriented toward the center, before paraffin embedding and hematoxylin and eosin (H\u0026amp;E) staining and immunohistochemistry (IHC) for Ki-67, CD31, PSMA, and HIF-1α.\u003c/p\u003e\u003cp\u003eTumor volume (TV) was calculated using caliper measurements as TV \u003cem\u003e(mm\u0026sup3;) = [length \u0026times; (width)\u0026sup2;]/2\u003c/em\u003e and doubling time (DT) as \u003cem\u003eDT\u0026thinsp;=\u0026thinsp;Ln2/a\u003c/em\u003e, where \u0026ldquo;a\u0026rdquo; is the slope of the exponential growth phase.\u003c/p\u003e\u003cp\u003eThe maximum tumor size permitted by the local ethics committee, (1500 mm\u003csup\u003e3\u003c/sup\u003e), was never reached by any mouse in this study.\u003c/p\u003e\n\u003ch3\u003eNuclear PET imaging\u003c/h3\u003e\n\u003cp\u003ePositron emission tomography / computed tomography (PET/CT) scans were performed using the IRIS PET/CT (Inviscan) scanner. Acquisitions were performed for 15 minutes, one hour after intravenous (i.v.) injection of 7.50\u0026thinsp;\u0026plusmn;\u0026thinsp;1.42 MBq of [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL (Curium France, Saint-Beauzire, France). Then, the mice were anesthetized in an induction chamber containing 2.5% isoflurane in air and followed by 1.5% isoflurane in air to maintain anesthesia using an animal anesthesia machine (Minerve). The energy window used was 250\u0026ndash;750 keV. Images were reconstructed according to the OSEM 3D algorithm including corrections for dead time, decay, and randoms. For \u003cem\u003ein vivo\u003c/em\u003e biodistribution, images were analyzed using InterView\u0026trade; Fusion Software (ver. 3.11.011). The volume of interest (VOI) of tumor, liver and muscle was delineated to calculate the tumor-to-liver ratio (TLR) and the tumor-to-muscle ratio (TMR) for each mouse. For ex vivo biodistribution, mice were anesthetized with isoflurane (5%) 1.5 hours p.i. Anesthesia was confirmed by a tail pinch, followed by cervical dislocation to euthanize the animals. The organs and tumors were then collected and weighed. Radioactivity was measured using a γ-counter (Wizard 1480, Perkin Elmer) and the gamma counting data were corrected for physical decay (109.8 minutes) and background. All uptake data were expressed as percent injected activity per gram (%IA/g).\u003c/p\u003e\u003cp\u003eThe experimental part of this study was performed on IVIA platform, Clermont-Ferrand, France (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.18145/ivia\u003c/span\u003e\u003cspan address=\"10.18145/ivia\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003eImmunohistochemistry and H\u0026E staining protocol\u003c/h3\u003e\n\u003cdiv class=\"Heading\"\u003eImmunohistochemistry and H\u0026amp;E staining protocol\u003c/div\u003e\u003cp\u003eFive \u0026micro;m sections were cut and adhered to poly-L-Lysine-coated glass microscope slides. The paraffin was then removed with two 15 minutes baths of SafeSolv (#00699464, VWR, France). The tissues were then rehydrated with successive ethanol baths at decreasing concentrations (100%, 100%, 95%, 70%, H\u003csub\u003e2\u003c/sub\u003eO; five minutes each).\u003c/p\u003e\u003cp\u003eFor the H\u0026amp;E protocol, nuclei were stained by a four-minute bath of hematoxylin (#C0303, Diapath, France), then rinsed with a four-minute bath of tap water. The sections were stained by eosin Y bath for four minutes and rinsed in the same manner before dehydration and mounting (described below).\u003c/p\u003e\u003cp\u003eFor IHC, heating-mediated antigen retrieval was performed using either a citrate buffer (10 mM sodium citrate tribasic (Sigma #066K0042); 0.05% Tween 20; pH 6) for Ki-67, cleaved-caspase3 and HIF-1α staining or a Tris-EDTA buffer (10 mM Base Trizma\u0026reg; (Sigma #T1503); 0.25 mM EDTA (Invitrogen, #15575-038); 0.05% Tween 20; pH 9) for CD31 staining. This was performed for 30 minutes at 95\u0026deg;C for both. The slides were cooled for 20 minutes, then rinsed in PBS 1X for three five-minute intervals. The tissues were blocked for one hour at room temperature (RT) in PBS 1X-BSA (1%)-Avidin (Vector laboratories, SP-2001). After rinsing, endogenous peroxidases were quenched using 0.3% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e in PBS 1X for 30 minutes at RT and rinsed again. Tumor sections were then incubated overnight at 4\u0026deg;C with the primary antibody in the staining solution (PBS 1X-BSA (1%)-Biotin (Vector laboratories, SP-2001)). We used the following antibodies i) anti-CD31 antibody (\u003cb\u003e1/2000\u003c/b\u003e, Abcam # ab182981) ii) anti-Ki-67 antibody (\u003cb\u003e1/100\u003c/b\u003e, Abcam # ab15580) iii) anti-PSMA antibody (\u003cb\u003e1/100\u003c/b\u003e, Cell Signaling Technology #12815) iv) anti-HIF-1α (\u003cb\u003e1/100\u003c/b\u003e, Cell Signaling Technology #48085). After rinsing, the tissue sections were stained with a biotinylated goat anti-rabbit IgG antibody (\u003cb\u003e1/500\u003c/b\u003e, Invitrogen # 656140) for one hour at RT. After rinsing, amplification was performed using Vectastain ABC Reagent (Vector laboratories, PK-4000) with a 30-minute incubation at RT. Revelation was performed after rinsing with a DAB kit (Vector laboratories, SK-4105). Tissue sections were rinsed and counterstained with a two-minute bath of hematoxylin and washed for two minutes in tap water. For H\u0026amp;E and IHC protocols, the tissues were dehydrated using successive ethanol baths at increasing concentrations (70%, 95%, 100%, SafeSolv, two minutes each prior to mounting with Shandon-Mount (Epredia, #1900333). Histological images were acquired using an Axio Scan.Z1 slide scanner and the ZEISS Colibri 7 camera. Antibody controls for PSMA, CD31 and Ki-67 stainings are shown in Fig S6 (Additional File 1).\u003c/p\u003e\u003cp\u003eImmunohistochemical staining was evaluated using the H-score method, which integrates both the intensity of staining and the percentage of stained cells. Staining intensity was categorized as follows: 0, no staining; 1, weak staining; 2, moderate staining and 3, strong staining. For PSMA, the H-score was calculated as a continuous variable (range 0\u0026ndash;300) using the following formula: \u003cem\u003eH-score\u0026thinsp;=\u0026thinsp;1\u0026times;(% cells with weak staining)\u0026thinsp;+\u0026thinsp;2\u0026times;(% cells with moderate staining)\u0026thinsp;+\u0026thinsp;3\u0026times;(% cells with strong staining)\u003c/em\u003e.\u003c/p\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003eStatistical Analysis\u003c/h2\u003e\u003cp\u003eStatistical analyses and graphical representations were performed using GraphPad Prism (version 10.3.1, GraphPad Software, San Diego, CA, USA). Variables were presented as mean with standard deviation. A Kruskal-Wallis test was performed to compare the doubling time of the MDA-MB-231\u003cem\u003ewt\u003c/em\u003e, MDA-MB-231\u003cem\u003ewt/\u003c/em\u003eMDA-MB-231\u003cem\u003epsma\u003c/em\u003e and MDA-MB-\u003cem\u003e231psma\u003c/em\u003e mouse models. The difference between the groups was tested using the Dunnett's multiple comparison test, and a p value of 0.05 or less was considered statistically significant.\u003c/p\u003e\u003c/div\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\u003ch2\u003eSite of inoculation, matrix and number of cells modulate tumor engraftment however PSMA remains unexpressed\u003c/h2\u003e\u003cp\u003eDifferent cell lines were tested and are summarized in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eList of Triple-negative breast cancer (TNBC) and endothelial cell line for preclinical \u003cem\u003ein vivo\u003c/em\u003e studies\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eType\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCell line\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eDisease\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eOrganism\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eSource\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eMutation\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eEndogenous PSMA expression\u003c/p\u003e\u003cp\u003e(by Flow Cytometry)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003eTriple-Negative Breast Cancer\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMDA-MB-231\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdenocarcinoma (pleural effusion) /\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eHomo sapiens\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eATCC\u003c/p\u003e\u003cp\u003e(HTB-26)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eCDKN2A/B, BRAF, KRAS, TERT, TP53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eNo\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMDA-MB-468\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdenocarcinoma (pleural effusion)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eHomo sapiens\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eATCC\u003c/p\u003e\u003cp\u003e(HTB-132)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eTP53, PTEN, RB1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eNo\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMDA-MB-436\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAdenocarcinoma\u003c/p\u003e\u003cp\u003e(pleural effusion)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eHomo sapiens\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eATCC\u003c/p\u003e\u003cp\u003e(HTB-130)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eBRCA1, FHIT, TP53, RB1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eNo\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBT-20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCarcinoma\u003c/p\u003e\u003cp\u003e(primary tumor)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eHomo sapiens\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eATCC\u003c/p\u003e\u003cp\u003e(HTB-19)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eCDKN2A, PIK3CA, RB1, TP53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eNo\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4T1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMimics human breast cancer, Stage IV\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eMus musculus\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eATCC\u003c/p\u003e\u003cp\u003e(CRL-2539)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eTP53, PIK3G\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eNo\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eEndothelial cells\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHUVEC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePrimary Human Umbilical Vein Endothelial Cells\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eHomo sapiens\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePromocell\u003c/p\u003e\u003cp\u003e(C-12208)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e/\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eNo\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe murine 4T1 cell line was tested for the syngeneic model, and the human TNBC cell lines for xenograft models. Primary human endothelial cell line HUVEC was tested for co-inoculation. Figure\u0026nbsp;1, summarizes the different strategies that were explored to improve tumor engraftment and PSMA expression in order to develop a preclinical TNBC murine model that expresses PSMA (Fig.\u0026nbsp;1).\u003c/p\u003e\u003cp\u003e\u003cb\u003eFigure\u0026nbsp;1: Flow Chart of Strategies Enhancing Engraftment, Growth, Vascularization and PSMA in TNBC Models.\u003c/b\u003e Murine models were developed by modifying the TNBC cell line, cell numbers, site of implantation, use of matrices and the co-inoculation with endothelial cells (HUVEC). For each model, PSMA expression was analyzed by immunohistochemistry. For few models, [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL PET images was harvested to study radiotracer accumulation and confirmed PSMA expression. TNBC: Triple-negative breast cancer\u003c/p\u003e\u003cp\u003ePrior to \u003cem\u003ein vivo\u003c/em\u003e experiments, all cell lines were tested negative for PSMA expression by flow cytometry (Additional file 1: Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e presents a curated selection of the most relevant murine models, highlighting key characteristics such as mouse strain, cell line, inoculation parameters, and tumor growth. The remaining 19 models are detailed in Supplementary Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e (Additional File 1).\u003c/p\u003e\u003cp\u003eTable 2 Characterization of syngeneic and xenograft murine models for triple-negative breast cancer development\u003c/p\u003e\u003cp\u003e\u003cimg 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\" width=\"724\" height=\"406\"\u003e\u003c/p\u003e\u003cp\u003e*MFP: Mammary fat pad.\u003c/p\u003e\u003cp\u003eTNBC: Triple-negative breast cancer. PBS: Phosphate buffer saline\u003c/p\u003e\u003cp\u003eFemale Balb/c mice inoculated with 10,000 4T1 cells in the MFP with PBS exhibited a tumor volume of 1,132\u0026thinsp;\u0026plusmn;\u0026thinsp;89 mm\u003csup\u003e3\u003c/sup\u003e at 25\u0026ndash;30 days post-inoculation (n\u0026thinsp;=\u0026thinsp;12) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eA).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eHematoxylin-eosin staining has revealed necrosis for each syngeneic model (5%-35% of the tumor surface between 13\u0026ndash;25 days p.i.) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). Ki-67 IHC showed a high proliferation. However, despite strong CD31 staining indicating neovascularization in the syngeneic model, no PSMA expression was detected at the same location as CD31 staining (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). Considering the tumor uptake of [\u0026sup1;⁸F]-DCFPyL, whole-body and axial, coronal and sagittal PET views at one hour post-injection (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eC, D) revealed physiological radiotracer accumulation in the salivary and lacrimal glands, as well as in the liver. There was a rapid elimination of activity from the blood pool, as well as significant renal excretion, with radiotracer accumulation in the bladder. No radiotracer uptake was observed in the tumor. The TLR was 0.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003e\u003cem\u003eEx vivo\u003c/em\u003e and \u003cem\u003ein vivo\u003c/em\u003e biodistribution of [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL on TNBC murine models\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eModels\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4T1, 1.10\u003csup\u003e4\u003c/sup\u003e, PBS, MFP (n\u0026thinsp;=\u0026thinsp;4)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eMDA-MB-468, 1.10\u003csup\u003e6\u003c/sup\u003e, Matrigel\u0026reg;, MFP (n\u0026thinsp;=\u0026thinsp;5)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eMDA-MB-231\u003cem\u003ewt\u003c/em\u003e, 1.10\u003csup\u003e6\u003c/sup\u003e, Matrigel\u0026reg;, MFP (n\u0026thinsp;=\u0026thinsp;5)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eMDA-MB-231\u003cem\u003ewt\u003c/em\u003e/MDA-MB-231\u003cem\u003epsma\u003c/em\u003e, 5.10\u003csup\u003e5\u003c/sup\u003e/5.10\u003csup\u003e5\u003c/sup\u003e, Matrigel\u0026reg;, MFP (n\u0026thinsp;=\u0026thinsp;6)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eMDA-MB-231\u003cem\u003epsma\u003c/em\u003e, 1.10\u003csup\u003e6\u003c/sup\u003e, Matrigel\u0026reg;, MFP (n\u0026thinsp;=\u0026thinsp;6)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"7\" rowspan=\"8\"\u003e\u003cp\u003e\u003cem\u003eEx vivo\u003c/em\u003e biodistribution\u003c/p\u003e\u003cp\u003e(IA%/g)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTumor\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e0.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e0.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e0.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e15.05\u0026thinsp;\u0026plusmn;\u0026thinsp;3.77\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e17.84\u0026thinsp;\u0026plusmn;\u0026thinsp;5.15\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBlood\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e0.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e0.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e0.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e0.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e0.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMammary Fat Pad\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e0.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e0.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e0.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e0.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e0.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04*\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSalivary gland\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e0.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e0.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.61\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e0.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e0.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e0.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05*\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLiver\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e1.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e1.036\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e1.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e1.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e1.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTail\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e2.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e0.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e0.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e0.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e0.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15*\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMuscle\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e0.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e0.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.61\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e0.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e0.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e0.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eKidney\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e143.70\u0026thinsp;\u0026plusmn;\u0026thinsp;23.59\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e59.21\u0026thinsp;\u0026plusmn;\u0026thinsp;23.99\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e61.98\u0026thinsp;\u0026plusmn;\u0026thinsp;13.88\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e50.37\u0026thinsp;\u0026plusmn;\u0026thinsp;12.59*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e57.59\u0026thinsp;\u0026plusmn;\u0026thinsp;18.09*\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003ePET imaging\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTLR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e0.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e0.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e9.04\u0026thinsp;\u0026plusmn;\u0026thinsp;2.51\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e10.26\u0026thinsp;\u0026plusmn;\u0026thinsp;4.28\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTMR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e1.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.73\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e1.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e1.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.87\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e22.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e32.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eThe groups are described as follows: Cell line, number of cells, solution of inoculation, site of inoculation.\u003c/p\u003e\u003cp\u003eValues are mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD. %IA/g: percentage of injected activity per gram. * n\u0026thinsp;=\u0026thinsp;4. TNBC: Triple-negative breast cancer. TLR: Tumor-to-liver ratio. TMR: Tumor-to-muscle ratio. PET: Positron emission tomography.\u003c/p\u003e\u003cp\u003eVariations in the inoculation site (ectopic/orthotopic) and the addition of a matrix (Geltrex\u0026reg;/Matrigel\u0026reg;) affected tumor morphology and neovascularization (see Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e and Additional file 1: Fig. S2), yet PSMA expression was universally absent.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThis sets the stage for a detailed examination of the most informative TNBC models. For the xenograft models, MDA-MB-231\u003cem\u003ewild-type\u003c/em\u003e (called MDA-MB-231\u003cem\u003ewt\u003c/em\u003e) and MDA-MB-468 cells were inoculated subcutaneously into the right flank in PBS (Additional file 1: Fig. S2) or in the MFP using Matrigel\u0026reg; (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e). At 23\u0026ndash;26 days p.i., tumor volumes were 246\u0026thinsp;\u0026plusmn;\u0026thinsp;49 mm\u003csup\u003e3\u003c/sup\u003e and 144\u0026thinsp;\u0026plusmn;\u0026thinsp;65 mm\u003csup\u003e3\u003c/sup\u003e for MDA-MB-231\u003cem\u003ewt\u003c/em\u003e and MDA-MB-468 inoculated with Matrigel\u0026reg;, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003eA, B.). IHC staining of MFP models revealed higher levels of neovascularization and necrosis than subcutaneous models when examined using H\u0026amp;E staining (30%-70% for MDA-MB-468 models and 50%-80% for MDA-MB-231\u003cem\u003ewt\u003c/em\u003e models, respectively, at 26-81- and 44-78-days post-inoculation (p.i.)). However, no PSMA expression was detected by IHC in any of the MDA-MB-231\u003cem\u003ewt\u003c/em\u003e or MDA-MB-468 models (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003eC, D.). In these TNBC models inoculated with Matrigel\u0026reg;, whole-body and axial, coronal and sagittal [\u0026sup1;⁸F]-DCFPyL PET views showed a similar distribution one hour post-injection of the radiopharmaceuticals compared to the 4T1 model, with lower uptake in the salivary and lacrimal glands (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003eE-H.) (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). These results were confirmed by an \u003cem\u003eex vivo\u003c/em\u003e biodistribution study showing an uptake in the salivary glands of 0.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04 IA%/g and 0.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.61 IA%/g compared to 0.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 IA%/g for 4T1, MDA-MB-231\u003cem\u003ewt\u003c/em\u003e and MDA-MB-468 models respectively. The calculated TLRs were 0.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11 and 0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18 for MDA-MB-231\u003cem\u003ewt\u003c/em\u003e and MDA-MB-468 models respectively (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cspan type=\"BoldUnderline\" class=\"BoldUnderline\" name=\"Emphasis\"\u003eCo-inoculation of HUVEC and TNBC cells failed to induce PSMA expression in the neovasculature.\u003c/span\u003e\u003c/p\u003e\u003cp\u003ePrimary HUVEC cells were co-inoculated with TNBC cells to promote human neovascularization and potentially induce PSMA expression.\u003c/p\u003e\u003cp\u003eMDA-MB-231\u003cem\u003ewt\u003c/em\u003e and MDA-MB-231\u003cem\u003ewt\u003c/em\u003e/HUVEC cells inoculated with Matrigel\u0026reg; exhibited tumor volumes of 303\u0026thinsp;\u0026plusmn;\u0026thinsp;64 mm\u003csup\u003e3\u003c/sup\u003e (n\u0026thinsp;=\u0026thinsp;6, 100% tumor engraftment) and 242\u0026thinsp;\u0026plusmn;\u0026thinsp;103 mm\u003csup\u003e3\u003c/sup\u003e (n\u0026thinsp;=\u0026thinsp;10, 100% tumor engraftment), respectively, at 27\u0026ndash;30 days p.i. (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003eA, Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003eA).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eIt should be mentioned that tumoral tissue evidenced proliferation (Ki-67 staining) and neovascularization (CD31 staining), but no PSMA expression. Additionally, no qualitative differences in neovascularization were observed between TNBC-only and TNBC/HUVEC models (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003eC and \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003eB).\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eValidation and characterization of the PSMA-positive TNBC murine model\u003c/h3\u003e\n\u003cp\u003eDue to the lack of syngeneic or xenograft TNBC models that express PSMA, MDA-MB-231 cells were modified by lentiviral transduction to overexpressed PSMA (MDA-MB-231\u003cem\u003epsma\u003c/em\u003e). To evaluate the effects of tumor PSMA expression and mimic intratumoral heterogeneity, female mice were inoculated with 1\u0026nbsp;million MDA-MB-231\u003cem\u003epsma\u003c/em\u003e or a 1:1 mixture of 500,000 MDA-MB-231\u003cem\u003ewt\u003c/em\u003e and 500,000 MDA-MB-231\u003cem\u003epsma\u003c/em\u003e cells both of which were suspended in Matrigel\u0026reg;. These groups were compared to mice inoculated with 1\u0026nbsp;million MDA-MB-231\u003cem\u003ewt\u003c/em\u003e with Matrigel\u0026reg;. At 28\u0026ndash;30 days post-inoculation, tumor volume were 208\u0026thinsp;\u0026plusmn;\u0026thinsp;61 mm\u003csup\u003e3\u003c/sup\u003e for MDA-MB-231\u003cem\u003epsma\u003c/em\u003e, 212\u0026thinsp;\u0026plusmn;\u0026thinsp;97 mm\u003csup\u003e3\u003c/sup\u003e for the MDA-MB-231\u003cem\u003ewt\u003c/em\u003e/MDA-MB-231\u003cem\u003epsma\u003c/em\u003e models and 305\u0026thinsp;\u0026plusmn;\u0026thinsp;65 mm\u003csup\u003e3\u003c/sup\u003e for MDA-MB-231\u003cem\u003ewt\u003c/em\u003e. Tumor doubling times (DT) of MDA-MB-231\u003cem\u003epsma\u003c/em\u003e, MDA-MB-231\u003cem\u003ewt\u003c/em\u003e/MDA-MB-231\u003cem\u003epsma\u003c/em\u003e and MDA-MB-231\u003cem\u003ewt\u003c/em\u003e were 20.2\u0026thinsp;\u0026plusmn;\u0026thinsp;8.3 days; 14.7\u0026thinsp;\u0026plusmn;\u0026thinsp;12.5 days; 24.9\u0026thinsp;\u0026plusmn;\u0026thinsp;6.6 days, respectively with no significant differences (\u003cem\u003eKruskal-Wallis test\u003c/em\u003e) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e5\u003c/span\u003eA.).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003ePET imaging showed physiological accumulation of [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL in excretory organs, including the bladder and kidneys for all models (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e5\u003c/span\u003eB-E). Unlike MDA-MB-231\u003cem\u003ewt\u003c/em\u003e, both PSMA-positive models demonstrated strong tracer uptake, with TLRs of 15.1\u0026thinsp;\u0026plusmn;\u0026thinsp;3.8 for the MDA-MB-231\u003cem\u003ewt\u003c/em\u003e/MDA-MB-231\u003cem\u003epsma\u003c/em\u003e model and 17.8\u0026thinsp;\u0026plusmn;\u0026thinsp;5.2 for MDA-MB-231\u003cem\u003epsma\u003c/em\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). No radiopharmaceutical uptake was observed in central tumor regions, which is consistent with necrosis that was confirmed by dissection and H\u0026amp;E staining (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e5\u003c/span\u003eF). Necrotic areas increased as tumor developed in both PSMA-positive models: 5\u0026ndash;40% (mean\u0026thinsp;=\u0026thinsp;20% \u0026plusmn; 16%) at 21\u0026ndash;34 days p.i. to 25\u0026ndash;60% (mean\u0026thinsp;=\u0026thinsp;51% \u0026plusmn; 14%) at 42\u0026ndash;55 days p.i. Higher PSMA expression was observed near hypoxic and apoptotic tumor cores. MDA-MB-231\u003cem\u003ewt\u003c/em\u003e/MDA-MB-231\u003cem\u003epsma\u003c/em\u003e and MDA-MB-231\u003cem\u003epsma\u003c/em\u003e models were characterized by a H-score: 180\u0026thinsp;\u0026plusmn;\u0026thinsp;28 and 192\u0026thinsp;\u0026plusmn;\u0026thinsp;10 respectively. No-PSMA-positive endothelial cells were detected in any models.\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eTNBC is the most aggressive breast cancer subtype. It has recently benefited from advances in antibody-drug conjugated and immunotherapy, as well as combination [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Nevertheless, the prognosis remains poor. The oncology therapeutic arsenal has recently expanded to include TRT. Syngeneic and xenograft models are essential tools for assessing therapeutic efficacy and tumor\u0026ndash;host interactions in a reproducible, controlled setting.\u003c/p\u003e\u003cp\u003eOut of 23 murine models characterized as PSMA-negative, some were limited by poor tumor growth (BT-20 line), indicating low tumorigenicity [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. These observations have led to the establishment of the first transfected model of TNBC with PSMA expression.\u003c/p\u003e\u003cp\u003eOne observation from this experimental work is the difficulty of reproducing existing models from the literature. Shirke \u003cem\u003eet al.\u003c/em\u003e reported three models inoculated in MFP: i) 4T1 cells in BALB/c mice (10 000 cells) ii-iii) MDA-MB-231 and MDA-MB-468 in BALB/c Nude mice (1.10\u003csup\u003e6\u003c/sup\u003e cells). Interestingly, in our study, female BALB/c mice inoculated with 10 000 4T1 cells in the mammary fat pads with PBS exhibited a tumor volume of 618\u0026thinsp;\u0026plusmn;\u0026thinsp;122 mm\u003csup\u003e3\u003c/sup\u003e versus 250 mm\u003csup\u003e3\u003c/sup\u003e for Shirke \u003cem\u003eet al.\u003c/em\u003e model at the same stage [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. The tumor volume of MDA-MB-468 cells inoculated in the MFP with PBS was not indicated in this study. In 2024, Heesch \u003cem\u003eet al.\u003c/em\u003e have published a model of Rj:NMRI-\u003cem\u003eFoxn1\u003c/em\u003e\u003csup\u003e\u003cem\u003enu/nu\u003c/em\u003e\u003c/sup\u003e inoculated with 5.10\u003csup\u003e6\u003c/sup\u003e MDA-MB-231 in the MFP that underwent [\u003csup\u003e177\u003c/sup\u003e]Lu-PSMA-I\u0026amp;T therapy. Unfortunately, the tumor volume was relative (normalized to the first day of treatment) so we could not compare the tumor development of our model. Despite four reproduced models described in the literature characterized by neovascular expression of PSMA, we were unable to find PSMA expression in our model as evidenced by IHC as well as [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL PET images (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eC; Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003eE-H; Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e5\u003c/span\u003eB-E). Heesch el al. demonstrated that hypoxia can induce PSMA expression in both tumoral and endothelial cell [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Hypoxic models were harvested (subcutaneously [\u003cspan additionalcitationids=\"CR25\" citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]) by Morgenroth et al. in 2019. MDA-MB-231 cells inoculated in the right flank have resulted in tumors with [\u003csup\u003e68\u003c/sup\u003eGa]Ga-PSMA-11 uptake by PET imaging (target\u003csub\u003emean\u003c/sub\u003e to background\u003csub\u003emean\u003c/sub\u003e ratio of 43.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9) [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e], while our subcutaneous models showed no PSMA expression .\u003c/p\u003e\u003cp\u003eThis lack of reproducibility is increasingly recognized as a major issue in biomedical research, with both scientific and ethical implications notably concerning the Reduction aspect of the 3Rs (Replacement, Reduction and Refinement) [\u003cspan additionalcitationids=\"CR28 CR29\" citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eSince we were unable to reproduce these models, we investigated how to induce PSMA expression in tumor neovascularization. In the non-prostatic cancers, PSMA is thought to increase the neovascularization and invasion \u003cem\u003evia\u003c/em\u003e nitric oxide production or the FAK-PAK pathway from laminin derivatives mediated by integrin β1 activation [\u003cspan additionalcitationids=\"CR32 CR33\" citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. We hypothesize that limited vascularization in the models may impair PSMA expression. Thus, orthotopic injection appeared as the first option to increase neovascularization, followed by the addition of matrix and human endothelial cells. Moreover, previous studies have demonstrated higher radiopharmaceutical uptake in orthotopic models than in ectopic models of the same volume, which may be due to a higher vascular perfusion [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Unfortunately, Matrigel\u0026reg;, Geltrex\u0026reg; and co-inoculation of TNBC cell lines with endothelial cells did not result in PSMA expression by IHC in any of the 23 models tested.\u003c/p\u003e\u003cp\u003eConsidering the heterogeneous expression of PSMA, analyzing a single 5 \u0026micro;m tissue section in IHC study is limiting because the analyzed section may not contain regions where the signal is present, which could lead to false-negative results. Since PSMA expression is heterogeneous within tumoral tissue [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e], the absence of PSMA expression in our study could be a consequence of the tumor intra-heterogeneity. Thus, nuclear imaging could be a relevant alternative method. The main limitations of nuclear imaging are the access of the radiopharmaceuticals, unlike IHC which is widely accessible and the limited resolution of the TEP camera which can miss low levels of radiotracer accumulation in cases of low PSMA expression. As the main aim of this study was to evaluate PSMA expression \u003cem\u003eex vivo\u003c/em\u003e, only a few mice in the selected groups were injected with PSMA-targeting radiopharmaceuticals. In our case, Tumor-to-Liver ratio was consistently under one, which would be considered a PSMA-negative tumor for therapeutic clinical selection [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. As the specificity of [\u003csup\u003e18\u003c/sup\u003eF]-DCPyL to murine PSMA has to be investigated, our results showed an [\u003csup\u003e18\u003c/sup\u003eF]-DCPyL uptake of 0.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 IA%/g in the salivary gland of BALB/c mice. These results suggest that the [\u003csup\u003e18\u003c/sup\u003eF]-DCPyL may target murine PSMA, a topic that has not yet been studied to the best of our knowledge. Given the inability to induce PSMA in tumor neovessels and its lack of expression in human TNBC cells, MDA-MB-231\u003cem\u003ewt\u003c/em\u003e cells were transfected \u003cem\u003evia\u003c/em\u003e lentiviral transduction to induce PSMA expression. Transfecting cancer cells to express PSMA has already been explored for Renal Cell Carcinoma (RCC) [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. To mimic the intra-tumoral heterogeneity observed clinically, MDA-MB-231\u003cem\u003epsma\u003c/em\u003e cells were implanted alone or together with MDA-MB-231\u003cem\u003ewt\u003c/em\u003e cells. Interestingly, we observed a higher PSMA expression near the hypoxic and apoptotic core. Further investigation is needed to understand the modulation of the transgene or PSMA in these zones. Despite PSMA expression in cancer cells, we were unable to demonstrate PSMA induction in tumor neovessel endothelial cells in this model. It would be interesting to transfect endothelial cells to create a model with neovascular PSMA expression [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. If validated, this model could provide a complementary approach to address questions related to the residence time of the radiopharmaceutical in PSMA-positive tumor neovascularization and its potential impact on therapeutic efficacy.\u003c/p\u003e\u003cp\u003ePET imaging with [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL showed comparable radiotracer uptake in tumors composed solely of MDA-MB-231\u003cem\u003epsma\u003c/em\u003e cells and in tumors containing a mixture of MDA-MB-231\u003cem\u003ewt\u003c/em\u003e and MDA-MB-231\u003cem\u003epsma\u003c/em\u003e cells. These results are consistent with IHC studies in which PSMA expression heterogeneity was observed on both models. For comparison, PC3-PIP (PSMA+, generated by lentiviral transduction of the PC-3 cell line [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]), prostate cancer xenografts exhibit [\u0026sup1;⁸F]-DCFPyL uptake of 44.2\u0026thinsp;\u0026plusmn;\u0026thinsp;9.7 and 84.3\u0026thinsp;\u0026plusmn;\u0026thinsp;12.3%IA/g at 1 h p.i. [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e] whereas our MDA-MB-231\u003cem\u003ewt\u003c/em\u003e/ MDA-MB-231\u003cem\u003epsma\u003c/em\u003e and MDA-MB-231\u003cem\u003epsma\u003c/em\u003e tumors showed an uptake of 15.05\u0026thinsp;\u0026plusmn;\u0026thinsp;3.77 and 17.84\u0026thinsp;\u0026plusmn;\u0026thinsp;5.15 respectively. This is consistent with clinical observations showing lower PSMA expression in TNBC than in mCRPC.\u003c/p\u003e\u003cp\u003eFinally, \u003cem\u003eex vivo\u003c/em\u003e biodistribution and dosimetric studies with the therapeutic radiopharmaceutical, [\u003csup\u003e177\u003c/sup\u003eLu]Lu-PSMA- should be conducted before initiating TRT studies. Our data suggest that early-stage treatment (\u0026lt;\u0026thinsp;tumors less than 250 mm\u0026sup3;) may help avoid necrotic cores and improve therapeutic outcomes.\u003c/p\u003e"},{"header":"CONCLUSIONS","content":"\u003cp\u003eBased on our experience and in accordance with literature reports, the TNBC tumor model and the endothelial cell and TNBC tumor co-inoculated models display faint or no PSMA expression. The reproducibility of these models is questionable. Here, to our knowledge, we present the first PSMA-transfected TNBC model exhibiting high PSMA expression \u003cem\u003evia\u003c/em\u003e IHC and high PSMA tracer accumulation \u003cem\u003evia\u003c/em\u003e PET in TNBC tumor cells. This new model guarantees high reproducibility and is suitable for preclinical evaluation of TRT.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003e%IA/g\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003ePercentage of injected activity per gram\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eATCC\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eAmerican type culture collection\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eBC\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eBreast cancer\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eCT\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eComputed tomography\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eDT\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eDoubling time\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eFBS\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eFetal bovine serum\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eH\u0026amp;E\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eHematoxylin-eosin\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eHU\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eHounsfield units\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eIHC\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eImmunohistochemistry\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003ei.v.\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eintra-venous\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eIVIA\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eIn vivo imaging Auvergne\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eMOI\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eMultiplicity of infection\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003emCRPC\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003emetastatic castration-resistant prostate cancer\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eMDA-MB-231\u003cem\u003ewt\u003c/em\u003e\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eMDA-MB-231\u003cem\u003ewild-type\u003c/em\u003e\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003emTNBC\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003emetastatic triple-negative breast cancer\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eMFP\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eMammary fat pad\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eOCT\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eOptimal cutting temperature\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003ePBS\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003ePhosphate buffer saline\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003ePET\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003ePositron emission tomography\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003ep.i.\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003epost inoculation\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003ePSMA\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eProstate specific membrane antigen\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eRT\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eRoom temperature\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eTLM\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eTumor-to-muscle ratio\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eTLR\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eTumor-to-liver ratio\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eTNBC\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eTriple-negative breast cancer\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eTRT\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eTargeted radionuclide therapy\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eTV\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eTumor volume\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003e\u003cu\u003eETHICS APPROVAL AND CONSENT TO PARTICIPATE\u0026nbsp;\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was conducted in accordance with ARRIVE guidelines (https://arriveguidelines.org) and approved protocols outlined by the Auvergne Local Ethics Committee C2E2A and the French Ministry of Education and Research (approval numbers 49893 and 23955). No human studies were conducted. The maximum tumor size permitted by the local ethics committee, (1500 mm\u003csup\u003e3\u003c/sup\u003e), was never reached by any mouse in this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eCONSENT FOR PUBLICATION\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eAVAILABILITY OF DATA AND MATERIALS\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analyzed during this study are included in this published article [and its supplementary information files]. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eCOMPETING INTERESTS\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests related to this work\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eFUNDING\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was supported by grants from the Ligue contre le cancer comit\u0026eacute; de l\u0026rsquo;Allier et du Cantal.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eAUTHORS\u0026apos; CONTRIBUTIONS\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBC and EJ performed \u003cem\u003ein vivo\u003c/em\u003e experiments, analyzed data, and interpreted results. LS, EB, SB, MA and AV participated \u003cem\u003ein vivo\u003c/em\u003e experiments. MK performed histological and immunohistochemical analysis. CMo and JD performed lentiviral transduction of TNBC cell lines. SL, AR, MT and SS performed (radio)chemical analysis. BC, EJ, FC drafted the first draft of the manuscript. EC, EMN, FPL, CMe, FC and EJ supervised and contributed intellectually to the work and carefully revised the manuscript. All authors read, revised and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eACKNOWLEDGEMENTS\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe would like to thank Christelle Soubeyrand-Damon and Anipath histopathology facility (Institute of Genetics, Reproduction and Development, UMR 6293 CNRS/U1103 INSERM/Clermont Auvergne University) for their technical assistance with the histochemical studies. We would also like to thank Christelle Blavignac and Cl\u0026eacute;mence Chapuis, Centre Imagerie Cellulaire Sante _UCA PARTNER, for their technical support and expertise in flow cytometry. We would like to thank CURIUM France (Saint-Beauzire, France) for supplying the [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL, that made it possible to perform PET/CT scans on mouse models.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe authors wish to acknowledge the IVIA platform (In Vivo Imaging Auvergne) at Clermont-Ferrand (France). We would like to thank Marjorie Maillet (master\u0026rsquo;s student) during her internship to this work.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eYin, L., Duan, J-J., Bian, X-W. \u0026amp; Yu, S. 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Chem.\u003c/em\u003e \u003cb\u003e27\u003c/b\u003e, 1655\u0026ndash;1662 (2016).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Targeted Radionuclide Therapy, Triple-Negative Breast Cancer, Prostate-Specific Membrane Antigen, Syngeneic model, Xenograft model, Orthotopic model, Lentiviral transduction","lastPublishedDoi":"10.21203/rs.3.rs-7904335/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7904335/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003eTriple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer, for which targeted therapies are emerging. Prostate-specific membrane antigen (PSMA), expressed mainly by the endothelial cells of the neovascularization of TNBC, or tumor cells, is an interesting target in TNBC that needs to be studied. This study aims to validate a reproductible, PSMA-positive murine model for Targeted Radionuclide Therapy (TRT) assessment.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eTwenty-five syngeneic or xenograft murine models were produced by modifying the TNBC cell line, the implantation site (ectopic and orthotopic), cell numbers and matrix (e.g Matrigel\u0026reg;) use. Tumor growth and engraftment were recorded. PSMA expression was evaluated histologically using immunohistochemistry (IHC) and PSMA-targeted positron emission tomography (PET)- computed tomography (CT) with [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL. The tumor-to-liver ratio (TLR) was used to quantify the uptake of the radiopharmaceutical. To address the absence of PSMA expression in native models, MDA-MB-231 cells were genetically modified \u003cem\u003evia\u003c/em\u003e lentiviral transduction to overexpress PSMA for \u003cem\u003ein vivo\u003c/em\u003e evaluation.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eDespite high cellular proliferation and extensive tumor neovascularization, immunohistochemical analyses revealed an absence of PSMA expression in all non-transfected models tested. This finding was further confirmed by [\u0026sup1;⁸F]-DCFPyL PET/CT imaging, which showed a TLR below 1, indicating negligible radiotracer uptake within tumoral tissues. In contrast, the MDA-MB-231\u003cem\u003epsma\u003c/em\u003e model achieved 100% tumor engraftment and a tumor volume of 208\u0026thinsp;\u0026plusmn;\u0026thinsp;61 mm\u003csup\u003e3\u003c/sup\u003e at 28 days post-injection (not significantly different from the MDA-MB-231\u003cem\u003ewt\u003c/em\u003e model). This model showed a TLR of 10.3\u0026thinsp;\u0026plusmn;\u0026thinsp;4.3 one hour after intravenous injection of [\u003csup\u003e18\u003c/sup\u003eF]-DCFPyL.\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e\u003cp\u003eThis study emphasizes the challenge of creating a reproducible murine model with PSMA expression. Nevertheless, the initial transfection of MDA-MB-231 cells to express PSMA resulted in the development of an innovative TNBC murine model that expresses PSMA. This model is reproducible and exhibits heterogeneous PSMA expression, and is representative of clinical observations. It is also suitable for evaluating PSMA-targeted TRT in TNBC.\u003c/p\u003e","manuscriptTitle":"From Hurdles to Breakthroughs: Building and Validating a PSMA-positive Triple-Negative Breast Cancer Mouse Model for Preclinical Targeted Radionuclide Therapy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-11 16:28:48","doi":"10.21203/rs.3.rs-7904335/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-11-26T19:11:13+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-26T16:19:38+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-24T04:33:58+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-21T02:49:47+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"103507849812019445530567235113431796051","date":"2025-11-04T14:41:48+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"188025571443284393625839298838779371899","date":"2025-11-01T14:57:25+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"179484051687334069894802290837455566993","date":"2025-10-31T00:50:13+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-10-30T13:20:39+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-10-30T13:11:45+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-10-30T12:50:02+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-10-27T15:56:07+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-10-27T15:51:12+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"05011d11-891e-41da-861b-5309a09633bc","owner":[],"postedDate":"November 11th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":57432789,"name":"Biological sciences/Cancer"},{"id":57432790,"name":"Health sciences/Oncology"}],"tags":[],"updatedAt":"2026-02-23T16:01:02+00:00","versionOfRecord":{"articleIdentity":"rs-7904335","link":"https://doi.org/10.1038/s41598-026-36724-7","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2026-02-16 15:57:32","publishedOnDateReadable":"February 16th, 2026"},"versionCreatedAt":"2025-11-11 16:28:48","video":"","vorDoi":"10.1038/s41598-026-36724-7","vorDoiUrl":"https://doi.org/10.1038/s41598-026-36724-7","workflowStages":[]},"version":"v1","identity":"rs-7904335","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7904335","identity":"rs-7904335","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}