Membrane-bound IL-7 Engineered TIL therapy for advanced ovarian cancer

Membrane-bound IL-7 Engineered TIL therapy for advanced ovarian cancer | 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 Case Report Membrane-bound IL-7 Engineered TIL therapy for advanced ovarian cancer Jing Guo, Guihai Ai, Chunyan Wang, Wei Huang, Yuliang Wu, Jihui Zhu, and 10 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4102406/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Ovarian cancer lacks effective therapy. Here, we reported three metastatic ovarian cancer patients administrated with a noval TIL therapy, which was designed glycosylphosphatidylinositol-anchored membrane-bound interleukin-7 (mbIL-7-GPI) to engineer TILs (mbIL-7-TIL) through piggyBac transposon system. In three advanced ovarian cancer patients, infusion of mbIL-7-TILs showed endurable toxicity and prolonged clinical response. After infusion, peripheral mbIL-7-TILs peaked around 5-7 days and then sharply decreased. We found that several clones of engineered T cell and subsequent clones of original T cells underwent significant proliferation in patient 1. Seven months post infusion, mbIL-7-TILs could be detected in tumor liquefactive necrosis of patient 2. These findings indicated that mbIL-7-TILs could efficiently home to tumor lesions and sustainably enhance antitumor immunity in situ, suggesting potent therapeutics to treat advanced OC. epithelial ovarian cancer adoptive cell therapy tumor-infiltrating lymphocytes gene-engineered Figures Figure 1 Figure 2 Introduction There is no effective therapy for ovarian cancer, although the development of targeted drugs. ACT with ex vivo expanded TILs has shown durable objective responses in patients with other solid tumors1, however, its effectiveness is still limited in treating ovarian cancer2. IL-7, known for its role in promoting immune cell proliferation, extending the lifespan of activated immune cells, and increasing the ratio of memory cells3. To improve TIL antitumor activity, we designed glycosylphosphatidylinositol-anchored membrane-bound interleukin-7 (mbIL-7-GPI) to engineer TILs (mbIL-7-TIL) through piggyBac transposon system (Figure S1). Here, we reported three participants, all diagnosed with advanced ovarian cancer, have achieved a partial response after infusion of mbIL-7-TIL according to RECIST criteria. Case presentation Patient characteristics All patients received intravenous infusion of genetically engineered mbIL-7-TIL following a 3-day lymphodepletion preconditioning (cyclophosphamide 20mg/kg/day every day and hydroxychloroquine 600mg on day one). This was followed by one-time infusion of PD-1 antibody (100 mg, sintilimab) and mbIL-7-TIL. Table 1 provides a summary of the patient characteristics, cell doses, and clinical findings of the three sequentially treated patients with metastatic OC in this trial. The patients were in their 30s, 40s, and 60s, respectively. All of them had undergone extensive pretreatment, including one patient who had previously received natural TIL therapy. The patients were discharged from the hospital within two weeks after infusion therapy. Table 1. Patient characteristic Value Patient 1 Patient 2 Patient 3 Baseline Histology Epithelial ovarian cancer High-grade serous ovarian cancer High-grade serous ovarian cancer ECOG 1 1 1 Sites of Disease pelvic, lung, liver liver, spleen peritoneum Clinical stages T T1 T3 T4 M N0 N0 N1 N M0 M0 M1 Target lesion sum of diameter (mm) 56 34 45 Mutation BRCA1/2 (-), PD-L (-) BRCA1/2 (-), PD-L (-) BRCA1/2 (-), PD-L (-) Prior Systemic Treatment Surgery yes yes yes Chemotherapy yes yes yes Radiotherapy yes no no Targeted drugs bevacizumab apatinib no PARP inhibitor niraparib no niraparib Endocrine therapy no no no Properties of the infused mbIL-7 TILs Infused Cells (×10 9 ) 1.91×10 9 1.96×10 9 1.67×10 9 CD8 + /CD3 + (%) 81% 59% 64% Expressed mbIL-7/ CD3 + (%) 26% 28% 45% Post-infusion of mbIL-7 TILs Follow up (days) 90* 215 167 Hospital stay (days) 12 12 12 Clinical response PR PR PR Adverse event (Grade) Fever 1 1 0 Leukopenia 4 2 2 Lymphopenia 4 3 3 Neutropenia 4 3 1 Anemia 2 4 3 Thrombocytopenia 0 0 0 mbIL-7, membrane-bound IL-7 TIL; *, died of covid-19 infection. Properties of infusion product All patients generated adequate mIL-7 TILs, with 26%, 28%, and 45% of cells showing positive transduction for mbIL-7 in 1.91×10 9 , 1.96×10 9 , and 1.67×10 9 cells, respectively (Table 1, Figure S1). In each cell culture, the phenotype of mIL-7-engineered TILs was compared to TILs treated identically but without transduction. The infused mIL-7 TILs phenotypically resembled memory T cells, as they were negative for CD45RA and partly expressed CCR7, CD62L, and CD69 (Figure S3). The transduced T cells increased and peaked five days after infusion and sharply decreased in the peripheral blood 14 days after cell transfer (Fig. 2A, 2C, 2E). The frequency of five distinct TIL clones, which were filtered based on their significant frequency in each patient's peripheral blood, persisted following cell infusion (Figure S1B, D, F, blue). Additionally, the frequency of endogenous TCR clones that were not shared by TIL clones in peripheral blood increased following cell infusion (Figure S1B, D, F, red). Clinical responses and safety following infusion of mIL-7 TILs All three participants achieved a partial response according to RECIST criteria, and tumor regression is ongoing at the latest follow-up (Table 1 and Figure 1). Notably, patient 1 had previously received conventional non-transduced TIL therapy, along with a low dosage of PD1 antibody in a previous clinical trial (NCT04766320) 4 , but experienced tumor progression within six months. However, with the use of mIL-7 TILs, there is an ongoing partial regression at three months in the metastatic disease to the lung and liver (Fig. 1A). Patient 2 had experienced pseudoprogression within five months after infusion of mIL-7 TILs in the metastatic lesions of liver and spleen (Fig. 1B). However, the lesions had necrosis after six months with the use of mIL-7 TILs, and the biopsy confirmed necrosis with high level of mIL-7 (Figure S4). Patient 3 had experienced partial response after 3 months, and the lesion had necrosis change based on image after six months with the use of mIL-7 TILs (Figure 1I). All patients who received the initial cell infusion showed favorable safety profiles (Table 1). Hematopoietic reconstitution occurred around two weeks with a similar pattern to conventional TIL therapy using the same lymphodepleting regimen (Figure S2). The concentrations of effector cytokines (e.g., IP-10) secreted into the supernatant after cell infusion remained stable, while the levels of cytokines (e.g., IL-6, IL-7, and IL-8) remained low after cell infusion (Fig. 2B, D, F). Discussion Administration of recombinant human IL-7 has been shown to induce T cell proliferation, increase T cell numbers, modulate peripheral T cell subsets, and enhance the diversity of the T cell receptor repertoire. these findings support the hypothesis that mbIL-7-TILs traffic to and reside in tumors, correlating with favorable clinical outcomes. The mbIL-7 molecules possess a glycosylphosphatidylinositol (GPI) structure, unlike conventional membrane-anchored cytokines incorporating a single transmembrane domain 9 . This GPI structure mediates clustering of IL-7 cytokines on the plasma membrane surface, enhancing the stimulatory capacity of mbIL-7-expressing TILs on surrounding immune cells while reducing stimulation on peripheral immune cells due to the strong homing ability of the genetically engineered T cells. Compared to genetically engineered TILs with an inducible gene encoding a secretory cytokine, which has been associated with severe dose-limiting toxicity 10 , engineered TILs with a GPI membrane-anchored structure could mitigate toxicity, as indicated by the low levels of cytokines in serum (Fig. 2 ). Notably, mbIL-7-TILs showed efficacy in patient 1, who had been refractory to natural TIL therapy. Data from Figs. 2 A, C, E demonstrate the extended survival of mbIL-7-TILs, with detectable levels maintained for at least 90 days. This prolonged presence of mbIL-7-TILs may be a contributing factor to the excellent duration of the clinical response (Fig. 1 ). Meanwhile, the level of tumor biomarker decreased in Patient 1 and Patient 3. However, Patient 2 exhibited a continuous increase in CA125 and HE4, likely a result of the tumor's large size (Figure S1 B, D, F). Importantly, the infusion of T cells engineered with mbIL-7 resulted in minimal toxic effects (Table 1 ). The expected transient toxic effects observed included fever resulting from the infusion product and myelosuppression, which are commonly associated with cyclophosphamide preconditioning therapy. As shown in Fig. 1 B, 1 F, and 1 J, the patients recovered from cytopenia within two weeks. Collectively, these findings suggest that the infusion of mbIL-7-TILs can effectively activate endogenous tumor-reactive T cells in the tumor microenvironment without causing serious adverse effects. It is important to note that the sample size in this study was limited to only three patients, as the clinical trial was still ongoing. However, the promising outcomes of this study provide encouragement to present our findings and give us the confidence to continue the trial in order to bring benefits to more patients. Declarations Ethical approval and consent to participate This study involves human participants was approved by the local ethic committee of Shanghai Tenth People's Hospital. Participants gave informed consent to participate in the study before taking part. Competing interests The authors declare that they have no competing interests. Acknowledgements We thank the nurses for their great work from Department of Obstetrics and Gynecology, Shanghai Tenth People's Hospital. We also thank the patients who participated in the clinical study. Authors’ contributions Concept or study design: Z.C., B.Z., and H.J.. Acquisition of data: J.G., G.A., C.W.,W.H., Y.W., J.Z., W.H., J.D., N.L., X.S., L.L., Y.G., C.L.. Analysis: J.G., G.A., C.W., Y.W., and B.Z.. Drafting the work or revising it critically for important intellectual content: J.G., Z.C., and H.J.. Funding This study was supported by National Natural Sciences Foundation of China (82103337, to J Guo; 82373269, to Z. Cheng); Science and Technology Commission of Shanghai Municipality (22XD1432200). Availability of data and materials Data of the three patients presented here were applicable. Conflict of interest: The authors declare no potential conflicts of interest. References Sarnaik AA, Hamid O, Khushalani NI, et al. Lifileucel, a Tumor-Infiltrating Lymphocyte Therapy, in Metastatic Melanoma. J Clin Oncol 2021;39:2656-66. Sarivalasis A, Morotti M, Mulvey A, Imbimbo M, Coukos G. Cell therapies in ovarian cancer. Ther Adv Med Oncol 2021;13:17588359211008399. Barata JT, Durum SK, Seddon B. Flip the coin: IL-7 and IL-7R in health and disease. Nat Immunol 2019;20:1584-93. Guo J, Luo N, Ai G, et al. Eradicating tumor in a recurrent cervical cancer patient with autologous tumor-infiltrating lymphocytes and a modified lymphodepleting regimen. J Immunother Cancer 2022;10. Ren H, Cao K, Wang M. A Correlation Between Differentiation Phenotypes of Infused T Cells and Anti-Cancer Immunotherapy. Front Immunol 2021;12:745109. Sackstein R, Schatton T, Barthel SR. T-lymphocyte homing: an underappreciated yet critical hurdle for successful cancer immunotherapy. Lab Invest 2017;97:669-97. Russo E, Santoni A, Bernardini G. Tumor inhibition or tumor promotion? The duplicity of CXCR3 in cancer. J Leukoc Biol 2020;108:673-85. House IG, Savas P, Lai J, et al. Macrophage-Derived CXCL9 and CXCL10 Are Required for Antitumor Immune Responses Following Immune Checkpoint Blockade. Clin Cancer Res 2020;26:487-504. Sharma P, Varma R, Sarasij RC, et al. Nanoscale organization of multiple GPI-anchored proteins in living cell membranes. Cell 2004;116:577-89. Zhang L, Morgan RA, Beane JD, et al. Tumor-infiltrating lymphocytes genetically engineered with an inducible gene encoding interleukin-12 for the immunotherapy of metastatic melanoma. Clin Cancer Res 2015;21:2278-88. Additional Declarations No competing interests reported. Supplementary Files SupplementaryMethods.docx Slide3.png Slide4.png Slide5.png Slide6.png Slide7.png Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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16:39:02","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4102406/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4102406/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":53016814,"identity":"e831ffb1-d9f7-4ca1-9353-65bbebc40fa7","added_by":"auto","created_at":"2024-03-19 16:06:37","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":225267,"visible":true,"origin":"","legend":"\u003cp\u003eClinical responses following infusion of mbIL-7 TIL. The contrast-enhanced computed tomographic scans.\u003c/p\u003e","description":"","filename":"Slide1.png","url":"https://assets-eu.researchsquare.com/files/rs-4102406/v1/9a8738fc943e474fc78a010d.png"},{"id":53016815,"identity":"be554068-d206-4beb-b0f2-0fb377ded339","added_by":"auto","created_at":"2024-03-19 16:06:38","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":121593,"visible":true,"origin":"","legend":"\u003cp\u003eA, C, E showed the transduced TIL in the peripheral blood as determined on PCR; B, D, F showed concentrations of the cytokines IL-7, IL-10, IL-2, 4,6, 8, 10, TNF, interferon-γ and IP-10 in the serum of the patient before cell therapy on day 0 and day 90 after cell therapy.\u003c/p\u003e","description":"","filename":"Slide2.png","url":"https://assets-eu.researchsquare.com/files/rs-4102406/v1/17b55714a45e5333c87dec0d.png"},{"id":53110023,"identity":"88c692e9-9e14-43e0-b6bb-b119385e50a2","added_by":"auto","created_at":"2024-03-20 17:20:21","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":547012,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4102406/v1/85e43537-5d9b-42e3-8ee4-b55bc7643d2e.pdf"},{"id":53016818,"identity":"c26caeae-751a-4294-a7ee-e16f518a2fb5","added_by":"auto","created_at":"2024-03-19 16:06:38","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":17854,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMethods.docx","url":"https://assets-eu.researchsquare.com/files/rs-4102406/v1/b20ff825646eee0cd54f6840.docx"},{"id":53016821,"identity":"4adfe74c-f40f-4161-93fb-b7713834feaf","added_by":"auto","created_at":"2024-03-19 16:06:38","extension":"png","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":138542,"visible":true,"origin":"","legend":"","description":"","filename":"Slide3.png","url":"https://assets-eu.researchsquare.com/files/rs-4102406/v1/b5f02d2c68b7f48aa29c4bd0.png"},{"id":53018024,"identity":"34c6180e-246c-4854-9fba-2eb09ae1b888","added_by":"auto","created_at":"2024-03-19 16:14:38","extension":"png","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":111717,"visible":true,"origin":"","legend":"","description":"","filename":"Slide4.png","url":"https://assets-eu.researchsquare.com/files/rs-4102406/v1/4533141b40912a154aefc5a9.png"},{"id":53016817,"identity":"8832680f-175e-47fe-a67f-544dd33f2625","added_by":"auto","created_at":"2024-03-19 16:06:38","extension":"png","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":127386,"visible":true,"origin":"","legend":"","description":"","filename":"Slide5.png","url":"https://assets-eu.researchsquare.com/files/rs-4102406/v1/013915d1c0a4a2305b49d182.png"},{"id":53016820,"identity":"f59167e9-2f08-421c-9a82-b256f9bfe0eb","added_by":"auto","created_at":"2024-03-19 16:06:38","extension":"png","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":10054,"visible":true,"origin":"","legend":"","description":"","filename":"Slide6.png","url":"https://assets-eu.researchsquare.com/files/rs-4102406/v1/4ccba6ff7a851cfaa14d2906.png"},{"id":53016823,"identity":"0ee2a0c8-d70f-4cbc-a292-4c2c29651991","added_by":"auto","created_at":"2024-03-19 16:06:38","extension":"png","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":147184,"visible":true,"origin":"","legend":"","description":"","filename":"Slide7.png","url":"https://assets-eu.researchsquare.com/files/rs-4102406/v1/f22c98b47ffadac1595ecc82.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Membrane-bound IL-7 Engineered TIL therapy for advanced ovarian cancer","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThere is no effective therapy for ovarian cancer, although the development of targeted drugs. ACT with ex vivo expanded TILs has shown durable objective responses in patients with other solid tumors1, however, its effectiveness is still limited in treating ovarian cancer2. IL-7, known for its role in promoting immune cell proliferation, extending the lifespan of activated immune cells, and increasing the ratio of memory cells3. To improve TIL antitumor activity, we designed glycosylphosphatidylinositol-anchored membrane-bound interleukin-7 (mbIL-7-GPI) to engineer TILs (mbIL-7-TIL) through piggyBac transposon system (Figure S1). Here, we reported three participants, all diagnosed with advanced ovarian cancer, have achieved a partial response after infusion of mbIL-7-TIL according to RECIST criteria.\u003c/p\u003e"},{"header":"Case presentation","content":"\u003cp\u003e\u003cstrong\u003ePatient characteristics\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll patients received intravenous infusion of genetically engineered\u0026nbsp;mbIL-7-TIL\u0026nbsp;following a 3-day lymphodepletion preconditioning (cyclophosphamide 20mg/kg/day every day and hydroxychloroquine 600mg on day one). This was followed by one-time infusion of PD-1 antibody (100 mg, sintilimab) and mbIL-7-TIL.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 1 provides a summary of the patient characteristics, cell doses, and clinical findings of the three sequentially treated patients with metastatic OC in this trial. The patients were in their 30s, 40s, and 60s, respectively. All of them had undergone extensive pretreatment, including one patient who had previously received natural TIL therapy. The patients were discharged from the hospital within two weeks after infusion therapy.\u003c/p\u003e\n\u003cp\u003eTable 1. Patient characteristic\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eValue\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003ePatient 1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003ePatient 2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003ePatient 3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eBaseline\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eHistology\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003eEpithelial ovarian cancer\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eHigh-grade serous ovarian cancer\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eHigh-grade serous ovarian cancer\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eECOG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eSites of Disease\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003epelvic, lung, liver\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eliver, spleen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eperitoneum\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eClinical stages\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003eT1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eT3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eT4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003eN0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eN0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eN1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003eM0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eM0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eM1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eTarget lesion sum of diameter (mm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eMutation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003eBRCA1/2 (-), PD-L (-)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eBRCA1/2 (-),\u0026nbsp;\u003c/p\u003e\n \u003cp\u003ePD-L (-)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eBRCA1/2 (-),\u0026nbsp;\u003c/p\u003e\n \u003cp\u003ePD-L (-)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003ePrior Systemic Treatment\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eSurgery\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003eyes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eyes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eyes\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eChemotherapy\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003eyes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eyes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eyes\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eRadiotherapy\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003eyes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eno\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eno\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eTargeted drugs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003ebevacizumab\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eapatinib\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eno\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003ePARP inhibitor\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003eniraparib\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eno\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eniraparib\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eEndocrine therapy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003eno\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eno\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003eno\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"48.97959183673469%\" colspan=\"2\"\u003e\n \u003cp\u003eProperties of the infused mbIL-7 TILs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eInfused Cells (\u0026times;10\u003csup\u003e9\u003c/sup\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e1.91\u0026times;10\u003csup\u003e9\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e1.96\u0026times;10\u003csup\u003e9\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e1.67\u0026times;10\u003csup\u003e9\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eCD8\u003csup\u003e+\u003c/sup\u003e/CD3\u003csup\u003e+\u003c/sup\u003e (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e81%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e59%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e64%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eExpressed mbIL-7/ CD3\u003csup\u003e+\u003c/sup\u003e (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e26%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e28%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e45%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003ePost-infusion of mbIL-7 TILs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eFollow up (days)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e90*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e215\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e167 \u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eHospital stay (days)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eClinical response\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003ePR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003ePR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003ePR\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eAdverse event (Grade)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eFever\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eLeukopenia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eLymphopenia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eNeutropenia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eAnemia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"29.591836734693878%\"\u003e\n \u003cp\u003eThrombocytopenia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"19.387755102040817%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.510204081632654%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003embIL-7, membrane-bound IL-7 TIL; *, died of covid-19 infection.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eProperties of\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003einfusion product\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll patients generated adequate mIL-7 TILs, with 26%, 28%, and 45% of cells showing positive transduction for mbIL-7 in 1.91\u0026times;10\u003csup\u003e9\u003c/sup\u003e, 1.96\u0026times;10\u003csup\u003e9\u003c/sup\u003e, and 1.67\u0026times;10\u003csup\u003e9\u003c/sup\u003e cells, respectively (Table 1, Figure S1).\u003c/p\u003e\n\u003cp\u003eIn each cell culture, the phenotype of mIL-7-engineered TILs was compared to TILs treated identically but without transduction. The infused mIL-7 TILs phenotypically resembled memory T cells, as they were negative for CD45RA and partly expressed CCR7, CD62L, and CD69 (Figure S3).\u003c/p\u003e\n\u003cp\u003eThe transduced T cells increased and peaked five days after infusion and sharply decreased in the peripheral blood 14 days after cell transfer (Fig. 2A, 2C, 2E). The frequency of five distinct TIL clones, which were filtered based on their significant frequency in each patient\u0026apos;s peripheral blood, persisted following cell infusion (Figure S1B, D, F, blue). Additionally, the frequency of endogenous TCR clones that were not shared by TIL clones in peripheral blood increased following cell infusion (Figure S1B, D, F, red).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical responses and safety following infusion of mIL-7 TILs\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll three participants achieved a partial response according to RECIST criteria, and tumor regression is ongoing at the latest follow-up (Table 1 and Figure 1). Notably, patient 1 had previously received conventional non-transduced TIL therapy, along with a low dosage of PD1 antibody in a previous clinical trial (NCT04766320)\u003csup\u003e4\u003c/sup\u003e, but experienced tumor progression within six months. However, with the use of mIL-7 TILs, there is an ongoing partial regression at three months in the metastatic disease to the lung and liver (Fig. 1A). Patient 2 had experienced pseudoprogression within five months after infusion of mIL-7 TILs in the metastatic lesions of liver and spleen (Fig. 1B). However, the lesions had necrosis after six months with the use of mIL-7 TILs, and the biopsy confirmed necrosis with high level of mIL-7 (Figure S4). Patient 3 had experienced partial response after 3 months, and the lesion had necrosis change based on image after six months with the use of mIL-7 TILs (Figure 1I).\u003c/p\u003e\n\u003cp\u003eAll patients who received the initial cell infusion showed favorable safety profiles (Table 1). Hematopoietic reconstitution occurred around two weeks with a similar pattern to conventional TIL therapy using the same lymphodepleting regimen (Figure S2).\u003c/p\u003e\n\u003cp\u003eThe concentrations of effector cytokines (e.g., IP-10) secreted into the supernatant after cell infusion remained stable, while the levels of cytokines (e.g., IL-6, IL-7, and IL-8) remained low after cell infusion (Fig. 2B, D, F).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eAdministration of recombinant human IL-7 has been shown to induce T cell proliferation, increase T cell numbers, modulate peripheral T cell subsets, and enhance the diversity of the T cell receptor repertoire. these findings support the hypothesis that mbIL-7-TILs traffic to and reside in tumors, correlating with favorable clinical outcomes.\u003c/p\u003e \u003cp\u003eThe mbIL-7 molecules possess a glycosylphosphatidylinositol (GPI) structure, unlike conventional membrane-anchored cytokines incorporating a single transmembrane domain\u003csup\u003e9\u003c/sup\u003e. This GPI structure mediates clustering of IL-7 cytokines on the plasma membrane surface, enhancing the stimulatory capacity of mbIL-7-expressing TILs on surrounding immune cells while reducing stimulation on peripheral immune cells due to the strong homing ability of the genetically engineered T cells. Compared to genetically engineered TILs with an inducible gene encoding a secretory cytokine, which has been associated with severe dose-limiting toxicity\u003csup\u003e10\u003c/sup\u003e, engineered TILs with a GPI membrane-anchored structure could mitigate toxicity, as indicated by the low levels of cytokines in serum (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Notably, mbIL-7-TILs showed efficacy in patient 1, who had been refractory to natural TIL therapy.\u003c/p\u003e \u003cp\u003eData from Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eA, C, E demonstrate the extended survival of mbIL-7-TILs, with detectable levels maintained for at least 90 days. This prolonged presence of mbIL-7-TILs may be a contributing factor to the excellent duration of the clinical response (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Meanwhile, the level of tumor biomarker decreased in Patient 1 and Patient 3. However, Patient 2 exhibited a continuous increase in CA125 and HE4, likely a result of the tumor's large size (Figure \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003eB, D, F).\u003c/p\u003e \u003cp\u003eImportantly, the infusion of T cells engineered with mbIL-7 resulted in minimal toxic effects (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The expected transient toxic effects observed included fever resulting from the infusion product and myelosuppression, which are commonly associated with cyclophosphamide preconditioning therapy. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003eB, \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003eF, and \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003eJ, the patients recovered from cytopenia within two weeks.\u003c/p\u003e \u003cp\u003eCollectively, these findings suggest that the infusion of mbIL-7-TILs can effectively activate endogenous tumor-reactive T cells in the tumor microenvironment without causing serious adverse effects.\u003c/p\u003e \u003cp\u003eIt is important to note that the sample size in this study was limited to only three patients, as the clinical trial was still ongoing. However, the promising outcomes of this study provide encouragement to present our findings and give us the confidence to continue the trial in order to bring benefits to more patients.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthical approval and consent to participate\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study involves human participants was approved by the local ethic committee of Shanghai Tenth People\u0026apos;s Hospital. Participants gave informed consent to participate in the study before taking part.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank the nurses for their great work from Department of Obstetrics and Gynecology, Shanghai Tenth People\u0026apos;s Hospital. We also thank the patients who participated in the clinical study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eConcept or study design: Z.C., B.Z., and H.J.. Acquisition of data: J.G., G.A., C.W.,W.H., Y.W., J.Z., W.H., J.D., N.L., X.S., L.L., Y.G., C.L.. Analysis: J.G., G.A., C.W., Y.W., and B.Z.. Drafting the work or revising it critically for important intellectual content: J.G., Z.C., and H.J..\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was supported by National Natural Sciences Foundation of China (82103337, to J Guo; 82373269, to Z. Cheng); Science and Technology Commission of Shanghai Municipality (22XD1432200).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData of the three patients presented here were applicable.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConflict of interest:\u003c/strong\u003e The authors declare no potential conflicts of interest.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eSarnaik AA, Hamid O, Khushalani NI, et al. Lifileucel, a Tumor-Infiltrating Lymphocyte Therapy, in Metastatic Melanoma. J Clin Oncol 2021;39:2656-66.\u003c/li\u003e\n\u003cli\u003eSarivalasis A, Morotti M, Mulvey A, Imbimbo M, Coukos G. Cell therapies in ovarian cancer. Ther Adv Med Oncol 2021;13:17588359211008399.\u003c/li\u003e\n\u003cli\u003eBarata JT, Durum SK, Seddon B. Flip the coin: IL-7 and IL-7R in health and disease. Nat Immunol 2019;20:1584-93.\u003c/li\u003e\n\u003cli\u003eGuo J, Luo N, Ai G, et al. Eradicating tumor in a recurrent cervical cancer patient with autologous tumor-infiltrating lymphocytes and a modified lymphodepleting regimen. J Immunother Cancer 2022;10.\u003c/li\u003e\n\u003cli\u003eRen H, Cao K, Wang M. A Correlation Between Differentiation Phenotypes of Infused T Cells and Anti-Cancer Immunotherapy. Front Immunol 2021;12:745109.\u003c/li\u003e\n\u003cli\u003eSackstein R, Schatton T, Barthel SR. T-lymphocyte homing: an underappreciated yet critical hurdle for successful cancer immunotherapy. Lab Invest 2017;97:669-97.\u003c/li\u003e\n\u003cli\u003eRusso E, Santoni A, Bernardini G. Tumor inhibition or tumor promotion? The duplicity of CXCR3 in cancer. J Leukoc Biol 2020;108:673-85.\u003c/li\u003e\n\u003cli\u003eHouse IG, Savas P, Lai J, et al. Macrophage-Derived CXCL9 and CXCL10 Are Required for Antitumor Immune Responses Following Immune Checkpoint Blockade. Clin Cancer Res 2020;26:487-504.\u003c/li\u003e\n\u003cli\u003eSharma P, Varma R, Sarasij RC, et al. Nanoscale organization of multiple GPI-anchored proteins in living cell membranes. Cell 2004;116:577-89.\u003c/li\u003e\n\u003cli\u003eZhang L, Morgan RA, Beane JD, et al. Tumor-infiltrating lymphocytes genetically engineered with an inducible gene encoding interleukin-12 for the immunotherapy of metastatic melanoma. Clin Cancer Res 2015;21:2278-88.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"epithelial ovarian cancer, adoptive cell therapy, tumor-infiltrating lymphocytes, gene-engineered","lastPublishedDoi":"10.21203/rs.3.rs-4102406/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4102406/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eOvarian cancer lacks effective therapy. Here, we reported three metastatic ovarian cancer patients administrated with a noval TIL therapy, which was designed glycosylphosphatidylinositol-anchored membrane-bound interleukin-7 (mbIL-7-GPI) to engineer TILs (mbIL-7-TIL) through piggyBac transposon system. In three advanced ovarian cancer patients, infusion of mbIL-7-TILs showed endurable toxicity and prolonged clinical response. After infusion, peripheral mbIL-7-TILs peaked around 5-7 days and then sharply decreased. We found that several clones of engineered T cell and subsequent clones of original T cells underwent significant proliferation in patient 1. Seven months post infusion, mbIL-7-TILs could be detected in tumor liquefactive necrosis of patient 2. These findings indicated that mbIL-7-TILs could efficiently home to tumor lesions and sustainably enhance antitumor immunity in situ, suggesting potent therapeutics to treat advanced OC.\u003c/p\u003e","manuscriptTitle":"Membrane-bound IL-7 Engineered TIL therapy for advanced ovarian cancer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-19 16:06:32","doi":"10.21203/rs.3.rs-4102406/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"83b463d4-4ef4-4c5f-9048-deb043b71119","owner":[],"postedDate":"March 19th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-03-20T17:12:14+00:00","versionOfRecord":[],"versionCreatedAt":"2024-03-19 16:06:32","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4102406","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4102406","identity":"rs-4102406","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}