{"paper_id":"ccf0c9fa-11f2-484d-91c3-60416635d00e","body_text":"Exosome-Derived lncRNA LIPE-AS1 Enhances Oocytes Maturation and Ameliorates Diminished Ovarian Reserve via the miR-330-5p/HDAC3 Axis\nJialing Li\nReproductive Medicine Center, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nNingxia Key Laboratory of Clinical and Pathogenic Microbiology, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorHua Guo\nDepartment of Gynecology, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorMiaomiao Tian\nNingxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorFeimiao Wang\nReproductive Medicine Center, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorJinmei Gao\nNingxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorJie Ma\nDepartment of Gynecology, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorCorresponding Author\nRong Hu\nReproductive Medicine Center, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nInstitute of Medical Sciences, General Hospital of Ningxia Medical University, Ningxia, China\nCorrespondence: Rong Hu ([email protected])\nSearch for more papers by this authorJialing Li\nReproductive Medicine Center, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nNingxia Key Laboratory of Clinical and Pathogenic Microbiology, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorHua Guo\nDepartment of Gynecology, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorMiaomiao Tian\nNingxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorFeimiao Wang\nReproductive Medicine Center, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorJinmei Gao\nNingxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorJie Ma\nDepartment of Gynecology, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nSearch for more papers by this authorCorresponding Author\nRong Hu\nReproductive Medicine Center, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, China\nInstitute of Medical Sciences, General Hospital of Ningxia Medical University, Ningxia, China\nCorrespondence: Rong Hu ([email protected])\nSearch for more papers by this authorABSTRACT\nDiminished ovarian reserve (DOR) is a multifactorial gynecological disorder that has emerged as a significant global health challenge. Currently, there are no effective preventive or therapeutic strategies for DOR. Exosome-derived long non-coding RNAs (lncRNA) in follicular fluid (FF) plays a crucial role in follicular development. We identified exosome-derived lncRNA LIPE-AS1 from the FF of DOR patients, which regulates histone deacetylase 3 (HDAC3) expression by competitively binding to miR-330-5p. Exosomes, as nanosized membrane vesicles, can deliver therapeutic agents in a targeted manner through ligand modification. In this study, we employed engineered exosomes combined with lncRNA for ovary-targeted therapy of DOR. First, we elucidated the role of lncRNA LIPE-AS1 in the pathogenesis of DOR. Next, we generated exosomes with high LIPE-AS1 expression (Exo-LIPE-AS1) using 293 T cells. Co-culture of Exo-LIPE-AS1 with oocytes from DOR models enhanced oocyte maturation and improve oocyte quality in vitro. Finally, we developed FSHβ-modified, LIPE-AS1-loaded exosomes (ExoFSHβ-LIPE-AS1), which demonstrated enhanced ovarian delivery efficiency in vivo. Consequently, ExoFSHβ-LIPE-AS1improved fertility outcomes in DOR models. Our findings demonstrate that exosomes serve as effective targeted vehicles for lncRNA LIPE-AS1, offering potential preventive and therapeutic benefits for DOR.\nConflicts of Interest\nThe authors declare no conflicts of interest.\nData Availability Statement\nAll datasets used and/or analyzed during this study are available from the corresponding author upon reasonable request.\nSupporting Information\n| Filename | Description |\n|---|---|\n| jbt70519-sup-0001-Fig_S1.tif1.4 MB | Fig S1. |\n| jbt70519-sup-0002-Fig_S2.tif292.7 KB | Fig S2. |\n| jbt70519-sup-0003-Fig_S3.tif116.5 KB | Fig S3. |\n| jbt70519-sup-0004-Fig_S4.tif716.4 KB | Fig S4. |\n| jbt70519-sup-0005-Fig_S5.tif12.5 MB | Fig S5. |\n| jbt70519-sup-0006-Fig_S6.tif706.5 KB | Fig S6. |\n| jbt70519-sup-0007-Supplementary_Table_1.docx12.3 KB | Supplementary Table 1. |\nPlease note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.\nReferences\n- 1A. Z. Steiner, D. Pritchard, F. Z. Stanczyk, et al., “Association Between Biomarkers of Ovarian Reserve and Infertility Among Older Women of Reproductive Age,” Journal of the American Medical Association 318 (2017): 1367–1376, https://doi.org/10.1001/jama.2017.14588.\n- 2L. Boucret, J. M. Chao de la Barca, C. Moriniere, et al., “Relationship Between Diminished Ovarian Reserve and Mitochondrial Biogenesis in Cumulus Cells,” Human Reproduction 30 (2015): 1653–1664, https://doi.org/10.1093/humrep/dev114.\n- 3S. J. Bunnewell, E. R. Honess, A. M. Karia, S. D. Keay, B. H. Al Wattar, and S. Quenby, “Diminished Ovarian Reserve in Recurrent Pregnancy Loss: A Systematic Review and Meta-Analysis,” Fertility and Sterility 113 (2020): 818–827.e3, https://doi.org/10.1016/j.fertnstert.2019.11.014.\n- 4Z. Zhou, D. Zheng, H. Wu, et al., “Epidemiology of Infertility in China: A Population-Based Study,” BJOG: An International Journal of Obstetrics & Gynaecology 125 (2018): 432–441, https://doi.org/10.1111/1471-0528.14966.\n- 5J. Rashtian and J. Zhang, “Luteal-Phase Ovarian Stimulation Increases the Number of Mature Oocytes in Older Women with Severe Diminished Ovarian Reserve,” Systems Biology in Reproductive Medicine 64 (2018): 216–219, https://doi.org/10.1080/19396368.2018.1448902.\n- 6B. C. Tarlatzis, “GnRH Antagonists in Ovarian Stimulation for IVF,” Human Reproduction Update 12 (2006): 333–340, https://doi.org/10.1093/humupd/dml001.\n- 7A. Revelli, L. D. Piane, S. Casano, E. Molinari, M. Massobrio, and P. Rinaudo, “Follicular Fluid Content and Oocyte Quality: From Single Biochemical Markers to Metabolomics,” Reproductive Biology and Endocrinology 7 (2009): 40, https://doi.org/10.1186/1477-7827-7-40.\n- 8R. J. Rodgers and H. F. Irving-Rodgers, “Formation of the Ovarian Follicular Antrum and Follicular Fluid,” Biology of Reproduction 82 (2010): 1021–1029, https://doi.org/10.1095/biolreprod.109.082941.\n- 9J. M. Chu, T. L. Yin, S. J. Zheng, J. Yang, B. F. Yuan, and Y. Q. Feng, “Metal Oxide-Based Dispersive Solid-Phase Extraction Coupled with Mass Spectrometry Analysis for Determination of Ribose Conjugates in Human Follicular Fluid,” Talanta 167 (2017): 506–512, https://doi.org/10.1016/j.talanta.2017.02.062.\n- 10K. A. Ahmed and J. Xiang, “Mechanisms of Cellular Communication Through Intercellular Protein Transfer,” Journal of Cellular and Molecular Medicine 15 (2011): 1458–1473, https://doi.org/10.1111/j.1582-4934.2010.01008.x.\n- 11C. Di Pietro, “Exosome-Mediated Communication in the Ovarian Follicle,” Journal of Assisted Reproduction And Genetics 33 (2016): 303–311, https://doi.org/10.1007/s10815-016-0657-9.\n- 12I. Ulitsky and D. P. Bartel, “LincRNAs: Genomics, Evolution, and Mechanisms,” Cell 154 (2013): 26–46, https://doi.org/10.1016/j.cell.2013.06.020.\n- 13B. Kleaveland, C. Y. Shi, J. Stefano, and D. P. Bartel, “A Network of Noncoding Regulatory RNAs Acts in the Mammalian Brain,” Cell 174 (2018): 350–362.e17, https://doi.org/10.1016/j.cell.2018.05.022.\n- 14S. Ghafouri-Fard, M. Moghadam, H. Shoorei, Z. Bahroudi, M. Taheri, and A. Taheriazam, “The Impact of Non-Coding RNAs on Normal Stem Cells,” Biomedicine & Pharmacotherapy = Biomedecine & Pharmacotherapie 142 (2021): 112050, https://doi.org/10.1016/j.biopha.2021.112050.\n- 15C. Zheng, S. Liu, Z. Qin, X. Zhang, and Y. Song, “LncRNA DLEU1 Is Overexpressed in Premature Ovarian Failure and Sponges miR-146b-5p to Increase Granulosa Cell Apoptosis,” Journal of Ovarian Research 14 (2021): 151, https://doi.org/10.1186/s13048-021-00905-x.\n- 16B. Liu, L. Liu, Z. Sulaiman, et al., “Comprehensive Analysis of lncRNA-miRNA-mRNA ceRNA Network and Key Genes in Granulosa Cells of Patients with Biochemical Primary Ovarian Insufficiency,” Journal Of Assisted Reproduction and Genetics 41 (2024): 15–29, https://doi.org/10.1007/s10815-023-02937-2.\n- 17H. I. Kim, J. Park, Y. Zhu, X. Wang, Y. Han, and D. Zhang, “Recent Advances in Extracellular Vesicles for Therapeutic Cargo Delivery,” Experimental & Molecular Medicine 56 (2024): 836–849, https://doi.org/10.1038/s12276-024-01201-6.\n- 18H. Chen, H. Yao, J. Chi, et al., “Engineered Exosomes as Drug and RNA Co-Delivery System: New Hope for Enhanced Therapeutics?,” Frontiers in Bioengineering and Biotechnology 11 (2023): 1254356, https://doi.org/10.3389/fbioe.2023.1254356.\n- 19P. F. Agris, R. H. Guenther, H. Sierzputowska-Gracz, et al., “Solution Structure of a Synthetic Peptide Corresponding to a Receptor Binding Region of Fsh (hFSH-β 33–53),” Journal of Protein Chemistry 11 (1992): 495–507, https://doi.org/10.1007/BF01025027.\n- 20P. Grasso, T. A. Santa-Coloma, and L. E. Reichert, “Synthetic Peptides Corresponding to Human Follicle-Stimulating Hormone (hFSH)-beta-(1-15) and hFSH-beta-(51-65) Induce Uptake of 45Ca++ by Liposomes: Evidence for Calcium-Conducting Transmembrane Channel Formation,” Endocrinology 128 (1991): 2745–2751, https://doi.org/10.1210/endo-128-6-2745.\n- 21X. Zhang, J. Chen, Y. Zheng, et al., “Follicle-Stimulating Hormone Peptide Can Facilitate Paclitaxel Nanoparticles to Target Ovarian Carcinoma In Vivo,” Cancer Research 69 (2009): 6506–6514, https://doi.org/10.1158/0008-5472.CAN-08-4721.\n- 22G. Yao, J. He, Y. Kong, et al., “Transcriptional Profiling of Long Noncoding RNAs and Their Target Transcripts in Ovarian Cortical Tissues From Women With Normal Menstrual Cycles and Primary Ovarian Insufficiency,” Molecular Reproduction and Development 86 (2019): 847–861, https://doi.org/10.1002/mrd.23158.\n- 23Y. Xiong, T. Liu, S. Wang, H. Chi, C. Chen, and J. Zheng, “Cyclophosphamide Promotes the Proliferation Inhibition of Mouse Ovarian Granulosa Cells and Premature Ovarian Failure by Activating the lncRNA-Meg3-p53-p66Shc Pathway,” Gene 596 (2017): 1–8, https://doi.org/10.1016/j.gene.2016.10.011.\n- 24E. B. Cordts, D. M. Christofolini, A. A. dos Santos, B. Bianco, and C. P. Barbosa, “Genetic Aspects of Premature Ovarian Failure: A Literature Review,” Archives of Gynecology and Obstetrics 283 (2011): 635–643, https://doi.org/10.1007/s00404-010-1815-4.\n- 25J. Bellver and J. Donnez, “Introduction,” Fertility and Sterility 111 (2019): 1033–1035, https://doi.org/10.1016/j.fertnstert.2019.04.043.\n- 26N. Tonti, T. Golia D'Augè, I. Cuccu, et al., “The Role of Tumor Biomarkers in Tailoring the Approach to Advanced Ovarian Cancer,” International Journal of Molecular Sciences 25 (2024): 11239, https://doi.org/10.3390/ijms252011239.\n- 27V. Di Donato, G. Caruso, T. Golia D'Augè, et al., “Prognostic Impact of Microscopic Residual Disease After Neoadjuvant Chemotherapy in Patients Undergoing Interval Debulking Surgery for Advanced Ovarian Cancer,” Archives of Gynecology and Obstetrics 311 (2025): 429–436, https://doi.org/10.1007/s00404-024-07775-w.\n- 28A. Thunen, D. La Placa, Z. Zhang, and J. E. Shively, “Role of LncRNA LIPE-AS1 in Adipogenesis,” Adipocyte 11 (2022): 11–27, https://doi.org/10.1080/21623945.2021.2013415.\n- 29R. G. Edwards, “Maturation In Vitro of Human Ovarian Oocytes,” Lancet 286 (1965): 926–929, https://doi.org/10.1016/s0140-6736(65)92903-x.\n10.1016/s0140-6736(65)92903-xGoogle Scholar\n- 30R. G. Edwards, B. D. Bavister, and P. C. Steptoe, “Early Stages of Fertilization In Vitro of Human Oocytes Matured In Vitro,” Nature 221 (1969): 632–635, https://doi.org/10.1038/221632a0.\n- 31R. G. Edwards, R. P. Donahue, T. A. Baramki, and H. W. Jones, “Preliminary Attempts to Fertilize Human Oocytes Matured In Vitro,” American Journal of Obstetrics and Gynecology 96 (1966): 192–200, https://doi.org/10.1016/0002-9378(66)90315-2.\n- 32I. J. Chamani and D. L. Keefe, “Epigenetics and Female Reproductive Aging,” Frontiers in Endocrinology 10 (2019): 473, https://doi.org/10.3389/fendo.2019.00473.\n- 33C. Simerly, M. Manil-Ségalen, C. Castro, et al., “Separation and Loss of Centrioles from Primordidal Germ Cells to Mature Oocytes in the Mouse,” Scientific Reports 8 (2018): 12791, https://doi.org/10.1038/s41598-018-31222-x.\n- 34K. I. Yamanaka, N. Aono, H. Yoshida, and E. Sato, “Cryopreservation and In Vitro Maturation of Germinal Vesicle Stage Oocytes of Animals for Application in Assisted Reproductive Technology,” Reproductive Medicine and Biology 6 (2007): 61–68, https://doi.org/10.1111/j.1447-0578.2007.00167.x.\n- 35A. M. Luciano and M. A. Sirard, “Successful In Vitro Maturation of Oocytes: A Matter of Follicular Differentiation,” Biology of Reproduction 98 (2018): 162–169, https://doi.org/10.1093/biolre/iox149.\n- 36G. Coticchio, M. Dal Canto, M. Mignini Renzini, et al., “Oocyte Maturation: Gamete-Somatic Cells Interactions, Meiotic Resumption, Cytoskeletal Dynamics and Cytoplasmic Reorganization,” Human Reproduction Update 21 (2015): 427–454, https://doi.org/10.1093/humupd/dmv011.\n- 37S. Sugimura, L. J. Ritter, M. L. Sutton-McDowall, D. G. Mottershead, J. G. Thompson, and R. B. Gilchrist, “Amphiregulin Co-Operates With Bone Morphogenetic Protein 15 to Increase Bovine Oocyte Developmental Competence: Effects on Gap Junction-Mediated Metabolite Supply,” MHR: Basic Science of Reproductive Medicine 20 (2014): 499–513, https://doi.org/10.1093/molehr/gau013.\n- 38G. N. Singina, E. N. Shedova, R. E. Uzbekov, and S. Uzbekova, “135 Effect of Different Concentrations of Follicular Fluid Exosome-Like Extracellular Vesicles on In Vitro Oocyte Maturation and Embryo Development in Cattle,” Reproduction, Fertility, and Development 34 (2021): 305–306, https://doi.org/10.1071/RDv34n2Ab135.\n- 39P. B. Ham and R. Raju, “Mitochondrial Function in Hypoxic Ischemic Injury and Influence of Aging,” Progress in Neurobiology 157 (2017): 92–116, https://doi.org/10.1016/j.pneurobio.2016.06.006.\n- 40T. Kiss, Á. Nyúl-Tóth, P. Balasubramanian, et al., “Nicotinamide Mononucleotide (NMN) Supplementation Promotes Neurovascular Rejuvenation in Aged Mice: Transcriptional Footprint of SIRT1 Activation, Mitochondrial Protection, Anti-Inflammatory, and Anti-Apoptotic Effects,” GeroScience 42 (2020): 527–546, https://doi.org/10.1007/s11357-020-00165-5.\n- 41D. Tiosano, J. A. Mears, and D. A. Buchner, “Mitochondrial Dysfunction in Primary Ovarian Insufficiency,” Endocrinology 160 (2019): 2353–2366, https://doi.org/10.1210/en.2019-00441.\n- 42S. Venkatesh, M. Kumar, A. Sharma, et al., “Oxidative Stress and ATPase6 Mutation Is Associated With Primary Ovarian Insufficiency,” Archives of Gynecology and Obstetrics 282 (2010): 313–318, https://doi.org/10.1007/s00404-010-1444-y.\n- 43M. Kumar, D. Pathak, S. Venkatesh, A. Kriplani, A. C. Ammini, and R. Dada, “Chromosomal Abnormalities & Oxidative Stress in Women With Premature Ovarian Failure (POF),” Indian Journal of Medical Research 135 (2012): 92–97, https://doi.org/10.4103/0971-5916.93430.\n- 44J. Tesarik, M. Galán-Lázaro, and R. Mendoza-Tesarik, “Ovarian Aging: Molecular Mechanisms and Medical Management,” International Journal of Molecular Sciences 22 (2021): 1371, https://doi.org/10.3390/ijms22031371.\n- 45N. G. Larsson, “Somatic Mitochondrial DNA Mutations in Mammalian Aging,” Annual Review of Biochemistry 79 (2010): 683–706, https://doi.org/10.1146/annurev-biochem-060408-093701.\n- 46W. T. Hung, X. Hong, L. K. Christenson, and L. K. McGinnis, “Extracellular Vesicles From Bovine Follicular Fluid Support Cumulus Expansion,” Biology of Reproduction 93 (2015): 117, https://doi.org/10.1095/biolreprod.115.132977.\n- 47T. A. Rodrigues, K. M. Tuna, A. A. Alli, et al., “Follicular Fluid Exosomes Act on the Bovine Oocyte to Improve Oocyte Competence to Support Development and Survival to Heat Shock,” Reproduction, Fertility, and Development 31 (2019): 888–897, https://doi.org/10.1071/RD18450.\n- 48W. T. Hung, R. Navakanitworakul, T. Khan, et al., “Stage-Specific Follicular Extracellular Vesicle Uptake and Regulation of Bovine Granulosa Cell Proliferation,” Biology of Reproduction 97 (2017): 644–655, https://doi.org/10.1093/biolre/iox106.\n- 49S. H. Lee, H. J. Oh, M. J. Kim, and B. C. Lee, “Canine Oviductal Exosomes Improve Oocyte Development via EGFR/MAPK Signaling Pathway,” Reproduction 160 (2020): 613–625, https://doi.org/10.1530/REP-19-0600.\n- 50M. Javadi, J. Soleimani Rad, M. Pashaiasl, M. S. G. Farashah, and L. Roshangar, “The Effects of Plasma-Derived Extracellular Vesicles on Cumulus Expansion and Oocyte Maturation in Mice,” Reproductive Biology 22 (2022): 100593, https://doi.org/10.1016/j.repbio.2021.100593.\n- 51L. Alvarez-Erviti, Y. Seow, H. Yin, C. Betts, S. Lakhal, and M. J. A. Wood, “Delivery of siRNA to the Mouse Brain by Systemic Injection of Targeted Exosomes,” Nature Biotechnology 29 (2011): 341–345, https://doi.org/10.1038/nbt.1807.\n- 52F. Momen-Heravi, S. Bala, T. Bukong, and G. Szabo, “Exosome-Mediated Delivery of Functionally Active miRNA-155 Inhibitor to Macrophages,” Nanomedicine: Nanotechnology, Biology and Medicine 10 (2014): 1517–1527, https://doi.org/10.1016/j.nano.2014.03.014.\n- 53Q. Han, Q. R. Xie, F. Li, et al., “Targeted Inhibition of SIRT6 via Engineered Exosomes Impairs Tumorigenesis and Metastasis in Prostate Cancer,” Theranostics 11 (2021): 6526–6541, https://doi.org/10.7150/thno.53886.","source_license":"CC0","license_restricted":false}