Reversal of Ovarian Aging in Mice Through AAV-mediated Oocyte Reprogramming in vivo

通过 AAV 介导的体内卵母细胞重编程逆转小鼠卵巢衰老

基本信息

批准号：
10723227
负责人：
Raymond M ANCHAN
金额：
$ 49.59万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10723227
关键词：
ATAC-seq Acceleration Age Aging Aneuploidy Assisted Reproductive Technology Bioinformatics Biological Process Biology Blindness Blood Capsid Caring Cell division Cells Chromosomes Clinical Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Complement Complex Congenital Abnormality Cytoplasm DNA Damage Development Down Syndrome Ectopic Expression Elderly Embryonic Development Entropy Epigenetic Process Evolution Exhibits Failure Female Fertility Fertilization Fertilization in Vitro Functional disorder Future Gene Delivery Gene Transfer Genetic Genetic Transcription Genomics Germ Cells Goals Hospitals Human Incidence Individual Infertility Knowledge Life Longevity Measures Mediating Meiosis Menopause Methodology Methods Methylation Modeling Molecular Target Mus Mutation Natural regeneration Oocytes Organism Outcome Ovarian Ovarian aging Ovary Pathway interactions Pattern Pregnancy loss RNA Recombinant adeno-associated virus (rAAV)Recurrence Role Somatic Cell Spontaneous abortion System Techniques Testing Time Transgenic Mice Variant Woman Work advanced maternal age age effect age related aged bisulfite sequencing cell injury effective therapy egg epigenetic drug epigenome epigenomics experimental study falls gene regulatory network genome-wide granulosa cell improved in vivo innovation mouse model multiple omics mutant novel ovarian dysfunction pluripotency pluripotency factor prevent programs reproductive reproductive function reproductive outcome reproductive senescence segregation somatic cell nuclear transfer therapy development transcriptomics translational potential trying to conceive

项目摘要

Project Summary Aging is a complex multifactorial biological process shared by all living organisms. In the ovary, it manifests as a gradual decline of normal reproductive function which culminates in menopause when the number of oocytes falls below a threshold. Menopause is preceded, from the mid-30s onward, by a dramatically increased incidence of oocyte dysfunction due to aging which results in meiotic segregation errors, aneuploidy and poor embryo development. This cellular dysfunction clinically presents as infertility, failure of assisted reproduction technologies (ART), (recurrent) pregnancy loss and birth defects. There are currently no effective treatments to improve oocyte function in women with advanced reproductive age and novel methodological strategies are critically needed. Many factors involved in the meiotic machinery in oocytes have been implicated in age- related oocyte dysfunction. However, the underlying cause of ovarian aging and why the female germline undergoes accelerated aging relative to somatic cells remains unknown. Time-dependent accumulation of cellular damage is widely considered to underlie aging and there is evidence that the rate of aging is controlled, at least to some extent, by genetic pathways conserved in evolution. Recent studies indicate that in somatic cells an erosion of the epigenetic landscape (epigenomic drift) resulting cellular dysfunction may be primarily responsible for aging. Interestingly, these studies show that ‘partial reprogramming’ through ectopic expression of a limited set of pluripotency factors can restore youthful epigenetic patterns and reverse signs of aging in somatic cells. However, the effect of partial reprogramming on the cell identity of meiotic cells, which exhibit unique gene regulatory networks and maintain features of pluripotency, is unknown. This proposal examines the global mechanism of ovarian aging, challenges its irreversibility and develops a novel pathway for the future development of treatments to slow or reverse ovarian aging. We will employ a transgenic mouse model to examine the hypothesis that partial reprogramming of the cumulus cell-oocyte complex can reverse ovarian aging and restore reproductive function. We will use integrative multi-omics to define the role of the methylation clock and epigenetic drift in ovarian aging and whether non-oocyte cells of the ovary contribute to oocyte dysfunction due to aging. We will investigate whether partial reprogramming can reverse age-related transcriptional and epigenetic changes in the oocyte-cumulus-complex to more youthful patterns and restore fertility in vivo without altering the cellular identity of oocytes. We will then examine the translational potential of this approach using a novel AAV delivery system with the ability to cross the blood- follicle barrier and target oocytes and granulosa cells in vivo. This work will increase our understanding of the biology of ovarian aging and provide novel molecular targets for the development of epigenetic drugs that reverse the decline in human reproductive ovarian function and reduce the miscarriages, recurrent pregnancy loss and birth defects associated with advanced reproductive age.

项目摘要衰老是所有生物体共享的复杂多因素生物学过程。在卵巢中，它表现为卵母细胞数量时，正常生殖功能的等级下降，最终导致更年期低于阈值。从30年代中期开始，更年期之前就大大增加了衰老引起的卵母细胞功能障碍的发生率，导致减数分裂偏析误差，非整倍性和差胚胎发育。该细胞功能障碍在临床上表现为不育，辅助繁殖失败技术（ART），（经常性）妊娠丧失和先天缺陷。目前没有有效的治疗方法提高具有高级生殖年龄和新颖方法论策略的女性的卵母细胞功能是至关重要的是。卵母细胞中的减数分裂机械涉及的许多因素已在年龄中暗示相关的卵母细胞功能障碍。但是，卵巢衰老的根本原因以及女性生殖线的原因相对于体细胞的加速衰老仍然未知。时间依赖的加速度细胞损伤被广泛认为是衰老的基础，并且有证据表明衰老率是控制的，至少在某种程度上，通过在进化中配置的遗传途径。最近的研究表明，在体细胞中细胞对细胞功能障碍的表观遗传景观（表观遗传学漂移）的侵蚀可能是主要的负责衰老。有趣的是，这些研究表明，“部分重编程”通过生态表达一组有限的多能因素可以恢复年轻的表观遗传模式和衰老的反向迹象体细胞。但是，部分重编程对显示的减数分裂细胞的细胞身份的影响独特的基因调节网络并保持多能性的特征是未知的。该提案研究了卵巢衰老的全球机制，挑战了其不可逆性，并发展了未来开发疗法的新途径，以减缓或反向卵巢衰老。我们将采用一个转基因小鼠模型，以检验以下假设，即积云细胞的部分重编程复合物可以逆转卵巢衰老并恢复复制功能。我们将使用综合的多媒体进行定义甲基化时钟和表观遗传漂移在卵巢衰老中的作用卵巢由于衰老而导致卵母细胞功能障碍。我们将调查部分重编程是否可以与年龄相关的转录和表观遗传变化的卵母细胞 - 复合物变为更年轻模式并恢复体内生育能力，而不会改变卵母细胞的细胞身份。然后，我们将检查这种方法使用新型的AAV递送系统的翻译潜力，能够越过血液卵泡屏障和靶卵母细胞和颗粒细胞在体内。这项工作将增加我们对卵巢衰老的生物学，并为发展表观遗传药物提供新的分子靶标扭转人类生殖卵巢功能的下降并减少流产，复发性怀孕与高级复制年龄相关的损失和出生缺陷。