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.

项目概要衰老是所有生物体共有的复杂的多因素生物过程，在卵巢中表现为：正常生殖功能逐渐衰退，当卵母细胞数量减少时，最终在更年期达到顶峰从 30 多岁中期开始，在进入更年期之前，该值会急剧增加。由于衰老导致卵母细胞功能障碍的发生率，导致减数分裂分离错误、非整倍体和不良这种细胞功能障碍在临床上表现为不孕、辅助生殖失败。技术（ART）、（复发性）流产和出生缺陷目前尚无有效的治疗方法。改善高龄妇女的卵母细胞功能和新的方法策略卵母细胞减数分裂机制中涉及的许多因素与年龄有关。然而，卵巢衰老的根本原因以及女性生殖系的原因。相对于体细胞的加速衰老仍然未知。细胞损伤被广泛认为是衰老的原因，并且有证据表明衰老速度是受到控制的，至少在某种程度上，通过进化中保守的遗传途径。细胞表观遗传景观的侵蚀（表观基因组漂移）导致的细胞功能障碍可能主要是这些研究表明，通过异位表达进行“部分重编程”是有意造成衰老的。一组有限的多能因子可以恢复年轻的表观遗传模式并逆转衰老迹象然而，部分重编程对减数分裂细胞的细胞特性的影响，这表现出独特的基因调控网络和维持多能性的特征，尚不清楚。该提案研究了卵巢衰老的整体机制，挑战其不可逆转性，并开发了一种我们将采用一种新的途径来开发减缓或逆转卵巢衰老的治疗方法。转基因小鼠模型检验卵丘细胞-卵母细胞部分重编程的假设复合物可以逆转卵巢衰老并恢复生殖功能。我们将使用综合多组学来研究。定义甲基化时钟和表观遗传漂移在卵巢衰老中的作用以及非卵母细胞是否我们将研究部分重编程是否会导致卵巢因衰老而导致卵母细胞功能障碍。逆转卵母细胞卵丘复合体中与年龄相关的转录和表观遗传变化，使其更年轻模式并在不改变卵母细胞的细胞特性的情况下恢复体内生育能力。这种方法的转化潜力是使用一种能够穿过血液的新型 AAV 递送系统这项工作将增加我们对体内卵泡屏障和靶卵母细胞和颗粒细胞的了解。卵巢衰老生物学并为表观遗传药物的开发提供新的分子靶点逆转人体卵巢生殖功能衰退，减少流产、复发妊娠与高龄生育相关的损失和出生缺陷。