Characterization of age-related changes in spermatogonial dedifferentiation

精原去分化的年龄相关变化的表征

• 批准号: 10215936
• 负责人: Erika A Bach
• 金额: $ 25.43万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10215936
• 关键词: Adhesions; Adult; Affect; Age; Aging; Back; Binding Proteins; Bioinformatics; Candidate Disease Gene; Cell Differentiation process; Cells; Characteristics; Complex; Cues; Cyst; DNA Polymerase II; Data; Data Analyses; Data Set; Development; Drops; Drosophila genus; Dystroglycan; Enhancers; Ensure; Extracellular Matrix; Fertility; Fostering; Gene Expression; Genes; Genetic; Genetic Transcription; Germ Cells; Goals; Human; Immunofluorescence Immunologic; In Situ Hybridization; Individual; Infertility; Intrinsic factor; Knowledge; Life; Male Infertility; Modeling; Molecular; Mus; N-terminal; Pathway interactions; Phosphotransferases; Process; Proteins; Publishing; RNA Binding; RNA Polymerase II; Recovery; Reporting; Research; Role; Signal Transduction; Spermatogenesis; Spermatogonia; Starvation; Stress; Supporting Cell; System; Techniques; Testing; Testis; Tissues; Up-Regulation; Work; age effect; age related; cell dedifferentiation; design; exhaustion; experimental study; fertility preservation; fly; genome-wide; germline stem cells; human disease; insight; knowledge base; male; middle age; novel therapeutics; offspring; overexpression; prevent; response; self-renewal; stem cells; tissue stem cells; tool; transcriptome; unpublished works

Project Summary The broad, long-term objectives of this application are to characterize how age-related intrinsic changes in gene expression and age-related extrinsic changes in the niche microenvironment impact the process of dedifferentiation. This proposal will examine these issues using spermatogonial dedifferentiation - in which differentiating germ cells revert to become spermatogonial stem cells - in the Drosophila testis. State of the art techniques will be used to detect genome-wide binding of RNA Polymerase II in dedifferentiating spermatogonia. The proposal will capitalize upon the powerful genetics and lineage tracing available in Drosophila, as well as the ability to unequivocally identify the niche, spermatogonial stem cells, spermatogonia and somatic support cells in the Drosophila testis. This proposal is supported by published results demonstrating that spermatogonial dedifferentiation is required to maintain a robust pool of spermatogonial stem cells over a lifetime and that this process requires Jun N-terminal kinase (JNK) signaling. This work is also supported by unpublished preliminary data showing that the rate of spermatogonial dedifferentiation declines steeply during aging. Additionally, this proposal is supported by unpublished results showing that the niche microenvironment changes substantially during aging: in old males, an ectopic extracellular matrix accumulates around the niche. In the first aim, targeted DamID will be used to obtain the genome-wide binding of RNA Pol II only in reverting spermatogonia in testes from young and old males. Analyses of these data sets will reveal genes that are activated in young but not old spermatogonia. In the second aim, one or two validated genes from Aim 1 will be used to test the model that candidate gene depletion in young testes should decrease dedifferentiation rates and its overexpression should increase reversion rates in old males. Experiments in the second aim will also test the hypothesis that decreased dedifferentiation in old males is a result of reverting cells not upregulating extracellular matrix-binding proteins. The studies in this proposal will increase the knowledge base about how aging affects dedifferentiation in more complex system and will foster new avenues of research into mechanisms and treatments for age-related male infertility.

项目摘要 该应用程序的广泛，长期目标是表征基因与年龄相关的内在变化的方式 利基微环境的表达和与年龄相关的外在变化影响 去分化。该建议将使用精子去分化来检查这些问题 - 其中 在果蝇睾丸中，区分生殖细胞恢复为精子干细胞。最先进的 技术将用于检测RNA聚合酶II的全基因组结合，以脱落精子。 该提案将利用果蝇可用的强大遗传学和谱系追踪以及 明确识别小众，精子干细胞，精子和躯体支持的能力 果蝇睾丸中的细胞。该建议得到了已发表的结果支持，表明精子量 需要去分化才能在一生中维持稳健的精子干细胞池，这是 过程需要Jun N末端激酶（JNK）信号传导。这项工作也得到了未发表的初步的支持 数据表明，精子去分化的速率在衰老期间急剧下降。另外，这个 未发表的结果支持提案，表明利基微环境发生了很大的变化 在衰老期间：在老男性中，异位细胞外基质在小众周围积聚。在第一个目标中，有针对性 DAMID将仅在睾丸中恢复精子中获得RNA POL II的全基因组结合 来自年轻男性和老年男性。对这些数据集的分析将揭示在年轻但不衰老的基因 精子。在第二个目标中，AIM 1的一个或两个经过验证的基因将用于测试模型 年轻睾丸中的候选基因耗竭应降低去分化率，其过表达应 提高老男性的回归率。第二个目标中的实验还将测试下降的假设 旧雄性的去分化是振兴细胞不上调细胞外基质结合蛋白的结果。 该提案中的研究将增加有关衰老如何影响更多衰老的知识基础 复杂的系统，并将促进与年龄相关的男性的机制和治疗方法研究的新途径 不育。