The Role of TAF4b in Spermatogonial Stem Cell Maintenance and Self-Renewal

TAF4b 在精原干细胞维持和自我更新中的作用

• 批准号: 8643093
• 负责人: Eric Gustafson
• 金额: $ 6万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8643093
• 关键词: Address; Affect; Age-Months; Binding; Binding Proteins; Biochemical; Bioinformatics; Biological; Biological Assay; Cell Culture Techniques; Cell Maintenance; Cell division; Cells; ChIP-seq; Complex; Data; Defect; Development; Developmental Biology; Diagnostic; Equilibrium; Etiology; Event; Exhibits; Female; Fertility; Gene Expression; Gene Expression Regulation; Gene Targeting; General Transcription Factors; Genes; Genetic Transcription; Genome; Germ Cells; Goals; Gonadal structure; Human; Invertebrates; Knockout Mice; Lead; Longevity; Luciferases; Maintenance; Male Infertility; Medicine; Methods; Modeling; Molecular; Mus; Nature; Organogenesis; Play; Process; Proteins; Proteomics; RNA Polymerase II; Regulation; Regulator Genes; Reporter Genes; Reproduction; Research Proposals; Research Training; Role; Sequence-Specific DNA Binding Protein; Spermatogonia; Stem cells; TATA-Box Binding Protein; Testing; Testis; Therapeutic; Tissues; Transcription Factor TFIIA; Transcriptional Regulation; Variant; Vertebrates; Wild Type Mouse; Work; base; cell type; cofactor; comparative; embryonic stem cell; human embryonic stem cell; insight; male; men; novel; pluripotency; postnatal; programs; promoter; protein function; public health relevance; reproductive; reproductive function; self-renewal; stem cell biology; stem cell division; tool

DESCRIPTION (provided by applicant): Understanding how stem cells balance self-renewal with differentiation is paramount in harnessing their therapeutic potential in medicine. While recent work has identified master transcriptional regulator genes controlling pluripotency in mouse and human embryonic stem (ES) cells, it is still unclear whether similar mechanisms direct cell fate decisions in tissue-specific unipotent stem cells, such as the spermatogonial stem cells (SSCs) found in mammalian testes. In contrast to the master regulator proteins in ES cells, recent works has discovered essential functions for cell type-specific or -enriched forms of the core transcription machinery in regulating gene expression. These are required to drive critical gene expression programs involved in diverse developmental events associated with multicellularity and organogenesis. The diversification of core transcription machinery is most notable in the germline where germ cell-specific or -enriched variants of the general transcription factors TFIIA and TFIID execute highly critical and selective functions in regulating reproduction and fertility. This new paradigm of germ cell-specific gene regulation is conserved between invertebrates and vertebrates, and may reflect an important mechanism for evolving germ cell-specific modes of gene regulation. However, the need to understand the molecular mechanisms underlying such exquisite regulation of fertility persists. The goal of this research proposal is to characterize the role of TAF4b in the establishment and maintenance of the self-renewing lineage of SSCs in the mouse testis. TAF4b is a gonad-enriched general transcription factor varient and both male and female Taf4b-deficient mice exhibit reproductive defects. Several lines of evidence suggest it plays an essential role in spermatogonia stem cell (SSC) self-renewal and longevity. One of the specific aims in this research proposal is to identify and characterize TAF4b-interacting protein cofactors to investigate their function in SSC regulation. One of the ways TAF4b may control SSC-specific gene expression is through direct interaction with other proteins that function to turn appropriate SSC genes on or off. This aim will test this hypothesis by identifying these proteins and analyzing their function in SSC gene expression. The second specific aim is to identify where TAF4b binds in the SSC genome and identify which genes TAF4b is directly targeting for regulation. This will provide mechanistic insight into how TAF4b determines which genes it will regulate, how SSC-specific gene regulatory programs are established and how these programs drive SSC self-renewal. The work proposed here, to uncover the mechanisms of TAF4b in regulating SSC maintenance and self-renewal, will reveal fundamental biological principles underlying both stem cell biology and reproductive functions in men required for fertility. These studies may reveal the etiologies behind unexplained male infertility and lead to advanced diagnostic and therapeutic tools to better address and manage male infertility.

描述（由申请人提供）：了解干细胞如何平衡自我更新与分化对于发挥其医学治疗潜力至关重要。虽然最近的工作已经确定了控制小鼠和人类胚胎干（ES）细胞多能性的主转录调节基因，但仍不清楚类似的机制是否指导组织特异性单能干细胞（例如在哺乳动物睾丸中发现的精原干细胞（SSC））中的细胞命运决定。与 ES 细胞中的主调节蛋白相反，最近的研究发现了细胞类型特异性或富集形式的重要功能。 调节基因表达的核心转录机制。这些是驱动参与与多细胞性和器官发生相关的多种发育事件的关键基因表达程序所必需的。核心转录机制的多样化在种系中最为显着，其中生殖细胞特异性或通用转录因子 TFIIA 和 TFIID 的富集变体在调节中执行高度关键和选择性的功能 繁殖和生育能力。这种生殖细胞特异性基因调控的新范式在无脊椎动物和脊椎动物之间是保守的，并且可能反映了进化生殖细胞特异性基因调控模式的重要机制。然而，仍然需要了解这种精细的生育调节背后的分子机制。本研究计划的目标是表征 TAF4b 在小鼠睾丸中 SSC 自我更新谱系的建立和维持中的作用。 TAF4b 是一种富含性腺的通用转录因子变体，Taf4b 缺陷的雄性和雌性小鼠均表现出生殖缺陷。多项证据表明它在精原干细胞（SSC）的自我更新和长寿中发挥着重要作用。本研究计划的具体目标之一是鉴定和表征 TAF4b 相互作用蛋白辅因子，以研究它们在 SSC 调节中的功能。 TAF4b 控制 SSC 特异性基因表达的方式之一是通过与其他蛋白质直接相互作用，这些蛋白质的功能是打开或关闭适当的 SSC 基因。该目标将通过鉴定这些蛋白质并分析它们在 SSC 基因表达中的功能来检验这一假设。第二个具体目标是确定 TAF4b 在 SSC 基因组中的结合位置，并确定 TAF4b 直接调控哪些基因。这将为 TAF4b 如何确定它将调节哪些基因、如何建立 SSC 特异性基因调控程序以及这些程序如何驱动 SSC 自我更新提供机制见解。这里提出的工作旨在揭示 TAF4b 调节 SSC 维持和自我更新的机制，将揭示干细胞生物学和男性生育所需生殖功能的基本生物学原理。这些研究可能揭示无法解释的男性不育症背后的病因，并带来先进的诊断和治疗工具，以更好地解决和管理男性不育症。