Transcriptional Programming in the Maintenance and Commitment of Nephron Progenitors

肾单位祖细胞维持和承诺中的转录编程

• 批准号: 10212379
• 负责人: Helena Bugacov
• 金额: $ 4.6万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10212379
• 关键词: Address; Adult; Affinity; Affinity Chromatography; Agonist; American; Automobile Driving; Binding; Binding Proteins; Biological Assay; Cell Culture System; Cell Culture Techniques; Cell Differentiation process; ChIP-seq; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Complex; Cultured Cells; Derivation procedure; Differentiated Gene; Embryonic Development; Enhancers; Equilibrium; Family; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Transcription; Guide RNA; Harvest; Immunoprecipitation; In Vitro; Kidney; Kidney Diseases; Knock-in Mouse; Laboratories; Lead; Link; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Modification; Molecular; Multiprotein Complexes; Mus; Mutation; Nephrology; Nephrons; Outcome; Pathway interactions; Play; Population; Process; Proteomics; Regulation; Role; Signal Pathway; Signal Transduction; Stimulus; Supporting Cell; Testing; Transcription Coactivator; Transcriptional Regulation; Up-Regulation; WNT Signaling Pathway; Western Blotting; base; beta catenin; body system; cell type; chromatin immunoprecipitation; curative treatments; experimental study; genetic regulatory protein; in vivo; inhibitor/antagonist; insight; loss of function; nephrogenesis; nephron progenitor; novel; novel therapeutic intervention; prevent; programs; regenerative; regenerative approach; regenerative therapy; repaired; self-renewal; stem cells

PROJECT ABSTRACT Approximately 4.9 million adults in the US suffer from kidney disease. Current therapies do not provide curative treatments for end state renal disease. Regenerative nephrology holds promise for developing novel therapeutic strategies. The functional unit of the kidney is the nephron, which is derived from a distinct progenitor cell population during embryonic development. A delicate balance of nephron progenitor cell (NPC) self-renewal and differentiation is required to generate a species-appropriate number of nephrons during the course of kidney development. The transcriptional regulator Six2, and transcriptional mediators of the Wnt/β-catenin signaling pathway, Lef/Tcf factors, play critical roles balancing the maintenance and commitment of NPCs. In this, Six2 is essential for NPC self-renewal and interacts with Lef/Tcf factors. β-catenin accumulation acts to switch the code of Lef/Tcf factor engagement at differentiation targets from a repressor to activator transcriptional signature. The overarching hypothesis tested in this proposal is that β-catenin accumulation switches the Lef/Tcf regulatory signature from an OFF to ON state. In AIM 1, I will perform loss of function experiments to unravel the mechanism of action of β-catenin concentration dependent Tcf/Lef factor engagement resulting in transcriptional changes. In AIM 2, I will perform multi-protein proteomic analysis to define multiprotein complexes surrounding β-catenin with immunoprecipitation and mass spectrometry-based affinity proteomics Mechanistic understanding of Wnt/β-catenin signaling in concert with identified binding partners will facilitate the derivation of nephron-like cell types in vitro and ultimately regenerative therapies for renal diseases.

项目摘要 美国大约有 490 万成年人患有肾脏疾病。目前的疗法无法提供治愈作用 终末期肾病的治疗。再生肾病学有望开发新的治疗方法 策略。肾脏的功能单位是肾单位，它源自一种独特的祖细胞 胚胎发育期间的种群。肾单位祖细胞 (NPC) 自我更新和 在肾脏发育过程中，需要分化才能产生适合物种数量的肾单位 发展。转录调节因子 Six2 和 Wnt/β-catenin 信号传导的转录介质 途径、Lef/Tcf 因子在平衡 NPC 的维持和承诺方面发挥着关键作用。在此，Six2 是 对于 NPC 自我更新至关重要，并与 Lef/Tcf 因子相互作用。 β-连环蛋白积累起到开关密码的作用 Lef/Tcf 因子在从阻遏物到激活物转录特征的分化目标上的参与。这 本提案测试的首要假设是 β-catenin 积累会切换 Lef/Tcf 调节 签名从关闭状态到开启状态。在 AIM 1 中，我将进行功能丧失实验来阐明其机制 β-连环蛋白浓度依赖性 Tcf/Lef 因子参与导致转录变化的作用。 在 AIM 2 中，我将进行多蛋白蛋白质组学分析来定义 β-连环蛋白周围的多蛋白复合物 通过免疫沉淀和基于质谱的亲和蛋白质组学机制的理解 Wnt/β-连环蛋白信号传导与已识别的结合伙伴相结合将促进肾单位样细胞的衍生 肾脏疾病的体外类型和最终的再生疗法。