Wilms tumor 1 (Wt1) mutation reveals mechanisms of cell lineage crosstalk in the developing kidney

肾母细胞瘤 1 (Wt1) 突变揭示了发育中肾脏中细胞谱系串扰的机制

• 批准号: 10665776
• 负责人: Keri Anne Drake
• 金额: $ 16.7万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10665776
• 关键词: Ablation; Affect; Apoptosis; Bioinformatics; Candidate Disease Gene; Cell Communication; Cell Cycle; Cell Differentiation process; Cell Lineage; Cells; Data; Defect; Development; Development Plans; Diphtheria Toxin; Disease; Ectopic Expression; Embryo; Environment; Foundations; Future; Gene Expression; Genes; Genetic; Genetic study; Genetically Engineered Mouse; Goals; Histology; In Vitro; K-Series Research Career Programs; Kidney; Lentivirus Vector; Ligands; Link; Maintenance; Mentors; Mentorship; Metanephric Diverticulum; Modeling; Molecular; Mutation; Nephroblastoma; Nephrology; Nephrons; Pathway interactions; Pattern; Play; Population; Proliferating; Recombinant Proteins; Regulation; Regulator Genes; Research Personnel; Rest; Role; Signal Pathway; Signal Transduction; Small Interfering RNA; Stromal Cells; Structure; Techniques; Testing; Training; Tumor Biology; United States National Institutes of Health; Validation; Vesicle; WT1 gene; Wilms Tumor Genes; Work; beta catenin; blastema; candidate identification; career; career development; clinically significant; experimental study; in vitro Assay; in vivo; insight; intercellular communication; loss of function; mouse model; mutant; mutant mouse model; nephrogenesis; nephron progenitor; novel; receptor; research and development; self-renewal; single nucleus RNA-sequencing; single-cell RNA sequencing; stem cell proliferation; stem cells; stromal progenitor; stromal progenitor population; tumor; tumorigenesis

PROJECT SUMMARY/ABSTRACT This NIH Mentored Career Development Award describes a three-year proposal providing the training and mentorship necessary for the candidate to develop into an independent investigator studying the genetic and molecular regulation of kidney development, with a specific focus on how mutations in Wilms tumor 1 (Wt1) affect cell lineage crosstalk in the developing kidney. To accomplish this, the candidate has developed a detailed career development plan integrating the outstanding mentorship and scientific environment at UT Southwestern with focused training in bioinformatics and in-vitro techniques to successfully transition to independence. Wt1, the first gene identified in Wilms tumor, regulates multiple aspects of normal renal development and has been previously shown to play a critical role in nephron progenitor cell (NPC) differentiation. Further examination of mutant kidneys with Wt1 loss-of-function specifically targeting the NPC lineage (Six2cre;Wt1c/c) shows additional effects on the nephrogenic stroma, with an expansion of stromal progenitor cells, disruptions in stromal patterning, and an increase in expression of several stromal markers previously shown to be upregulated in Wilms tumors, leading to the hypothesis that signals from the NPCs and/or early nephron structures non-autonomously regulate stromal development. The testing of this hypothesis will interrogate how NPC-to-stromal crosstalk directs stromal progenitor maintenance and/or differentiation in normal kidney development, which will be accomplished through two proposed aims. In aim 1, experiments utilizing genetically engineered mouse models are directed at determining how signals from nephron lineage derived cells regulate the nephrogenic stroma. To do this, stromal proliferation, apoptosis, and patterning will be examined in mutant mouse models targeting different populations of the nephron lineage (ie: self-renewing NPCs vs. early nephron structures) in comparison to the effects on the stroma identified in Six2cre;Wt1c/c mutant kidneys. Aim 2 will investigate the mechanism(s) of NPC-to-stromal crosstalk, using bioinformatics to identify candidate receptor/ligand interactions and in-vitro assays to functionally validate these pathways by determining if disrupting specific signaling pathways results in stromal defects in control kidneys, and likewise, if restored signaling pathways can rescue normal stromal differentiation in mutant kidney explants. Given that advances in renal development research have significantly contributed to the understanding of Wilms tumor, and vice versa, with the study of Wilms tumor biology leading to advances in developmental nephrology, the aims outlined in this proposal have the potential to further uncover how intricate reciprocal signaling interactions regulate progenitor cell proliferation and/or differentiation in normal kidney development and provide the foundation for future work examining how disruptions in progenitor cell crosstalk between the nephron and stromal lineages may alter the embryonic microenvironment and predispose to the development of Wilms tumor.

项目总结/摘要 这个NIH指导职业发展奖描述了一个为期三年的建议，提供培训， 辅导必要的候选人发展成为一个独立的研究人员的遗传和 肾脏发育的分子调控，特别关注肾母细胞瘤1（Wt 1）中的突变 影响发育中的肾脏中的细胞谱系串扰。为了做到这一点，候选人已经制定了一个 详细的职业发展计划，结合UT的优秀导师和科学环境 西南部的生物信息学和体外技术的重点培训，以成功过渡到 独立Wt 1是在肾母细胞瘤中发现的第一个基因，调节正常肾功能的多个方面。 在肾单位祖细胞（NPC）的发育中起着关键作用， 分化特异性靶向NPC的Wt 1功能丧失突变肾的进一步检查 细胞系（Six 2cre; Wt 1c/c）对肾源性间质表现出额外的影响， 祖细胞，基质模式的破坏，以及几种基质标志物表达的增加 此前已证明其在肾母细胞瘤中上调，这导致了来自NPC的信号的假设 和/或早期肾单位结构非自主地调节基质发育。测试这个 假设将询问NPC-基质串扰如何指导基质祖细胞维持和/或 在正常肾脏发育中的分化，这将通过两个提出的目标来实现。在目标1中， 利用基因工程小鼠模型的实验旨在确定来自 肾单位谱系衍生的细胞调节肾发生基质。为了做到这一点，基质增殖，凋亡， 将在靶向肾单位谱系的不同群体的突变小鼠模型（即： 自我更新的NPC与早期肾单位结构），与 Six 2cre; Wt 1c/c突变肾。目的2将研究NPC与基质串扰的机制，使用 生物信息学来鉴定候选受体/配体相互作用和体外测定来功能性地验证这些 通过确定破坏特定信号传导途径是否导致对照肾中的基质缺陷， 同样，如果恢复的信号通路可以挽救突变肾中的正常基质分化， 外植体鉴于肾脏发育研究的进展显著促进了 了解肾母细胞瘤，反之亦然，与研究肾母细胞瘤生物学导致进展， 发展肾脏学，在这个建议中概述的目标有可能进一步揭示如何复杂的 相互的信号传导相互作用调节正常肾脏中的祖细胞增殖和/或分化 发展，并提供基础，为未来的工作，研究如何中断祖细胞串扰 肾单位和间质谱系之间的相互作用可能改变胚胎微环境， Wilms肿瘤的发展