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.

项目概要/摘要 该 NIH 指导职业发展奖描述了一项为期三年的提案，提供培训和 候选人发展成为研究遗传和疾病的独立研究者所必需的指导 肾脏发育的分子调控，特别关注肾母细胞瘤 1 (Wt1) 的突变 影响发育中肾脏的细胞谱系串扰。为了实现这一目标，候选人开发了一个 详细的职业发展计划融合了 UT 优秀的指导和科学环境 西南航空在生物信息学和体外技术方面进行了重点培训，成功过渡到 独立。 Wt1 是在肾母细胞瘤中发现的第一个基因，调节正常肾脏的多个方面 发育，之前已被证明在肾单位祖细胞 (NPC) 中发挥关键作用 差异化。进一步检查 Wt1 功能丧失的突变肾脏，专门针对 NPC 谱系（Six2cre；Wt1c/c）显示出对肾源性基质的额外影响，伴随着基质的扩张 祖细胞、基质模式破坏以及多种基质标志物表达增加 先前显示在肾母细胞瘤中表达上调，从而得出以下假设：来自 NPC 的信号 和/或早期肾单位结构非自主调节基质发育。对此的测试 假设将询问 NPC 与基质的串扰如何指导基质祖细胞的维持和/或 正常肾脏发育的分化，这将通过两个拟议目标来实现。在目标 1 中， 利用基因工程小鼠模型进行的实验旨在确定来自 肾单位谱系衍生细胞调节肾源性基质。为此，基质增殖、细胞凋亡和 将在针对不同肾单位谱系群体的突变小鼠模型中检查模式（即： 自我更新的 NPC 与早期肾单位结构）相比，对基质的影响在 Six2cre；Wt1c/c 突变肾。目标 2 将研究 NPC 与基质串扰的机制，使用 生物信息学来识别候选受体/配体相互作用，并进行体外测定以对其进行功能验证 通过确定破坏特定信号通路是否会导致对照肾脏的基质缺陷来确定通路， 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极大地促进了 对肾母细胞瘤的了解，反之亦然，对肾母细胞瘤生物学的研究导致了以下领域的进步 发育肾病学，该提案中概述的目标有可能进一步揭示复杂的 相互信号相互作用调节正常肾脏中的祖细胞增殖和/或分化 开发并为未来研究祖细胞串扰如何受到干扰的工作奠定基础 肾单位和基质谱系之间的相互作用可能会改变胚胎微环境并导致 维尔姆斯瘤的发展。