Bridging the gap of late gestation human nephrogenesis using a non-human primate model

项目摘要 早产儿肾单位数量少，患慢性肾脏疾病(CKD)的风险高 以及成年后的终末期肾病。大多数肾单位是在妊娠后期通过一种鲜为人知的 这一过程称为侧支肾生成(LBN)。作为妊娠晚期人类肾脏发育的直接研究 非人灵长类动物模型(恒河猴)最近被确定为适合桥接的模型 这种知识鸿沟。长期目标是将LBN的分子发现应用于非人类灵长类动物 旨在延长早产儿肾脏发生的治疗方法的发展模式。这个 中心假设是信号环境的变化涉及到发育中的肾脏的组成部分(肾单位 造血祖细胞(NPC)、输尿管芽(UB)和间质)导致LBN在晚期持续数周 怀孕了。这项拟议研究的基本原理是，遗传上易驯化的非人类灵长类动物模型 系统可用于检验假设并应用旨在改善人类肾单位的治疗干预措施 天赋。中心假说将通过使用最新的分子技术来检验，以了解 支持非人灵长类LBN的分子机制，包括单细胞RNA测序，激光 利用RNA测序、单核RNA测序和单核ATAC捕获显微切割 测序。生后2日龄猴肾单侧输尿管的形态观察 茎具有与LBN一致的侧枝，表明常见的绒猴可能在遗传上用作 易驯服的灵长类模型。预期结果包括发育中最大的灵长类动物的组装 肾脏分子数据集和基因和途径的识别丰富和调控网络活跃 在LBN期间，以及确定一个遗传易受控制的模型来研究LBN，以了解为什么早产 导致人类早期停止肾脏生成，以及如何延长这一过程。这些结果预计将产生 分子鉴定对目前对妊娠晚期人类肾脏发生的认识有积极影响 针对早产儿的途径和潜在的治疗干预措施。除了中概述的目标之外 这项提议，职业发展计划包括分子生物学和生物信息学的说教培训，通过 生物信息学证书课程，最新分子技术和多学科的湿法实验室经验 OMIC平台，以及通过一个选定的咨询委员会向独立过渡的职业发展 并在K08获奖期间提交R01。

Project Summary Premature infants have low nephron number (endowment) and are at high risk for chronic kidney disease (CKD) and end stage renal disease as adults. Most nephrons are added late in gestation through a poorly understood process called lateral branch nephrogenesis (LBN). As direct study of late gestation human kidney development is difficult, the non-human primate model (rhesus macaque) was recently identified as a suitable model to bridge this knowledge gap. The long-term goal is to apply the molecular findings of LBN in the non-human primate model towards development of therapeutic methods aimed at extending nephrogenesis in preterm infants. The central hypothesis is that a shift in the signaling milieu involving components of the developing kidney (nephron progenitor cells (NPC), ureteric bud (UB), and stroma) results in sustaining LBN over multiple weeks in late gestation. The rationale for this proposed research is that the genetically tractable non-human primate model system can be used to test hypotheses and apply therapeutic interventions aimed at improving human nephron endowment. The central hypothesis will be tested by using the latest molecular technologies to understand the molecular mechanism sustaining LBN in the non-human primate, including single-cell RNA sequencing, laser capture microdissection with RNA sequencing, single-nucleus RNA sequencing, and single-nucleus ATAC sequencing. Preliminary morphologic study on the postnatal day two marmoset kidney identified a single ureteric stalk with lateral branches consistent with LBN, suggesting the common marmoset could be used as a genetically tractable primate model. Expected outcomes include assembly of the largest primate late gestation developing kidney molecular dataset and identification of genes and pathways enriched and regulatory networks active during LBN, as well as identification of a genetically tractable model to study LBN to understand why prematurity leads to early cessation of nephrogenesis in humans, and how to extend it. These results are expected to have a positive impact on the current understanding of late gestation human nephrogenesis by identifying molecular pathways and potential therapeutic interventions for those born prematurely. In addition to the aims outlined in this proposal, career development plans include didactic training in molecular biology and bioinformatics through the Certificate program in Bioinformatics, wet-lab experience with the latest molecular technologies and multi- omic platforms, and career advancement through a selected advisory committee for transition to independence and submission of R01 during the K08 award period.