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

使用非人类灵长类动物模型弥合妊娠晚期人类肾发生的差距

• 批准号: 10347725
• 负责人: Meredith Schuh
• 金额: $ 16.73万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10347725
• 关键词: 3-Dimensional; Adult; Advisory Committees; Age; Anatomy; Animals; Archives; Award; Binding Sites; Bioinformatics; Biological Models; Birth; CRISPR/Cas technology; Callithrix; Callithrix jacchus jacchus; Career Mobility; Cell Nucleus; Cells; Chronic Kidney Failure; Coupled; Data; Data Set; Development Plans; End stage renal failure; Endowment; Environment; Fetal Kidney; Freezing; Future; Gene set enrichment analysis; Genes; Genetic; Genetic Transcription; Genomics; Gestational Age; Goals; Health; Human; In Situ Hybridization; Kidney; Knowledge; Lateral; Life; Macaca mulatta; Maps; Metanephric Diverticulum; Methods; Modeling; Molecular; Molecular Biology; Morphology; Mus; Nephrons; Outcome; Pathway interactions; Population; Pregnancy; Premature Birth; Premature Infant; Primates; Process; RNA; Research; Rhesus; Risk; Sampling; Second Pregnancy Trimester; Signal Transduction; Small Nuclear RNA; System; TWIST1 gene; Technology; Testing; Therapeutic; Therapeutic Intervention; Third Pregnancy Trimester; Tissues; Training; XCL1 gene; age related; career development; cell stroma; certificate program; critical period; druggable target; epigenome; gene regulatory network; genetic manipulation; high risk; improved; laboratory experience; laser capture microdissection; multiple omics; nephrogenesis; nephron progenitor; nonhuman primate; novel; periviable; postnatal; postnatal period; premature; prevent; single-cell RNA sequencing; stem cells; therapeutic development; transcription factor; transcriptome; transcriptome sequencing; transcriptomics

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.

项目摘要 早产婴儿的肾单位数量较低（end赋），并且患慢性肾脏疾病（CKD）的风险很高 和成年后的末期肾脏疾病。大多数肾单位在妊娠后期通过鲜为人知的 过程称为侧支肾病（LBN）。作为妊娠晚期人肾脏发育的直接研究 很难，最近将非人类灵长类动物模型（恒河猕猴）确定为桥梁的合适模型 这个知识差距。长期目标是将LBN的分子发现应用于非人类灵长类动物 开发旨在扩展早产儿肾病的治疗方法的模型。这 中心假设是涉及发育肾脏成分的信号传导环境的转移 祖细胞（NPC），输尿管芽（UB）和基质）导致lbn持续多个星期的lbn 妊娠。这项拟议的研究的理由是遗传上可触犯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肾肾经马曲的治疗干预措施 捐赠。中心假设将通过使用最新的分子技术来检验 非人类灵长类动物中维持LBN的分子机制，包括单细胞RNA测序，激光 用RNA测序，单核RNA测序和单核ATAC捕获显微解剖 测序。产后两摩斯特肾脏的初步形态学研究确定了单个输尿管 与LBN一致的横向分支的茎，这表明可以用作遗传学 可拖动的灵长类动物模型。预期的结果包括最大的灵长类动物晚期妊娠的组装 肾脏分子数据集以及富含基因和途径的富含和调节网络的识别 在LBN期间，以及鉴定可遗传障碍模型以研究LBN以了解为什么早产 导致早期停止人类的肾脏形成，以及如何扩展肾脏。这些结果预计将有 通过鉴定分子来对当前对妊娠的人肾病的理解产生积极影响 过早出生的人的途径和潜在的治疗干预措施。除了概述的目的 这项建议，职业发展计划包括通过 生物信息学证书计划，最新分子技术的湿lab体验和多种 通过选定的咨询委员会过渡到独立的咨询委员会的OMIC平台和职业发展 并在K08奖励期内提交R01。