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 早产儿肾单位数量低（禀赋），患慢性肾病（CKD）的风险高 和终末期肾病。大多数肾单位是在妊娠后期通过一种不太清楚的 称为侧分支肾发生（LBN）的过程。作为妊娠晚期人类肾脏发育的直接研究 是困难的，非人灵长类动物模型（恒河猴）最近被确定为一个合适的模型，以桥梁 这种知识差距。长期目标是将LBN的分子发现应用于非人类灵长类动物 旨在开发旨在延长早产儿肾发生的治疗方法的模型。的 中心假设是涉及发育中的肾脏（肾单位）成分的信号环境发生变化 在晚期膀胱移行细胞癌中，LBN（包括前体细胞（NPC）、输尿管芽（UB）和间质）导致LBN持续多周。 怀孕这项拟议研究的基本原理是，遗传上易于处理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 捐赠。中心假设将通过使用最新的分子技术来测试，以了解 在非人灵长类动物中维持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 测序对出生后第二天绒猴肾脏的初步形态学研究确定了一个单一的输尿管 茎与侧枝与LBN一致，这表明普通绒猴可以作为遗传学上的 温顺的灵长类动物模型预期成果包括组装最大的灵长类动物妊娠后期发展 肾脏分子数据集和鉴定基因和途径丰富和调控网络活跃 在LBN期间，以及确定一个遗传上易于处理的模型来研究LBN，以了解为什么早产 导致人类肾脏发生的早期停止，以及如何延长它。这些结果预计将有 通过鉴定分子标记，对目前对妊娠晚期人类肾发生的理解产生了积极影响。 途径和潜在的治疗干预措施，为早产儿。除了《公约》概述的目标外， 该建议，职业发展计划包括分子生物学和生物信息学的教学培训， 生物信息学证书课程，最新分子技术的湿实验室经验和多 经济平台，并通过一个选定的咨询委员会促进向独立过渡的职业发展 并在K08授标期间提交R01。