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

基本信息

批准号：
10689659
负责人：
Meredith Schuh
金额：
$ 16.58万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
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/10689659
关键词：
3-Dimensional ATAC-seq Adult Advisory Committees Age Anatomy Animals 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 Kidney Diseases 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 System TWIST1 gene Technology Testing Therapeutic Therapeutic Intervention Third Pregnancy Trimester Tissues Training With laterality 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 nucleus RNA-sequencing single-cell RNA sequencing stem cells therapeutic development transcription factor transcriptome transcriptome sequencing transcriptomics

项目摘要

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.

项目概要早产儿的肾单位数量（禀赋）较低，患慢性肾脏病 (CKD) 的风险较高和成人终末期肾病。大多数肾单位是在妊娠后期通过一种知之甚少的方式添加的称为侧支肾发生（LBN）的过程。作为妊娠晚期人类肾脏发育的直接研究很难，非人类灵长类动物模型（恒河猴）最近被确定为一个合适的模型来弥补这种知识差距。长期目标是将 LBN 的分子发现应用于非人类灵长类动物开发旨在延长早产儿肾发生的治疗方法的模型。这中心假设是涉及发育中肾脏（肾单位）成分的信号环境的转变祖细胞 (NPC)、输尿管芽 (UB) 和基质）导致晚期 LBN 维持数周妊娠。这项研究的基本原理是，遗传上易处理的非人类灵长类动物模型系统可用于测试假设并应用旨在改善人类肾单位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，以了解非人类灵长类动物维持 LBN 的分子机制，包括单细胞 RNA 测序、激光通过 RNA 测序、单核 RNA 测序和单核 ATAC 捕获显微切割测序。产后第二天狨猴肾脏的初步形态学研究发现单个输尿管茎具有与 LBN 一致的侧枝，表明普通狨猴可以用作遗传驯服的灵长类动物模型。预期成果包括最大的灵长类动物妊娠晚期发育的组装肾脏分子数据集以及基因和通路的鉴定丰富且调控网络活跃 LBN 期间，以及识别遗传易处理的模型来研究 LBN，以了解早产的原因导致人类肾发生提前停止，以及如何延长它。这些结果预计将有通过识别分子对当前对妊娠晚期人类肾发生的理解产生积极影响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该提案中，职业发展计划包括分子生物学和生物信息学方面的教学培训生物信息学证书课程、最新分子技术和多技术的湿实验室经验 Omic 平台，以及通过选定的咨询委员会实现职业发展以过渡到独立并在K08奖励期间提交R01。