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Renal vascular remodeling and arteriogenesis: cues from smooth muscle progenitor cells

肾血管重塑和动脉生成：来自平滑肌祖细胞的线索

基本信息

批准号：
10320039
负责人：
Ondine B Cleaver
金额：
$ 35.25万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10320039
关键词：
Ablation Acute Renal Failure with Renal Papillary Necrosis Address Adult Affect American Blood Vessels Blood flow Bypass Cell Differentiation process Cell Lineage Cell model Cells Coupling Cues Data Defect Development Dialysis procedure Embryo End stage renal failure Endothelial Cells Endothelium Event Exhibits FOXC1 gene Genetic Goals Growth Health Homeostasis Hypertension Hypoxia Impairment In Vitro Invaded Kidney Kidney Failure Laboratories Laminin Metanephric Diverticulum Molecular Mus Muscle satellite cell NTN1 gene Nephrons Organ Organogenesis Pattern Pharmacology Phenotype Proteins Regulation Renal Replacement Therapy Renal Tissue Reporting Role Signal Transduction Smooth Muscle Smooth Muscle Myocytes Stromal Cells Survival Rate System Testing Time Transgenic Mice Trees Vascular Smooth Muscle Vascular remodeling angiogenesis cell type in vitro testing in vivo kidney vascular structure mouse model mutant neogenin nephrogenesis nephron progenitor overexpression postnatal receptor renal artery renal hypoxia stem cells transcription factor

项目摘要

SUMMARY Approximately 500,000 Americans have end-stage renal disease. Although organ function can be supplemented using dialysis, the 10-year survival rate is just over 10%. Ex vivo organogenesis has the potential to meet this demand by providing functional tissue for renal replacement therapy. However to effectively generate functional kidneys in the laboratory, all cell types within the kidney and their functional ontogeny and coupling must be understood. Renal blood vessels are one such component that is critical to the health and homeostasis of the kidney. Building the kidney's intricate arterial network requires synchronized actions of endothelial cells (ECs) and vascular smooth muscle cells (vSMCs) in a sequence that is poorly understood. vSMCs arise in the developing kidney from Foxd1+ nephrogenic stromal cells (NSCs). Ablation of Foxd1 or Foxd1+ cells cause similar defects in renal arteries suggesting Foxd1+ NSC encoded signals may govern renal arteriogenesis. We have found that the secreted laminin-like protein Netrin-1 (Ntn1) is expressed by Foxc1+ NSCs in the embryonic kidney. To date, the role of Ntn1 during kidney development has not been reported. Genetic ablation of Ntn1 from Foxd1+ NSCs (hereafter denoted as Ntn1NSCKO) blocks formation of a perfused renal arterial tree during development. In addition, we find Ntn1NSCKO display an aberrant vSMCs-associated cortical vasculature and increased kidney hypoxia. As development proceeds, Ntn1NSCKO also exhibit reduced ureteric bud (UB) branching, and postnatal Ntn1NSCKO mice display delayed nephrogenesis with ~30% fewer glomeruli. Our preliminary data identifies a critical role for Ntn1 in kidney development, but the mechanism(s) of this regulation is unknown. In this proposal, we address this and ask which receptor is needed for kidney formation: stromal-neogenin1, NPC-Unc5b or endothelial-Unc5b? We hypothesize that NSC-derived Ntn1 cell-autonomously directs vSMC lineage specification via its receptor neogenin1 (Neo1) and the transcription factor Klf4, to instruct renal arterial tree assembly, upon which nephron development indirectly depends. We will test this hypothesis by carrying out following aims: 1) We will test the cell autonomy of NSC-derived Ntn1 signaling during renal arterial assembly, by asking how the stroma responds upon absence of Ntn1, where ectopic SMCs come from in the mutants, and which Ntn1 receptors (hence which cell types) are required for arterial development (which recapitulate the Ntn1 phenotype when ablated). 2) We will examine if aberrant nephrogenesis is caused directly via Ntn1 loss in NPCs, or indirectly due to failure of renal arteriogenesis. We will characterize failed nephrogenesis in the Ntn1 mutant kidneys; we will use in vitro systems to see what cell types respond to Ntn1 (NPCs versus ECs); we will test whether late deletion of Ntn1 recapitulates the Ntn1 mutant phenotype, as it bypasses failed arteriogenesis and associated hypoxia; and lastly we will test the role of hypoxia on NPCs directly. 3) We will examine whether Klf4 is required for stromal differentiation and renal arteriogenesis, and whether it signals downstream of Ntn1.

总结大约50万美国人患有终末期肾病。虽然器官功能可以如果辅以透析治疗，10年存活率仅略高于10%。离体器官形成有可能以通过提供用于肾替代治疗的功能组织来满足这种需求。为了有效地产生实验室中的功能性肾脏，肾脏内的所有细胞类型及其功能性个体发育和偶联必须被理解。肾血管就是这样一个组成部分，它对健康和体内平衡至关重要。肾脏的构建肾脏复杂的动脉网络需要内皮细胞的同步动作 (ECs)和血管平滑肌细胞（vSMC）的顺序知之甚少。vSMC出现在从Foxd 1+肾源性基质细胞（NSC）发育肾脏。Foxd 1或Foxd 1+细胞的消融导致肾动脉中的类似缺陷表明Foxd 1 + NSC编码的信号可能支配肾动脉生成。我们已经发现，分泌的层粘连蛋白样蛋白Netrin-1（Ntn 1）由Foxc 1 + NSCs在神经干细胞中表达。胚胎肾迄今为止，Ntn 1在肾脏发育过程中的作用尚未报道。从Foxd 1 + NSC中基因切除Ntn 1（下文称为Ntn 1 NSCKO）阻断了灌注的肾动脉树在发展过程中。此外，我们发现Ntn 1 NSCKO显示异常的vSMC相关的皮质血管和肾脏缺氧增加。随着开发的进行，Ntn 1 NSCKO也表现出减少输尿管芽（UB）分支和出生后Ntn 1 NSCKO小鼠显示肾发生延迟，肾小球我们的初步数据确定了Ntn 1在肾脏发育中的关键作用，但其机制这一规定是未知的。在这个提议中，我们解决了这个问题，并询问肾脏需要哪种受体。形成：基质-新生素1、NPC-Unc 5 b或内皮-Unc 5 b？我们假设神经干细胞来源的Ntn 1细胞通过其自身的功能自主地指导vSMC谱系特化。受体再生素1（Neo 1）和转录因子Klf 4，以指导肾动脉树组装，肾单位的发育间接依赖于哪个。我们将通过实现以下目标来检验这一假设： 1)我们将测试肾动脉组装过程中神经干细胞来源的Ntn 1信号传导的细胞自主性，基质响应于Ntn 1的缺失，其中异位SMC来自突变体，并且Ntn 1 受体（因此是哪种细胞类型）是动脉发育所必需的（概括了Ntn 1表型当消融时）。2)我们将检查异常的肾发生是否直接通过NPC中的Ntn 1缺失引起，或者间接由于肾动脉生成的失败。我们将描述Ntn 1突变体中肾发生失败的特征，肾脏;我们将使用体外系统来观察哪些细胞类型对Ntn 1有反应（NPC与EC）;我们将测试 Ntn 1的晚期缺失是否重演了Ntn 1突变表型，因为它绕过了失败的动脉生成，最后，我们将直接测试缺氧对NPC的作用。3)我们将研究Klf 4是否是间质分化和肾动脉形成所必需的，以及它是否在Ntn 1下游发出信号。