Defining the molecular landscape of hyperfiltration-mediated glomerular injury using kidney allografts as a model system

摘要： 在过去的二十年里，肾移植的长期存活率没有显著提高。积累 数据支持进行性肾小球疾病导致晚期肾移植失败的假设， 滤过介导的肾小球损伤是一个假定的驱动因素。此外，超滤也参与了这一进程- 糖尿病和肥胖症等肾脏疾病的锡永，这两种疾病都已达到流行病的程度。怎么- 然而，这种肾脏疾病进展的共同机制的分子基础仍然不清楚， 这是这项建议的科学依据。 我的长期目标是了解导致晚期同种异体移植物丢失的疾病机制，重点是延长 同种异体移植物寿命。本申请的总体目标是阐明 超滤引发并驱动肾同种异体移植物中的足细胞脱离过程。我们还将测试 平行机制在高滤过糖尿病患者的独立队列中起作用。 为了实现这一目标，我的中心假设是，超滤导致一个特征分子， 肾小球中的肾小球足迹，驱动足细胞应激和加速脱离。为了确定潜在的 超滤的分子机制，批量和单细胞RNA测序技术的组合将 用于鉴定肾小球细胞特异性基因特征以及与肾小球疾病相关的细胞之间的相互作用。 肾小球基底膜是已知的驱动足细胞脱离。为了实现非侵入性监测， 为了检测足细胞损失，我们将使用尿颗粒足细胞分离试验，并使用 过滤分数研究。我们将使用三个具体目标来检验中心假设： 目标1。定义肾小球对超滤的转录反应。 目标2.定义驱动足细胞加速脱离的肾小球转录谱，并识别 超滤对这段关系的影响 目的3：明确同种异体移植物超滤与足细胞应力和脱离的关系。 这项研究是创新的，因为它使用人类肾脏移植作为模型系统，专注于超滤 作为通过最新技术分析的肾脏疾病进展的常见机制。拟议 研究是重要的，因为确定了肾脏疾病之间疾病进展的共同途径， 可能导致新的靶向治疗剂和非侵入性监测策略的发展。Ulti 因此，这些知识对于减缓肾脏疾病进展至关重要，无论其病因如何。

ABSTRACT: Long-term kidney allograft survival has not improved significantly over the last two decades. Accumulating data supports the hypothesis that progressive glomerular disease drives late kidney allograft failure, with hyper- filtration mediated glomerular injury being a putative driver. In addition, hyperfiltration is implicated in the progres- sion of kidney diseases such as in diabetes and obesity, both of which have reached epidemic proportions. How- ever, the molecular underpinnings of this shared mechanism of kidney disease progression remain unclear and forms the scientific basis of this proposal. My long-term goal is to understand disease mechanisms driving late allograft loss, with a focus on prolonging allograft lifespan. The overall objective of this application is to elucidate the molecular mechanisms by which hyperfiltration initiates and drives the podocyte detachment process in kidney allografts. We will also test whether parallel mechanisms are operating in an independent cohort of hyperfiltering diabetic patients. Towards achieving this objective my central hypothesis is that hyperfiltration leads to a characteristic molec- ular footprint in the glomerulus that drives podocyte stress and accelerated detachment. To identify the underlying molecular mechanism of hyperfiltration, a combination of bulk- and single cell RNA-sequencing technology will be used to identify glomerular cell-specific gene signatures as well as interactions between cells associated with the glomerular basement membrane that are known to drive podocyte detachment. To enable non-invasive mon- itoring of podocyte loss, we will use urine pellet podocyte detachment assays and measure hyperfiltration using filtration fraction studies. We will test the central hypothesis using three specific aims: Aim 1. Define the glomerular transcriptional response to hyperfiltration. Aim 2. Define the glomerular transcriptional profile that drives accelerated podocyte detachment and identify the effect of hyperfiltration on this relationship. Aim 3: Define the relationship of allograft hyperfiltration with podocyte stress and detachment. The research is innovative as it uses human kidney allografts as a model system to focus on hyperfiltration as a common mechanism of kidney disease progression analyzed by state-of-the-art technologies. The proposed research is significant as the identification of shared pathways of disease progression between kidney diseases could lead to the development of novel targeted therapeutic agents and non-invasive monitoring strategies. Ulti- mately, such knowledge will be crucial to slow down kidney disease progression regardless of its etiology.