The role of maladaptive VEGFR2 signaling in renal stroma for chronic kidney disease

肾间质中适应不良的 VEGFR2 信号传导在慢性肾脏病中的作用

• 批准号: 10739504
• 负责人: Takuto Chiba
• 金额: $ 12.14万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-24 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10739504
• 关键词: Acute Renal Failure with Renal Papillary Necrosis; Age; Attenuated; Blood Vessels; Cell Separation; Cells; Chronic Kidney Failure; Cisplatin; Coculture Techniques; Critical Illness; Data; Dependovirus; Disease; Disease Progression; Disease model; Endothelial Cells; Endothelium; Expenditure; Faculty; Fatty Acids; Female; Genes; Heart Arrest; Heterogeneity; Home; Human; In Situ Hybridization; In Vitro; Inflammation; Inflammatory; Injury; Injury to Kidney; KDR gene; Kidney; Kidney Diseases; Knock-out; Knockout Mice; Mediating; Medicare; Medicine; Mentored Research Scientist Development Award; Mentors; Mesenchymal; Methods; Mitochondria; Modeling; Molecular; Mus; Myofibroblast; Nature; PDH kinase; PTPRJ gene; Pathway interactions; Patients; Pericytes; Phenotype; Platelet-Derived Growth Factor; Play; Population; Predictive Factor; Process; Production; Reperfusion Injury; Reporter; Research; Respiration; Risk; Role; Signal Transduction; Small Interfering RNA; Source; Tamoxifen; Testing; Therapeutic; Thrombospondin 1; Universities; Validation; Volatile Fatty Acids; adeno-associated viral vector; cell injury; chemotherapeutic agent; etomoxir; experimental study; fatty acid metabolism; fatty acid oxidation; inhibitor; innovation; kidney cell; loss of function; male; mouse model; pharmacologic; preservation; programs; recruit; renal ischemia; repaired; sex; side effect; single-cell RNA sequencing; therapeutic evaluation; therapy development; transcriptome sequencing

PROJECT SUMMARY/ ABSTRACT A dire consequence of acute kidney injury (AKI) is a dramatically increased risk to develop chronic kidney disease (CKD). CKD accounts for 6.7% of Medicare expenses. Understanding the mechanisms by which AKI progresses to CKD is essential for developing therapies, for which none currently exist. Renal microvasculature, including pericytes and endothelial cells, are damaged in AKI, leading to recruitment of inflammatory cells which contributes to progression to CKD. However, cellular, and molecular mechanisms that drive this process are largely unknown. Pericytes are a heterogeneous mesenchymal population and have been identified as a major source of myofibroblasts that drives CKD. Understanding the molecular mechanisms that mediates maladaptive endothelial-pericyte crosstalk leading to exacerbated and prolonged inflammation could drive therapeutic exploitation of this phenomenon. Previously, it has been shown that the systemic blockade of Vascular endothelial growth factor receptor 2 (VEGF-R2) blocks CKD progression. My preliminary data, knocking out VegfR2 in renal stromally derived cells (RSC) (termed VegfR2RSC-/-) that includes pericytes, confirms the protective nature of this loss-of-function. I found that, in CKD models by renal ischemia/ reperfusion injury (IRI) as well as by cisplatin, VegfR2RSC-/- mice have attenuated CKD progression, along with having mitigated inflammation and preserved vascular function. My bulk RNA-sequencing analysis with isolated RSCs demonstrates that inflammatory pathways are activated while short-chain fatty acid metabolism pathways are suppressed during AKI-to-CKD transition. Mechanistically, VegfR2RSC-/- kidneys (1) have reduced expression of a pro-inflammatory signaling axis of Thrombospondin-1 (TSP1)/ CD148, and (2) have increased expression of fatty acid metabolism associated genes contributing to the enhanced protection. To home in on the timing of the protection, I have generated a tamoxifen-inducible RSC-specific VegfR2 knockout (iVegfR2RSC-/-) mouse. I found that, after pre-treatment of tamoxifen, iVegfR2RSC-/- mice are significantly protected against AKI. Together, these data informed my overarching hypothesis that renal pericyte-specific VEGF-R2 signaling dysregulates pericyte- endothelial crosstalk stimulating inflammation to exacerbate CKD. I propose the following aims to test this: Aim 1 will test the hypothesis that pericyte-specific VEGF-R2 signaling exacerbates AKI-to-CKD transition. Aim 2 will test the hypothesis that pericyte VEGF-R2 signaling mediates maladaptive pericyte-endothelial crosstalk to exacerbate inflammation, promoting AKI-to-CKD. Aim 3 will test the hypothesis that inhibiting VegfR2 signaling in renal pericytes enhances vascular repair, mitigates inflammation, and blocks progression to CKD after AKI. For the K01 Award, I enlisted innovative mentors. The University of Pittsburgh has an extraordinary number of faculty with research programs focused on AKI, and on vascular medicine. Successful completion of these studies will shed light on how renal pericyte specific VEGF-R2 signaling exacerbates CKD progression.

项目摘要/摘要 急性肾脏损伤（AKI）的可怕后果是发展慢性肾脏疾病的风险大大增加 （CKD）。 CKD占医疗保险费用的6.7％。了解AKI进步的机制 CKD对于开发疗法至关重要，目前不存在该疗法。肾脏微脉管系统，包括 周细胞和内皮细胞在AKI中受损，导致炎症细胞的募集 有助于向CKD发展。但是，驱动此过程的细胞和分子机制是 在很大程度上未知。周细胞是异构间充质种群，已被确定为主要 驱动CKD的肌纤维细胞来源。了解介导不良适应性的分子机制 内皮 - 盛行的串扰导致恶化和长时间的炎症可能会导致治疗性 这种现象的剥削。以前，已经显示出血管的全身性阻滞 内皮生长因子受体2（VEGF-R2）阻止CKD进展。我的初步数据，淘汰 肾脏基质衍生细胞（RSC）（称为vegfr2rsc - / - ）的VEGFR2确认 这种功能丧失的保护性质。我发现，在肾脏缺血/再灌注损伤（IRI）的CKD模型中 以及通过顺铂，VEGFR2RSC - / - 小鼠减弱了CKD的进展，并缓解了 炎症和保留的血管功能。我的大量RNA测序分析与孤立的RSC 证明炎症途径被激活，而短链脂肪酸代谢途径是 在AKI到CKD过渡期间被抑制。从机械上讲，VEGFR2RSC - / - 肾脏（1）的表达降低 血小板素-1（TSP1）/ CD148和（2）的促炎信号传导轴的表达增加 脂肪酸代谢相关的基因有助于增强保护。在家中 保护，我已经生成了他莫昔芬诱导的RSC特异性VEGFR2敲除（IVEGFR2RSC - / - ）鼠标。我找到了 在他莫昔芬预处理后，IVEGFR2RSC - / - 小鼠受到AKI的显着保护。在一起，这些 数据告诉我我的总体假设，即肾周细胞特异性VEGF-R2信号传导不稳定周围 内皮串扰刺激炎症加剧CKD。我建议以下目的测试以下目的：目标 1将检验以下假设：周细胞特异性VEGF-R2信号传导加剧了AKI至CKD转变。目标2将 测试周围vegf-r2信号传导介导不良适应性周细胞内皮串扰的假设 加剧炎症，促进AKI至CKD。 AIM 3将检验抑制VEGFR2信号的假设 在肾周中，周细胞增强血管修复，减轻炎症，并阻止AKI后的CKD进展。 为了获得K01奖，我招募了创新的导师。匹兹堡大学的数量很大 研究计划的教师侧重于AKI和血管医学。这些成功完成 研究将阐明肾周细胞特异性VEGF-R2信号传导如何加剧CKD的进展。