A new paradigm of glomerular immune cell homing

肾小球免疫细胞归巢的新范例

基本信息

批准号：
10608895
负责人：
JANOS PETI-PETERDI
金额：
$ 47.5万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
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：
2022-12-15 至 2026-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10608895
关键词：
Affect Anatomy Anti-Inflammatory Agents Antibodies Antiinflammatory Effect Blood Vessels CD3 Antigens CD44 gene CD8-Positive T-Lymphocytes CD8B1 gene Cd68 Cell Culture Techniques Cell Separation Cell physiology Cells Cellular Infiltration Chemotaxis Complement Cytometry Development Diabetic Nephropathy Disease Extracellular Matrix FRAP1 gene Functional disorder Future Genes Genetic Glomerulonephritis Health Home Homing Immune In Vitro Inflammation Inflammatory Inflammatory Response Injury Kidney Kidney Diseases Knock-out Label Laboratories Leucocytic infiltrate Lupus Nephritis Macula densa Maintenance Microcirculation Modeling Molecular Mus PTPRC gene Pathway interactions Pattern Physiological Play Population Positioning Attribute Property Protein Biosynthesis Proteins Proteomics Role Serum Signaling Protein Streptozocin Stromal Cell-Derived Factor 1 Structure Testing Therapeutic Time Tissues Transgenic Mice Work cell growth cell type chemokine comorbidity cytokine density diabetic gain of function glomerular endothelium hemodynamics immunoregulation improved in vivo inhibitor kidney cell kidney cortex loss of function microscopic imaging migration mortality multiphoton microscopy nephrotoxicity non-diabetic novel therapeutic intervention optogenetics paracrine pre-clinical single-cell RNA sequencing systemic inflammatory response tool transcriptome transcriptomics translational therapeutics vascular bed

项目摘要

SUMMARY This study aims to investigate a radically new paradigm of glomerular immune cell homing and to uncover the underlying cell and molecular mechanisms and their role in physiological and renal inflammatory disease conditions including diabetic kidney disease (DKD) and lupus nephritis (LN). As an additional translational aim, these studies will inform the repurposing of current treatments for kidney disease and the development of new anti-inflammatory therapies. Circulating and kidney resident immune cells are known to preferentially home in and around the glomerulus compared to other vascular beds in both normal and inflammatory states, suggesting the presence of unique immunomodulatory mechanisms in the glomerular microcirculation that protect the kidney filter. However, the underlying cell and molecular mechanisms have been largely unknown. The focus of the proposed studies is a new function of the cells of the macula densa (MD) which are strategically positioned at the glomerular vascular entrance and traditionally known to regulate renal and glomerular hemodynamics. Our laboratory recently identified that MD cells have the highest rate of protein synthesis among all kidney cell types and they secrete a variety of paracrine-acting angiogenic, cell growth and patterning, and extracellular matrix signaling proteins. Preliminary work identified the high MD-enrichment of several pathways and genes in the inflammatory response and cellular infiltration by leukocytes (e.g., Cxcl12, Cxcl14, Ptgs2, Ptges, Anxa1, Ccn1, Mif) and observed the MD-centric glomerular density and migration of CD44+ and CD8+ T cells in inflammatory diseases including lupus nephritis. Our central hypothesis is that MD cells drive the preferential glomerular homing of immune cells by the release of paracrine-acting pro-inflammatory factors including chemokines and cytokines. This project will use comprehensive experimental approaches including new transgenic mice with conditional, inducible and optogenetic tools (MD-mTORgof/lof, MD-Ai27, MD-Ai39, CCN1-knockout (KO), CCN1- GFP mice), models of renal and systemic inflammation (NTS, STZ-DKD, NZM.2328 LN models), in vivo MPM imaging, MD transcriptome analysis, in vitro cell culture and proteomics, and pre-clinical therapeutic translation. The specific aims are to (1) Examine the global immunomodulatory role and mechanisms of MD cells under physiological and renal inflammatory disease conditions, (2) Elucidate the renal anti-inflammatory role of CCN1, and (3) Test the anti-inflammatory effects of SGLT2 inhibitors (SGLT2i). These newly identified MD mechanisms may be targeted and will inform the development of future anti-inflammatory therapeutic strategies.

概括这项研究旨在研究肾小球免疫细胞寄居的根本新范式，并发现潜在的细胞和分子机制及其在生理和肾脏炎症性疾病中的作用包括糖尿病肾病（DKD）和狼疮肾炎（LN）在内的疾病。作为另一个翻译目标，这些研究将为当前肾脏疾病治疗的重新利用和新的新疗法提供了信息抗炎疗法。已知循环和肾脏常驻免疫细胞在与正常状态和炎症状态下其他血管床相比，肾小球周围在保护肾脏的肾小球微循环中存在独特的免疫调节机制筛选。但是，基本的细胞和分子机制在很大程度上未知。重点拟议的研究是黄斑densa（MD）细胞的新功能，该功能在战略上位于肾小球血管入口，传统上已知可以调节肾脏和肾小球血流动力学。我们的实验室最近确定MD细胞在所有肾细胞类型中的蛋白质合成率最高他们分泌各种旁分泌作用的血管生成，细胞生长和图案，以及细胞外基质信号蛋白。初步工作确定了多种途径和基因的高度MD增强白细胞（例如CXCL12，CXCL14，PTGS2，PTGES，PTGES，ANXA1，CCN1，CCN1，CCN1，CCN1，CCN1， MIF）并观察到以MD为中心的肾小球密度以及CD44+和CD8+ T细胞在炎症中的迁移包括狼疮肾炎在内的疾病。我们的中心假设是MD细胞驱动优先的肾小球通过释放旁分泌作用促炎性因子（包括趋化因子和细胞因子。该项目将使用全面的实验方法，包括新的转基因小鼠条件，诱导和光遗传学工具（MD-MTORGOF/LOF，MD-AI27，MD-AI39，CCN1-KNOCKOUT（KO），CCN1-- GFP小鼠），肾脏和全身炎症的模型（NTS，STZ-DKD，NZM.2328 LN型号），体内MPM 成像，MD转录组分析，体外细胞培养和蛋白质组学以及临床前治疗翻译。具体目的是（1）检查MD细胞下的全球免疫调节作用和机制生理和肾脏炎症性疾病，（2）阐明CCN1的肾脏抗炎作用（3）测试SGLT2抑制剂（SGLT2I）的抗炎作用。这些新确定的MD机制可能是针对性的，并将为未来的抗炎治疗策略提供信息。