A new paradigm of glomerular immune cell homing

肾小球免疫细胞归巢的新范例

• 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
• 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

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.

总结 本研究旨在研究肾小球免疫细胞归巢的一种全新模式，并揭示肾小球免疫细胞归巢的机制。 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）和狼疮性肾炎（LN）的病症。作为附加的平移目标， 这些研究将为目前肾脏疾病治疗方法的再利用和新药物的开发提供信息。 抗炎治疗。已知循环和肾脏驻留免疫细胞优先归巢于 与正常和炎症状态下的其他血管床相比， 肾小球微循环中存在保护肾脏的独特免疫调节机制 滤波然而，基本的细胞和分子机制在很大程度上是未知的。的重点 提出的研究是致密斑（MD）细胞的一种新功能， 肾小球血管入口，并且传统上已知调节肾和肾小球血液动力学。我们 一个实验室最近发现，在所有肾细胞类型中，MD细胞具有最高的蛋白质合成速率 它们分泌多种旁分泌作用的血管生成、细胞生长和图案化以及细胞外基质， 信号蛋白初步的工作确定了高MD富集的几个途径和基因， 炎症反应和白细胞的细胞浸润（例如，Cxcl12、Cxcl14、Ptgs2、Ptges、Anxa1、Ccn1， Mif），并观察炎症性肾小球疾病中以MD为中心的CD 44+和CD 8 + T细胞的密度和迁移。 包括狼疮性肾炎在内的疾病。我们的中心假设是MD细胞驱动优先肾小球 通过释放旁分泌作用的促炎因子（包括趋化因子）和 细胞因子该项目将使用综合实验方法，包括新的转基因小鼠， 条件性、诱导性和光遗传学工具（MD-mTORgof/1 of、MD-Ai 27、MD-Ai 39、CCN 1-敲除（KO）、CCN 2-敲除（KO）、CCN 3-敲除（KO）、CCN 2-敲除（KO）、C2-敲除（ GFP小鼠）、肾脏和全身炎症模型（NTS、STZ-DKD、NZM. 2328 LN模型）、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。 具体的目的是（1）检查MD细胞的整体免疫调节作用和机制， 生理和肾脏炎性疾病状况，（2）阐明CCN 1的肾脏抗炎作用， （3）检测SGLT 2抑制剂（SGLT 2 i）的抗炎作用。这些新发现的MD机制 可能是有针对性的，并将为未来的抗炎治疗策略的发展提供信息。