Targeting Podocyte-Endothelial Cell Crosstalk as a FSGS Therapy

靶向足细胞-内皮细胞串扰作为 FSGS 疗法

• 批准号: 10635547
• 负责人: Stuart James Shankland
• 金额: $ 77.52万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10635547
• 关键词: Actins; Acute; Address; Adult; Applications Grants; Biological Markers; CASP1 gene; Cell Count; Cells; Chronic; Chronic Kidney Failure; Cicatrix; Cytoskeleton; Data; Data Analyses; Devices; Disease; Disease Outcome; Disease Progression; Disease remission; End stage renal failure; Endothelial Cells; Endothelium; Epithelial Cells; Event; Exhibits; Experimental Designs; Focal and Segmental Glomerulosclerosis; Functional disorder; Genes; Genetic; Glycocalyx; Goals; Grant; Health; Human; Inflammasome; Inflammatory; Injury; Knowledge; Ligands; Link; Measures; Mediating; Mediator; Mesenchymal; Mus; Mutant Strains Mice; Neptune; Outcome; Paracrine Communication; Parietal; Pathogenesis; Pathology; Patient-Focused Outcomes; Patients; Phenotype; Physiological; Process; Public Health; Publishing; Receptor Cell; Receptor Signaling; Renal glomerular disease; Research; Risk; Signal Transduction; Testing; Therapeutic; Transgenic Mice; Translations; Validation; Vascular Endothelial Growth Factors; alpha 2-Glucoproteins; cell injury; cell type; design; gain of function; genetic approach; glomerular endothelium; human data; improved; in silico; in vivo; injured; innovation; loss of function; novel therapeutic intervention; overexpression; paracrine; podocyte; prevent; protein expression; receptor; senescence; single nucleus RNA-sequencing; slit diaphragm; theories; therapeutic target; transcriptomics; translational impact

PROJECT SUMMARY/ABSTRACT The scope of the problem is that 50% of patients with focal segmental glomerulosclerosis (FSGS), the leading cause of primary proteinuric glomerular disease in adults, progress to chronic kidney disease. Thus, developing new treatment strategies to improve FSGS patient outcomes is of utmost importance. Yet, there is a large unmet need in our mechanistic understanding of disease progression. The traditional view in FSGS is that podocytes are central to its pathology and that they are the first cell type to be injured. A more contemporary view is an expansion of this paradigm and includes injury to neighboring glomerular cells. This grant will focus on glomerular endothelial cells (GEnCs). The rationale is that (1) in FSGS, GEnCs decrease in number, loose their glycocalyx and their fenestrae widen; (2) patients with GEnC damage have the highest rates of disease progression; (3) GEnC injury scores for all glomerular diseases are highest in FSGS; (4) GEnC genes/biomarkers are linked to the lowest remission rates and poor long-term outcomes of FSGS. Yet the understanding of how podocytes cause secondary GEnC injury is very incomplete. To begin to close the knowledge gap, we undertook an in silico approach to identify paracrine ligand-receptor signaling networks between podocytes and GEnCs. Surprisingly, we discovered that experimental and human FSGS results in a senescence associated secretory phenotype (SASP) and an activated inflammasome phenotype in non-aged podocytes, both of which are characterized by the secretion of distinct classes of signaling mediators. Our preliminary data shows that inhibiting the NLRP3 inflammasome in podocytes improves GEnC health. Our specific aims will test two hypotheses: Specific Aim #1 tests the hypothesis that SASP and inflammasome activation from injured podocytes are responsible for GEnC damage. Specific Aim #2 tests the hypothesis that interfering with the paracrine injury signals from injured podocytes is a therapeutic target to prevent GEnC dys- function in FSGS. The innovative experimental approaches we will use include: (i) In vivo loss-of-function validation using mutant mice, in which we can reduce/inhibit the SASP or inflammasome phenotypes in podocytes in the setting of experimental FSGS, and then measure the impact on GEnC health; (ii) in vivo gain- of-function validation using transgenic mice, in which we can forcibly induce either a SASP or inflammasome phenotype in podocytes to understand the impact on GEnCs; (iii) Quality-by-Design utilizing a systematic, high complexity approach based on the Design-of-Experiment theory and Multivariate Data Analysis to address the contribution of the podocyte secretome on GEnC health and function. As such, this proposal is highly significant for its short-term impact on understanding the crosstalk between podocytes and GEnCs in FSGS, and for its long-term impact in developing new therapeutic strategies to lower the risk and magnitude for secondary GEnC damage in patients with FSGS.

项目摘要/摘要 问题的范围是50%的局灶性节段性肾小球硬化(FSGS)患者，主要是 成人原发蛋白尿性肾小球疾病的病因，进展为慢性肾脏疾病。因此， 开发新的治疗策略以改善FSGS患者的预后是至关重要的。然而，有一种 在我们对疾病进展的机械性理解中，一个巨大的未得到满足的需求。FSGS的传统观点是 足细胞是其病理的中心，它们是第一种受损的细胞类型。A更多 当代观点是这一范式的扩展，包括对邻近肾小球细胞的损伤。这 格兰特将专注于肾小球内皮细胞(GEnCs)。其基本原理是(1)在FSGS中，GEnc减少 (2)Genc损伤的患者最多 疾病进展率；(3)所有肾小球疾病的Genc损伤评分在FSGS中最高；(4) 基因/生物标记物与FSGS的最低缓解率和不良的长期结果有关。还没有 足细胞是如何引起继发性膝部损伤的，目前的认识还很不完全。要开始关闭 知识缺口，我们采用了一种电子方法来识别旁分泌配体-受体信号网络。 足细胞和GEC之间。令人惊讶的是，我们发现实验和人类的FSGS导致了 非老年人衰老相关分泌表型(SASP)和激活的炎症体表型 足细胞，两者的特征都是分泌不同类别的信号介质。我们的 初步数据显示，抑制足细胞中的NLRP3炎症体可以改善基因组健康。我们的 特殊目的将检验两个假设：特殊目的#1检验SASP和炎症性小体的假设 受损足细胞的激活是导致Genc损伤的原因。具体目标#2检验了以下假设 干扰受损足细胞旁分泌损伤信号是预防遗传疾病的治疗靶点。 在FSGS中的功能。我们将使用的创新实验方法包括：(I)体内功能丧失 使用突变小鼠进行验证，在其中我们可以减少/抑制SASP或炎症体表型 在实验性FSGS的设置下，然后测量对Genc健康的影响；(Ii)在体内获得- 使用转基因小鼠进行功能验证，在其中我们可以强制诱导SASP或炎症小体 足细胞的表型以了解对GEnC的影响；(Iii)利用系统的、高的 基于实验设计理论和多元数据分析的复杂性方法 足细胞分泌组对基因健康和功能的贡献。因此，这项建议是高度 对于理解FSGS中足细胞和GEC之间的串扰具有重要的短期影响， 以及它在开发新的治疗策略以降低风险和严重程度方面的长期影响 FSGS患者的继发性遗传损伤。