Characterization and therapeutic targeting of neutrophil serine proteases in ANCA vasculitis

ANCA 血管炎中性粒细胞丝氨酸蛋白酶的表征和治疗靶向

• 批准号: 437811195
• 负责人: Professor Dr. Ralph Kettritz
• 金额: --
• 依托单位: Experimental and Clinical Research Center (ECRC)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/437811195?language=en
• 关键词: Characterization; therapeutic; targeting; neutrophil; serine

Anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitides (AAV) are necrotizing small-vessel inflammatory diseases that can affect every organ, frequently manifesting in the kidneys as necrotizing crescentic glomerulonephritis with rapidly-progressive renal failure. ANCA bind to either proteinase 3 (PR3) or myeloperoxidase (MPO) on the surface of neutrophils and monocytes that exclusively express these autoantigens. ANCA activate these cells that adhere to and damage the endothelium leading to systemic vasculitis. We characterized several injury mechanisms some of which involve enzymatically active neutrophil serine proteases (NSPs). The NSP family consist of PR3, human neutrophil elastase (HNE) and cathepsin G (CatG) and low-abundant NSP4. PR3 is a unique NSP for it provides the major ANCA antigen. NSPs are generated from inactive zymogen proforms by cathepsin C (CatC) that removes an N-terminal dipeptide. Loss-of-function mutations in CatC prevents NSP maturation and leads to zymogen degradation. Pharmacological CatC inhibitors have the potential to abrogate NSP maturation thereby eliminating NSP proteins and proteolytic activity. Thus, pharmacological CatC inhibition provides possibly a novel treatment strategy in AAV. With this proposal, we will (i) systematically characterize NSP proteins and proteolytical activity in human and murine neutrophils and monocytes, (ii) target NSPs by pharmacological CatC inhibition to characterize NSP-mediated damage mechanisms using stem cell differentiation models and endothelial cells, and (iii) test the hypothesis that pharmacological CatC inhibition protects from ANCA-induced vasculitis using murine AAV disease models. Our findings will improve understanding ANCA-induced disease mechanisms that link NSPs to vasculitis. In addition, the study could encourage clinical trials exploring pharmacological CatC inhibition as a new treatment for AAV.

抗中性粒细胞胞浆抗体 (ANCA) 相关血管炎 (AAV) 是一种坏死性小血管炎症性疾病，可影响每个器官，通常在肾脏中表现为坏死性新月体肾小球肾炎并伴有快速进展的肾功能衰竭。 ANCA 与专门表达这些自身抗原的中性粒细胞和单核细胞表面的蛋白酶 3 (PR3) 或髓过氧化物酶 (MPO) 结合。 ANCA 激活这些粘附并损伤内皮的细胞，导致系统性血管炎。我们表征了几种损伤机制，其中一些涉及具有酶活性的中性粒细胞丝氨酸蛋白酶（NSP）。 NSP 家族由 PR3、人中性粒细胞弹性蛋白酶 (HNE) 和组织蛋白酶 G (CatG) 以及低丰度的 NSP4 组成。 PR3 是一种独特的 NSP，因为它提供了主要的 ANCA 抗原。 NSP 是通过组织蛋白酶 C (CatC) 从无活性的酶原原形生成的，可去除 N 末端二肽。 CatC 的功能缺失突变会阻止 NSP 成熟并导致酶原降解。药理学 CatC 抑制剂有可能阻止 NSP 成熟，从而消除 NSP 蛋白和蛋白水解活性。因此，药理学 CatC 抑制可能为 AAV 提供一种新的治疗策略。 通过这项提议，我们将（i）系统地表征人和小鼠中性粒细胞和单核细胞中的 NSP 蛋白和蛋白水解活性，（ii）通过药理学 CatC 抑制来靶向 NSP，以使用干细胞分化模型和内皮细胞来表征 NSP 介导的损伤机制，以及（iii）使用小鼠测试药理学 CatC 抑制可预防 ANCA 诱导的血管炎的假设 AAV 疾病模型。我们的研究结果将增进对 ANCA 诱发的将 NSP 与血管炎联系起来的疾病机制的理解。此外，该研究还可以鼓励临床试验探索药理学 CatC 抑制作为 AAV 的新疗法。