Mechanistic Studies on Meprin Metalloproteases and Meprin-Substrate Interactions in Tissue Injury

Meprin 金属蛋白酶和 Meprin-底物相互作用在组织损伤中的机制研究

• 批准号: 10199258
• 负责人: Elimelda Moige Ongeri
• 金额: $ 30万
• 依托单位: NORTH CAROLINA AGRI & TECH ST UNIV
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10199258
• 关键词: Actins; Alzheimer' s Disease; Amyloid beta-Protein Precursor; Anti-Inflammatory Agents; Area; Award; CCL2 gene; Chronic; Complex; Coupled; Cyclic AMP-Dependent Protein Kinases; Cytoskeletal Proteins; Development; Diabetes Mellitus; Diabetic Nephropathy; Diagnostic; Disease; Disease Progression; E-Cadherin; Early Diagnosis; Endothelial Cells; Enzymes; Extracellular Matrix; Extracellular Matrix Proteins; Fibrosis; Functional disorder; Genes; Genetic Predisposition to Disease; Hypoxia; In Vitro; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Injury to Kidney; Interleukin-1 beta; Interleukin-18; Interleukin-6; Kidney Diseases; Knockout Mice; Knowledge; Leukocytes; Malignant Neoplasms; Mediating; Mediator of activation protein; Mentors; Meprin; Metabolism; Metalloproteases; Minority Groups; Molecular; Molecular Biology; Neurodegenerative Disorders; Organ; Physiological; Protein Isoforms; Protein Kinase C; Proteins; Proteomics; Proximal Kidney Tubules; Pulmonary Fibrosis; Renal carcinoma; Research; Severities; Signal Pathway; Signal Transduction; Signaling Protein; Single Nucleotide Polymorphism; Skin; Small Intestines; Substrate Interaction; TREM2 gene; Therapeutic; Tight Junctions; Transcend; Underrepresented Minority; biomarker development; brush border membrane; cancer cell; cytokine; diabetic; in vivo; insight; macrophage; monocyte; mouse model; occludin; osteosarcoma; podocyte; precision medicine; programs; response; therapeutic target; tissue injury; tool; translational study; villin

PROJECT SUMMARY/ ABSTRACT. My research program focuses on understanding the cellular and molecular mechanisms underlying the progression of tissue injury mediated by meprin metalloproteases. Meprins comprise of two subunits, α and β, which form two protein isoforms, meprin A (α-α or α-β) and meprin B (β-β) with distinct and overlapping substrates. Meprins are most abundantly expressed in the brush-border membranes of proximal kidney tubules and small intestines. Meprins are also expressed in leukocytes (monocytes and macrophages), podocytes, skin, endothelial cells, and cancer cells. Meprins have been implicated in the pathophysiology of inflammatory- and fibrosis-associated diseases that include kidney disease, inflammatory bowel disease, lung fibrosis, neurodegenerative disease (e.g. Alzheimer’s disease), and cancer. Single nucleotide polymorphisms (SNPs) in the meprin β gene were shown to associate with severity of certain diseases such as diabetic kidney disease and cancer. My research group uses a combination of molecular biology and proteomic approaches to identify meprin substrates and characterize the interactions between meprin isoforms and their substrates. These are coupled with in vivo studies with meprin knockout mouse models to determine how meprin activity impacts the progression of disease. Known meprin substrates include extracellular matrix (ECM) proteins, modulators of inflammation (e.g. proinflammatory cytokines [IL-1β, IL-6, IL-18, MCP-1; and anti-inflammatory proteins Ac-SDKP), cell signaling proteins (e.g. protein kinase A and protein kinase C), mediators of the hypoxia response (e.g. osteosarcoma-9), tight junction proteins (e.g. claudin 5, occludin, E-cadherin, and Z0-1) cytoskeletal proteins (e.g. villin and actin) and proteins that contribute to plaques in AD (e.g. amyloid precursor protein and triggering receptor expressed on myeloid cells 2). The diversity of meprins substrates suggests that complex mechanisms are involved under different conditions and in different organs. It’s important to gain understanding of these mechanisms to facilitate development of diagnostic and therapeutic tools. For the five year period of this proposal, we will conduct studies in three areas; (i) to determine how SNPS in the meprin β gene impact its interactions with substrates and physiological sheddases, (ii) determine how meprin interactions with substrates modulate signaling pathways and impact responses in hypoxia, inflammation, and ECM metabolism, and (iii) to evaluate the use of meprin and meprin cleavage products as biomarkers for development of diagnostic tools for early detection of disease. The proposed research will transcend basic (in vitro and in vivo) to gain insights on the basis for genetic predispositions associated with meprins. Translational studies are also proposed to apply this knowledge in development of diagnostic tools applicable to diabetic kidney injury and Alzheimer’s disease (AD), an important step in advancing precision medicine. Furthermore, this award will facilitate mentoring of trainees from underrepresented minority populations and thus promote diversity of the biomedical workforce.

项目摘要/摘要我的研究项目侧重于了解细胞和 meprin 金属蛋白酶介导的组织损伤进展的分子机制。 Meprins 由两个亚基 α 和 β 组成，形成两种蛋白质异构体：meprin A（α-α 或 α-β）和 meprin 具有不同且重叠底物的 B (β-β) 在刷状缘中表达最丰富。 近端肾小管和小肠的膜也在白细胞中表达。 （单核细胞和巨噬细胞）、足细胞、皮肤、内皮细胞和癌细胞。 与炎症和纤维化相关疾病（包括肾脏疾病）的病理生理学有关 疾病、炎症性肠病、肺纤维化、神经退行性疾病（例如阿尔茨海默病），以及 meprin β 基因中的单核苷酸多态性 (SNP) 被证明与癌症的严重程度相关。 我的研究小组结合使用了某些疾病，例如糖尿病肾病和癌症。 分子生物学和蛋白质组学方法来识别 meprin 底物并表征相互作用 meprin 异构体及其底物之间的研究与 meprin 敲除的体内研究相结合。 用于确定 meprin 活性如何影响疾病进展的小鼠模型。 包括细胞外基质 (ECM) 蛋白、炎症调节剂（例如促炎细胞因子 [IL-1β、 IL-6、IL-18、MCP-1；和抗炎蛋白 Ac-SDKP)、细胞信号蛋白（例如蛋白激酶 A 和 蛋白激酶 C）、缺氧反应介质（例如骨肉瘤-9）、紧密连接蛋白（例如 密蛋白 5、occludin、E-钙粘蛋白和 Z0-1）细胞骨架蛋白（例如绒毛蛋白和肌动蛋白）以及 导致 AD 中的斑块（例如淀粉样前体蛋白和骨髓细胞上表达的触发受体 2).meprins底物的多样性表明在不同的条件下涉及复杂的机制 了解这些机制对于促进不同的条件和不同的器官非常重要。 在本提案的五年期间，我们将进行诊断和治疗工具的开发。 三个领域的研究；(i) 确定 meprin β 基因中的 S​​NPS 如何影响其与底物的相互作用 和生理脱落酶，(ii) 确定 meprin 与底物的相互作用如何调节信号传导 缺氧、炎症和 ECM 代谢的途径和影响反应，以及 (iii) 评估 meprin 和 meprin 裂解产物作为生物标志物，用于开发早期检测的诊断工具 拟议的研究将超越基础（体外和体内）以获得有关疾病的见解。 还建议应用与 meprins 相关的遗传倾向。 开发适用于糖尿病肾损伤和阿尔茨海默病的诊断工具的知识 （AD），这是推进精准医学的重要一步。此外，该奖项将促进对精准医学的指导。 来自代表性不足的少数群体的学员，从而促进生物医学劳动力的多样性。