Blocking Latent TGF-β2 Activation as a Therapeutic Strategy for Renal Fibrosis

阻断潜在的 TGF-β2 激活作为肾纤维化的治疗策略

基本信息

批准号：
10215977
负责人：
Viet Quoc Le
金额：
$ 15.53万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10215977
关键词：
Acute Renal Failure with Renal Papillary Necrosis Affinity Antibodies Aristolochic Acids Binding Binding Proteins Biological Assay Biology Blocking Antibodies Blood Platelets Boston Capital Cardiovascular Diseases Cell Cycle Arrest Cell Line Cells Chronic Kidney Failure Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Complement Complement Factor B Complex Crystallization Crystallography DNA Insertion Elements DNA Repair Data Deposition Deuterium Development Diabetic Nephropathy Disease model Drug Kinetics Drug Targeting Education Electron Microscopy End stage renal failure Endothelial Cells Ensure Environment Epithelial Cells Epitopes Etiology Exhibits Expression Profiling Extracellular Matrix Failure Fellowship Fibroblasts Fibrosis Follow-Up Studies Foundations Functional disorder Future Genetic Goals Grant Growth Growth Factor Hospitals Human Hydrogen Immunofluorescence Microscopy Immunoglobulin G Infiltration Inflammatory Infrastructure Injury to Kidney Institutes Institution Integrin Inhibition Integrins Ischemia Kidney Kidney Diseases Knock-in Mouse LRRC32 gene Libraries Link M cell Mediating Mentors Mentorship Modality Modeling Molecular Molecular Biology Molecular Conformation Monitor Mus Mutation Myofibroblast National Institute of Diabetes and Digestive and Kidney Diseases Negative Staining Pathway interactions Pediatric Hospitals Pericytes Pharmaceutical Preparations Property Protein Biochemistry Proteins Regulatory T-Lymphocyte Renal function Reperfusion Therapy Reporter Research Research Personnel Resolution Resources Role Signal Transduction Structure Surface System Techniques Testing Therapeutic Therapeutic antibodies Training Transforming Growth Factor Beta 2 Transforming Growth Factor beta Receptors Transforming Growth Factors Treatment Efficacy Ureteral obstruction Vision Vocational Guidance Woman X-Ray Crystallography Yeasts antibody libraries base career career development cell type diabetic rat dimer drug development drug discovery extracellular global health improved in vitro activity in vitro testing in vivo innovation insertion/deletion mutation insight kidney fibrosis lead candidate medical schools model building mortality mouse model nanobodies novel strategies novel therapeutic intervention novel therapeutics prevent programs response skills structural biology success symposium therapeutic development therapeutic target

项目摘要

ABSTRACT. Transforming growth factor beta2 (TGF-β2) is an important therapeutic target for renal fibrosis, the principal cause of end-stage renal failure in chronic kidney disease (CKD). TGF-β2 triggers renal fibrosis in vivo and, in response to kidney injury, is upregulated in renal myofibroblasts, pericytes, and proximal tubule epithelial cells—cell types that mediate kidney fibrosis. Earlier studies showed that an antibody to mature TGF-β2 arrested renal fibrosis in a rat model of diabetic kidney disease, but further therapeutic development was not followed up. In vivo, TGF-β2 exists mainly as a latent pro-complex (proTGF-β2) in which prodomains are noncovalently bound to the growth factor. Secreted proTGF-β2 is stored in different extracellular milieus where it undergoes activation, i.e. release of the growth factor (mature TGF-β2), to initiate signaling. Preliminary data point to αVβ6-dependent and -independent mechanisms of proTGF-β2 activation as different modalities that can be therapeutically targeted for renal fibrosis. Aim 1 of this proposal is to develop new antibodies that specifically target the prodomain and block proTGF-β2 activation as a novel therapeutic strategy for renal fibrosis. Antibodies will be selected from an innovative yeast display antibody library, screened for activation-blocking activity in vitro, and tested for therapeutic efficacy in mouse models of acute kidney injury. Aim 2 is to determine high-resolution crystal structures of proTGF-β2 to define the mechanism underlying latency and facilitate drug development by uncovering new strategies to prevent activation. The candidate has assembled an exceptional team of mentors and advisors with expertise in renal pathophysiology, drug discovery, and structural biology to ensure the success of the project. The team will provide career guidance and training in techniques essential for the candidate’s future independent career at the interface of structural biology, drug discovery, renal fibrosis, and CKD. The candidate will receive extensive training in 1) X-ray crystallography, 2) antibody discovery, 3) renal pathophysiology, 4) immunofluorescence microscopy, and 5) mouse models of acute kidney injury and renal fibrosis. These skills will extend the candidate’s already versatile foundation in genetics, molecular biology, protein biochemistry, and structural biology. Boston Children’s Hospital and surrounding institutions (e.g., Harvard Medical School) constitute a robust training environment with unparalleled intellectual capital and remarkable infrastructure, which include cutting-edge yeast display platforms for antibody discovery at the Institute for Protein Innovation and unparalleled resources and expertise in the Renal Division at Brigham and Women’s Hospital, that will enhance the candidate’s growth and support his proposed research. Career development will be accomplished through direct mentorship, education through fellowship training offices, and attendance of conferences. The results of this proposal will establish the foundation of the candidate’s future research programs as an independent investigator in renal biology. The candidate plans to apply for the NIDDK Small Grant Program if available to K awardees and an R01 to facilitate his transition to independence.

摘要：转化生长因子β2（TGF-β2）是肾纤维化的重要治疗靶点。 TGF-β2 是慢性肾病 (CKD) 终末期肾衰竭的主要原因，可引发体内肾纤维化。并且，作为对肾损伤的反应，肾肌成纤维细胞、周细胞和近端小管上皮细胞中的表达上调细胞——早期介导肾纤维化的细胞类型，研究表明成熟 TGF-β2 的抗体可以抑制。糖尿病肾病大鼠模型中的肾纤维化，但没有跟进进一步的治疗开发。在体内，TGF-β2 主要作为潜在的前复合物 (proTGF-β2) 存在，其中前结构域非共价结合分泌的 proTGF-β2 储存在不同的细胞外环境中，并在那里进行激活，即释放生长因子（成熟 TGF-β2），启动信号传导，初步数据表明αVβ6 依赖性。 -proTGF-β2 激活的独立机制作为可以治疗的不同方式该提案的目标 1 是开发专门针对肾纤维化的新抗体。前结构域和阻断 proTGF-β2 激活将作为肾纤维化的新治疗策略。从创新的酵母展示抗体库中筛选，在体外筛选激活阻断活性，并且在急性肾损伤小鼠模型中测试治疗效果，目的 2 是确定高分辨率。 proTGF-β2 的晶体结构可定义潜在潜伏期机制并促进药物开发发现防止激活的新策略。候选人组建了一支出色的导师团队。以及具有肾脏病理生理学、药物发现和结构生物学专业知识的顾问，以确保该团队将提供对项目至关重要的职业指导和技术培训。候选人未来在结构生物学、药物发现、肾纤维化等领域的独立职业生涯 CKD。候选人将接受 1) X 射线晶体学、2) 抗体发现、3) 肾脏方面的广泛培训。病理生理学，4) 免疫荧光显微镜，5) 急性肾损伤和肾损伤的小鼠模型这些技能将扩展候选人在遗传学、分子生物学、蛋白质生物化学和结构生物学。波士顿儿童医院及周边机构（例如，哈佛医学院）构建了一个强大的培训环境，拥有无与伦比的智力资本和卓越的基础设施，其中包括用于抗体发现的尖端酵母展示平台布莱根大学蛋白质创新研究所以及肾脏科无与伦比的资源和专业知识妇女医院，这将促进候选人的成长并支持他拟议的职业生涯。发展将通过直接指导、通过奖学金培训办公室进行的教育以及该提案的出席情况将为候选人的未来奠定基础。候选人计划申请 NIDDK 作为肾脏生物学独立研究者的研究项目。小额赠款计划（如果适用于 K 获奖者和 R01），以促进他向独立过渡。