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：
10591547
负责人：
Viet Quoc Le
金额：
$ 15.38万
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/10591547
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 Complex Crystallization Crystallography DNA Damage DNA Insertion Elements 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 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 Transforming Growth Factor beta Receptors Treatment Efficacy Ureteral obstruction Vocational Guidance Woman X-Ray Crystallography Yeasts antibody libraries career career development cell type diabetic rat dimer drug development drug discovery extracellular global health improved in vitro activity 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 repaired response skills structural biology success symposium therapeutic development therapeutic evaluation 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.

抽象的。转化生长因子β2（TGF-β2）是肾纤维化的重要治疗靶点，慢性肾脏疾病（CKD）终末期肾衰竭的主要原因。 TGF-β2在体内触发肾纤维化并且，为了应对肾脏损伤，在肾脏肌纤维细胞，周细胞和近端细胞上皮进行了更新细胞 - 介导肾纤维化的细胞类型。较早的研究表明，对成熟TGF-β2的抗体停滞了糖尿病肾脏疾病大鼠模型中的肾纤维化，但没有跟踪进一步的治疗发育。在体内，TGF-β2主要作为潜在的促复合物（ProtgF-β2）存在，其中prodomain属于无共价结合分泌的Protgf-β2存储在不同的细胞外环境中，在那里经历激活，即生长因子的释放（成熟的TGF-β2），以启动信号传导。初步数据指向αVβ6依赖性 ProtgF -β2激活的非依赖性机制是可以在治疗上进行的不同模态针对肾纤维化。该提案的目的1是开发专门针对的新抗体 Prodomain和阻断ProtgF-β2活化是一种新型的肾纤维化治疗策略。抗体将是从创新的酵母显示抗体库中选择，在体外筛选以进行激活阻滞活性，然后在急性肾脏损伤的小鼠模型中测试了热效率。目标2是确定高分辨率 ProtgF-β2的晶体结构来定义潜伏的潜在机制，并通过揭示防止激活的新策略。候选人组建了一个杰出的导师团队以及具有肾脏病理学，药物发现和结构生物学专业知识的顾问，以确保项目的成功。团队将提供职业指导和培训，以对候选人在结构生物学，药物发现，肾纤维化和 CKD。候选人将在1）X射线晶体学中接受广泛的培训，2）抗体发现，3）肾脏病理生理学，4）免疫荧光显微镜和5）急性肾脏损伤和肾脏的小鼠模型纤维化。这些技能将扩大候选人在遗传学，分子生物学上的多功能基础，蛋白质生物化学和结构生物学。波士顿儿童医院和周边机构（例如哈佛医学院）构成了一个无与伦比的智力资本和非凡的基础设施，其中包括用于抗体发现的尖端酵母显示平台蛋白质创新与无与伦比的资源和专业知识研究所妇女医院将增强候选人的成长并支持他的拟议研究。职业开发将通过直接指导，通过奖学金培训办公室进行教育以及会议的出席。该提案的结果将确定候选人未来的基础作为肾脏生物学独立研究者的研究计划。候选人计划申请NIDDK 小型赠款计划（如果是K获奖者的可用，则可以使用R01，以促进他向独立的过渡。