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Characterization of Tagged Type IV Collagen

标记的 IV 型胶原蛋白的表征

基本信息

批准号：
10724541
负责人：
Douglas Gould
金额：
$ 44.41万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-15 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10724541
关键词：
Address Affinity Chromatography Alleles Alzheimer&apos s Disease Amino Acids Astrocytes Basement membrane Biochemical Biological Process Biology Blood Vessels Cells Cerebral small vessel disease Cerebrovascular Disorders Cerebrovascular system Characteristics Circular Dichroism Collagen Collagen Type IV Congenital Disorders Defect Dementia Deposition Development Disease Electron Microscopy Embryo Endoplasmic Reticulum Event Extracellular Matrix Fibroblasts Fluorescence Fluorescence Microscopy Genetic Goals Heterogeneity Homeostasis Human Hybrids Impaired cognition Impairment In Vitro Kidney Knowledge Label Mediating Methods Microscopy Molecular Mouse Strains Mus Muscle Mutant Strains Mice Mutation Organ Outcome Pathogenicity Pathologic Pathology Physiologic pulse Post-Translational Protein Processing Property Protein Isoforms Proteins Reporter Research Resources SWI1 Signal Transduction Sorting System Testing Tissues Validation Vascular Endothelial Cell Visualization Western Blotting age related biophysical properties cell behavior cell type cerebrovascular cerebrovascular health cognitive change crosslink genome wide association study genome-wide in vivo inducible Cre innovation insight mutant novel pup recombinase spatiotemporal tool transcriptomics vascular cognitive impairment and dementia

项目摘要

PROJECT SUMMARY Mutations in type IV collagen alpha 1 (COL4A1) and alpha 2 (COL4A2) are an important cause of monogenic cerebral small vessel disease and are also significantly associated with cerebral small vessel disease hallmarks in large-scale genome wide association studies underscoring their importance for general cerebrovascular health in humans. The broad goal of our research is to understand the biological functions COL4A1 and COL4A2 in the extracellular matrix and determine the mechanisms by which they cause genetic and acquired cerebrovascular disease and disfunction. The specific purpose of this proposal is to validate a potentially powerful tool that will enable unveiling of tissue specific and temporal changes in collagen deposition and degradation. The inability to distinguish cell type specific or temporal changes to the extracellular matrix constitutes a significant obstacle for understanding a fundamental aspect of tissue biology. To address this barrier, we developed a mouse line in which COL4A1 is fused with switchable fluorescent proteins for isoform-specific visualization and biochemical tags for differential affinity purification. This simple but powerful tool will enable unprecedented characterization of spatial, temporal, biochemical and biophysical parameters of ECM that are currently impossible to achieve. If the detailed validations outlined in this proposal are successful, we will have developed a transformative tool that will provide significant insight into a fundamental aspect of tissue biology and that can be applied to every organ of the body in normal development or in pathological settings.

项目概要 IV 型胶原蛋白 α 1 (COL4A1) 和 α 2 (COL4A2) 突变是单基因遗传性疾病的重要原因脑小血管疾病，也与脑小血管疾病特征显着相关在大规模全基因组关联研究中强调了它们对一般脑血管健康的重要性在人类中。我们研究的总体目标是了解 COL4A1 和 COL4A2 在细胞外基质并确定它们引起遗传性和获得性脑血管病的机制疾病和功能障碍。该提案的具体目的是验证一个潜在的强大工具，该工具将能够揭示胶原蛋白沉积和降解的组织特异性和时间变化。无法区分细胞外基质的细胞类型特异性或时间变化构成了理解组织生物学基本方面的重大障碍。为了解决这个障碍，我们开发了一种小鼠品系，其中 COL4A1 与可切换荧光蛋白融合，以实现异构体特异性用于差异亲和纯化的可视化和生化标签。这个简单但功能强大的工具将能够对 ECM 的空间、时间、生化和生物物理参数进行了前所未有的表征目前不可能实现。如果本提案中概述的详细验证成功，我们将开发了一种变革性工具，将为组织生物学的基本方面提供重要的见解这可以应用于正常发育或病理情况下的身体每个器官。