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Defining and Modulating Mechanisms of Collagen Proteostasis

胶原蛋白稳态的定义和调节机制

基本信息

批准号：
10183166
负责人：
Matthew Donald Shoulders
金额：
$ 33.01万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-11 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10183166
关键词：
Address Amino Acid Sequence Animals Basement membrane Biochemical Biogenesis Biological Biology C-terminal Cartilage Categories Cells Cellular Stress Chemicals Chronic Code Collagen Collagen Diseases Collagen Gene Collagen Type I Complex Cysteine Data Defect Development Disease Endoplasmic Reticulum Extracellular Matrix Failure Functional disorder Goals Homeostasis Homo Individual Knowledge Lead Life Link Maps Mass Spectrum Analysis Medical Modeling Modification Molecular Molecular Chaperones Mutation National Institute of Arthritis and Musculoskeletal and Skin Diseases Nature Outcome Pathologic Pathology Pathway interactions Patients Pharmacology Phenotype Process Production Property Proteins Proteomics Public Health Quality Control Research Research Personnel Role Skin System Testing Therapeutic Therapeutic Intervention Tissues Validation Variant Work autosomal dominant mutation base bone cell motility comparative insight intercellular communication new therapeutic target polypeptide prevent protein complex protein folding protein misfolding proteostasis response scaffold targeted treatment trafficking wound healing

项目摘要

As the primary proteinaceous component of bone, skin, cartilage, basement membranes, and more, collagen serves as the molecular scaffold for animal life. Owing to the highly hierarchical nature of the extracellular matrix, the properties of collagenous supramolecular scaffolds are fundamentally defined by the complex intracellular process of collagen folding and quality control. Unsurprisingly, therefore, defects in intracellular collagen proteostasis engender diverse diseases known as the collagenopathies. These defects are most commonly caused by autosomal dominant mutations in collagen genes, and can be variously ascribed to three primary issues: (1) Escape of misfolded or dysfunctional collagen strands into the extracellular matrix; (2) Insufficient secretion of properly folded collagen; and/or (3) Intracellular accumulation of misfolding collagen molecules that leads to chronic cell dysfunction. All three of these defects are associated with a failure of the endoplasmic reticulum's (ER's) proteostasis network (a highly integrated system of chaperones, quality control mechanisms, and secretory machineries) to properly solve the collagen production problem, particularly in the context of mutations that lead to disease. Elucidating molecular mechanisms of collagen proteostasis in the ER is therefore of paramount importance to enable the development of disease- modifying therapies. To this end, the current proposal aims to answer three key questions: (1) Can collagen proteostasis defects by rescued by rational chemical biologic modulation of the ER proteostasis network? (2) How is a misfolding collagen strand identified by the ER quality control machinery? (3) How is collagen assembly, which is the critical first step in collagen folding, regulated both for wild-type collagen and for misfolding, disease-causing collagen variants? In Specific Aim 1, state-of-the-art chemical biology strategies targeted at the ER proteostasis network and the unfolded protein response are deployed to test the hypothesis that disease-associated collagen proteostasis defects can be resolved by proteostasis network modulation. In Specific Aim 2, the mechanisms of collagen quality control (which are known to exist but remain ill-defined) will be studied in detail, both for wild-type and a range of misfolding collagen variants. This Aim involves mass spectrometry-based quantitative comparative interactomics to detect such mechanisms, followed by biochemical validation and characterization. In Specific Aim 3, the molecular code for collagen assembly will be defined, and strategies to address assembly defects via the proteostasis network will be pursued. Insights obtained using this combination of biochemical and cell and chemical biological experimental strategies are expected to have a positive and ultimately translatable impact, because they are highly likely to yield new targets for therapeutic intervention in diverse collagenopathies that are not accessible by other experimental approaches.

作为骨骼、皮肤、软骨、基底膜等的主要蛋白质成分，胶原蛋白是动物生命的分子支架。由于组织的高度等级性质细胞外基质，胶原超分子支架的特性基本上由胶原蛋白折叠和质量控制的复杂细胞内过程。因此，不足为奇的是，细胞内胶原蛋白稳态导致多种疾病，称为胶原病。这些缺陷最常见的是由胶原蛋白基因的常染色体显性突变引起的，并且可以有多种原因归因于三个主要问题：（1）错误折叠或功能失调的胶原蛋白链逃逸到细胞外基质； (2)正确折叠的胶原蛋白分泌不足；和/或 (3) 细胞内积累错误折叠的胶原蛋白分子会导致慢性细胞功能障碍。所有这三个缺陷都是相关的内质网 (ER) 蛋白质稳态网络（高度集成的系统）出现故障伴侣、质量控制机制和分泌机制）妥善解决胶原蛋白的产生问题，特别是在导致疾病的突变的背景下。阐明其分子机制因此，内质网中的胶原蛋白稳态对于疾病的发展至关重要。修改疗法。为此，当前提案旨在回答三个关键问题：（1）胶原蛋白可以内质网蛋白质稳态网络的合理化学生物调节可以挽救蛋白质稳态缺陷吗？ (2) ER 质量控制机器如何识别错误折叠的胶原蛋白链？（三）胶原蛋白如何组装是胶原蛋白折叠的关键第一步，对野生型胶原蛋白和错误折叠、致病的胶原蛋白变异？在具体目标 1 中，最先进的化学生物学策略针对 ER 蛋白质稳态网络，并部署未折叠的蛋白质反应来检验假设与疾病相关的胶原蛋白稳态缺陷可以通过蛋白稳态网络调节来解决。在具体目标 2，胶原蛋白质量控制机制（已知存在但仍不明确）将对野生型和一系列错误折叠胶原蛋白变体进行详细研究。这个目标涉及质量基于光谱测定的定量比较相互作用组学来检测此类机制，然后生化验证和表征。在具体目标 3 中，胶原蛋白组装的分子代码为定义，并将寻求通过蛋白质稳态网络解决组装缺陷的策略。见解使用生物化学和细胞和化学生物实验策略的结合获得的结果是预计将产生积极且最终可转化的影响，因为它们极有可能产生新的其他实验无法达到的多种胶原蛋白病的治疗干预目标接近。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Using CRISPR/Cas9 to generate a heterozygous COL2A1 p.R719C iPSC line (MCRIi019-A-6) model of human precocious osteoarthritis.

使用 CRISPR/Cas9 生成人类早熟骨关节炎杂合 COL2A1 p.R719C iPSC 系 (MCRIi019-A-6) 模型。

DOI：
10.1016/j.scr.2023.103020
发表时间：
2023
期刊：
Stem cell research
影响因子：
1.2
作者：
Yammine,KathrynM;MirdaAbularach,Sophia;Sampurno,Lisa;Bateman,JohnF;Lamandé,ShireenR;Shoulders,MatthewD
通讯作者：
Shoulders,MatthewD

Genetic Engineering by DNA Recombineering.

DOI：
10.1002/cpch.70
发表时间：
2019-09-01
期刊：
Current protocols in chemical biology
影响因子：
0
作者：
Papa, Louis J 3rd;Shoulders, Matthew D
通讯作者：
Shoulders, Matthew D

A cysteine-based molecular code informs collagen C-propeptide assembly.

DOI：
10.1038/s41467-018-06185-2
发表时间：
2018-10-11
期刊：
Nature communications
影响因子：
16.6
作者：
DiChiara AS;Li RC;Suen PH;Hosseini AS;Taylor RJ;Weickhardt AF;Malhotra D;McCaslin DR;Shoulders MD
通讯作者：
Shoulders MD

Adapting Secretory Proteostasis and Function Through the Unfolded Protein Response.