Global Measurements of Protein Folding Stability for Characterization of Aging and Disease

用于表征衰老和疾病的蛋白质折叠稳定性的全局测量

• 批准号: 10022495
• 负责人: Michael C Fitzgerald
• 金额: $ 30.33万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-23 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10022495
• 关键词: Aging; Biological; Biological Assay; Biological Process; Biology; Brain; Cell Line; Chemicals; Clinical; Colorectal; Colorectal Cancer; Development; Disease; Drug resistance; Gene Expression; Gene Expression Profiling; Genome; Left; Lip structure; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Metastatic Neoplasm to the Liver; Methodology; Methods; Modification; Molecular; Molecular Conformation; Mouse Protein; Mus; Organoids; Pathway interactions; Patients; Physiological Processes; Pilot Projects; Property; Proteins; Proteolysis; Proteome; Proteomics; Research; Resistance; Sampling; Specimen; Techniques; Thermodynamics; Tissue-Specific Gene Expression; Work; Xenograft procedure; aged; base; brain tissue; clinically relevant; colon cancer cell line; design; drug sensitivity; drug-sensitive; human disease; mouse model; novel; oxaliplatin; predictive test; protein biomarkers; protein expression; protein folding; protein function; therapeutic target; tumor; Aging; Biological; Biological Assay; Biological Process; Biology; Brain; Cell Line; Chemicals; Clinical; Colorectal; Colorectal Cancer; Development; Disease; Drug resistance; Gene Expression; Gene Expression Profiling; Genome; Left; Lip structure; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Metastatic Neoplasm to the Liver; Methodology; Methods; Modification; Molecular; Molecular Conformation; Mouse Protein; Mus; Organoids; Pathway interactions; Patients; Physiological Processes; Pilot Projects; Property; Proteins; Proteolysis; Proteome; Proteomics; Research; Resistance; Sampling; Specimen; Techniques; Thermodynamics; Tissue-Specific Gene Expression; Work; Xenograft procedure; aged; base; brain tissue; clinically relevant; colon cancer cell line; design; drug sensitivity; drug-sensitive; human disease; mouse model; novel; oxaliplatin; predictive test; protein biomarkers; protein expression; protein folding; protein function; therapeutic target; tumor

ABSTRACT Genome- and proteome-based gene expression profiling studies have been widely used over the past decade to characterize biological states including those associated with normal biological process (e.g., aging) and with disease (e.g., cancer). While such differential gene expression profiling studies can help identify the cellular pathways and physiological processes associated with a biological state they often fail to produce a complete understanding of the biological state because gene expression levels are not directly tied to protein function. This has limited the number of useful protein biomarkers and therapeutic targets discovered from such studies, and left gaps in our understanding of disease states. Proposed here is the development and application of thermodynamic measurements of protein stability to characterize different biological states including those associated with aging and cancer. Such thermodynamic measurements of protein stability are expected to be more closely related to protein function than are protein expression levels, and thus produce more useful protein biomarkers and therapeutic targets of disease and generate a better understanding of the molecular basis of disease. The proposed work will further develop and utilize three mass spectrometry-based proteomics methods, termed SPROX, LiP and PP, to characterize the protein folding and stability changes associated with a fundamental biological process, aging, and a disease, colorectal cancer. The specific aims of this work are: (1) to interface SPROX and LiP with middle- down proteomics methods to enable proteoform specific folding and stability measurments; (2) to utilize the middle down SPROX and LiP workflows developed in (1) to make proteoform specific thermodynamic stability measurements on mouse brain proteins derived from a mouse model of aging; (3) to utilize the SPROX, LiP, and PP techniques to identify protein biomarker with altered folding and stability in patient-derived colorectal cancer cell lines with different sensitivities to oxaliplatin treatment; (4) to utilize the protein biomarkers discovered in (3) to develop a clinically relevant assay for predicting oxaliplatin resistance/sensitivity in colorectal cancer.

抽象的 基于基因组和蛋白质组基因表达分析研究已被广泛使用 在过去的十年中，要表征生物态，包括与正常相关的生物状态 生物过程（例如，衰老）和疾病（例如癌症）。而这种差异基因 表达分析研究可以帮助识别细胞途径和生理过程 与生物状态相关，他们常常无法完全理解 生物状态是因为基因表达水平与蛋白质功能无直接相关。这就是 限制从此类发现的有用蛋白质生物标志物和治疗靶标的数量 研究，并在我们对疾病状态的理解中留下了差距。这里提出的是发展 并应用蛋白质稳定性的热力学测量以表征不同 生物状态，包括与衰老和癌症相关的生物状态。这样的热力学 预计蛋白质稳定性的测量与蛋白质功能更紧密相关 而不是蛋白质表达水平，因此产生更有用的蛋白质生物标志物， 疾病的治疗靶标，并更好地了解 疾病。 拟议的工作将进一步发展并利用三个基于质谱的工作 蛋白质组学方法称为sprox，唇和PP，以表征蛋白质折叠和 稳定性变化与基本生物学过程，衰老和疾病相关， 结直肠癌。这项工作的具体目的是：（1）与中间的接口和嘴唇 降低蛋白质组学方法以实现蛋白质成型特异性折叠和稳定性测量； （2） 利用（1）中开发的中间向下sprox和唇部工作流程制作蛋白质成型 衍生自小鼠的小鼠脑蛋白上的特定热力学稳定性测量 衰老模型； （3）利用sprox，唇和PP技术鉴定蛋白质生物标志物 患者来源的结直肠癌细胞系的折叠和稳定性改变了不同 奥沙利铂治疗的敏感性； （4）利用（3）中发现的蛋白质生物标志物 开发临床相关的测定法，以预测结直肠癌的奥沙利铂耐药性/敏感性 癌症。