Impact of pathogenic missense mutations on the ARID domain of ARID1a

致病性错义突变对 ARID1a ARID 结构域的影响

基本信息

批准号：
10654137
负责人：
Hugo Sanabria
金额：
$ 44.31万
依托单位：
CLEMSON UNIVERSITY
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：
2023-04-12 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10654137
关键词：
ARID Domain ARID1A gene Affinity Amino Acids Binding Bioinformatics Biomedical Research Biophysics Breast Cancer Patient Cell Proliferation Cholangiocarcinoma Chromatin Chromatin Remodeling Factor Classification Clinical Trials Complex Computer Models DNA DNA Binding DNA Binding Domain DNA Repair DNA biosynthesis DNA-Protein Interaction Databases Environment Fluorescence Fluorescence Resonance Energy Transfer Fostering Foundations Frameshift Mutation Future Gene Expression Genes Genetic Transcription Goals Health Health Benefit Helix-Turn-Helix Motifs Libraries Malignant Neoplasms Malignant neoplasm of ovary Measures Missense Mutation Modeling Molecular Mechanisms of Action Mutate Mutation Names Non-Small-Cell Lung Carcinoma Nuclear Outcome Ovarian Pancreas Pathogenicity Pathway interactions Patients Physical condensation Positioning Attribute Prevention Program Development Property Protein Family Proteins Research Research Training Science Solid Neoplasm Spectrum Analysis Structural Models Structure Structure-Activity Relationship Sucrose System Talents Testing The Cancer Genome Atlas Therapeutic Tumor Suppressor Genes Tumor Suppressor Proteins Universities Uterus Work alpha helix anticancer research biobank biophysical analysis career career development design experience experimental study fighting graduate student improved insight interest loss of function loss of function mutation member mutant novel therapeutics personalized medicine physical science programs protein folding resistance mutation screening single molecule temporal measurement translational oncology undergraduate student

项目摘要

SUMMARY The AT-rich interacting domain-containing protein 1a (ARID1a), also named BAF250a, p270, or hOSA1, is a vital component of the Switch/Sucrose Non-Fermentable (SWI/SNF) chromatin remodeling complex. ARID1a, as part of the SWI/SNF complex, is responsible for crucial nuclear activities, including regulating transcription, DNA synthesis, DNA damage repair, and cell proliferation. Consequently, ARID1a is classified as a tumor suppressor gene and it is frequently mutated in solid tumor malignancies cancers, amounting to ~6% of every cancer and ~45% of all ovarian cancers. Loss of function mutations in ARID1a due to frameshift, stop-gain, or missense mutations leads to dysregulation of many gene pathways, including the prevention of tumor suppressor activities. Although the loss of function is evident in stop-gain and frameshift mutations, the impact of pathogenic missense mutations is subtler and more difficult to understand or predict. We hypothesize that pathogenic missense mutations impact the protein stability, DNA binding affinity, and structural dynamics, perturbing its function. Specifically, we seek to understand the effects of pathogenic missense mutations in the ARID domain of ARID1a, which is responsible for its direct interaction with DNA. Therefore, the broad goal of this research program is to understand the effects of pathogenic missense mutations in the ARID domain of ARID1a. The long-term implications of this program are the development of a unique framework that will pave the way for directly probing identified pathogenic missense mutations for future guided patient-specific screening therapeutic approaches. Our proposed study integrates biophysical studies, computational approaches, and single-molecule spectroscopy to characterize the stability, DNA binding, and structural dynamics of the ARID domain. Our studies will give insights into the correlation between missense pathogenic mutations and the structure-dynamics- function relationship. The PI and our research team are uniquely positioned to pursue the following specific aims: (1) to determine the impact of pathogenic missense mutations on the stability of the ARID domain; (2) to determine the impact of pathogenic missense mutations in the binding affinity of ARID domain to DNA; and (3) to determine the changes in the structure and dynamics of ARID domain as perturbed by pathogenic missense mutations. This interdisciplinary project will engage undergraduates in research to foster their interest and career development in both physical sciences and cancer research. The expected outcome will be a structural model of ARID1a’s ARID domain interaction with DNA, a library of pathogenic mutations ranked by the impact on stability and affinity, a system to improve pathogenic predictors, and the foundation for developing novel therapeutics and personalized medicine in the fight against cancer.

概括富含 AT 的相互作用结构域蛋白 1a (ARID1a)，也称为 BAF250a、p270 或 hOSA1，是一种重要的 Switch/蔗糖不可发酵 (SWI/SNF) 染色质重塑复合物的组成部分，作为 ARID1a 的一部分。 SWI/SNF 复合体的组成部分，负责重要的核活动，包括调节转录、DNA 经过合成、DNA 损伤修复和细胞增殖测试，ARID1a 被归类为肿瘤抑制因子。该基因在实体瘤恶性肿瘤中经常发生突变，约占每种癌症的 6% 约 45% 的卵巢癌由于移码、停止增益或错义而导致 ARID1a 功能突变丧失。突变导致许多基因途径失调，包括抑制肿瘤抑制活性。尽管在停止增益和移码突变中功能丧失很明显，但致病性错义的影响突变更加微妙，更难以理解或预测。突变会影响蛋白质稳定性、DNA 结合亲和力和结构动力学，从而扰乱其功能。具体来说，我们试图了解 ARID1a 的 ARID 结构域中致病性错义突变的影响，它负责与 DNA 直接相互作用。因此，该研究计划的总体目标是了解 ARID1a ARID 结构域致病性错义突变的长期影响。该计划的意义在于开发一个独特的框架，为直接探索铺平道路确定了致病性错义突变，用于未来指导患者特异性筛查治疗方法。我们提出的研究整合了生物物理学研究、计算方法和单分子我们的研究通过光谱学来表征 ARID 结构域的稳定性、DNA 结合和结构动力学。将深入了解错义致病突变与结构动力学之间的相关性 PI 和我们的研究团队具有独特的优势，可以实现以下具体目标： (1) 确定致病性错义突变对 ARID 结构域稳定性的影响；确定致病性错义突变对 ARID 结构域与 DNA 结合亲和力的影响；以及 (3) 确定 ARID 结构域在致病性错义干扰下的结构和动态变化这个跨学科项目将让本科生参与研究，以培养他们的兴趣和职业生涯。物理科学和癌症研究的发展预期结果将是一个结构模型。 ARID1a 的 ARID 结构域与 DNA 的相互作用，这是一个致病性突变库，按对 DNA 的影响进行排序稳定性和亲和力，改善致病预测因子的系统，以及开发新的基础抗击癌症的疗法和个性化医疗。