PBRM1 bromodomain missense mutations in ccRCC vascular signaling

ccRCC 血管信号传导中的 PBRM1 溴结构域错义突变

基本信息

批准号：
10604440
负责人：
Karina Lynn Bursch
金额：
$ 4.98万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
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：
2023-05-01 至 2028-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10604440
关键词：
ATAC-seq Address Affect Angiogenesis Inhibitors Angiogenesis Pathway Benign Binding Binding Sites Biochemical Biochemistry Biological Assay Biological Markers Biophysics Blood Vessels Bromodomain Cell Line Cell Proliferation Cellular Assay Chromatin Chromatin Remodeling Factor Clear cell renal cell carcinoma Clinical Clinical Trials Complex DNA DNA Binding DNA-Binding Proteins Data Databases Disease Drug Exposure Early Diagnosis Education Epigenetic Process Escherichia coli Family Fellowship Gene Targeting Genes Genetic Transcription Genomics Goals Growth Histone Acetylation Histones Hypoxia Inducible Factor Immune checkpoint inhibitor In Vitro Knowledge Laboratories Length Link Lysine Malignant Neoplasms Mediating Missense Mutation Modality Mutate Mutation Neoplasms in Vascular Tissue Nuclear Nuclear Proteins Nucleic Acids Operative Surgical Procedures Outcome Output Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Physicians Predisposition Production Proteins RNA Reader Recombinants Regulation Renal Cell Carcinoma Reporting Role SMARCA4 gene Scientist Signal Pathway Signal Transduction Site-Directed Mutagenesis Sucrose Tail Testing Toxic effect Transcriptional Regulation Treatment-related toxicity VHL gene Validation Vascular blood supply alternative treatment angiogenesis blood vessel development cancer type chromatin modification genetic signature genome sciences improved improved outcome in silico inhibitor lens lentivirally transduced loss of function loss of function mutation member molecular dynamics mutant mutational status neglect normoxia patient response polybromo precision medicine predictive marker prevent promoter protein function protein structure response side effect standard of care targeted treatment transcription factor transcriptome sequencing tumor ubiquitin ligase unnecessary treatment

项目摘要

PROJECT SUMMARY Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cell carcinoma. Although outcomes are favorable with early detection and surgery, approximately 30% of patients present with metastatic disease. This highlights the need for an improved biochemical understanding of and targeted therapeutic options for ccRCC. ccRCC is a uniquely vascular cancer due to mutations in the von Hippel-Lindau (VHL) gene, which encodes the substrate recognition subunit of a ubiquitin ligase that degrades hypoxia- inducible factor-α (HIF-α) transcription factors in normoxia. VHL loss results in nuclear HIF-α accumulation and aberrant expression of downstream pro-angiogenic gene targets in a likely HIF-2α-biased manner. Thus, ccRCC tumors are uniquely susceptible to anti-angiogenic therapies, which though effective, are associated with substantial undesirable toxicities for patients. Recent clinical trials have demonstrated that mutations in Polybromo-1 (PBRM1), an epigenetic acetyl-lysine reader protein commonly mutated in ccRCC, correlate with improved outcomes for anti-angiogenic therapies. However, these studies presume that all PBRM1 mutations are loss-of-function and neglect to consider that ~15% of ccRCC-related PBRM1 mutations are missense mutations, generating full-length proteins with a spectrum of undetermined structural and functional impacts. ccRCC-associated PBRM1 missense mutations cluster in bromodomains (BDs), the functional domains of PBRM1 that bind acetyl-lysine residues in histones and other nuclear proteins and nucleic acids. Therefore, a better knowledge of the biochemical impacts of PBRM1 BD missense mutations in ccRCC is critical to promote PBRM1 as a predictive biomarker for patient response to anti-antiangiogenic therapies, so that non-responding patients can avoid unnecessary therapeutic toxicity and seek alternative treatments. This fellowship will elucidate the biochemical impacts of ccRCC-associated PBRM1 BD missense mutations on protein stability and acetylated histone binding function and determine the role of PBRM1 in the regulation of the HIF-2α and angiogenesis pathways. Aim 1 will identify which PBRM1 BD missense mutations are biochemically benign or deleterious for protein structure and function by integrating computational prediction with biophysical assay validation. Aim 2 will determine the effects of PBRM1 BD missense mutations on HIF-2α and angiogenesis pathway regulation by assessing HIF-2α transcriptional output, HIF-2α promoter accessibility, and angiogenic potential in ccRCC cellular assays. This fellowship addresses knowledge gaps in the biochemical basis of ccRCC pathogenesis with the overall goal to promote the most effective use of standard ccRCC treatment modalities. Successful completion of this fellowship will further precision medicine approaches for the treatment of ccRCC by advancing PBRM1 mutational status as a nuanced and predictive biomarker to prevent unnecessary patient drug exposures.

项目摘要透明细胞肾细胞癌（CCRCC）是肾细胞癌最常见的亚型。虽然结局对早期检测和手术是有利的，大约30％的患者转移性疾病。这凸显了需要对生化理解的提高和针对性 CCRCC的治疗选择。由于von Hippel-Lindau突变，CCRC是一种独特的血管癌（VHL）基因，它编码泛素连接酶的底物识别亚基，该亚基降解了缺氧 - 常氧中的诱导因子-α（HIF-α）转录因子。 VHL损失导致核HIF-α积累和下游促血管生成基因靶标的异常表达以HIF-2α偏置的方式表达。那， CCRCC肿瘤易受抗血管生成疗法的唯一影响，尽管有效对患者有很大的不良毒性。最近的临床试验表明 Polybromo-1（PBRM1），一种通常在CCRCC中突变的表观遗传学乙酰赖氨酸读蛋白，与改善抗血管生成疗法的结果。但是，这些研究认为所有PBRM1突变是否有功能丧失和忽视，认为约有15％的与CCRC相关的PBRM1突变是错过的突变，产生具有一系列不确定的结构和功能影响的全长蛋白。 CCRCS相关的PBRM1错义突变突变集群（BDS），该功能域（BDS）的功能域结合乙酰赖氨酸的PBRM1保留在组蛋白，其他核蛋白和核酸中。因此，更好地了解CCRCC中PBRM1 BD错义突变的生化影响对于促进至关重要 PBRM1作为患者对抗抗抗病毒疗法的反应的预测生物标志物，因此无反应患者可以避免不必要的治疗毒性并寻求替代治疗。这个奖学金将阐明了CCRCC相关的PBRM1 BD错义突变对蛋白质稳定性的生化影响和乙酰化的组蛋白结合功能，并确定PBRM1在HIF-2α和血管生成途径。 AIM 1将确定哪些PBRM1 BD错义突变是生化良性或通过将计算预测与生物物理试验整合到蛋白质结构和功能的有害蛋白质结构和功能验证。 AIM 2将确定PBRM1 BD错义突变对HIF-2α和血管生成的影响通过评估HIF-2α转录输出，HIF-2α启动子的可及性和血管生成来调节途径调节 CCRCC细胞测定中的潜力。该奖学金以生化基础的知识差距解决 CCRCC发病机理的总体目标是促进最有效使用标准CCRCC治疗方式。成功完成此奖学金将进一步精确医学方法通过推进PBRM1突变状态作为细微差别和预测的生物标志物来治疗CCRCC 不必要的患者药物暴露。