Role of Transcription Factor ADP-ribosylation in Breast Cancer Biology

转录因子 ADP-核糖基化在乳腺癌生物学中的作用

• 批准号: 10209984
• 负责人: WILLIAM Lee KRAUS
• 金额: $ 41.09万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10209984
• 关键词: ADP ribosylation; ADP-Ribosylation Factors; Address; Affect; Biochemical; Biological; Biological Assay; Biology; Breast Cancer Cell; Cancer Biology; Cause of Death; Cell Culture Techniques; Cells; Clinical; DNA Binding; DNA Repair; DNA Repair Pathway; Data; Diagnosis; Enhancers; Enzymes; Estrogen Receptor alpha; Exhibits; FDA approved; Family; Gene Activation; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genetic Enhancer Element; Genetic Transcription; Genomics; Growth; Malignant Neoplasms; Mediating; Methods; Modeling; Molecular; Monitor; Mus; Mutate; Mutation; Neoplasm Metastasis; Nuclear; Outcome; Patients; Pharmaceutical Preparations; Phenotype; Play; Poly(ADP-ribose) Polymerases; Polymerase; Post-Translational Protein Processing; Process; Proteins; Proteome; Proteomics; Regulation; Role; Sampling; Site; Testing; Therapeutic; Therapeutic Intervention; Transcriptional Regulation; Xenograft procedure; analog; base; brca gene; cancer cell; cancer heterogeneity; cancer subtypes; cancer type; cell type; clinically relevant; cohort; computational pipelines; drug discovery; epigenomics; improved; inhibitor/antagonist; insight; malignant breast neoplasm; migration; molecular subtypes; response; transcription factor; treatment response; tumor

Project Summary/Abstract The molecular heterogeneity of cancers poses a major hurdle for treatment and drug discovery efforts. Previous studies have addressed this challenge by characterizing distinct molecular subtypes of specific cancers (e.g., breast cancers), based on cell type-specific patterns of gene expression. To interrogate the molecular underpinnings of cancer subtypes, the Kraus Lab has developed a robust and multi-faceted computational pipeline that integrates data from various genomic assays to define a Total Functional Score of Enhancer Elements (TFSEE) for each subtype. One outcome of this method is the identification of cancer subtype- enriched transcription factors (TFs) that promote subtype-specific enhancer formation and drive downstream transcriptional outcomes. In breast cancers, several TFSEE-identified subtype-specific TFs are uniquely required for the growth of the cognate breast cancer subtype, but do not affect the proliferation or viability of other subtypes. Recent studies have also shown that ADP-ribosylation (ADPRylation), a post-translational modification of proteins, varies dramatically across the different subtypes of breast cancers. ADPRylation is mediated by the Poly(ADP-ribose) polymerase (PARP) family of enzymes, including PARP-1, a nuclear enzyme which is the target of FDA-approved PARP inhibitor drugs. PARPs are well known for the roles in DNA repair, but recent studies suggest an important BRCA1/2-independent role in transcriptional regulation as well. In preliminary analyses, we have identified a cohort of cancer-related TFs that are ADPRylated in breast cancers. The long-term objective of these studies is to achieve a better understanding of the molecular and biochemical mechanisms underlying the regulation of breast cancer subtype-specific TFs by ADPRylation, as well as the responses of distinct breast cancer subtypes to PARP inhibitors. Our hypothesis is that ADPRylation of subtype-specific TFs dictates their function and may influence the response of breast cancer cells to PARP inhibitors. We have proposed a project that will use an integrated set of biochemical, molecular, cell-based, mouse-based, genomic, and proteomic assays to test our overarching and specific mechanistic hypotheses. Specifically, we will: (1) Identify TFs that are ADPRylated in breast cancers (Aim 1), (2) Determine how ADPRylation of TFs affects their molecular and biochemical functions (Aim 2), and (3) Determine the effects of TF ADPRylation on the responses of breast cancer cells to clinically used PARP inhibitors (Aim 3). These studies will take advantage of the expertise of the PI’s lab in PARPs, ADPRylation, enhancer function, and gene regulation in cancer. Although focused initially on breast cancers, our results should be broadly applicable across a variety of cancer types. Our integrative approach using ‘omics’ and functional assays will provide new insights into the regulation of TF ADP-ribosylation in breast cancers that will serve as a model for how to explore PARP function and ADP-ribosylation in cancer cells. The use of mouse-based models and patient samples will allow us to explore the clinical relevance of our mechanistic results. We anticipate that our studies will suggest new avenues for the therapeutic potential of PARP inhibitors in cancers beyond DNA repair pathways.

项目摘要/摘要 癌症的分子异质性为治疗和药物发现工作带来了一个主要障碍。 先前的研究通过表征特定癌症的不同分子亚型来解决这一挑战 （例如，乳腺癌），基于基因表达的细胞类型特异性模式。询问分子 癌症亚型的基础，克劳斯实验室开发了强大且多方面的计算 将各种基因组测定数据集成数据以定义增强子的总功能评分的管道 每个亚型的元素（TFSEE）。该方法的一个结果是鉴定癌症亚型 富集的转录因子（TFS）促进亚型特异性增强子形成并下游驱动 转录结果。在乳腺癌中，几个TFSEE识别的亚型特异性TF是独特的 同源乳腺癌亚型生长所必需的，但不影响 其他亚型。最近的研究还表明，ADP-核糖基化（Adprylation），一种翻译后 蛋白质的修饰，各种在乳腺癌的不同亚型中急剧。 Adprylation是 由聚（ADP-核糖）聚合酶（PARP）酶介导的酶，包括PARP-1，一种核酶 这是FDA批准的PARP抑制剂药物的靶标。 PARP以DNA修复中的作用而闻名， 但是最近的研究表明，在转录调控中也具有重要的BRCA1/2独立作用。在 初步分析，我们已经确定了与癌症相关的TF的队列，这些TF在乳腺癌中被丙烯蛋白。 这些研究的长期目标是更好地了解分子和 通过adprylation调节乳腺癌亚型特异性TF的生化机制，AS 以及不同乳腺癌亚型对PARP抑制剂的反应。我们的假设是Adprylation 亚型特异性TF决定其功能，并可能影响乳腺癌细胞对PARP的反应 抑制剂。我们提出了一个项目，该项目将使用一组集成的生化，分子，基于细胞的项目 基于小鼠的基因组和蛋白质组学测定，以测试我们的总体和特定机械假设。 具体而言，我们将：（1）识别在乳腺癌中丙烯的TF（AIM 1），（2）确定如何 TF的adprylation会影响其分子和生化功能（AIM 2），（3）确定 TF对乳腺癌细胞对临床使用的PARP抑制剂的反应的TF adprylation（AIM 3）。这些研究 将利用PI，Adprylation，Enhancer功能和基因的PI实验室的专业知识 癌症的调节。尽管最初专注于乳腺癌，但我们的结果应广泛适用 多种癌症类型。我们使用“ OMICS”和功能测定的综合方法将提供新的见解 在乳腺癌中的TF ADP-核糖基化调节中，该模型将作为探索PARP的模型 癌细胞中的功能和ADP-核糖基化。使用基于鼠标的模型和患者样品将允许 我们探索机械结果的临床相关性。我们预计我们的研究将暗示新的 DNA修复途径以外的癌症中PARP抑制剂的治疗潜力的途径。