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.

项目概要/摘要 癌症的分子异质性给治疗和药物发现工作带来了主要障碍。 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），基于细胞类型特异性的基因表达模式。询问分子 Kraus 实验室以癌症亚型为基础，开发了一种强大且多方面的计算模型 整合来自各种基因组测定的数据以定义增强器的总功能评分的管道 每个子类型的元素 (TFSEE)。该方法的一个成果是识别癌症亚型 富集的转录因子 (TF)，可促进亚型特异性增强子形成并驱动下游 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 是同源乳腺癌亚型生长所需的，但不影响乳腺癌亚型的增殖或生存能力 其他亚型。最近的研究还表明，ADP-核糖基化（ADPRylation）是一种翻译后蛋白 蛋白质的修饰在乳腺癌的不同亚型中存在显着差异。 ADPR化是 由聚（ADP-核糖）聚合酶 (PARP) 家族酶介导，包括 PARP-1（一种核酶） 这是 FDA 批准的 PARP 抑制剂药物的目标。 PARP 因其在 DNA 修复中的作用而闻名， 但最近的研究表明，BRCA1/2 在转录调控中也发挥着重要的独立作用。在 通过初步分析，我们已经确定了一组在乳腺癌中发生 ADPR 化的癌症相关 TF。 这些研究的长期目标是更好地了解分子和 ADPRylation 调节乳腺癌亚型特异性 TF 的生化机制，如 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 的数量决定了它们的功能，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，将使用一套集成的生化、分子、细胞、 基于小鼠的基因组和蛋白质组检测来测试我们的总体和具体机制假设。 具体来说，我们将：(1) 识别乳腺癌中 ADPRylated 的 TF（目标 1），(2) 确定如何 TF 的 ADPR 化会影响其分子和生化功能（目标 2），以及 (3) 确定 TF ADPRylation 对乳腺癌细胞对临床使用的 PARP 抑制剂反应的影响（目标 3）。这些研究 将利用 PI 实验室在 PARP、ADPRylation、增强子功能和基因方面的专业知识 癌症中的调节。尽管最初专注于乳腺癌，但我们的结果应该广泛适用于 多种癌症类型。我们使用“组学”和功能分析的综合方法将提供新的见解 乳腺癌中 TF ADP-核糖基化的调节，这将作为如何探索 PARP 的模型 癌细胞中的功能和 ADP-核糖基化。基于小鼠的模型和患者样本的使用将允许 我们探索我们的机制结果的临床相关性。我们预计我们的研究将提出新的建议 PARP 抑制剂在 DNA 修复途径之外的癌症治疗潜力的途径。