Novel Mechanisms of MED12-TGFbeta-mediated resistance to PARP Inhibitors in BRCA-deficient Cancer Cells

MED12-TGFbeta 介导的 BRCA 缺陷癌细胞对 PARP 抑制剂耐药的新机制

• 批准号: 10536964
• 负责人: Lindsey Marie Pale
• 金额: $ 3.34万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10536964
• 关键词: Address; Antineoplastic Agents; Architecture; Aspergillus Nuclease S1; BARD1 gene; BRCA deficient; BRCA1 gene; BRCA2 gene; Binding; Biological Assay; Breast Cancer Cell; Bromodeoxyuridine; Cancer Patient; Cells; Chemoresistance; Cisplatin; Clinic; Clustered Regularly Interspaced Short Palindromic Repeats; Complement; Complex; Cryoelectron Microscopy; DNA Repair; DNA Repair Gene; DNA replication fork; Data; Development; Disease remission; Double Strand Break Repair; Drug resistance; Exposure to; Fiber; Gene Expression; Genetic Determinism; Genetic Screening; Genome Stability; Genomic Instability; Genomics; Goals; Hela Cells; Knock-out; Knowledge; Laboratories; Light; Maintenance; Malignant Neoplasms; Malignant neoplasm of ovary; Mediating; Mediator of activation protein; Medicine; Mentorship; Oncology; PALB2 gene; Pathway interactions; Patients; Peptides; Pharmaceutical Preparations; Phenotype; Play; Publications; Publishing; Research; Resistance; Resources; Role; Sampling; Scientist; Signal Pathway; Small Interfering RNA; Structure; Techniques; Testing; Therapeutic; Transforming Growth Factor beta; Work; alkalinity; anticancer research; cancer cell; cancer therapy; clinically relevant; college; experience; genome-wide; graduate student; homologous recombination; hydroxyurea; inhibitor; leukemia; malignant breast neoplasm; mutant; novel; novel therapeutic intervention; particle; protein complex; protein expression; repaired; resistance mechanism; response; restoration; skills; therapy resistant

Project Summary This F31 application focuses on characterizing a novel resistance mechanism for PARP inhibitors. I am requesting support for developing a project which I have previously initiated and published on. This proposal addresses a fundamental gap in knowledge and could significantly impact cancer therapeutic strategies and cancer patient survival. Moreover, the resources this application would provide would greatly impact my development as a graduate student and independent scientist. This proposal would allow me the opportunity to be exposed to state-of-the-art laboratory techniques including genomic sequencing and cryo-EM. This would provide me with a complex mix of critical skills that will be important for my development into a well-rounded scientist, and for which are highly attainable due to the support and resources provided by my mentorship team and at Penn State College of Medicine. In this proposal, I focus on elucidating the mechanisms by which MED12 and TGFb mediate resistance to PARP inhibitors. I previously showed loss of MED12 confers resistance to PARP inhibitors in BRCA-deficient cells via activation of the TGFb pathway. This resistance was underscored by an increase in homologous recombination DNA repair efficiency and replication fork stability following TGFb pathway activation in BRCA-deficient cells. Here, I hope to expand on this research to further define the mechanism behind this chemoresistance, to reduce this gap in knowledge, and to gain experience and expertise in the rapidly growing field of structural oncology in cancer research. To investigate the role of MED12 and TGFb in chemoresistance further, I will explore three aims: 1) Investigate specific interactions between MED12 with the Mediator complex and TGFbR2 and the impact of these interactions on chemosensitivity in BRCA-deficient cells, 2) determine the role of ssDNA gap suppression in MED12-TGFb-mediated chemoresistance and genomic stabilization, and 3) investigate the impact of TGFb activation on the structure of mutant BRCA1 in patient-derived HCC1937 breast cancer cells. Achieving these aims will expand my knowledge in the field of DNA damage and repair and expose me to cutting-edge new techniques in the field of structural oncology. Thus, this work will advance my personal goals, and significantly impact the field by providing new information on chemosensitivity in patients harboring BRCA-mutant cancers.

项目摘要 这个F31的应用集中于描述一种新的PARP抑制剂的耐药机制。我是 请求支持我之前发起并发布的一个项目的开发。这项建议 解决了知识上的根本差距，并可能显著影响癌症治疗策略和 癌症患者的存活率。此外，此应用程序提供的资源将极大地影响我的 作为研究生和独立科学家的发展。这项提议将使我有机会 接触最先进的实验室技术，包括基因组测序和冷冻-EM。这将会 为我提供复杂的关键技能组合，这些技能对于我发展成为全面发展的 科学家，由于我的指导团队提供的支持和资源，这是非常有可能实现的 在宾夕法尼亚州立医学院。 在这项建议中，我着重阐明MED12和TGFb介导对PARP的抗性的机制 抑制剂。我之前发现MED12的缺失通过以下途径在BRCA缺陷的细胞中赋予对PARP抑制剂的耐药性 激活TGFb途径。同源重组的增加突显了这种耐药性。 BRCA缺陷细胞中TGFb通路激活后的DNA修复效率和复制叉稳定性。 在这里，我希望扩大这项研究，以进一步确定这种化疗耐药性背后的机制，以减少 这一知识差距，并在快速增长的结构肿瘤学领域获得经验和专业知识 癌症研究。 为了进一步研究MED12和TGFb在化疗耐药中的作用，我将探索三个目的：1)研究 MED12与介体复合体和TGFBR2的特异性相互作用及其影响 BRCA缺陷细胞对化疗敏感性的相互作用，2)决定了单链DNA缺口抑制在 MED12-TGFb介导的化疗耐药和基因组稳定，以及3)研究TGFb的影响 患者来源的HCC1937乳腺癌细胞中突变的BRCA1结构的激活。实现这些目标 AIMS将扩大我在DNA损伤和修复领域的知识，并使我接触到尖端的新 结构肿瘤学领域的技术。因此，这项工作将推进我的个人目标，并显著 通过提供有关BRCA突变癌症患者化疗敏感性的新信息，影响该领域。