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 该F31申请的重点是表征PARP抑制剂的新型耐药机制。我是 请求支持开发一个我以前发起并发表的项目。这个建议 解决了知识的根本差距，并可能对癌症治疗策略产生重大影响， 癌症患者生存率。此外，此应用程序将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。这一提议将使我有机会 接触最先进的实验室技术，包括基因组测序和冷冻电镜。这将 为我提供了一个复杂的关键技能组合，这将是我的发展成为一个全面的重要 科学家，由于我的导师团队提供的支持和资源， 以及宾夕法尼亚州立大学医学院。 在这个提议中，我着重阐明MED 12和TGF β介导PARP抗性的机制 抑制剂的我以前曾表明，MED 12的缺失通过以下途径使BRCA缺陷细胞对PARP抑制剂产生耐药性： TGF β通路的激活。这种抗性通过同源重组的增加而得到强调 BRCA缺陷细胞中TGF β途径激活后的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学抗性背后的机制，以减少 这种知识的差距，并获得经验和专业知识，在快速增长的结构肿瘤学领域， 癌症研究。 为了进一步研究MED 12和TGF β在化疗耐药中的作用，我将探索三个目标：1）研究 MED 12与介体复合物和TGFbR 2之间的特异性相互作用以及这些相互作用的影响 BRCA缺陷细胞中化学敏感性的相互作用，2）确定ssDNA间隙抑制在BRCA缺陷细胞中的作用， MED 12-TGF β介导的化学抗性和基因组稳定性，以及3）研究TGF β 在患者来源的HCC 1937乳腺癌细胞中突变BRCA 1的结构上的激活。实现这些 目的将扩大我在DNA损伤和修复领域的知识，并使我接触到尖端的新技术。 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，并显着 通过提供关于携带BRCA突变型癌症患者的化疗敏感性的新信息来影响该领域。