PPP6C Regulation of ERK Signaling in Melanoma

PPP6C 对黑色素瘤中 ERK 信号传导的调节

• 批准号: 9539242
• 负责人: Eunice Cho
• 金额: $ 2.92万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-01 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9539242
• 关键词: Anchorage-Independent Growth; Automobile Driving; BRAF gene; Biochemical; Biological Assay; CRISPR/Cas technology; Catalytic Domain; Cell Line; Cells; Clinical; Complex; Cultured Cells; Data; Down-Regulation; Drug resistance; Ensure; Genetic; Genomics; Goals; Growth Factor; In Vitro; In complete remission; Lesion; Light; MEK inhibition; MEKs; Malignant - descriptor; Malignant Neoplasms; Melanoma Cell; Mitogen-Activated Protein Kinases; Molecular; Mutate; Mutation; NF1 gene; Oncogenic; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Phosphoric Monoester Hydrolases; Phosphotransferases; Predisposition; Protein Dephosphorylation; Protein Phosphatase 2A Regulatory Subunit PR53; Regulation; Resistance; Role; Signal Pathway; Signal Transduction; Skin Cancer; Therapeutic; Treatment Efficacy; Work; base; clinical efficacy; exome sequencing; experimental study; genome editing; improved; inhibitor/antagonist; insight; interest; knock-down; loss of function; loss of function mutation; melanocyte; melanoma; member; mouse model; mutant; novel; protein phosphatase 6; response; small hairpin RNA; targeted sequencing; targeted treatment; tumor progression; tumorigenic; Anchorage-Independent Growth; Automobile Driving; BRAF gene; Biochemical; Biological Assay; CRISPR/Cas technology; Catalytic Domain; Cell Line; Cells; Clinical; Complex; Cultured Cells; Data; Down-Regulation; Drug resistance; Ensure; Genetic; Genomics; Goals; Growth Factor; In Vitro; In complete remission; Lesion; Light; MEK inhibition; MEKs; Malignant - descriptor; Malignant Neoplasms; Melanoma Cell; Mitogen-Activated Protein Kinases; Molecular; Mutate; Mutation; NF1 gene; Oncogenic; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Phosphoric Monoester Hydrolases; Phosphotransferases; Predisposition; Protein Dephosphorylation; Protein Phosphatase 2A Regulatory Subunit PR53; Regulation; Resistance; Role; Signal Pathway; Signal Transduction; Skin Cancer; Therapeutic; Treatment Efficacy; Work; base; clinical efficacy; exome sequencing; experimental study; genome editing; improved; inhibitor/antagonist; insight; interest; knock-down; loss of function; loss of function mutation; melanocyte; melanoma; member; mouse model; mutant; novel; protein phosphatase 6; response; small hairpin RNA; targeted sequencing; targeted treatment; tumor progression; tumorigenic

PROJECT SUMMARY/ABSTRACT Melanoma, the deadliest form of skin cancer, is characterized by aberrant hyperactivation of the ERK mitogen- activated protein kinase (MAPK) signaling pathway. Inhibitors of BRAF and MEK, both members of this pathway, have shown clinical efficacy and are used in combination to treat BRAF mutant melanoma, the most common melanoma genomic subtype. However, not all BRAF mutant melanomas respond to these inhibitors, and those that do respond eventually acquire resistance. Better understanding the molecular mechanisms underlying the response and resistance to BRAF and MEK inhibitors will bring us closer to developing durable therapeutic strategies. The main goal of this project is to establish a novel regulatory role for PPP6C, the catalytic subunit of protein phosphatase 6 (PPP6C), in the ERK signaling network. This will allow for understanding how observed genetic lesions in PPP6C alter this oncogenic signaling pathway, likely driving malignant transformation as well as drug resistance in melanoma. Our interest in PPP6C is based on 1) preliminary work observing downregulation of PPP6C promotes resistance to MEK inhibition and increases ERK activation and 2) whole exome and targeted sequencing projects identifying PPP6C is mutated in 7-12% of melanomas. This proposal will examine the effects of PPP6C silencing on ERK activation and inhibitor sensitivity across a panel of melanoma patient-derived cells to confirm a role for PPP6C in regulating ERK signaling. The specific signaling mechanisms underlying PPP6C regulation of ERK signaling will be determined by cellular and biochemical approaches investigating how PPP6C directly or indirectly regulates MEK activation. Regulation of ERK signaling and MEK/BRAF inhibitor susceptibility by cancer-associated PPP6C mutations will be examined in cultured cells and in a mouse model of melanoma. To assess the tumorigenic potential of PPP6C loss, PPP6C knockdown melanocytes will be evaluated for growth factor independent proliferation and anchorage independent growth. The findings from these studies will provide insight into how to optimize the use of clinical MEK and BRAF inhibitors in melanoma and identify opportunities to improve the efficacy of targeted therapies.

项目摘要/摘要 黑色素瘤是皮肤癌最致命的形式，其特征是ERK促丝分裂原的异常过度激活 活化的蛋白激酶（MAPK）信号通路。 BRAF和MEK的抑制剂，这两个成员 途径，已显示出临床功效，并被用于治疗BRAF突变型黑色素瘤，最多。 常见的黑色素瘤基因组亚型。但是，并非所有BRAF突变黑色素瘤都对这些抑制剂做出反应， 那些确实做出反应的人最终获得了抵抗。更好地了解分子机制 对BRAF和MEK抑制剂的反应和抵抗力的基础将使我们更接近发展耐用的 治疗策略。该项目的主要目标是为PPP6C建立新的调节作用 ERK信号网络中蛋白质磷酸酶6（PPP6C）的催化亚基。这将允许 了解PPP6C中观察到的遗传病变如何改变这种致癌信号通路，可能驱动 黑色素瘤中的恶性转化以及耐药性。我们对PPP6C的兴趣基于1） 观察PPP6C下调的初步工作促进了对MEK抑制的抵抗力并增加 ERK激活和2）识别PPP6C的整个外显子组和靶向测序项目被突变为7-12％ 黑色素瘤。该建议将检查PPP6C沉默对ERK激活和抑制剂的影响 黑色素瘤患者衍生细胞的敏感性确认PPP6C在调节ERK中的作用 信号。将确定ERK信号调节的特定信号传导机制 通过细胞和生化方法研究PPP6C如何直接或间接调节MEK 激活。通过癌症相关的PPP6C调节ERK信号传导和MEK/BRAF抑制剂易感性 将在培养细胞和黑色素瘤小鼠模型中检查突变。评估肿瘤性 PPP6C损失的潜力，PPP6C敲低黑素细胞将被评估为生长因子独立 增殖和独立生长。这些研究的发现将提供有关如何 优化黑色素瘤中临床MEK和BRAF抑制剂的使用，并确定改善的机会 靶向疗法的功效。