Mechanistic, prognostic, and therapeutic impact of the mitochondrial fission GTPase DRP1 in melanoma

项目总结 Madhavika N.Serasinghe博士是伊坎医学院肿瘤学系的讲师 在西奈山(ISMMS)。她在生物化学、分子和细胞生物学方面有很强的背景，并具有广泛的 有线粒体生物学方面的研究经验。在博士后学习期间，作为一名讲师，她研究了 Ras-MAPK介导的致癌转化中的线粒体生物学，特别关注黑色素瘤。房协- MAPK通路在癌症中经常通过受体酪氨酸激酶Ras GTP酶的突变而过度激活 (如RASG12V)或BRAF激酶(如BRAFV600E)。线粒体分裂蛋白Drp1是RASG12V所必需的 小鼠胚胎成纤维细胞的转化和磷酸化DRP1(DRP1S616℗)是一种预测因子 BRAFV600E阳性黑色素瘤进展的生物标志物。在提议的项目中，Serasinghe博士假设Drp1 是BRAFV600E转化黑素细胞所必需的，而由Drp1介导的线粒体分裂在 在克服癌基因诱导的衰老和色素痣向黑色素瘤进展中的重要作用。在《目标1》中，她将 研究DRp1介导的线粒体生物学改变在恶性转化中的机制影响 原代人黑素细胞慢病毒BRAFV600E表达/PTEN基因敲除模型的建立她的第二个目标将集中在 将她的发现转化为临床相关背景。与治疗性抗体中心合作 在ISMMS的开发中，她将开发一种人类特异的单抗，作为预测分析的基础 确定需要定期临床监测以早期发现和处理黑色素瘤的患者亚群。 作为目标2的一部分，她将开发对黑色素瘤具有潜在治疗价值的drp1特异性小分子抑制剂 和其他癌症。与ISMMS实验治疗研究所合作，利用结构指导 虚拟筛选2000万化合物库的方法，她将选择和验证潜在的活页夹使用 体外和基于细胞的Drp1活性和结合分析。这些翻译目标将为塞拉辛格博士提供亲身实践 在药物发现过程和生物标记物分析开发方面的经验，并使她获得新的技能 和技术专长，为未来的翻译研究。这些研究预测的临床方面将使她能够 获得更多基础研究的工作台到床边翻译。Serasinghe博士的近期职业目标是 在曾傑瑞奇普克博士的指导下开发一个独立的研究项目，获得联邦资金，并建立 一家学术癌症研究所的实验室作为终身教职员工进行高质量的癌症研究。 研究。在ISMMS，她将得到极好的指导，出色的科学和职业发展培训，以及 发展多学科合作的机会，以推进她的研究范围。Serasinghe医生的长期 职业目标是成为黑色素瘤研究的领导者，并致力于发现创新的预防方法， 针对黑色素瘤的预后和治疗方法。NCI-K22奖项将为Serasinghe博士提供宝贵的 支持实现她的研究和职业目标，为癌症研究做出重要贡献，并支持使命 美国国家卫生研究院和国家情报局。

PROJECT SUMMARY Dr. Madhavika N. Serasinghe is an Instructor in the Department of Oncological Sciences at the Icahn School of Medicine at Mount Sinai (ISMMS). She has a strong background in biochemistry, molecular and cell biology, and extensive research experience in mitochondrial biology. During her post-doctoral studies and as an Instructor she studied the role of mitochondrial biology in oncogenic RAS-MAPK mediated transformation, with special focus on melanoma. The RAS- MAPK pathway is frequently hyper-activated in cancer through mutations in the receptor tyrosine kinases, RAS GTPase (e.g. RASG12V), or the BRAF kinase (e.g. BRAFV600E). The mitochondrial fission protein DRP1 is requisite for RASG12V mediated transformation of mouse embryonic fibroblasts, and phosphorylated DRP1 (DRP1S616℗) serves as a predictive biomarker for BRAFV600E positive melanoma progression. In the proposed project Dr. Serasinghe hypothesizes that DRP1 is required for melanocyte transformation by BRAFV600E, and that mitochondrial fission mediated by DRP1 plays an important role in overcoming oncogene induced senescence and progression of nevi to melanoma. In aim 1, she will examine the mechanistic impact of DRP1 mediated changes to mitochondrial biology on malignant transformation using a lentiviral BRAFV600E expression /PTEN knockdown model of primary human melanocytes. Her second aim will focus on translating her findings to a clinically relevant context. In collaboration with the Center for Therapeutic Antibody Development at ISMMS, she will develop a human specific monoclonal antibody as a basis for a prognostic assay to determine subsets of patients who require regular clinical monitoring for early detection and management of melanoma. As a part of aim 2, she will develop DRP1 specific small molecule inhibitors with potential therapeutic value in melanoma and other cancers. In collaboration with the Experimental Therapeutics Institute at ISMMS, utilizing structure-guided approaches to virtually screen a library of >20 million compounds, she will select and validate potential binders using in vitro and cell-based DRP1 activity and binding assays. These translational aims will provide Dr. Serasinghe hands-on experience in the drug discovery process as well as biomarker assay development, and allow her to acquire new skill sets and technical expertise for future translational research. The projected clinical aspects of these studies will enable her to gain further exposure to bench-to-bedside translation of basic research. Dr. Serasinghe’s immediate career goal is to develop an independent research program under the mentorship of Dr. Jerry Chipuk, obtain federal funding, and establish a laboratory at an academic cancer research institute as a tenure track faculty member to conduct high quality cancer research. At ISMMS, she will receive excellent mentorship, exceptional scientific and career development training, and opportunities to develop multidisciplinary collaborations to advance the scope of her research. Dr. Serasinghe’s long term career goal is to become a leader in melanoma research, and dedicate her efforts to discovering innovative preventative, prognostic, and therapeutic modalities against melanoma. The NCI-K22 award will provide Dr. Serasinghe valuable support to achieve her research and career goals, make important contributions to cancer research, and support the mission of the NIH and NCI.