Copper reduction as a novel therapy in BRAF-mutant positive cancers

简介(申请人提供)：我作为癌症生物学家的培训始于北卡罗来纳大学教堂山分校(UNC-CH)，最初是我在卡罗莱纳夏季研究奖学金期间在Chning J.Der博士实验室的本科生，后来是Adrienne D.Cox博士实验室的研究生。目前我是杜克大学Christopher M.Counter博士实验室的博士后研究员。这些研究经验使我在研究肿瘤发生、获取蛋白质功能的生化策略以及癌症的体内模型方面掌握了广泛的分子和细胞生物学技术，更重要的是，坚定了我在未来追求学术教师职位的愿望。然而，通过K01奖励资助机制提供的额外指导阶段将为我提供受保护的时间，以便与同事和合作者网络一起制定我自己的独立研究计划，提高我的演讲和管理技能，并在“教授之路”上取得进展。具体地说，我正在积极推进这项申请中提出的一个创新研究项目，该项目将通过将我的研究扩展到肿瘤发生的体内模型的临床前试验来增加我的研究技能，让我有机会在杜克大学内部和国家科学会议上展示我的研究，并促进新研究途径的发展，一旦我成为一名独立研究员，这些途径可以在校外拨款的支持下探索。环境：药理学和癌症生物学系以及杜克大学癌症研究所(DCI)内部的科学界，DCI和医学院(SOM)提供的资源和设施，杜克大学博士后服务办公室(OPS)提供的支持和资源，以及我的导师Christopher M.Counter博士的经验、承诺和支持，在我癌症生物学生涯的这个指导阶段，以及我从一名指导人员过渡到独立研究员的过程中，都将是我宝贵的财富。简而言之，关于我的职业发展和成为教授的途径，Counter博士的实验室为我提供了很好的设备来从事我的研究兴趣，并不断得到R01拨款的资助，这些拨款导致在 高影响力的期刊，并为我提供定期展示我的研究和论文的机会。此外，Counter博士和我的导师委员会成员Pendergast博士、Thiele博士和Wang博士在博士后研究员培训方面拥有丰富的经验，他们将就我的研究和科学手稿给我建设性的反馈，并帮助我铺平职业道路，特别是在向独立研究员过渡的一年。老年退休金计划将有助于我继续接受负责任的研究行为(RCR)培训，并提供广泛的服务，包括模拟面试、实践学术工作讲座和研讨会，如有效的学术求职策略、实验室管理和财务管理。此外，Counter博士还担任NCI指定的DCI肿瘤生物学项目的联合负责人，该项目由来自广泛的基础和临床部门的30个实验室组成，专注于癌症研究，并是DCI黑色素瘤疾病现场小组的成员，该小组旨在将基础研究发现转化为研究人员发起的第一阶段试验。我有很多机会在他们每周或每月的工作进展会议上展示我的研究成果。总而言之，杜克大学现有的研究环境，特别是在药理学和癌症生物学系以及DCI内部，是开展我的研究项目和职业发展的理想选择。研究项目：如上所述，我博士后培训的两个主要目标是增加我对体内肿瘤发生模型的接触，并激发我对基础研究发现取得进展的兴趣。为此，我提出的研究项目源于我们之前的观察，即铜(Cu)的流入通过铜-MEK1相互作用促进其底物ERK1/2的MEK1磷酸化。黑色素瘤和广泛的其他癌症，如甲状腺和结肠癌，在丝氨酸/苏氨酸激酶BRAF中存在激活突变，通常对针对突变BRAF的小分子激酶抑制剂产生显著反应。然而，它们迅速产生抗药性，只产生短期的临床反应。因此，多种抑制MAPK信号的方法的结合为更有效地治疗BRAF突变阳性癌症带来了巨大的希望。我们目前在《自然》杂志上进行的审查工作表明，高亲和力铜转运蛋白CTR1的基因缺失减少了BRAFV600E转化细胞的肿瘤生长，用于治疗威尔逊病的铜络合剂不仅在BRAFV600E转化细胞上产生了类似的结果，而且在对BRAF抑制剂维莫拉非尼耐药的细胞上也产生了类似的结果。综上所述，这些结果表明，铜螯合疗法可以改变治疗BRAF突变阳性癌症的目的。因此，为了解决与我的研究兴趣相关的癌症生物学中一个新颖而有意义的研究问题，并继续发展我在癌症生物学方面的技能，我提出了专注于单独使用或与BRAF抑制剂Vemurafenib联合使用铜的药理螯合作用的科学目标，在由固有活性BRAFV600E和Pten丢失驱动的黑色素瘤的GEMM中。此外，我还将在BRAF突变的甲状腺和结肠癌中研究CTR1基因消融和铜螯合作用，这些突变也依赖于MAPK通路的持续激活。这些研究努力将巩固我作为癌症生物学家的培训，并引发新的研究问题，这些问题将成为我追求癌症生物学学术教职的基础。

DESCRIPTION (provided by applicant): My training as a cancer biologist began at the University of North Carolina at Chapel Hill (UNC- CH), first as an undergrad in the lab of Channing J. Der, Ph.D., during my Carolina Summer Research Fellowship, and later as a graduate student in the lab of Adrienne D. Cox, Ph.D. Currently I am a post-doctoral fellow in the lab of Christopher M. Counter, Ph.D., at Duke University. These research experiences have trained me in a wide array of molecular and cellular biology techniques to study tumorigenesis, biochemical strategies to access protein function, and in vivo models of cancer, and more importantly, firmly solidified my desire to pursue an academic faculty position in the future. However, the additional mentored phase provided through the K01 award mechanism of funding will provide me with the protected time to develop my own independent research program with a network of colleagues and collaborators, improve my presentation and management skills, and progress along the "path to professorship". Specifically, I am actively pursuing an innovative research project proposed in this application that will add to my research skills by expanding my research to preclinical trials in in vivo models of tumorigenesis, give me the opportunity to present my research within Duke University and at national scientific conferences, and foster the development of novel research avenues that can be explored with extramural grant support once I've become an independent researcher. Environment: The scientific community within the Department of Pharmacology and Cancer Biology and Duke Cancer Institute (DCI), the resources and facilities provided by the DCI and School of Medicine (SOM), the support and resources provided by the Office of Postdoctoral Services (OPS) at Duke University, and the experience, commitment and support from my mentor, Christopher M. Counter, Ph.D., will be valuable assets during this mentored phase of my career in cancer biology and as I transition from a mentored to independent researcher. Briefly, in regards to my career development and path to professorship, Dr. Counter's lab is well equipped for me to undertake my research interests, is continually funded by R01 grants that result in publication of high profile papers in high impact journals, and provides me with opportunities to regularly present my research and papers. In addition, Dr. Counter and the members of my mentorship committee, Drs. Pendergast, Thiele, and Wang, have vast experience in the training of postdoctoral fellows and will give me constructive feedback on my research and scientific manuscripts, along with helping pave my career path, especially in the transition year to an independent investigator. The OPS will be instrumental in my continued training in the Responsible Conduct of Research (RCR) and in providing a wide spectrum of services, including mock interviews, practice academic job talks, and workshops such as effective strategies for the academic job search, managing a lab, and financial stewardship. Additionally, Dr. Counter serves as co-leader of the NCI- designated DCI Tumor Biology Program, comprised of 30 labs from a broad spectrum of basic and clinical departments focusing on cancer research, and is a member of the DCI Melanoma Disease-Site Group, which aims to translate basic research discoveries to investigator-initiated phase I trials. I have many opportunities to present my research at their weekly or monthly work-in-progress meetings. Taken together, the research environment that exists at Duke University, specifically within the Department of Pharmacology and Cancer Biology and DCI, is ideal to carry out my research project and career development. Research Project: As mentioned above, two major goals of my post-doctoral training are to increase my exposure to in vivo models of tumorigenesis and to engage my interest in seeing basic research discoveries move forward. To that end, my proposed research project evolved from our previous observations that copper (Cu) influx enhances MEK1 phosphorylation of its substrates ERK1/2 through a Cu-MEK1 interaction. Melanomas and a wide spectrum of other cancers, like thyroid and colon cancer, that harbor activating mutations in the serine/threonine kinase BRAF, often respond dramatically to small molecule kinase inhibitors against mutant BRAF. However, they rapidly develop resistance, yielding only short-term clinical responses. Thus, combinations of multiple approaches to inhibit MAPK signaling hold great promise for more effective treatment of BRAF mutation-positive cancers. Our current work under review at the journal Nature shows that genetic loss of the high affinity Cu transporter Ctr1 reduced the tumor growth of BRAFV600E-transformed cells, and that a Cu chelator used in the treatment of Wilson's disease yielded a similar result not only on BRAFV600E-transformed cells, but also on cells resistant to a BRAF inhibitor, vemurafenib. Taken together, these results suggest that Cu-chelation therapy could be repurposed for the treatment of BRAF mutation- positive cancers. Thus in order to address a novel and significant research question in cancer biology that is relevant to my research interests and to continue to develop my skills in cancer biology, I am proposing scientific aims focused on the use of pharmacological chelation of Cu either alone or in combination with the BRAF inhibitor, vemurafenib, in a GEMM of melanoma driven by constitutively active BrafV600E and loss of Pten. In addition, I will investigate Ctr1 genetic ablation and Cu chelation in BRAF-mutant thyroid and colon cancers that are also reliant on sustained MAPK pathway activation. These research endeavors will solidify my training as a cancer biologist and spark novel research questions that will become the foundation for my pursuit of an academic faculty position in cancer biology.