Dissecting clonal diversity in melanoma to overcome therapy resistance andmetastasis

项目摘要/摘要 转移性黑色素瘤在临床上具有挑战性。尽管标准护理组合BRAF和MEK 抑制剂(BRAFi/Meki)治疗BRAF突变型黑色素瘤有效率高 复发在大多数情况下会发生。我们实验室和其他现场人员的工作已经确定了亚群 本质上耐BRAFi/Meki的黑色素瘤细胞是高度转移的，是治疗复发的驱动因素。 这些“持续性”黑色素瘤细胞表现出类似于神经脊干细胞的分子和生物学特性 (NCSC)，包括高侵袭性、可塑性和自我更新能力。我们的长期目标是澄清 调节NCSC样黑色素瘤细胞可塑性和侵袭性的主要分子机制。 我们的初步研究发现，发育中的LPAR1-YAP1轴对黑色素瘤的侵袭性至关重要。 该项目的主要目标之一将评估LPAR1-YAP1活动是否代表可操作的 易损毁NCSC样细胞的转移潜能和内在抵抗力。拟议的目标将 利用跨过诱导性基因编辑技术、谱系的多学科系统生物学方法 用时间转移性患者来源的异种黑色素瘤(Met-PDX)分析NCSC样黑色素瘤细胞 模型、scRNAseq、RNA FISH和高级生物信息学，目的是将获得的知识转化为 为晚期癌症患者提供新的治疗策略。 另一个主要目标是建立一个独立的研究计划，进行多学科和 合作研究。为了确保完成拟议的工作，我和我的委员会确定了四个 我将通过持续的培训以及关于单细胞分析的教学课程来加强核心能力 和生物信息学方法。我还将通过研讨会参加职业培训，内容包括拨款撰写、 手稿准备和提交、负责任的研究进行、利益冲突、实验室/预算 我的导师们的管理和指导。凭借K01奖项提供的保护时间和稳定性， 除了我将接受的全面培训外，我相信我将能够建立一个独立的 研究计划，并成功获得R01资金。 我的短期职业目标是： 1.获得一个独立的教职员工职位，扩大我的科学同事网络。 2.确定NCSC样黑色素瘤细胞的克隆多样性和分子脆弱性。 3.通过我的导师/咨询委员会继续发展我的科学能力，并获得R01资金。 我的长期职业目标是： 1.根据取得的新成果，开发正在进行的项目，拓展新方向和临床前研究。 2.成功续签R01资金并获得替代资金来源，继续职业发展。 3.将发现转化为临床，培训未来的指导对象，并维持一个富有成效的研究计划。

Project Summary/Abstract Metastatic melanoma is challenging to clinically address. Although standard of care combination BRAF and MEK inhibitor (BRAFi/MEKi) therapy has high response rates in patients with BRAF-mutant melanoma, therapy relapse occurs in most cases. Work from our laboratory and others in the field has identified subpopulations of intrinsically BRAFi/MEKi-resistant melanoma cells that are highly metastatic and drivers of therapy relapse. These “persistent” melanoma cells display molecular and biologic properties akin to neural crest stem cells (NCSC), including high invasiveness, plasticity and self-renewal capacity. Our long-term goal is to elucidate the dominant molecular mechanisms that mediate the plasticity and aggressiveness of NCSC-like melanoma cells. Our preliminary studies identify a developmental LPAR1-YAP1 axis as critical for melanoma aggressiveness. One of the main objectives of this project will assess whether LPAR1-YAP1 activity represents an actionable vulnerability to ablate the metastatic potential and intrinsic resistance of NCSC-like cells. The proposed aims will leverage a multidisciplinary systems-biology approach spanning inducible genetic-editing techniques, lineage analysis of NCSC-like melanoma cells through time, metastatic patient-derived xenograft (Met-PDX) melanoma models, scRNAseq, RNA FISH and advanced bioinformatics, with the goal of translating the gained knowledge into novel therapeutic strategies for patients with advanced cancer. Another main goal is to establish an independent research program conducting multidisciplinary and collaborative research. To ensure the completion of the proposed work, my committee and I have identified four core competencies I will strengthen with continuous training, as well as didactic courses on single cell analysis and bioinformatic approaches. I will also participate in career training through workshops covering grant writing, manuscript preparation and presentation, responsible conduct of research, conflicts of interest, lab/budget management and guidance from my mentors. With the protected time and stability provided by the K01 award, as well as the comprehensive training I will receive, I am confident that I will be able to establish an independent research program and successfully acquire R01 funding. My short-term career goals are to: 1. Secure an independent faculty position and expand my scientific network of colleagues. 2. Define clonal diversity and molecular vulnerabilities of NCSC-like melanoma cells. 3. Continue to develop my scientific capabilities through my mentor/advisory committee and attain R01 funding. My long-term career goals are to: 1. Develop ongoing projects, expand into new directions and preclinical studies based on new results obtained. 2. Successfully renew R01 funding and attain alternate sources of funding, continue career development. 3. Translate discoveries clinically, train future mentees, and maintain a productive research program.