Dissecting clonal diversity in melanoma to overcome therapy resistance andmetastasis

项目概要/摘要 转移性黑色素瘤在临床上具有挑战性。尽管标准治疗组合BRAF和MEK 抑制剂（BRAFi/MEKi）治疗在BRAF突变型黑色素瘤患者中具有高应答率， 大多数情况下会复发。我们的实验室和该领域其他人的工作已经确定了 在一些实施方案中，本发明涉及具有内在BRAFi/MEKi抗性的黑色素瘤细胞，其是高度转移性的并且是治疗复发的驱动因素。 这些“持久性”黑色素瘤细胞显示出类似于神经嵴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。我们的长期目标是阐明 主导的分子机制，介导NCSC样黑色素瘤细胞的可塑性和侵略性。 我们的初步研究确定了发育中的LPAR 1-YAP 1轴对黑色素瘤的侵袭性至关重要。 该项目的主要目标之一是评估LPAR 1-YAP 1活动是否代表一种可操作的 易消除NCSC样细胞的转移潜能和内在抗性。拟议的目标将 利用跨诱导型基因编辑技术、谱系 通过时间分析NCSC样黑素瘤细胞，转移性患者来源的异种移植物（Met-PDX）黑素瘤 模型，scRNAseq，RNA FISH和先进的生物信息学，目的是将获得的知识 用于晚期癌症患者的新治疗策略。 另一个主要目标是建立一个独立的研究计划， 合作研究。为确保完成拟议的工作，我和我的委员会确定了四个 我将通过持续培训以及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。我还将参加职业培训，通过研讨会涵盖赠款写作， 手稿准备和演示，负责任的研究行为，利益冲突，实验室/预算 管理和指导。凭借K 01奖项提供的保护时间和稳定性， 以及我将接受的全面培训，我相信我将能够建立一个独立的 研究计划，并成功获得R 01资助。 我的短期职业目标是： 1.确保一个独立的教师职位，并扩大我的同事的科学网络。 2.定义NCSC样黑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R 01资金。 我的长期职业目标是： 1.开发正在进行的项目，根据新的结果扩展到新的方向和临床前研究。 2.成功更新R 01资金并获得替代资金来源，继续职业发展。 3.临床翻译发现，培训未来的学员，并保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.