Understanding and Overcoming Resistance to BRAF/MEK Kinase Inhibitors in Melanoma

了解并克服黑色素瘤对 BRAF/MEK 激酶抑制剂的耐药性

• 批准号: 10381269
• 负责人: Meenhard F Herlyn
• 金额: $ 4.62万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10381269
• 关键词: Administrative Supplement; Automobile Driving; BRAF gene; Bar Codes; Biology; Cells; Computational Biology; Development; Disease; Dose; Drug resistance; Event; Experimental Designs; Goals; Grant; Immunotherapy; MAP3K1 gene; Malignant Neoplasms; Methods; Molecular; Parents; Pathway interactions; Patients; Pharmaceutical Preparations; Population; Property; RNA; RNA Computations; Recurrent disease; Regimen; Relapse; Resistance; Statistical Data Interpretation; Techniques; Work; cellular imaging; kinase inhibitor; melanoma; new therapeutic target; response; targeted treatment; theories; therapy resistant; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT Targeted immunotherapies against BRAFV600E/K-positive melanoma cancers have been efficacious at eradicating disease in >80% of patients. Nevertheless, the existence of populations of pre-resistant rare cells has posed a challenge to the efficacy of melanoma-targeted therapies, often leading to a disease relapse in the majority of patients. In the parent R01 grant, we proposed a multi-faceted approach combining new experimental techniques, statistical analysis, and theory to understand the origin of these rare, transient drug-resistant cell states and devise strategies to control them. In the current administrative supplement, our goal is to dissect the primordial cellular pathways governing cellular reprogramming that convert the transient state of resistance in the rare single cells into the permanent state of drug resistance that drive relapse. This work is related to the parent R01, but explores new concepts and experimental designs. By using a low dose exposure regimen, followed by high dose exposures, we seek to investigate whether a selective or an adaptive cellular response underlies the mechanism resulting in the conversion to permanent drug resistance. In a selection response, the permanently resistant cells share the same final resistant properties as the transient initial population. In an adaptive response, however, the final resistant properties of the permanently resistant cells could be different, most likely greater, than that of the initial transient cells. We will employ a multi-pronged approach consisting of single cell imaging, single cell barcoding, single cell RNA FISH, RNA sequencing and computational biology techniques to elucidate the primordial cellular/molecular regulators driving cellular reprogramming that results in permanently resistant cells. The mechanisms revealed by these studies will be important for the development of new targeted therapies that will be able to eradicate the relapse caused by therapy resistant cells.

项目概要/摘要 针对 BRAFV600E/K 阳性黑色素瘤的靶向免疫疗法已 能有效根除>80%患者的疾病。尽管如此，人口的存在 预先耐药的稀有细胞对黑色素瘤靶向治疗的功效提出了挑战， 往往导致大多数患者疾病复发。在母公司 R01 拨款中，我们建议 结合新实验技术、统计分析和理论的多方面方法 了解这些罕见的、短暂的耐药细胞状态的起源并制定策略 控制他们。在当前的行政补充中，我们的目标是剖析原始细胞 控制细胞重编程的途径，将瞬时抵抗状态转变为 罕见的单细胞进入永久耐药状态，从而导致复发。这项工作是相关的 与母版 R01 不同，但探索了新概念和实验设计。通过使用低剂量 暴露方案，然后是高剂量暴露，我们试图研究是否有选择性 或者适应性细胞反应是导致转化为机制的基础 永久性耐药性。在选择反应中，永久抵抗细胞共享 与瞬时初始群体相同的最终抗性特性。在适应性反应中， 然而，永久耐药细胞的最终耐药特性可能会有所不同，大多数 可能比最初的瞬时细胞更大。我们将采取多管齐下的方式 包括单细胞成像、单细胞条形码、单细胞 RNA FISH、RNA 测序 和计算生物学技术来阐明原始细胞/分子调节因子 驱动细胞重编程，导致细胞永久耐药。机制 这些研究揭示的结果对于开发新的靶向疗法非常重要 将能够根除治疗耐药细胞引起的复发。