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

项目摘要/摘要 针对BRAFV600E/K阳性黑色素瘤癌症的靶向免疫疗法已被 80%的患者在根除疾病方面有效。然而，人口的存在 耐药稀有细胞的出现对黑色素瘤靶向治疗的疗效构成了挑战， 往往会导致一种疾病在大多数患者中复发。在母公司R01拨款中，我们建议 结合新的实验技术、统计分析和理论的多方面方法 为了了解这些罕见的、短暂的耐药细胞状态的起源并制定策略 控制他们。在目前的行政副刊中，我们的目标是剖析原始细胞 控制细胞重新编程的通路，这些通路转换细胞中的瞬时阻力状态 罕见的单个细胞进入永久耐药状态，从而导致复发。这项工作是相关的 为父R01，但探索新的概念和实验设计。通过使用低剂量 暴露方案，然后是高剂量暴露，我们试图调查是否有选择性地 或者适应性细胞反应是导致转化为 永久抗药性。在选择响应中，永久抵抗的细胞共享 与瞬时初始种群相同的最终抗性特性。在适应性响应中， 然而，永久抗性细胞的最终抗性可能是不同的，大多数 可能比最初的瞬变细胞更大。我们会多管齐下。 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 推动细胞重新编程，导致永久抵抗细胞。其作用机制 这些研究揭示的将对开发新的靶向治疗具有重要意义 将能够根除由耐药细胞引起的复发。

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.