Project 1: Tumor Cell Intrinsic Determinants of Early Dissemination in Melanoma

项目1概要 越来越多的数据表明，表型和转录可塑性是重要的特征，有助于 癌细胞的转移进展、向性和对治疗的反应。我们和其他人提供了 令人信服的证据表明，在原发性黑色素瘤中存在瘤内异质性，这表明它是一种早期的恶性肿瘤。 对肿瘤发生很重要。此外，最近的报告表明，黑色素瘤细胞可以转换， 其中一些与更具侵略性的生物学有关， 转录可塑性在这种表型变异。然而，肿瘤细胞内在特征的演变 与黑色素瘤进展相关的疾病仍然在很大程度上未被表征。这些研究评估了 涉及黑色素瘤转移的转录组学和表观遗传学特征通常集中在个体 分子特征局限于特定的细胞群，并且未能整合转移性肿瘤。 整个过程。我们假设，转录异质性的出现，引起不同的 原发性皮肤黑色素瘤的细胞状态是驱动早期转移性播散的关键步骤， 具有特定的转录程序，具有大部分的转移潜力。项目1的目标是 研究转移的分子驱动因素的时间和功能， 转录异质性和细胞状态的转移轨迹。我们将在新的基因 工程小鼠模型，反映人类疾病以及临床注释的患者样品。使用 一种创新的方法，整合了遗传条形码，单细胞RNA测序，空间转录组学， 和高度多重免疫组织化学，我们将评估特定转录细胞状态的存在， 在小鼠和人原发性黑色素瘤中，并使用动物模型评估它们对转移潜能的影响。 模型和统计学与已知患者结局相关（目标1）。然后我们将研究 通过测试单个候选基因对它们的出现的贡献， 维持及其对转移潜力的影响（目标2）。 本研究项目将利用核心B和C的病理、技术和分析资源。 与项目2和3的整合将剖析黑色素瘤细胞之间不断发展的双向串扰， 它们的微环境最终导致肿瘤转移了解特定细胞的贡献 早期传播的潜在状态和分子程序可以提高我们对患者进行亚分类的能力 根据转移风险诊断原发性黑色素瘤，并确定哪些患者受益最大 加强监测或辅助治疗。我们的研究也可能为新的 预防或靶向转移的治疗方法。最后，这项研究的结果可能适用于其他 其中早期播散导致转移潜能增加和预后更差的肿瘤类型。

PROJECT 1 SUMMARY Increasing data suggest that phenotypic and transcriptional plasticity are important features that contribute to metastatic progression, tropism, and response to therapy of cancer cells. We and others have provided compelling evidence that intratumoral heterogeneity exists in primary melanomas, suggesting it is an early feature important for tumorigenesis. Further, recent reports demonstrate that melanoma cells can switch between phenotypic states, some of which are associated with more aggressive biology, implicating transcriptional plasticity in this phenotypic variation. However, the evolution of tumor cell intrinsic features associated with melanoma progression remains largely uncharacterized. The studies that have evaluated transcriptomic and epigenetic features involved in melanoma metastasis have typically focused on individual molecular features restricted to a specific cell population, and have fallen short of integrating the metastatic process as a whole. We hypothesize that emergence of transcriptional heterogeneity that gives rise to distinct cell states in primary cutaneous melanoma is a critical step driving early metastatic dissemination, and that cells with a specific transcriptional program harbor the bulk of metastatic potential. The goal of Project 1 is to investigate the timing and function of molecular drivers of metastasis, the emergence and evolution of transcriptional heterogeneity and metastatic trajectories of cell states. We will do this in novel genetically engineered mouse models that reflect human disease as well as in clinically annotated patient samples. Using an innovative approach that integrates genetic bar coding, single cell RNA sequencing, spatial transcriptomics, and highly-multiplexed immunohistochemistry, we will assess the presence of specific transcriptional cell states within murine and human primary melanomas, and assess their influence on metastatic potential using animal models and statistical correlates to known patient outcomes (Aim 1). We will then study the mechanisms driving these transcriptional cell states by testing the contribution of individual candidate genes to their emergence and maintenance and their impact on metastatic potential (Aim 2). This research project will leverage the pathological, technological, and analytical resources of Cores B and C. Integration with Projects 2 and 3 will dissect the evolving, bidirectional crosstalk between melanoma cells and their microenvironment that culminates in a metastasizing tumor. Understanding the contribution of specific cell states and the molecular programs underlying early dissemination can improve our ability to subclassify patients diagnosed with primary melanoma by their risk of metastasis and identify those patients who could benefit most from increased surveillance or adjuvant therapies. Our studies might also provide a rationale for novel therapeutic approaches to prevent or target metastasis. Finally, findings from this research may apply to other tumor types wherein early dissemination results on increased metastatic potential and worse outcomes.