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

项目1总结 越来越多的数据表明，表型和转录可塑性是导致 癌细胞的转移进展、趋向性和治疗反应。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肿瘤内的异质性，表明这是一种早期的 对肿瘤发生很重要的特征。此外，最近的报告表明，黑色素瘤细胞可以切换 在表型状态之间，其中一些与更具攻击性的生物学相关，暗示 这种表型变异中的转录可塑性。然而，肿瘤细胞内在特征的进化 与黑色素瘤进展相关的疾病在很大程度上仍不明确。已经评估过的研究 涉及黑色素瘤转移的转录和表观遗传学特征通常集中在个体 分子特征仅限于特定的细胞群体，并且未能整合转移的 作为一个整体的过程。我们假设转录异质性的出现导致了不同的 原发皮肤黑色素瘤的细胞状态是推动早期转移扩散的关键步骤，而细胞 有特定的转录程序，就有大部分的转移潜能。项目1的目标是 研究转移的分子驱动因素的时间和功能，肿瘤的出现和演变 细胞状态的转录异质性和转移轨迹。我们将在小说中从基因上做到这一点 反映人类疾病和临床注解患者样本的转基因小鼠模型。vbl.使用 一种创新的方法，它集成了遗传条码、单细胞RNA测序、空间转录 和高度多元化的免疫组织化学，我们将评估特定转录细胞状态的存在 在小鼠和人的原发黑色素瘤中，并使用动物评估它们对转移潜能的影响 模型和统计与已知的患者结局相关(目标1)。然后我们将研究驱动机制 这些转录细胞状态通过测试单个候选基因对它们的出现和 维持及其对转移潜能的影响(目标2)。 该研究项目将利用B和C岩芯的病理、技术和分析资源。 与项目2和项目3的集成将剖析黑色素瘤细胞和 最终导致肿瘤转移的微环境。了解特定细胞的贡献 早期传播的状态和分子计划可以提高我们对患者进行细分的能力 根据转移风险诊断为原发性黑色素瘤，并确定哪些患者受益最大 来自更多的监测或辅助治疗。我们的研究也可能为小说提供理论基础 预防或靶向转移的治疗方法。最后，这项研究的发现可能适用于其他 早期扩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.