Role of the smooth muscle layer in bladder cancer biology and progression: a systems and experimental approach

项目总结 癌症是最常见的癌症。在发生在中空器官的肿瘤中，有两个主要事件 在肿瘤的进展过程中，基底膜的破坏和光滑的侵袭 肌肉层。在膀胱癌中，肌肉侵犯与淋巴转移和远处转移有关 转移和预后不良。尽管人们对肿瘤微环境有广泛的兴趣，但实际上 关于肿瘤细胞和平滑肌细胞(SMC)是如何相互作用的，目前还一无所知。初步 观察结果支持我们的中心假设，即平滑肌层可能促进，而不是 抑制肿瘤进展，并在一定程度上影响肿瘤细胞的表型- 依赖的态度。我们的目标是通过自上而下的系统来解决这一重大的知识差距 和自下而上的实验方法。在目标1中，我们建议将高吞吐量和 细胞-细胞创新计算分析的空间分辨单细胞表达谱 肌肉浸润性膀胱癌患者标本相互作用鉴定细胞 在肌肉侵袭前沿调节肿瘤细胞表型的相互作用。特别强调将 被置于询问肿瘤细胞-SMC串扰的这个界面上，尽管所有的自分泌和 旁分泌相互作用将以不偏不倚的方式在系统层面进行分析。在目标2中，我们 建议使用共培养系统来机械解剖肿瘤细胞-SMC串扰，包括 鉴定SMC诱导的肿瘤细胞转录变化，确定这种交叉- 谈话是由接触或可溶因素促成的，并确定构成这一点的候选途径 对下游验证的影响。这些方法的融合将揭示出 SMC激活具有预后和/或预测价值，并可能导致发现新的 肌肉浸润性膀胱癌的治疗靶点。我们希望发现一个尚未探索的角色 研究肿瘤微环境在膀胱癌表型和进展中的作用。

PROJECT SUMMARY Carcinomas are the most common cancers. In tumors arising in hollow organs, two major events during tumor progression are disruption of the basement membrane and invasion of the smooth muscle layer. In bladder cancer, muscle-invasion is associated with lymph node and distant metastases and poor prognosis. Despite extensive interest in the tumor microenvironment, virtually nothing is known about how tumor cells and smooth muscle cells (SMC) interact. Preliminary observations support our central hypothesis that the smooth muscle layer may promote, rather than restrain, tumor progression and that it can impact on the tumor cell phenotype in a context- dependent manner. We aim to address this significant knowledge gap through a top-down systems and bottom-up experimental approach. In Aim 1, we propose combining high-throughput and spatially-resolved single cell expression profiling with innovative computational analyses of cell-cell interactions in samples from patients with muscle-invasive bladder cancer to identify cellular interactions that modulate tumor cell phenotype at the muscle-invasive front. Special emphasis will be placed on interrogating tumor cell-SMC crosstalk at this interface, although all autocrine and paracrine interactions will be analyzed at a systems level in an unbiased fashion. In Aim 2, we propose using co-culture systems to mechanistically dissect tumor cell-SMC crosstalk, including identifying transcriptomic changes induced by SMC on tumor cells, determining whether this cross- talk is mediated by contact or soluble factors, and identifying candidate pathways that underlie this effect for downstream validation. The amalgamation of these approaches will reveal markers of SMC activation with prognostic and/or predictive value and may lead to the discovery of new therapeutic targets for muscle-invasive bladder cancer. We expect to uncover a yet unexplored role of the tumor microenvironment in bladder cancer phenotypes and progression.