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

项目摘要 癌症是最常见的癌症。在发生于中空器官的肿瘤中， 在肿瘤进展过程中，基底膜的破坏和光滑的 肌肉层在膀胱癌中，肌肉浸润与淋巴结和远处转移有关。 转移和预后差。尽管对肿瘤微环境有广泛的兴趣， 对于肿瘤细胞和平滑肌细胞（SMC）如何相互作用还一无所知。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 抑制肿瘤的进展，并且它可以影响肿瘤细胞的表型， 依赖的方式。我们的目标是通过一个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，我们提出将高通量和 空间分辨的单细胞表达谱分析与细胞间的创新计算分析 来自肌肉浸润性膀胱癌患者的样本中的相互作用，以识别细胞 在肌肉侵袭前沿调节肿瘤细胞表型的相互作用。将特别强调 被放置在询问肿瘤细胞-SMC串扰在这个接口，虽然所有的自分泌和 旁分泌的相互作用将在系统水平上以无偏见的方式进行分析。在目标2中， 提出使用共培养系统来机械地解剖肿瘤细胞-SMC串扰，包括 鉴定肿瘤细胞上SMC诱导的转录组学变化，确定这种交叉- 谈话是由接触或可溶性因素介导的，并确定潜在的候选途径 影响下游验证。这些方法的合并将揭示 SMC激活具有预后和/或预测价值，并可能导致发现新的 肌层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 膀胱癌表型和进展中的肿瘤微环境。

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.