Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA, USA.
Department of Medicine and Surgery, Section of Anatomic Pathology and Histology, University of Perugia, Perugia, Italy.
Cell Rep Methods. 2022 Aug 22;2(8):100271. doi: 10.1016/j.crmeth.2022.100271.
Clonal evolution and lineage plasticity are key contributors to tumor heterogeneity and response to treatment in cancer. However, capturing signal transduction events in coexisting clones remains challenging from a technical perspective. In this study, we developed and tested a signal-transduction-based workflow to isolate and profile coexisting clones within a complex cellular system like non-small cell lung cancers (NSCLCs). Cooccurring clones were isolated under immunohistochemical guidance using laser-capture microdissection, and cell signaling activation portraits were measured using the reverse-phase protein microarray. To increase the translational potential of this work and capture druggable vulnerabilities within different clones, we measured expression/activation of a panel of key drug targets and downstream substrates of FDA-approved or investigational agents. We isolated intermixed clones, including poorly represented ones (<5% of cells), within the tumor microecology and identified molecular characteristics uniquely attributable to cancer cells that undergo lineage plasticity and neuroendocrine transdifferentiation in NSCLCs.
克隆进化和谱系可塑性是肿瘤异质性和癌症治疗反应的关键因素。然而，从技术角度来看，捕捉共存克隆中的信号转导事件仍然具有挑战性。在这项研究中，我们开发并测试了一种基于信号转导的工作流程，以分离和分析非小细胞肺癌（NSCLC）等复杂细胞系统中的共存克隆。使用激光捕获显微切割，在免疫组织化学指导下分离共发生的克隆，并使用反相蛋白微阵列测量细胞信号激活图谱。为了增加这项工作的转化潜力并捕获不同克隆中的可用药弱点，我们测量了一组关键药物靶点和 FDA 批准或研究性药物的下游底物的表达/激活情况。我们在肿瘤微生态中分离了混合克隆，包括代表性较低的克隆（<5%的细胞），并确定了 NSCLC 中经历谱系可塑性和神经内分泌转分化的癌细胞特有的分子特征。
