Characterization of circulating tumor cells in patients with metastatic bladder cancer utilizing functionalized microfluidics

作者：Zeqi Niu, Molly Kozminsky, Kathleen C. Day, Luke J. Broses, Marian L. Henderson, Christopher Patsalis, Rebecca Tagett, Zhaoping Qin, S. Blumberg, Zachery R. Reichert, Sofía D. Merajver, Aaron M. Udager, Phillip L. Palmbos, Sunitha Nagrath, Mark L. Day · 发表于：Neoplasia · 年份：2024 · DOI：10.1016/j.neo.2024.101036 · 被引用次数：11 · 研究领域：Cancer Cells and Metastasis、Bladder and Urothelial Cancer Treatments、Tissue Engineering and Regenerative Medicine

Assessing the molecular profiles of bladder cancer (BC) from patients with locally advanced or metastatic disease provides valuable insights, such as identification of invasive markers, to guide personalized treatment. Currently, most molecular profiling of BC is based on highly invasive biopsy or transurethral tumor resection. Liquid biopsy takes advantage of less-invasive procedures to longitudinally profile disease. Circulating tumor cells (CTCs) isolated from blood are one of the key analytes of liquid biopsy. In this study, we developed a protein and mRNA co-analysis workflow for BC CTCs utilizing the graphene oxide (GO) microfluidic chip. The GO chip was conjugated with antibodies against both EpCAM and EGFR to isolate CTCs from 1 mL of blood drawn from BC patients. Following CTC capture, protein and mRNA were analyzed using immunofluorescent staining and ion-torrent-based whole transcriptome sequencing, respectively. Elevated CTC counts were significantly associated with patient disease status at the time of blood draw. We found a count greater than 2.5 CTCs per mL was associated with shorter overall survival. The invasive markers EGFR, HER2, CD31, and ADAM15 were detected in CTC subpopulations. Whole transcriptome sequencing showed distinct RNA expression profiles from patients with or without tumor burden at the time of blood draw. In patients with advanced metastatic disease, we found significant upregulation of metastasis-related and chemotherapy-resistant genes. T...