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

Zeqi Niu; Molly Kozminsky; Kathleen C. Day; Luke J. Broses; Marian L. Henderson; Christopher Patsalis; Rebecca Tagett; Zhaoping Qin; Sarah Blumberg; Zachery R. Reichert; Sofia D. Merajver; Aaron M. Udager; Phillip L. Palmbos; Sunitha Nagrath; Mark L. Day

Neoplasia: An International Journal for Oncology Research (Nov 2024)

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

Zeqi Niu,
Molly Kozminsky,
Kathleen C. Day,
Luke J. Broses,
Marian L. Henderson,
Christopher Patsalis,
Rebecca Tagett,
Zhaoping Qin,
Sarah Blumberg,
Zachery R. Reichert,
Sofia D. Merajver,
Aaron M. Udager,
Phillip L. Palmbos,
Sunitha Nagrath,
Mark L. Day

Affiliations

Zeqi Niu: Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; Biointerface Institute, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA
Molly Kozminsky: Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; Biointerface Institute, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA
Kathleen C. Day: Department of Urology, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA
Luke J. Broses: Department of Urology, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA
Marian L. Henderson: Department of Internal Medicine, Hematology Oncology Division, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA
Christopher Patsalis: Department of Internal Medicine, Hematology Oncology Division, University of Michigan, Ann Arbor, MI 48109, USA; Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA
Rebecca Tagett: Bioinformatics Core, Michigan Medicine, University of Michigan, Ann Arbor, MI, USA
Zhaoping Qin: Department of Pathology, University of Michigan, Ann Arbor, MI 48109, USA
Sarah Blumberg: Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA
Zachery R. Reichert: Department of Internal Medicine, Hematology Oncology Division, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA
Sofia D. Merajver: Department of Internal Medicine, Hematology Oncology Division, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA
Aaron M. Udager: Department of Pathology, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA
Phillip L. Palmbos: Department of Internal Medicine, Hematology Oncology Division, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA
Sunitha Nagrath: Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; Biointerface Institute, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA; Co-corresponding author at: Department of Chemical Engineering, University of Michigan, Ann Arbor,MI 48109, USA.
Mark L. Day: Department of Urology, University of Michigan, Ann Arbor, MI 48109, USA; Rogel Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA; Corresponding author at: Department of Urology, Rogel Cancer Center University of Michigan, USA.

Journal volume & issue: Vol. 57
p. 101036

Abstract

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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. This methodology demonstrates the capability of GO chip-based assays to identify tumor-related RNA signatures, highlighting the prognostic potential of CTCs in metastatic BC patients.

Published in Neoplasia: An International Journal for Oncology Research

ISSN: 1522-8002 (Print); 1476-5586 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Medicine: Internal medicine: Neoplasms. Tumors. Oncology. Including cancer and carcinogens
Website: http://www.journals.elsevier.com/neoplasia/

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