Automation, live-cell imaging, and endpoint cell viability for prostate cancer drug screens

Rolando D Z Lyles; Maria J Martinez; Benjamin Sherman; Stephan Schürer; Kerry L Burnstein

doi:10.1371/journal.pone.0287126

Automation, live-cell imaging, and endpoint cell viability for prostate cancer drug screens

PLoS One. 2023 Oct 10;18(10):e0287126. doi: 10.1371/journal.pone.0287126. eCollection 2023.

Authors

Rolando D Z Lyles^{1

2}, Maria J Martinez^{2

3}, Benjamin Sherman^{2

3}, Stephan Schürer^{2

3}, Kerry L Burnstein^{2

3}

Affiliations

¹ Sheila and David Fuente Graduate Program in Cancer Biology, University of Miami Miller School of Medicine, Miami, Florida, United States of America.
² Sylvester Comprehensive Cancer Center, University of Miami, Miami, Florida, United States of America.
³ Department of Molecular & Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, Florida, United States of America.

Abstract

Androgen deprivation therapy (ADT) is the standard of care for high risk and advanced prostate cancer; however, disease progression from androgen-dependent prostate cancer (ADPC) to lethal and incurable castration-resistant prostate cancer (CRPC) and (in a substantial minority of cases) neuroendocrine prostate cancer (NEPC) is common. Identifying effective targeted therapies is challenging because of acquired resistance to established treatments and the vast heterogeneity of advanced prostate cancer (PC). To streamline the identification of potentially active prostate cancer therapeutics, we have developed an adaptable semi-automated protocol which optimizes cell growth and leverages automation to enhance robustness, reproducibility, and throughput while integrating live-cell imaging and endpoint viability assays to assess drug efficacy in vitro. In this study, culture conditions for 72-hr drug screens in 96-well plates were established for a large, representative panel of human prostate cell lines including: BPH-1 and RWPE-1 (non-tumorigenic), LNCaP and VCaP (ADPC), C4-2B and 22Rv1 (CRPC), DU 145 and PC3 (androgen receptor-null CRPC), and NCI-H660 (NEPC). The cell growth and 72-hr confluence for each cell line was optimized for real-time imaging and endpoint viability assays prior to screening for novel or repurposed drugs as proof of protocol validity. We demonstrated effectiveness and reliability of this pipeline through validation of the established finding that the first-in-class BET and CBP/p300 dual inhibitor EP-31670 is an effective compound in reducing ADPC and CRPC cell growth. In addition, we found that insulin-like growth factor-1 receptor (IGF-1R) inhibitor linsitinib is a potential pharmacological agent against highly lethal and drug-resistant NEPC NCI-H660 cells. This protocol can be employed across other cancer types and represents an adaptable strategy to optimize assay-specific cell growth conditions and simultaneously assess drug efficacy across multiple cell lines.

Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Androgen Antagonists / therapeutic use
Androgens / metabolism
Automation
Cell Line, Tumor
Cell Survival
Humans
Male
Prostatic Neoplasms, Castration-Resistant* / metabolism
Receptors, Androgen / metabolism
Reproducibility of Results

Substances

Androgens
Androgen Antagonists
Receptors, Androgen

Grants and funding

Award Recipient: KLB Grant Number: 9BC13 Grant Title: Data-Driven Identification of Novel Precision Drug Combination Therapies for Prostate Cancer Sponsor: Florida Department of Health Program: William G. "Bill" Bankhead, Jr., and David Coley Cancer Research Program Website: https://www.floridahealth.gov/provider-and-partner-resources/research/funding-opportunity-announcements/coleyfoa.html No, the funders did not and will not have a role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Award Recipient: KLB Grant Number: No number assigned Grant Title: Targeting KIF20A a critical promoter of castration resistant prostate cancer in robust pre-clinical models Sponsor: The Mike Slive Foundation Program: Prostate Cancer Research Pilot Grant Program Website: https://mikeslivefoundation.org/ No, the funders did not and will not have a role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Award Recipient: KLB Grant Number: No number assigned Grant Title: Internal funds supporting Dr. Kerry Burnstein, Associate Director, Education and Training Sponsor: Sylvester Comprehensive Cancer Center at the University of Miami Program: Professorship Website: https://umiamihealth.org/sylvester-comprehensive-cancer-center/about-sylvester/leadership/leadership No, the funders did not and will not have a role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.