eprintid: 21905 rev_number: 23 userid: 3671 importid: 2442 dir: disk0/00/02/19/05 datestamp: 2014-06-18 20:29:17 lastmod: 2019-01-22 16:55:02 status_changed: 2014-06-18 20:29:17 type: article metadata_visibility: show item_issues_count: 0 eprint_status: archive creators_name: Joy, ME creators_name: Vollmer, LL creators_name: Hulkower, K creators_name: Stern, AM creators_name: Peterson, CK creators_name: Boltz, RC creators_name: Roy, P creators_name: Vogt, A creators_email: mej29@pitt.edu creators_email: llv4@pitt.edu creators_email: creators_email: STERNAM@pitt.edu creators_email: creators_email: rcb56@pitt.edu creators_email: par19@pitt.edu creators_email: avogt@pitt.edu creators_id: MEJ29 creators_id: LLV4 creators_id: creators_id: STERNAM creators_id: creators_id: RCB56 creators_id: PAR19 creators_id: AVOGT contributors_type: http://www.loc.gov/loc.terms/relators/EDT contributors_name: Weaver, Alissa M. title: A high-content, multiplexed screen in human breast cancer cells identifies profilin-1 inducers with anti-migratory activities ispublished: pub divisions: sch_med_Computational_Systems_Biology divisions: sch_med_Pathology divisions: sch_eng_bio full_text_status: public abstract: Profilin-1 (Pfn-1) is a ubiquitously expressed actin-binding protein that is essential for normal cell proliferation and migration. In breast cancer and several other adenocarcinomas, Pfn-1 expression is downregulated when compared to normal tissues. Previous studies from our laboratory have shown that genetically modulating Pfn-1 expression significantly impacts proliferation, migration, and invasion of breast cancer cells in vitro, and mammary tumor growth, dissemination, and metastatic colonization in vivo. Therefore, small molecules that can modulate Pfn-1 expression could have therapeutic potential in the treatment of metastatic breast cancer. The overall goal of this study was to perform a multiplexed phenotypic screen to identify compounds that inhibit cell motility through upregulation of Pfn-1. Screening of a test cassette of 1280 compounds with known biological activities on an Oris™ Pro 384 cell migration platform identified several agents that increased Pfn-1 expression greater than two-fold over vehicle controls and exerted anti-migratory effects in the absence of overt cytotoxicity in MDA-MB-231 human breast cancer cells. Concentration-response confirmation and orthogonal follow-up assays identified two bona fide inducers of Pfn-1, purvalanol and tyrphostin A9, that confirmed in single-cell motility assays and Western blot analyses. SiRNA-mediated knockdown of Pfn-1 abrogated the inhibitory effect of tyrphostin A9 on cell migration, suggesting Pfn-1 is mechanistically linked to tyrphostin A9's anti-migratory activity. The data illustrate the utility of the high-content cell motility assay to discover novel targeted anti-migratory agents by integrating functional phenotypic analyses with target-specific readouts in a single assay platform. © 2014 Joy et al. date: 2014-02-10 date_type: published publication: PLoS ONE volume: 9 number: 2 refereed: TRUE centers: cen_other_drugdiscoveryinst centers: cen_other_mageewomensresearchinst id_number: 10.1371/journal.pone.0088350 citation: Joy, ME and Vollmer, LL and Hulkower, K and Stern, AM and Peterson, CK and Boltz, RC and Roy, P and Vogt, A (2014) A high-content, multiplexed screen in human breast cancer cells identifies profilin-1 inducers with anti-migratory activities. PLoS ONE, 9 (2). document_url: http://d-scholarship-dev.library.pitt.edu/21905/1/journal.pone.0088350.pdf document_url: http://d-scholarship-dev.library.pitt.edu/21905/8/licence.txt