Proteomic modulation in breast tumors after metformin exposure

results from a “window of opportunity” trial

K. Kalinsky, T. Zheng, H. Hibshoosh, X. Du, P. Mundi, J. Yang, S. Refice, Sheldon M. Feldman, B. Taback, E. Connolly, K. D. Crew, M. A. Maurer, D. L. Hershman

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Purpose: Reverse Phase Protein Array (RPPA) is a high-throughput antibody-based technique to assess cellular protein activity. The goal of this study was to assess protein marker changes by RPPA in tumor tissue from a pre-surgical metformin trial in women with operable breast cancer (BC). Methods: In an open-label trial, metformin 1500-mg PO daily was administered prior to resection in 35 non-diabetic patients with stage 0–III BC, body mass index ≥25 kg/m2. For RPPA, formalin-fixed paraffin-embedded (FFPE) samples were probed with 160 antibodies. Paired and two-sample t-tests were performed (p ≤ 0.05). Multiple comparisons were adjusted for by fixing the false discovery rate at 25 %. We evaluated whether pre- and post-metformin changes of select markers by RPPA were identified by immunohistochemistry (IHC) in these samples. We also assessed for these changes by western blot in metformin-treated BC cell lines. Results: After adjusting for multiple comparisons in the 32 tumors from metformin-treated patients vs. 34 untreated historical controls, 11 proteins were significantly different between cases vs. controls: increases in Raptor, C-Raf, Cyclin B1, Cyclin D1, TRFC, and Syk; and reductions in pMAPKpT202,Y204, JNKpT183,pT185, BadpS112, PKC.alphapS657, and SrcpY416. Cyclin D1 change after metformin by IHC was not observed. In cell lines, reductions in JNKpT183 and BadpS112 were seen, with no change in Cyclin D1 or Raptor. Conclusions: These results suggest that metformin modulates apoptosis/cell cycle, cell signaling, and invasion/motility. These findings should be assessed in larger metformin trials. If confirmed, associations between these changes and BC clinical outcome should be evaluated. Clinicaltrials.gov identifier: NCT00930579.

Original languageEnglish (US)
Pages (from-to)180-188
Number of pages9
JournalClinical and Translational Oncology
Volume19
Issue number2
DOIs
StatePublished - Feb 1 2017
Externally publishedYes

Fingerprint

Metformin
Proteomics
Breast Neoplasms
Protein Array Analysis
Cyclin D1
Raptors
Immunohistochemistry
Cyclin B1
Cell Line
Proteins
Antibodies
Paraffin
Formaldehyde
Neoplasms
Cell Cycle
Body Mass Index
Western Blotting
Apoptosis

Keywords

  • Breast Cancer
  • Cyclin D1
  • Immunohistochemistry
  • Metformin
  • Pre-surgical
  • Reverse Phase Protein Array

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Proteomic modulation in breast tumors after metformin exposure : results from a “window of opportunity” trial. / Kalinsky, K.; Zheng, T.; Hibshoosh, H.; Du, X.; Mundi, P.; Yang, J.; Refice, S.; Feldman, Sheldon M.; Taback, B.; Connolly, E.; Crew, K. D.; Maurer, M. A.; Hershman, D. L.

In: Clinical and Translational Oncology, Vol. 19, No. 2, 01.02.2017, p. 180-188.

Research output: Contribution to journalArticle

Kalinsky, K, Zheng, T, Hibshoosh, H, Du, X, Mundi, P, Yang, J, Refice, S, Feldman, SM, Taback, B, Connolly, E, Crew, KD, Maurer, MA & Hershman, DL 2017, 'Proteomic modulation in breast tumors after metformin exposure: results from a “window of opportunity” trial', Clinical and Translational Oncology, vol. 19, no. 2, pp. 180-188. https://doi.org/10.1007/s12094-016-1521-1
Kalinsky, K. ; Zheng, T. ; Hibshoosh, H. ; Du, X. ; Mundi, P. ; Yang, J. ; Refice, S. ; Feldman, Sheldon M. ; Taback, B. ; Connolly, E. ; Crew, K. D. ; Maurer, M. A. ; Hershman, D. L. / Proteomic modulation in breast tumors after metformin exposure : results from a “window of opportunity” trial. In: Clinical and Translational Oncology. 2017 ; Vol. 19, No. 2. pp. 180-188.
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abstract = "Purpose: Reverse Phase Protein Array (RPPA) is a high-throughput antibody-based technique to assess cellular protein activity. The goal of this study was to assess protein marker changes by RPPA in tumor tissue from a pre-surgical metformin trial in women with operable breast cancer (BC). Methods: In an open-label trial, metformin 1500-mg PO daily was administered prior to resection in 35 non-diabetic patients with stage 0–III BC, body mass index ≥25 kg/m2. For RPPA, formalin-fixed paraffin-embedded (FFPE) samples were probed with 160 antibodies. Paired and two-sample t-tests were performed (p ≤ 0.05). Multiple comparisons were adjusted for by fixing the false discovery rate at 25 {\%}. We evaluated whether pre- and post-metformin changes of select markers by RPPA were identified by immunohistochemistry (IHC) in these samples. We also assessed for these changes by western blot in metformin-treated BC cell lines. Results: After adjusting for multiple comparisons in the 32 tumors from metformin-treated patients vs. 34 untreated historical controls, 11 proteins were significantly different between cases vs. controls: increases in Raptor, C-Raf, Cyclin B1, Cyclin D1, TRFC, and Syk; and reductions in pMAPKpT202,Y204, JNKpT183,pT185, BadpS112, PKC.alphapS657, and SrcpY416. Cyclin D1 change after metformin by IHC was not observed. In cell lines, reductions in JNKpT183 and BadpS112 were seen, with no change in Cyclin D1 or Raptor. Conclusions: These results suggest that metformin modulates apoptosis/cell cycle, cell signaling, and invasion/motility. These findings should be assessed in larger metformin trials. If confirmed, associations between these changes and BC clinical outcome should be evaluated. Clinicaltrials.gov identifier: NCT00930579.",
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AU - Du, X.

AU - Mundi, P.

AU - Yang, J.

AU - Refice, S.

AU - Feldman, Sheldon M.

AU - Taback, B.

AU - Connolly, E.

AU - Crew, K. D.

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N2 - Purpose: Reverse Phase Protein Array (RPPA) is a high-throughput antibody-based technique to assess cellular protein activity. The goal of this study was to assess protein marker changes by RPPA in tumor tissue from a pre-surgical metformin trial in women with operable breast cancer (BC). Methods: In an open-label trial, metformin 1500-mg PO daily was administered prior to resection in 35 non-diabetic patients with stage 0–III BC, body mass index ≥25 kg/m2. For RPPA, formalin-fixed paraffin-embedded (FFPE) samples were probed with 160 antibodies. Paired and two-sample t-tests were performed (p ≤ 0.05). Multiple comparisons were adjusted for by fixing the false discovery rate at 25 %. We evaluated whether pre- and post-metformin changes of select markers by RPPA were identified by immunohistochemistry (IHC) in these samples. We also assessed for these changes by western blot in metformin-treated BC cell lines. Results: After adjusting for multiple comparisons in the 32 tumors from metformin-treated patients vs. 34 untreated historical controls, 11 proteins were significantly different between cases vs. controls: increases in Raptor, C-Raf, Cyclin B1, Cyclin D1, TRFC, and Syk; and reductions in pMAPKpT202,Y204, JNKpT183,pT185, BadpS112, PKC.alphapS657, and SrcpY416. Cyclin D1 change after metformin by IHC was not observed. In cell lines, reductions in JNKpT183 and BadpS112 were seen, with no change in Cyclin D1 or Raptor. Conclusions: These results suggest that metformin modulates apoptosis/cell cycle, cell signaling, and invasion/motility. These findings should be assessed in larger metformin trials. If confirmed, associations between these changes and BC clinical outcome should be evaluated. Clinicaltrials.gov identifier: NCT00930579.

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