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Antioxidant functions of DHHC3 suppress anti-cancer drug activities

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Abstract

Ablation of protein acyltransferase DHHC3 selectively enhanced the anti-cancer cell activities of several chemotherapeutic agents, but not kinase inhibitors. To understand why this occurs, we used comparative mass spectrometry-based palmitoyl-proteomic analysis of breast and prostate cancer cell lines, ± DHHC3 ablation, to obtain the first comprehensive lists of candidate protein substrates palmitoylated by DHHC3. Putative substrates included 22–28 antioxidant/redox-regulatory proteins, thus predicting that DHHC3 should have antioxidant functions. Consistent with this, DHHC3 ablation elevated oxidative stress. Furthermore, DHHC3 ablation, together with chemotherapeutic drug treatment, (a) elevated oxidative stress, with a greater than additive effect, and (b) enhanced the anti-growth effects of the chemotherapeutic agents. These results suggest that DHHC3 ablation enhances chemotherapeutic drug potency by disabling the antioxidant protections that contribute to drug resistance. Affirming this concept, DHHC3 ablation synergized with another anti-cancer drug, PARP inhibitor PJ-34, to decrease cell proliferation and increase oxidative stress. Hence, DHHC3 targeting can be a useful strategy for selectively enhancing potency of oxidative stress-inducing anti-cancer drugs. Also, comprehensive identification of DHHC3 substrates provides insight into other DHHC3 functions, relevant to in vivo tumor growth modulation.

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Availability of data and materials

Datasets generated and analyzed during the current study will be available as supplemental tables, and data with additional details will be available in the appropriate repository.

Abbreviations

DHHC3:

Enzyme #3 in a family containing Asp–His–His–Cys motif

NPC1:

NPC intracellular cholesterol transporter 1

PalmPISC:

Palmitoyl protein identification and site characterization

PARP:

Poly (ADP-ribose) polymerase

PRDX4:

Peroxiredoxin 4

SILAC:

Stable isotope labeling by amino acids in cell culture

TMEM192:

Transmembrane protein 192

TMX1 and TMX3:

Thioredoxin related transmembrane proteins 1 and 3

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Funding

This study was supported by NIH grant CA42368 to M Hemler and C Sharma, DoD grant W81XWH-11-1-0112 to M Hemler, and DoD grant W81XWH-11-1-0113 to M Freeman. The funding body played no role in the design of the study, collection, analysis, and interpretation of data, or in writing the manuscript.

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Authors and Affiliations

  1. Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA

    Chandan Sharma & Martin E. Hemler

  2. Division of Cancer Biology and Therapeutics, Departments of Surgery and Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA, 90048, USA

    Wei Yang & Michael R. Freeman

  3. Department of Pathology and Precision Vaccines Program, Boston Children’s Hospital, Boston, MA, 02215, USA

    Hanno Steen

  4. Dana-Farber Cancer Institute, Rm SM-520, 450 Brookline Ave, Boston, MA, 02215, USA

    Chandan Sharma

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  1. Chandan Sharma
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  2. Wei Yang
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  3. Hanno Steen
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  4. Michael R. Freeman
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  5. Martin E. Hemler
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Contributions

WY and HS carried out mass spectrometry experiments, assisted by CS. CS carried out all other experiments. MF helped to direct the project. CS prepared figures and tables and wrote the manuscript, with assistance from MEH.

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Correspondence to Chandan Sharma or Martin E. Hemler.

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Sharma, C., Yang, W., Steen, H. et al. Antioxidant functions of DHHC3 suppress anti-cancer drug activities. Cell. Mol. Life Sci. 78, 2341–2353 (2021). https://doi.org/10.1007/s00018-020-03635-3

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  • DOI: https://doi.org/10.1007/s00018-020-03635-3

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