WDR5 supports colon cancer cells by promoting methylation of H3K4 and suppressing DNA damage

Beth K. Neilsen, Binita Chakraborty, Jamie L. McCall, Danielle E. Frodyma, Richard L. Sleightholm, Kurt W. Fisher, Robert E Lewis

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background: KMT2/MLL proteins are commonly overexpressed or mutated in cancer and have been shown to support cancer maintenance. These proteins are responsible for methylating histone 3 at lysine 4 and promoting transcription and DNA synthesis; however, they are inactive outside of a multi-protein complex that requires WDR5. WDR5 has been implicated in cancer for its role in the COMPASS complex and its interaction with Myc; however, the role of WDR5 in colon cancer has not yet been elucidated. Methods: WDR5 expression was evaluated using RT-qPCR and western blot analysis. Cell viability and colony forming assays were utilized to evaluate the effects of WDR5 depletion or inhibition in colon cancer cells. Downstream effects of WDR5 depletion and inhibition were observed by western blot. Results: WDR5 is overexpressed in colon tumors and colon cancer cell lines at the mRNA and protein level. WDR5 depletion reduces cell viability in HCT116, LoVo, RKO, HCT15, SW480, SW620, and T84 colon cancer cells. Inhibition of the WDR5:KMT2/MLL interaction using OICR-9429 reduces cell viability in the same panel of cell lines albeit not to the same extent as RNAi-mediated WDR5 depletion. WDR5 depletion reduced H3K4Me3 and increased phosphorylation of H2AX in HCT116, SW620, and RKO colon cancer cells; however, OICR-9429 treatment did not recapitulate these effects in all cell lines potentially explaining the reduced toxicity of OICR-9429 treatment as compared to WDR5 depletion. WDR5 depletion also sensitized colon cancer cells to radiation-induced DNA damage. Conclusions: These data demonstrate a clear role for WDR5 in colon cancer and future studies should examine its potential to serve as a therapeutic target in cancer. Additional studies are needed to fully elucidate if the requirement for WDR5 is independent of or consistent with its role within the COMPASS complex. OICR-9429 treatment was particularly toxic to SW620 and T84 colon cancer cells, two cell lines without mutations in WDR5 and KMT2/MLL proteins suggesting COMPASS complex inhibition may be particularly effective in tumors lacking KMT2 mutations. Additionally, the ability of WDR5 depletion to amplify the toxic effects of radiation presents the possibility of targeting WDR5 to sensitize cells to DNA-damaging therapies.

Original languageEnglish (US)
Article number673
JournalBMC Cancer
Volume18
Issue number1
DOIs
StatePublished - Jun 20 2018

Fingerprint

Colonic Neoplasms
Methylation
DNA Damage
Cell Line
Neoplasms
Cell Survival
Poisons
Proteins
Western Blotting
Mutation
Radiation Effects
Therapeutics
RNA Interference
Genetic Therapy
Histones
Lysine
Colon
Maintenance
Phosphorylation
Radiation

Keywords

  • Colon cancer
  • H3K4Me3
  • OICR-9429
  • WDR5
  • γH2AX

ASJC Scopus subject areas

  • Oncology
  • Genetics
  • Cancer Research

Cite this

Neilsen, B. K., Chakraborty, B., McCall, J. L., Frodyma, D. E., Sleightholm, R. L., Fisher, K. W., & Lewis, R. E. (2018). WDR5 supports colon cancer cells by promoting methylation of H3K4 and suppressing DNA damage. BMC Cancer, 18(1), [673]. https://doi.org/10.1186/s12885-018-4580-6

WDR5 supports colon cancer cells by promoting methylation of H3K4 and suppressing DNA damage. / Neilsen, Beth K.; Chakraborty, Binita; McCall, Jamie L.; Frodyma, Danielle E.; Sleightholm, Richard L.; Fisher, Kurt W.; Lewis, Robert E.

In: BMC Cancer, Vol. 18, No. 1, 673, 20.06.2018.

Research output: Contribution to journalArticle

Neilsen, BK, Chakraborty, B, McCall, JL, Frodyma, DE, Sleightholm, RL, Fisher, KW & Lewis, RE 2018, 'WDR5 supports colon cancer cells by promoting methylation of H3K4 and suppressing DNA damage', BMC Cancer, vol. 18, no. 1, 673. https://doi.org/10.1186/s12885-018-4580-6
Neilsen BK, Chakraborty B, McCall JL, Frodyma DE, Sleightholm RL, Fisher KW et al. WDR5 supports colon cancer cells by promoting methylation of H3K4 and suppressing DNA damage. BMC Cancer. 2018 Jun 20;18(1). 673. https://doi.org/10.1186/s12885-018-4580-6
Neilsen, Beth K. ; Chakraborty, Binita ; McCall, Jamie L. ; Frodyma, Danielle E. ; Sleightholm, Richard L. ; Fisher, Kurt W. ; Lewis, Robert E. / WDR5 supports colon cancer cells by promoting methylation of H3K4 and suppressing DNA damage. In: BMC Cancer. 2018 ; Vol. 18, No. 1.
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AU - Chakraborty, Binita

AU - McCall, Jamie L.

AU - Frodyma, Danielle E.

AU - Sleightholm, Richard L.

AU - Fisher, Kurt W.

AU - Lewis, Robert E

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N2 - Background: KMT2/MLL proteins are commonly overexpressed or mutated in cancer and have been shown to support cancer maintenance. These proteins are responsible for methylating histone 3 at lysine 4 and promoting transcription and DNA synthesis; however, they are inactive outside of a multi-protein complex that requires WDR5. WDR5 has been implicated in cancer for its role in the COMPASS complex and its interaction with Myc; however, the role of WDR5 in colon cancer has not yet been elucidated. Methods: WDR5 expression was evaluated using RT-qPCR and western blot analysis. Cell viability and colony forming assays were utilized to evaluate the effects of WDR5 depletion or inhibition in colon cancer cells. Downstream effects of WDR5 depletion and inhibition were observed by western blot. Results: WDR5 is overexpressed in colon tumors and colon cancer cell lines at the mRNA and protein level. WDR5 depletion reduces cell viability in HCT116, LoVo, RKO, HCT15, SW480, SW620, and T84 colon cancer cells. Inhibition of the WDR5:KMT2/MLL interaction using OICR-9429 reduces cell viability in the same panel of cell lines albeit not to the same extent as RNAi-mediated WDR5 depletion. WDR5 depletion reduced H3K4Me3 and increased phosphorylation of H2AX in HCT116, SW620, and RKO colon cancer cells; however, OICR-9429 treatment did not recapitulate these effects in all cell lines potentially explaining the reduced toxicity of OICR-9429 treatment as compared to WDR5 depletion. WDR5 depletion also sensitized colon cancer cells to radiation-induced DNA damage. Conclusions: These data demonstrate a clear role for WDR5 in colon cancer and future studies should examine its potential to serve as a therapeutic target in cancer. Additional studies are needed to fully elucidate if the requirement for WDR5 is independent of or consistent with its role within the COMPASS complex. OICR-9429 treatment was particularly toxic to SW620 and T84 colon cancer cells, two cell lines without mutations in WDR5 and KMT2/MLL proteins suggesting COMPASS complex inhibition may be particularly effective in tumors lacking KMT2 mutations. Additionally, the ability of WDR5 depletion to amplify the toxic effects of radiation presents the possibility of targeting WDR5 to sensitize cells to DNA-damaging therapies.

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KW - H3K4Me3

KW - OICR-9429

KW - WDR5

KW - γH2AX

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