Article | Published:

Translational Therapeutics

Bcl-2-dependent synthetic lethal interaction of the IDF-11774 with the V0 subunit C of vacuolar ATPase (ATP6V0C) in colorectal cancer

British Journal of Cancervolume 119pages13471357 (2018) | Download Citation

Background

The IDF-11774, a novel clinical candidate for cancer therapy, targets HSP70 and inhibits mitochondrial respiration, resulting in the activation of AMPK and reduction in HIF-1α accumulation.

Methods

To identify genes that have synthetic lethality to IDF-11774, RNA interference screening was conducted, using pooled lentiviruses expressing a short hairpin RNA library.

Results

We identified ATP6V0C, encoding the V0 subunit C of lysosomal V-ATPase, knockdown of which induced a synergistic growth-inhibitory effect in HCT116 cells in the presence of IDF-11774. The synthetic lethality of IDF-11774 with ATP6V0C possibly correlates with IDF-11774-mediated autolysosome formation. Notably, the synergistic effect of IDF-11774 and the ATP6V0C inhibitor, bafilomycin A1, depended on the PIK3CA genetic status and Bcl-2 expression, which regulates autolysosome formation and apoptosis. Similarly, in an experiment using conditionally reprogramed cells derived from colorectal cancer patients, synergistic growth inhibition was observed in cells with low Bcl-2 expression.

Conclusions

Bcl-2 is a biomarker for the synthetic lethal interaction of IDF-11774 with ATP6V0C, which is clinically applicable for the treatment of cancer patients with IDF-11774 or autophagy-inducing anti-cancer drugs.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Leung, A. W., de Silva, T., Bally, M. B. & Lockwood, W. W. Synthetic lethality in lung cancer and translation to clinical therapies. Mol. Cancer 15, 61 (2016).

  2. 2.

    Srivas, R. et al. A network of conserved synthetic lethal interactions for exploration of precision cancer therapy. Mol. Cell 63, 514–525 (2016).

  3. 3.

    Diehl, P., Tedesco, D. & Chenchik, A. Use of RNAi screens to uncover resistance mechanisms in cancer cells and identify synthetic lethal interactions. Drug Discov. Today Technol. 11, 11–18 (2014).

  4. 4.

    Baratta, M. G. et al. An in-tumor genetic screen reveals that the BET bromodomain protein, BRD4, is a potential therapeutic target in ovarian carcinoma. Proc. Natl Acad. Sci. USA 112, 232–237 (2015).

  5. 5.

    Schonherr, M. et al. Genome-wide RNAi screen identifies protein kinase Cb and new members of mitogen-activated protein kinase pathway as regulators of melanoma cell growth and metastasis. Pigment. Cell. Melanoma Res. 27, 418–430 (2014).

  6. 6.

    Prahallad, A. et al. Unresponsiveness of colon cancer to BRAF(V600E) inhibition through feedback activation of EGFR. Nature 483, 100–103 (2012).

  7. 7.

    Liu-Sullivan, N. et al. Pooled shRNA screen for sensitizers to inhibition of the mitotic regulator polo-like kinase (PLK1). Oncotarget 2, 1254–1264 (2011).

  8. 8.

    Khorashad, J. S. et al. shRNA library screening identifies nucleocytoplasmic transport as a mediator of BCR-ABL1 kinase-independent resistance. Blood 125, 1772–1781 (2015).

  9. 9.

    Mendes-Pereira, A. M. et al. Genome-wide functional screen identifies a compendium of genes affecting sensitivity to tamoxifen. Proc. Natl Acad. Sci. USA 109, 2730–2735 (2012).

  10. 10.

    Dai, B. et al. KEAP1-dependent synthetic lethality induced by AKT and TXNRD1 inhibitors in lung cancer. Cancer Res. 73, 5532–5543 (2013).

  11. 11.

    Olsen, M. H., Nielsen, H., Dalton, S. O. & Johansen, C. Cancer incidence and mortality among members of the Danish resistance movement deported to German concentration camps: 65-Year follow-up. Int. J. Cancer 136, 2476–2480 (2015).

  12. 12.

    Siegel, R., Desantis, C. & Jemal, A. Colorectal cancer statistics, 2014. CA Cancer J. Clin. 64, 104–117 (2014).

  13. 13.

    Cancer Genome Atlas N. Comprehensive molecular characterization of human colon and rectal cancer. Nature 487, 330–337 (2012).

  14. 14.

    Liao, X. et al. Prognostic role of PIK3CA mutation in colorectal cancer: cohort study and literature review. Clin. Cancer Res. 18, 2257–2268 (2012).

  15. 15.

    Cartwright, T. H. Treatment decisions after diagnosis of metastatic colorectal cancer. Clin. Colorectal Cancer 11, 155–166 (2012).

  16. 16.

    Goldberg, R. M. Therapy for metastatic colorectal cancer. Oncologist 11, 981–987 (2006).

  17. 17.

    Ban, H. S. et al. The novel hypoxia-inducible factor-1alpha inhibitor IDF-11774 regulates cancer metabolism, thereby suppressing tumor growth. Cell Death Dis. 8, e2843 (2017).

  18. 18.

    Ban, H. S. et al. Identification of targets of the HIF-1 inhibitor IDF-11774 using alkyne-conjugated photoaffinity probes. Bioconjug. Chem. 27, 1911–1920 (2016).

  19. 19.

    Kim, B. K. et al. p300 cooperates with c-Jun and PARP-1 at the p300 binding site to activate RhoB transcription in NSC126188-mediated apoptosis. Biochim. Biophys. Acta 1839, 364–p373 (2014).

  20. 20.

    Palechor-Ceron, N. et al. Radiation induces diffusible feeder cell factor(s) that cooperate with ROCK inhibitor to conditionally reprogram and immortalize epithelial cells. Am. J. Pathol. 183, 1862–1870 (2013).

  21. 21.

    Liu, X. et al. ROCK inhibitor and feeder cells induce the conditional reprogramming of epithelial cells. Am. J. Pathol. 180, 599–607 (2012).

  22. 22.

    Im, J. Y. et al. DNA damage induced apoptosis suppressor (DDIAS) is upregulated via ERK5/MEF2B signaling and promotes beta-catenin-mediated invasion. Biochim. Biophys. Acta 1859, 1449–1458 (2016).

  23. 23.

    Chou, T. C. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol. Rev. 58, 621–681 (2006).

  24. 24.

    Yoshii, S. R., Mizushima, N. Monitoring and measuring autophagy. Int. J. Mol. Sci. 18, 1865 (2017).

  25. 25.

    Tsuboyama, K. et al. The ATG conjugation systems are important for degradation of the inner autophagosomal membrane. Science 354, 1036–1041 (2016).

  26. 26.

    Im, J. Y. et al. DNA damage-induced apoptosis suppressor (DDIAS), a novel target of NFATc1, is associated with cisplatin resistance in lung cancer. Biochim. Biophys. Acta 1863, 40–49 (2016).

  27. 27.

    Perez-Sayans, M., Garcia-Garcia, A., Reboiras-Lopez, M. D. & Gandara-Vila, P. Role of V-ATPases in solid tumors: importance of the subunit C (Review). Int. J. Oncol. 34, 1513–1520 (2009).

  28. 28.

    Mangieri, L. R. et al. ATP6V0C knockdown in neuroblastoma cells alters autophagy-lysosome pathway function and metabolism of proteins that accumulate in neurodegenerative disease. PLoS. ONE 9, e93257 (2014).

  29. 29.

    Wu, Y. T. et al. Dual role of 3-methyladenine in modulation of autophagy via different temporal patterns of inhibition on class I and III phosphoinositide 3-kinase. J. Biol. Chem. 285, 10850–10861 (2010).

  30. 30.

    Kimura, S., Noda, T. & Yoshimori, T. Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3. Autophagy 3, 452–460 (2007).

  31. 31.

    Ragazzoni, Y. et al. The thiazole derivative CPTH6 impairs autophagy. Cell Death Dis. 4, e524 (2013).

  32. 32.

    Corcelle, E. et al. Control of the autophagy maturation step by the MAPK ERK andp38: lessons from environmental carcinogens. Autophagy 3, 57–p39 (2007).

  33. 33.

    Marino, G., Niso-Santano, M., Baehrecke, E. H. & Kroemer, G. Self-consumption: the interplay of autophagy and apoptosis. Nat. Rev. Mol. Cell Biol. 15, 81–94 (2014).

  34. 34.

    Liu, E. Y. & Ryan, K. M. Autophagy and cancer--issues we need to digest. J. Cell. Sci. 125(Pt 10), 2349–2358 (2012).

  35. 35.

    Decuypere, J. P., Parys, J. B. & Bultynck, G. Regulation of the autophagic bcl-2/beclin 1 interaction. Cells 1, 284–312 (2012).

  36. 36.

    Yang, Z. J., Chee, C. E., Huang, S. & Sinicrope, F. A. The role of autophagy in cancer: therapeutic implications. Mol. Cancer Ther. 10, 1533–1541 (2011).

  37. 37.

    Selvakumaran, M., Amaravadi, R. K., Vasilevskaya, I. A. & O’Dwyer, P. J. Autophagy inhibition sensitizes colon cancer cells to antiangiogenic and cytotoxic therapy. Clin. Cancer Res. 19, 2995–3007 (2013).

  38. 38.

    Min, H. et al. Bortezomib induces protective autophagy through AMP-activated protein kinase activation in cultured pancreatic and colorectal cancer cells. Cancer Chemother. Pharmacol. 74, 167–76 (2014).

  39. 39.

    Amaravadi, R. K. et al. Autophagy inhibition enhances therapy-induced apoptosis in a Myc-induced model of lymphoma. J. Clin. Invest. 117, 326–336 (2007).

  40. 40.

    Fais, S., De Milito, A., You, H. & Qin, W. Targeting vacuolar H+-ATPases as a new strategy against cancer. Cancer Res. 67, 10627–10630 (2007).

  41. 41.

    Mauvezin, C. & Neufeld, T. P. Bafilomycin A1 disrupts autophagic flux by inhibiting both V-ATPase-dependent acidification and Ca-P60A/SERCA-dependent autophagosome–lysosome fusion. Autophagy 11, 1437–1438 (2015).

  42. 42.

    Xu, J. et al. Expression and functional role of vacuolar H(+)-ATPase in human hepatocellular carcinoma. Carcinogenesis 33, 2432–2440 (2012).

  43. 43.

    Lu, X. et al. The growth and metastasis of human hepatocellular carcinoma xenografts are inhibited by small interfering RNA targeting to the subunit ATP6L of proton pump. Cancer Res. 65, 6843–6849 (2005).

  44. 44.

    Wan, G., Mahajan, A., Lidke, D. & Rajput, A. Bcl-2 together with PI3K p110alpha regulates cell morphology and cell migration. Cell death Dis. 6, e2006 (2015).

  45. 45.

    Marquez, R. T. & Xu, L. Bcl-2:Beclin 1 complex: multiple, mechanisms regulating autophagy/apoptosis toggle switch. Am. J. Cancer Res. 2, 214–221 (2012).

Download references

Acknowledgements

This work was supported by the KRIBB Initiative program (KGM4751713), National Research Foundation (NRF) (NRF-2017R1A2B2011936 and NRF-2018R1A5A2023127) and Health Technology R&D (HI13C2162).

Authors contributions

B-K.K. designed and performed experiments and wrote part of manuscript. S.W.N. provided an initial idea and designed experiments. B.S.M. provided cancer patient-derived cell lines. HSB, J-Y.I. and J.P. performed experiment. K.L. synthesized IDF-11774. JL helped shRNA screening experiment. S-Y.K. and MK analyzed the hi-seq data of shRNA screening. H.L. and S.P. reviewed manuscript. M.W. advised all experiment and wrote a manuscript.

Author information

Author notes

  1. These authors contributed equally: Bo-Kyung Kim, Soon Woo Nam, Byung Soh Min

Affiliations

  1. Personalized Genomic Medicine Research Center, KRIBB, Daejeon, 34141, Korea

    • Bo-Kyung Kim
    • , Joo-Young Im
    • , Youngjoo Lee
    • , Seon-Young Kim
    • , Mirang Kim
    •  & Misun Won
  2. The Catholic University of Korea, Incheon St Mary’s Hospital, 56 Dongsuro Bupyunggu, Incheon, 06591, Korea

    • Soon Woo Nam
  3. Serverance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, 03722, Korea

    • Byung Soh Min
    •  & Soonmyung Paik
  4. Metabolic Regulation Research Center, KRIBB, Daejeon, 34141, Korea

    • Hyun Seung Ban
  5. College of Pharmacy, Dongguk University-Seoul, Goyang, 410-820, Korea

    • Kyeong Lee
  6. Drug Discovery Team, ILDONG Pharmaceutical Co. Ltd., Hwaseong, Hwaseong, 445-811, Korea

    • Joon-Tae Park
    •  & Hongsub Lee
  7. Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon, Korea, Daejeon, 34113, Korea

    • Misun Won

Authors

  1. Search for Bo-Kyung Kim in:

  2. Search for Soon Woo Nam in:

  3. Search for Byung Soh Min in:

  4. Search for Hyun Seung Ban in:

  5. Search for Soonmyung Paik in:

  6. Search for Kyeong Lee in:

  7. Search for Joo-Young Im in:

  8. Search for Youngjoo Lee in:

  9. Search for Joon-Tae Park in:

  10. Search for Seon-Young Kim in:

  11. Search for Mirang Kim in:

  12. Search for Hongsub Lee in:

  13. Search for Misun Won in:

Ethics approval

All animal experimental protocols were approved by the Bioethics Committee of the Korea Research Institute of Bioscience and Biotechnology.

Competing interests

The authors declare no competing interests.

Note

This work is published under the standard license to publish agreement. After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution 4.0 International (CC BY 4.0).

Corresponding author

Correspondence to Misun Won.

Electronic supplementary material

About this article

Publication history

Received

Revised

Accepted

Published

Issue Date

DOI

https://doi.org/10.1038/s41416-018-0289-1