Macroautophagy is dispensable for growth of KRAS mutant tumors and chloroquine efficacy

Proc Natl Acad Sci U S A. 2016 Jan 5;113(1):182-7. doi: 10.1073/pnas.1515617113. Epub 2015 Dec 17.

Abstract

Macroautophagy is a key stress-response pathway that can suppress or promote tumorigenesis depending on the cellular context. Notably, Kirsten rat sarcoma (KRAS)-driven tumors have been reported to rely on macroautophagy for growth and survival, suggesting a potential therapeutic approach of using autophagy inhibitors based on genetic stratification. In this study, we evaluated whether KRAS mutation status can predict the efficacy to macroautophagy inhibition. By profiling 47 cell lines with pharmacological and genetic loss-of-function tools, we were unable to confirm that KRAS-driven tumor lines require macroautophagy for growth. Deletion of autophagy-related 7 (ATG7) by genome editing completely blocked macroautophagy in several tumor lines with oncogenic mutations in KRAS but did not inhibit cell proliferation in vitro or tumorigenesis in vivo. Furthermore, ATG7 knockout did not sensitize cells to irradiation or to several anticancer agents tested. Interestingly, ATG7-deficient and -proficient cells were equally sensitive to the antiproliferative effect of chloroquine, a lysosomotropic agent often used as a pharmacological tool to evaluate the response to macroautophagy inhibition. Moreover, both cell types manifested synergistic growth inhibition when treated with chloroquine plus the tyrosine kinase inhibitors erlotinib or sunitinib, suggesting that the antiproliferative effects of chloroquine are independent of its suppressive actions on autophagy.

Keywords: ATG7; KRAS; autophagy; cancer; chloroquine.

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Autophagy / drug effects*
  • Autophagy / genetics
  • Autophagy-Related Protein 7
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Transformation, Neoplastic / drug effects*
  • Cell Transformation, Neoplastic / genetics
  • Cell Transformation, Neoplastic / pathology
  • Chloroquine / pharmacology*
  • Drug Resistance, Neoplasm / genetics*
  • Erlotinib Hydrochloride / pharmacology
  • Gene Knockout Techniques
  • Humans
  • Indoles / pharmacology
  • Mutation
  • Protein Kinase Inhibitors / pharmacology
  • Proto-Oncogene Proteins p21(ras) / genetics*
  • Pyrroles / pharmacology
  • Radiation Tolerance / genetics
  • Sunitinib
  • Ubiquitin-Activating Enzymes / genetics

Substances

  • Antineoplastic Agents
  • Indoles
  • KRAS protein, human
  • Protein Kinase Inhibitors
  • Pyrroles
  • Chloroquine
  • Erlotinib Hydrochloride
  • Proto-Oncogene Proteins p21(ras)
  • ATG7 protein, human
  • Autophagy-Related Protein 7
  • Ubiquitin-Activating Enzymes
  • Sunitinib