Effect of early-stage autophagy inhibition in BRAF V600E autophagy-dependent brain tumor cells

Cell Death Dis. 2019 Sep 12;10(9):679. doi: 10.1038/s41419-019-1880-y.

Abstract

Autophagy is a multistage process. Progress within the field has led to the development of agents targeting both early (initiation) and late (fusion) stages of this process. The specific stage of autophagy targeted may influence cancer treatment outcomes. We have previously shown that central nervous system (CNS) tumors with the BRAFV600E mutation are autophagy dependent, and late-stage autophagy inhibition improves the response to targeted BRAF inhibitors (BRAFi) in sensitive and resistant cells. Drugs directed toward initiation of autophagy have been shown to reduce tumor cell death in some cancers, but have not been assessed in CNS tumors. We investigated early-stage inhibition for autophagy-dependent CNS tumors. BRAFi-sensitive and resistant AM38 and MAF794 cell lines were evaluated for the response to pharmacologic and genetic inhibition of ULK1 and VPS34, two crucial subunits of the autophagy initiation complexes. Changes in autophagy were monitored by western blot and flow cytometry. Survival was evaluated in short- and long-term growth assays. Tumor cells exhibited a reduced autophagic flux with pharmacologic and genetic inhibition of ULK1 or VPS34. Pharmacologic inhibition reduced cell survival in a dose-dependent manner for both targets. Genetic inhibition reduced cell survival and confirmed that it was an autophagy-specific effect. Pharmacologic and genetic inhibition were also synergistic with BRAFi, irrespective of RAFi sensitivity. Inhibition of ULK1 and VPS34 are potentially viable clinical targets in autophagy-dependent CNS tumors. Further evaluation is needed to determine if early-stage autophagy inhibition is equal to late-stage inhibition to determine the optimal clinical target for patients.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Aminopyridines / pharmacology
  • Autophagy / drug effects*
  • Autophagy / genetics
  • Benzamides / pharmacology
  • Blotting, Western
  • Brain Neoplasms / genetics
  • Brain Neoplasms / metabolism*
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cell Survival / genetics
  • Central Nervous System Neoplasms / genetics
  • Central Nervous System Neoplasms / metabolism*
  • Flow Cytometry
  • Humans
  • Mutation / genetics
  • Proto-Oncogene Proteins B-raf / genetics
  • Proto-Oncogene Proteins B-raf / metabolism*
  • Pyrimidines / pharmacology

Substances

  • 1-((2-((2-chloropyridin-4yl)amino)-4'-(cyclopropylmethyl)-(4,5'-bipyrimidin)-2'-yl)amino)-2-methylpropan-2-ol
  • Aminopyridines
  • Benzamides
  • Pyrimidines
  • SBI-0206965
  • Proto-Oncogene Proteins B-raf