Phenotypic Screen with TSC-Deficient Neurons Reveals Heat-Shock Machinery as a Druggable Pathway for mTORC1 and Reduced Cilia

Cell Rep. 2020 Jun 23;31(12):107780. doi: 10.1016/j.celrep.2020.107780.

Abstract

Tuberous sclerosis complex (TSC) is a neurogenetic disorder that leads to elevated mechanistic targeting of rapamycin complex 1 (mTORC1) activity. Cilia can be affected by mTORC1 signaling, and ciliary deficits are associated with neurodevelopmental disorders. Here, we examine whether neuronal cilia are affected in TSC. We show that cortical tubers from TSC patients and mutant mouse brains have fewer cilia. Using high-content image-based assays, we demonstrate that mTORC1 activity inversely correlates with ciliation in TSC1/2-deficient neurons. To investigate the mechanistic relationship between mTORC1 and cilia, we perform a phenotypic screen for mTORC1 inhibitors with TSC1/2-deficient neurons. We identify inhibitors of the heat shock protein 90 (Hsp90) that suppress mTORC1 through regulation of phosphatidylinositol 3-kinase (PI3K)/Akt signaling. Pharmacological inhibition of Hsp90 rescues ciliation through downregulation of Hsp27. Our study uncovers the heat-shock machinery as a druggable signaling node to restore mTORC1 activity and cilia due to loss of TSC1/2, and it provides broadly applicable platforms for studying TSC-related neuronal dysfunction.

Keywords: 17-AGG; Hsp27; Hsp90; TSC; autism; brain; cilia; ciliopathy; mTOR.

Publication types

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

MeSH terms

  • Aging / metabolism
  • Animals
  • Benzoquinones / pharmacology
  • Brain / pathology
  • Cilia / metabolism*
  • Down-Regulation / drug effects
  • HSP27 Heat-Shock Proteins / metabolism
  • HSP90 Heat-Shock Proteins / metabolism
  • Heat-Shock Response* / drug effects
  • Humans
  • Lactams, Macrocyclic / pharmacology
  • Mechanistic Target of Rapamycin Complex 1 / metabolism*
  • Mice, Knockout
  • Neurons / drug effects
  • Neurons / metabolism*
  • Phenotype
  • Phosphatidylinositol 3-Kinases / metabolism
  • Proto-Oncogene Proteins c-akt / metabolism
  • Rats
  • Sirolimus / pharmacology
  • Time Factors
  • Tuberous Sclerosis Complex 1 Protein / metabolism*
  • Tuberous Sclerosis Complex 2 Protein / metabolism*
  • Up-Regulation / drug effects

Substances

  • Benzoquinones
  • HSP27 Heat-Shock Proteins
  • HSP90 Heat-Shock Proteins
  • Lactams, Macrocyclic
  • Tuberous Sclerosis Complex 1 Protein
  • Tuberous Sclerosis Complex 2 Protein
  • tanespimycin
  • Mechanistic Target of Rapamycin Complex 1
  • Proto-Oncogene Proteins c-akt
  • Sirolimus