Purkinje cells derived from TSC patients display hypoexcitability and synaptic deficits associated with reduced FMRP levels and reversed by rapamycin

Mol Psychiatry. 2018 Nov;23(11):2167-2183. doi: 10.1038/s41380-018-0018-4. Epub 2018 Feb 15.

Abstract

Accumulating evidence suggests that cerebellar dysfunction early in life is associated with autism spectrum disorder (ASD), but the molecular mechanisms underlying the cerebellar deficits at the cellular level are unclear. Tuberous sclerosis complex (TSC) is a neurocutaneous disorder that often presents with ASD. Here, we developed a cerebellar Purkinje cell (PC) model of TSC with patient-derived human induced pluripotent stem cells (hiPSCs) to characterize the molecular mechanisms underlying cerebellar abnormalities in ASD and TSC. Our results show that hiPSC-derived PCs from patients with pathogenic TSC2 mutations displayed mTORC1 pathway hyperactivation, defects in neuronal differentiation and RNA regulation, hypoexcitability and reduced synaptic activity when compared with those derived from controls. Our gene expression analyses revealed downregulation of several components of fragile X mental retardation protein (FMRP) targets in TSC2-deficient hiPSC-PCs. We detected decreased expression of FMRP, glutamate receptor δ2 (GRID2), and pre- and post-synaptic markers such as synaptophysin and PSD95 in the TSC2-deficient hiPSC-PCs. The mTOR inhibitor rapamycin rescued the deficits in differentiation, synaptic dysfunction, and hypoexcitability of TSC2 mutant hiPSC-PCs in vitro. Our findings suggest that these gene expression changes and cellular abnormalities contribute to aberrant PC function during development in TSC affected individuals.

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

  • Adult
  • Autism Spectrum Disorder / complications
  • Autism Spectrum Disorder / metabolism
  • Cerebellar Diseases / metabolism
  • Cerebellum / metabolism
  • Child
  • Child, Preschool
  • Female
  • Fragile X Mental Retardation Protein / drug effects
  • Fragile X Mental Retardation Protein / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / metabolism
  • Mechanistic Target of Rapamycin Complex 1 / genetics
  • Models, Biological
  • Purkinje Cells / metabolism*
  • Purkinje Cells / pathology
  • Sirolimus / pharmacology
  • Synapses / metabolism
  • Synapses / physiology
  • TOR Serine-Threonine Kinases / metabolism
  • Tuberous Sclerosis / metabolism*
  • Tuberous Sclerosis / physiopathology
  • Tuberous Sclerosis Complex 1 Protein
  • Tuberous Sclerosis Complex 2 Protein
  • Tumor Suppressor Proteins / genetics

Substances

  • FMR1 protein, human
  • Tuberous Sclerosis Complex 1 Protein
  • Tuberous Sclerosis Complex 2 Protein
  • Tumor Suppressor Proteins
  • Fragile X Mental Retardation Protein
  • Mechanistic Target of Rapamycin Complex 1
  • TOR Serine-Threonine Kinases
  • Sirolimus