Mutations in GFAP Disrupt the Distribution and Function of Organelles in Human Astrocytes

Cell Rep. 2018 Oct 23;25(4):947-958.e4. doi: 10.1016/j.celrep.2018.09.083.

Abstract

How mutations in glial fibrillary acidic protein (GFAP) cause Alexander disease (AxD) remains elusive. We generated iPSCs from two AxD patients and corrected the GFAP mutations to examine the effects of mutant GFAP on human astrocytes. AxD astrocytes displayed GFAP aggregates, recapitulating the pathological hallmark of AxD. RNA sequencing implicated the endoplasmic reticulum, vesicle regulation, and cellular metabolism. Corroborating this analysis, we observed enlarged and heterogeneous morphology coupled with perinuclear localization of endoplasmic reticulum and lysosomes in AxD astrocytes. Functionally, AxD astrocytes showed impaired extracellular ATP release, which is responsible for attenuated calcium wave propagation. These results reveal that AxD-causing mutations in GFAP disrupt intracellular vesicle regulation and impair astrocyte secretion, resulting in astrocyte dysfunction and AxD pathogenesis.

Keywords: Alexander disease; CRISPR; endoplasmic reticulum; iPSC; lysosome.

Publication types

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

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Alexander Disease / metabolism
  • Alexander Disease / pathology
  • Animals
  • Astrocytes / cytology
  • Astrocytes / metabolism*
  • Calcium Signaling
  • Cell Differentiation
  • Endoplasmic Reticulum / metabolism
  • Glial Fibrillary Acidic Protein / genetics*
  • Humans
  • Lysosomes / metabolism
  • Mice
  • Mutation / genetics*
  • Organelles / metabolism*
  • Protein Aggregates
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism

Substances

  • Glial Fibrillary Acidic Protein
  • Protein Aggregates
  • RNA, Messenger
  • Adenosine Triphosphate