DYT1 Dystonia Patient-Derived Fibroblasts Have Increased Deformability and Susceptibility to Damage by Mechanical Forces

Navjot Kaur Gill; Chau Ly; Paul H Kim; Cosmo A Saunders; Loren G Fong; Stephen G Young; G W Gant Luxton; Amy C Rowat

doi:10.3389/fcell.2019.00103

DYT1 Dystonia Patient-Derived Fibroblasts Have Increased Deformability and Susceptibility to Damage by Mechanical Forces

Front Cell Dev Biol. 2019 Jun 26:7:103. doi: 10.3389/fcell.2019.00103. eCollection 2019.

Authors

Navjot Kaur Gill¹, Chau Ly^{1

2}, Paul H Kim³, Cosmo A Saunders⁴, Loren G Fong³, Stephen G Young^{3

5

6}, G W Gant Luxton^{4

7}, Amy C Rowat¹

Affiliations

¹ Department of Integrative Biology and Physiology, University of California, Los Angeles, Los Angeles, CA, United States.
² Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA, United States.
³ Department of Medicine, University of California, Los Angeles, Los Angeles, CA, United States.
⁴ Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN, United States.
⁵ Department of Human Genetics, University of California, Los Angeles, Los Angeles, CA, United States.
⁶ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, United States.
⁷ Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN, United States.

Abstract

DYT1 dystonia is a neurological movement disorder that is caused by a loss-of-function mutation in the DYT1/TOR1A gene, which encodes torsinA, a conserved luminal ATPases-associated with various cellular activities (AAA+) protein. TorsinA is required for the assembly of functional linker of nucleoskeleton and cytoskeleton (LINC) complexes, and consequently the mechanical integration of the nucleus and the cytoskeleton. Despite the potential implications of altered mechanobiology in dystonia pathogenesis, the role of torsinA in regulating cellular mechanical phenotype, or mechanotype, in DYT1 dystonia remains unknown. Here, we define the deformability of mouse fibroblasts lacking functional torsinA as well as human fibroblasts isolated from DYT1 dystonia patients. We find that the deletion of torsinA or the expression of torsinA containing the DYT1 dystonia-causing ΔE302/303 (ΔE) mutation results in more deformable cells. We observe a similar increased deformability of mouse fibroblasts that lack lamina-associated polypeptide 1 (LAP1), which interacts with and stimulates the ATPase activity of torsinA in vitro, as well as with the absence of the LINC complex proteins, Sad1/UNC-84 1 (SUN1) and SUN2, lamin A/C, or lamin B1. Consistent with these findings, we also determine that DYT1 dystonia patient-derived fibroblasts are more compliant than fibroblasts isolated from unafflicted individuals. DYT1 dystonia patient-derived fibroblasts also exhibit increased nuclear strain and decreased viability following mechanical stretch. Taken together, our results establish the foundation for future mechanistic studies of the role of cellular mechanotype and LINC-dependent nuclear-cytoskeletal coupling in regulating cell survival following exposure to mechanical stresses.

Keywords: LINC complex; cell mechanical properties; lamins; mechanotype; nuclear envelope; torsinA.

Abstract

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