Transcriptome analysis using patient iPSC-derived skeletal myocytes: Bet1L as a new molecule possibly linked to neuromuscular junction degeneration in ALS

Exp Neurol. 2021 Nov:345:113815. doi: 10.1016/j.expneurol.2021.113815. Epub 2021 Jul 24.

Abstract

Amyotrophic lateral sclerosis (ALS) is a fatal neuromuscular disease in which patients gradually become paralyzed due to loss of motor function. Many genetically inheritable mutations have been linked to ALS; however, the majority of ALS patients are considered sporadic. Therefore, there is a need for a common therapy that is effective for all ALS patients. Although there is evidence of the disease beginning in the periphery at the neuromuscular junction (NMJ), the specific processes involved in skeletal muscle and at the NMJ are still largely unknown. To study common disease mechanisms in ALS skeletal muscle, we performed RNA sequencing of skeletal myocytes differentiated from induced pluripotent stem cells (iPSCs) derived from familial ALS (with C9ORF72, SOD1, or TARDBP mutations) and sporadic ALS patients. Compared to healthy control lines, the myocytes from all ALS lines showed downregulation of four genes: BET1L, DCX, GPC3, and HNRNPK. We next measured the expression levels of these four genes in hind limb muscle samples from a rat model of familial ALS (SOD1G93A transgenic) and found that only the Bet1L gene, which encodes Bet1 Golgi Vesicular Membrane Trafficking Protein Like, was commonly downregulated. Bet1L protein appeared to be localized to the basal lamina of the NMJ, with decreased expression over time in SOD1G93A transgenic rats. Importantly, the expression levels began to decrease early in the disease process. Our results indicate that loss of Bet1L at the NMJ could be of interest for better understanding ALS disease progression.

Keywords: Amyotrophic lateral sclerosis (ALS); Bet1L; Familial; Induced pluripotent stem cells; Neuromuscular junction; SOD1(G93A) transgenic rat; Sporadic.

Publication types

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

MeSH terms

  • Adult
  • Amyotrophic Lateral Sclerosis / genetics
  • Amyotrophic Lateral Sclerosis / metabolism*
  • Amyotrophic Lateral Sclerosis / pathology
  • Animals
  • Cell Differentiation / physiology
  • Female
  • Gene Expression Profiling / methods*
  • Humans
  • Induced Pluripotent Stem Cells / metabolism*
  • Male
  • Middle Aged
  • Muscle Fibers, Skeletal / metabolism*
  • Muscle Fibers, Skeletal / pathology
  • Muscle, Skeletal / metabolism
  • Muscle, Skeletal / pathology
  • Neuromuscular Junction / metabolism*
  • Neuromuscular Junction / pathology
  • Qc-SNARE Proteins / deficiency*
  • Qc-SNARE Proteins / genetics
  • Rats
  • Rats, Sprague-Dawley
  • Rats, Transgenic
  • Sequence Analysis, RNA / methods

Substances

  • BET1L protein, human
  • Qc-SNARE Proteins