Pathogenic DPAGT1 variants in limb‐girdle congenital myasthenic syndrome (LG‐CMS) associated with tubular aggregates and ORAI1 hypoglycosylation

Abstract

AbstractAimsLimb‐girdle congenital myasthenic syndrome (LG‐CMS) is a genetically heterogeneous disorder characterised by muscle weakness and fatigability. The LG‐CMS gene DPAGT1 codes for an essential enzyme of the glycosylation pathway, a posttranslational modification mechanism shaping the structure and function of proteins. In DPAGT1‐related LG‐CMS, decreased glycosylation of the acetylcholine receptor (AChR) reduces its localization at the neuromuscular junction (NMJ) and results in diminished neuromuscular transmission. LG‐CMS patients also show tubular aggregates on muscle biopsy, but the origin and potential contribution of the aggregates to disease development are not understood. Here, we describe two LG‐CMS patients with the aim of providing a molecular diagnosis and to shed light on the pathways implicated in tubular aggregate formation.MethodsFollowing clinical examination of the patients, we performed next‐generation sequencing (NGS) to identify the genetic causes, analysed the biopsies at the histological and ultrastructural levels, investigated the composition of the tubular aggregates and performed experiments on protein glycosylation.ResultsWe identified novel pathogenic DPAGT1 variants in both patients and pyridostigmine treatment quantitatively improved muscle force and function. The tubular aggregates contained proteins of the sarcoplasmic reticulum (SR) and structurally conformed to the aggregates observed in tubular aggregate myopathy (TAM). TAM arises from overactivation of the plasma membrane calcium channel ORAI1, and functional studies on muscle extracts from our LG‐CMS patients evidenced abnormal ORAI1 glycosylation.ConclusionsWe expand the genetic variant spectrum of LG‐CMS and provide a genotype/phenotype correlation for pathogenic DPAGT1 variants. The discovery of ORAI1 hypoglycosylation in our patients highlights a physiopathological link between LG‐CMS and TAM.