Phenotypical and Myopathological Consequences of Compound Heterozygous Missense and Nonsense Variants in SLC18A3

Abstract

Background: Presynaptic forms of congenital myasthenic syndromes (CMS) due to pathogenic variants in SLC18A3 impairing the synthesis and recycling of acetylcholine (ACh) have recently been described. SLC18A3 encodes the vesicular ACh transporter (VAChT), modulating the active transport of ACh at the neuromuscular junction, and homozygous loss of VAChT leads to lethality. Methods: Exome sequencing (ES) was carried out to identify the molecular genetic cause of the disease in a 5-year-old male patient and histological, immunofluorescence as well as electron- and CARS-microscopic studies were performed to delineate the muscle pathology, which has so far only been studied in VAChT-deficient animal models. Results: ES unraveled compound heterozygous missense and nonsense variants (c.315G>A, p.Trp105* and c.1192G>C, p.Asp398His) in SLC18A3. Comparison with already-published cases suggests a more severe phenotype including impaired motor and cognitive development, possibly related to a more severe effect of the nonsense variant. Therapy with pyridostigmine was only partially effective while 3,4 diaminopyridine showed no effect. Microscopic investigation of the muscle biopsy revealed reduced fibre size and a significant accumulation of lipid droplets. Conclusions: We suggest that nonsense variants have a more detrimental impact on the clinical manifestation of SLC18A3-associated CMS. The impact of pathogenic SLC18A3 variants on muscle fibre integrity beyond the effect of denervation is suggested by the build-up of lipid aggregates. This in turn implicates the importance of proper VAChT-mediated synthesis and recycling of ACh for lipid homeostasis in muscle cells. This hypothesis is further supported by the pathological observations obtained in previously published VAChT-animal models.

Presynaptic Congenital Myasthenic Syndromes: Understanding Clinical Phenotypes through In vivo Models

“…Clinical onset of SLC18A3 CMS occurs in the neonatal period, and is often associated with apnoeic episodes, hypotonia, variable degrees of joint contractures, and feeding difficulties [10,47,48]. However, four patients also displayed prenatal alterations [49,50]. Ptosis, ophthalmoplegia, mild facial weakness and generalized fatigable weakness are other common symptoms.…”

“…A decremental response to RNS is usually detected. Pyridostigmine improves clinical manifestations, and 3,4-Diaminopyridine (3,4-DAP) and ephedrine may be added in some cases [10,48,50]. Intriguingly, a mild reduction of left ventricular systolic function was rescued after treatment with AChEI in one patient [10] (Table 1).…”