Protein signature of human skin fibroblasts allows the study of the molecular etiology of rare neurological diseases

Abstract Background The elucidation of pathomechanisms leading to the manifestation rare (genetically caused) neurological diseases including neuromuscular (NMD) represents an important step toward understanding genesis respective disease and might help define starting points for (new) therapeutic intervention concepts. However, these “discovery studies” are often limited by availability human biomaterial. Moreover, given that results next-generation-sequencing approaches frequently result in identification ambiguous variants, testing their pathogenicity is crucial but also depending on patient-derived material. Methods Human skin fibroblasts were used generate a spectral library using pH8-fractionation followed nano LC-MS/MS. Afterwards, Allgrove-patient derived subjected data independent acquisition approach. In addition, proteomic signature enriched nuclear protein fraction was studied. Proteomic findings confirmed immunofluorescence muscle biopsy from same patient cellular lipid homeostasis cause Allgrove syndrome analysed fluorescence (BODIPY-staining) coherent anti-Stokes Raman scattering (CARS) microscopy. Results To systematically address question if serve as valuable biomaterial (molecular) studies NMD, we generated cataloguing 8280 proteins variety such linked genetic forms motoneuron diseases, congenital myasthenic syndromes, neuropathies disorders. silico-based pathway analyses revealed expression diversity involved contraction decisive neuronal function maintenance suggesting suitability study etiology NMD. Based findings, next aimed further demonstrate this vitro model NMD use case: Dysregulation paradigmatic could be protein-functions symptoms known disease. investigation composition allowed protein-dysregulations according with structural perturbations observed biopsy. BODIPY-staining CARS microscopy suggest altered storage part underlying etiology. Conclusions Our combined reveal may system molecular exemplified unbiased fashion.