Diego Lopergolo
Novel biomarkers for limb-girdle muscular dystrophy associated to CAPN3 mutation
Autori
- DIEGO LOPERGOLO (DEPARTMENT OF MEDICINE, SURGERY AND NEUROSCIENCES, UNIVERSITY OF SIENA, SIENA, ITALY. – MEDICAL GENETICIST)
- SARA AGUTI (DEPARTMENT OF MEDICINE, SURGERY AND NEUROSCIENCES, UNIVERSITY OF SIENA, SIENA, ITALY. – )
- GIAN NICOLA GALLUS (DEPARTMENT OF MEDICINE, SURGERY AND NEUROSCIENCES, UNIVERSITY OF SIENA, SIENA, ITALY. – )
- SILVIA BIANCHI (DEPARTMENT OF MEDICINE, SURGERY AND NEUROSCIENCES, UNIVERSITY OF SIENA, SIENA, ITALY. – )
- SIMONA SALVATORE (DEPARTMENT OF MEDICINE, SURGERY AND NEUROSCIENCES, UNIVERSITY OF SIENA, SIENA, ITALY. – )
- ANNA RUBEGNI (MOLECULAR MEDICINE FOR NEURODEGENERATIVE AND NEUROMUSCULAR DISEASE UNIT, IRCCS STELLA MARIS FOUNDATION, PISA, ITALY – )
- GIANNA BERTI (DEPARTMENT OF MEDICINE, SURGERY AND NEUROSCIENCES, UNIVERSITY OF SIENA, SIENA, ITALY. – )
- PATRIZIA FORMICHI (DEPARTMENT OF MEDICINE, SURGERY AND NEUROSCIENCES, UNIVERSITY OF SIENA, SIENA, ITALY. – )
- NICOLA DE STEFANO (DEPARTMENT OF MEDICINE, SURGERY AND NEUROSCIENCES, UNIVERSITY OF SIENA, SIENA, ITALY. – )
- ALESSANDRO MALANDRINI (DEPARTMENT OF MEDICINE, SURGERY AND NEUROSCIENCES, UNIVERSITY OF SIENA, SIENA, ITALY. – )
Presentatore
DIEGO LOPERGOLO
Modalità
Oral Communication
Abstract
“Background: LGMD encompasses a large group of muscular dystrophies. CAPN3 mutations are associated with autosomal recessive LGMD-1 and autosomal dominant LGMD-4. Diagnosis is currently often based on invasive methods requiring muscle biopsy: in most of the cases, Western blotting (WB) analysis from muscle is essential, as muscle samples are currently the only known tissues to express the full-length CAPN3 isoform.
Methods: We analyzed CAPN3 in a cohort including 60 probable LGMD patients. Selected patients underwent a complete neurological examination, muscle biopsy, and skin biopsies. The amount of CAPN3 was evaluated by WB analysis in muscle and skin tissues. The total RNA isolated from muscle, fibroblast and urine was processed for qualitative analysis. The expression of CAPN3 was investigated by qRT-PCR. The CAPN3 3D structure has been analyzed using PyMOL.
Results: Seven different CAPN3 mutations were detected, of which two were novel. After sequencing CAPN3 transcripts from fibroblast and urine, we detected different CAPN3 isoforms surprisingly including the full-length transcript. We found comparable protein levels from fibroblasts and muscle tissue; in particular, patients harboring a novel CAPN3 mutation showed a 30% reduction in protein compared to controls from both tissues.
Conclusions: Our findings showed for the first time the presence of the CAPN3 full-length transcript in urine and skin samples. Moreover, the comparable CAPN3 protein levels between muscle and skin samples allow us to hypothesize the use of skin biopsy and probably of urine samples as an alternative less invasive method to assess the amount of CAPN3.”