Duchenne muscular dystrophy (DMD) is an incurable disease caused by out-of-frame DMD gene deletions while in frame deletions lead to the milder Becker muscular dystrophy (BMD). In the last decade several antisense oligonucleotides drugs have been developed to induce a partially functional internally deleted dystrophin, similar to that produced in BMD, and expected to ameliorate the disease course. The pattern of dystrophin expression and functionality in dystrophinopathy patients is variable due to multiple factors, such as molecular functionality of the dystrophin and its distribution. To benchmark the success of therapeutic intervention, a clear understanding of dystrophin expression patterns in dystrophinopathy patients is vital. Recently, several groups have used innovative techniques to quantify dystrophin in muscle biopsies of children but not in patients with milder BMD. This study reports on dystrophin expression using both Western blotting and an automated, high-throughput, image analysis platform in DMD, BMD, and intermediate DMD/BMD skeletal muscle biopsies. Our results found a significant correlation between Western blot and immunofluorescent quantification indicating consistency between the different methodologies. However, we identified significant inter- and intradisease heterogeneity of patterns of dystrophin expression in patients irrespective of the amount detected on blot, due to variability in both fluorescence intensity and dystrophin sarcolemmal circumference coverage. Our data highlight the heterogeneity of the pattern of dystrophin expression in BMD, which will assist the assessment of dystrophin restoration therapies.

Torelli, S., Scaglioni, D., Sardone, V., Ellis, M. J., Domingos, J., Jones, A., Feng, L., Chambers, D., Eastwood, D. M., Leturcq, F., Yaou, R. B., Urtizberea, A., Sabouraud, P., Barnerias, C., Stojkovic, T., Ricci, E., Beuvin, M., Bonne, G., Sewry, C. A., Willis, T., Kulshrestha, R., Tasca, G., Phadke, R., Morgan, J. E., Muntoni, F., High-Throughput Digital Image Analysis Reveals Distinct Patterns of Dystrophin Expression in Dystrophinopathy Patients, <<JOURNAL OF NEUROPATHOLOGY AND EXPERIMENTAL NEUROLOGY>>, 2021; 80 (10): 955-965. [doi:10.1093/jnen/nlab088] [http://hdl.handle.net/10807/197069]

High-Throughput Digital Image Analysis Reveals Distinct Patterns of Dystrophin Expression in Dystrophinopathy Patients

Ricci, Enzo;Tasca, Giorgio;
2021

Abstract

Duchenne muscular dystrophy (DMD) is an incurable disease caused by out-of-frame DMD gene deletions while in frame deletions lead to the milder Becker muscular dystrophy (BMD). In the last decade several antisense oligonucleotides drugs have been developed to induce a partially functional internally deleted dystrophin, similar to that produced in BMD, and expected to ameliorate the disease course. The pattern of dystrophin expression and functionality in dystrophinopathy patients is variable due to multiple factors, such as molecular functionality of the dystrophin and its distribution. To benchmark the success of therapeutic intervention, a clear understanding of dystrophin expression patterns in dystrophinopathy patients is vital. Recently, several groups have used innovative techniques to quantify dystrophin in muscle biopsies of children but not in patients with milder BMD. This study reports on dystrophin expression using both Western blotting and an automated, high-throughput, image analysis platform in DMD, BMD, and intermediate DMD/BMD skeletal muscle biopsies. Our results found a significant correlation between Western blot and immunofluorescent quantification indicating consistency between the different methodologies. However, we identified significant inter- and intradisease heterogeneity of patterns of dystrophin expression in patients irrespective of the amount detected on blot, due to variability in both fluorescence intensity and dystrophin sarcolemmal circumference coverage. Our data highlight the heterogeneity of the pattern of dystrophin expression in BMD, which will assist the assessment of dystrophin restoration therapies.
2021
Inglese
Torelli, S., Scaglioni, D., Sardone, V., Ellis, M. J., Domingos, J., Jones, A., Feng, L., Chambers, D., Eastwood, D. M., Leturcq, F., Yaou, R. B., Urtizberea, A., Sabouraud, P., Barnerias, C., Stojkovic, T., Ricci, E., Beuvin, M., Bonne, G., Sewry, C. A., Willis, T., Kulshrestha, R., Tasca, G., Phadke, R., Morgan, J. E., Muntoni, F., High-Throughput Digital Image Analysis Reveals Distinct Patterns of Dystrophin Expression in Dystrophinopathy Patients, <<JOURNAL OF NEUROPATHOLOGY AND EXPERIMENTAL NEUROLOGY>>, 2021; 80 (10): 955-965. [doi:10.1093/jnen/nlab088] [http://hdl.handle.net/10807/197069]
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10807/197069
Citazioni
  • ???jsp.display-item.citation.pmc??? 2
  • Scopus 6
  • ???jsp.display-item.citation.isi??? 5
social impact