Innovative Computerized Dystrophin Quantification Method Based on Spectral Confocal Microscopy
Por:
Codina-Bergadà A, Roldan-Molina M, Natera-de Benito D, Ortez-Gonzalez CI, Planas R, Matalonga L, Cuadras-Palleja D, Carrera-García L, Exposito-Escudero JM, Márquez-Pereira JM, Jimenez-Mallebrera C, M Porta J, Nascimento-Osorio A and Jou-Munoz C
Publicada:
1 abr 2023
Ahead of Print:
28 mar 2023
Resumen:
Several clinical trials are working on drug development for Duchenne and Becker muscular dystrophy (DMD and BMD) treatment, and, since the expected increase in dystrophin is relatively subtle, high-sensitivity quantification methods are necessary. There is also a need to quantify dystrophin to reach a definitive diagnosis in individuals with mild BMD, and in female carriers. We developed a method for the quantification of dystrophin in DMD and BMD patients using spectral confocal microscopy. It offers the possibility to capture the whole emission spectrum for any antibody, ensuring the selection of the emission peak and allowing the detection of fluorescent emissions of very low intensities. Fluorescence was evaluated first on manually selected regions of interest (ROIs), proving the usefulness of the methodology. Later, ROI selection was automated to make it operator-independent. The proposed methodology correctly classified patients according to their diagnosis, detected even minimal traces of dystrophin, and the results obtained automatically were statistically comparable to the manual ones. Thus, spectral imaging could be implemented to measure dystrophin expression and it could pave the way for detailed analysis of how its expression relates to the clinical course. Studies could be further expanded to better understand the expression of dystrophin-associated protein complexes (DAPCs).
Filiaciones:
Codina-Bergadà A:
Neuromuscular Unit, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Biobank, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Institut de Recerca Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Roldan-Molina M:
Institut de Recerca Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Confocal Microscopy and Cellular Imaging Unit, Genetic and Molecular Medicine Department, Pediatric Institute for Rare Diseases, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Natera-de Benito D:
Neuromuscular Unit, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Institut de Recerca Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Ortez-Gonzalez CI:
Neuromuscular Unit, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Institut de Recerca Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Planas R:
Institut de Robòtica i Informàtica Industrial, Technical University of Catalonia (UPC) and the Spanish Council for Scientific Research (CSIC) Llorens i Artigas 4-6, 08028 Barcelona, Spain
Matalonga L:
CNAG-CRG Centro Nacional de Análisis Genomico (CNAG)-Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Baldiri Reixac 4, 08028 Barcelona, Spain
Cuadras-Palleja D:
Institut de Recerca Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Statistics Department, Fundació Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Carrera-García L:
Neuromuscular Unit, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Institut de Recerca Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Exposito-Escudero JM:
Neuromuscular Unit, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Institut de Recerca Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Márquez-Pereira JM:
Biobank, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Jimenez-Mallebrera C:
Neuromuscular Unit, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Institut de Recerca Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
M Porta J:
Institut de Robòtica i Informàtica Industrial, Technical University of Catalonia (UPC) and the Spanish Council for Scientific Research (CSIC) Llorens i Artigas 4-6, 08028 Barcelona, Spain
Nascimento-Osorio A:
Neuromuscular Unit, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Institut de Recerca Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Jou-Munoz C:
Neuromuscular Unit, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Biobank, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Institut de Recerca Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
Pathology Department, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain
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