Aim: Detection of drug-induced dystrophin in patient muscle biopsy is a surrogate outcome measure for Duchenne muscular dystrophy. We sought to establish and validate an orthogonal approach to measurement of dystrophin protein and RNA in muscle biopsies. Materials & methods: Validated methods were developed for dystrophin western blotting, mass spectrometry, immunostaining and reverse transcriptase PCR of biopsy mRNA using muscle biopsy standards. Results: Both western blotting and mass spectrometry validated methods demonstrated good linearity, and acceptable precision and accuracy with a lower limit of quantitation at 1%. Immunostaining and reverse transcriptase PCR methods were shown to be reliable. Conclusion: The described orthogonal approach is sufficient to support measures of dystrophin as a surrogate outcome in a clinical trial.
Aim: Detection of drug-induced dystrophin in patient muscle biopsy is a surrogate outcome measure for Duchenne muscular dystrophy. We sought to establish and validate an orthogonal approach to measurement of dystrophin protein and RNA in muscle biopsies. Materials & methods: Validated methods were developed for dystrophin western blotting, mass spectrometry, immunostaining and reverse transcriptase PCR of biopsy mRNA using muscle biopsy standards. Results: Both western blotting and mass spectrometry validated methods demonstrated good linearity, and acceptable precision and accuracy with a lower limit of quantitation at 1%. Immunostaining and reverse transcriptase PCR methods were shown to be reliable. Conclusion: The described orthogonal approach is sufficient to support measures of dystrophin as a surrogate outcome in a clinical trial.
Authors: Vahid Farrokhi; Jason Walsh; Joe Palandra; Joanne Brodfuehrer; Teresa Caiazzo; Jane Owens; Michael Binks; Srividya Neelakantan; Florence Yong; Pinky Dua; Caroline Le Guiner; Hendrik Neubert Journal: Gene Ther Date: 2021-11-05 Impact factor: 4.184
Authors: Paula R Clemens; Vamshi K Rao; Anne M Connolly; Amy D Harper; Jean K Mah; Edward C Smith; Craig M McDonald; Craig M Zaidman; Lauren P Morgenroth; Hironori Osaki; Youhei Satou; Taishi Yamashita; Eric P Hoffman Journal: JAMA Neurol Date: 2020-08-01 Impact factor: 18.302