1. Orthogonal analysis of dystrophin protein and mRNA as a surrogate outcome for drug development
- Author
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Kitipong Uaesoontrachoon, Romain Beauvois, Taishi Yamashita, Jordan Warford, Youhei Satou, Swati Mummidivarpu, Joyce Rowsell, Kevork Mekhssian, Eric P. Hoffman, Amanda Mullen, Anahita Keyhani, Maja Malbasic, William Ross, Hélène Montpetit, Hironori Osaki, Alexandra MacKinnon, Yetrib Hathout, Marina Moraca, Kanneboyina Nagaraju, Molly Praest, Mark Matyas, and Sadish Srinivassane
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,Duchenne muscular dystrophy ,Biopsy ,Clinical Biochemistry ,Blotting, Western ,Mass Spectrometry ,Dystrophin ,03 medical and health sciences ,0302 clinical medicine ,Drug Discovery ,medicine ,Humans ,RNA, Messenger ,Muscle biopsy ,biology ,medicine.diagnostic_test ,business.industry ,Surrogate endpoint ,Biochemistry (medical) ,Exons ,medicine.disease ,Exon skipping ,Reverse transcription polymerase chain reaction ,030104 developmental biology ,biology.protein ,business ,030217 neurology & neurosurgery ,Immunostaining - Abstract
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.
- Published
- 2019