Dystrophin quantification: biological and translational research implications

Karen Anthony, Virginia Arechavala-Gomeza, Laura E Taylor, Adeline Vulin, Yuuki Kaminoh, Silvia Torelli, Lucy Feng, Narinder Janghra, Gisele Bonne, Maud Beuvin, Rita Barresi, Matt Henderson, Steven Laval, Afrodite Lourbakos, Giles Campion, Volker Straub, Thomas Voit, Caroline A Sewry, Jennifer E Morgan, Kevin M FlaniganFrancesco Muntoni

Research output: Contribution to JournalArticle

Abstract

Objective: We formed a multi-institution collaboration in order to compare dystrophin quantification methods, reach a consensus on the most reliable method, and report its biological significance in the context of clinical trials. Methods: Five laboratories with expertise in dystrophin quantification performed a data-driven comparative analysis of a single reference set of normal and dystrophinopathy muscle biopsies using quantitative immunohistochemistry and Western blotting. We developed standardized protocols and assessed inter- and intralaboratory variability over a wide range of dystrophin expression levels. Results: Results from the different laboratories were highly concordant with minimal inter- and intralaboratory variability, particularly with quantitative immunohistochemistry. There was a good level of agreement between data generated by immunohistochemistry and Western blotting, although immunohistochemistry was more sensitive. Furthermore, mean dystrophin levels determined by alternative quantitative immunohistochemistry methods were highly comparable. Conclusions: Considering the biological function of dystrophin at the sarcolemma, our data indicate that the combined use of quantitative immunohistochemistry and Western blotting are reliable biochemical outcome measures for Duchenne muscular dystrophy clinical trials, and that standardized protocols can be comparable between competent laboratories. The methodology validated in our study will facilitate the development of experimental therapies focused on dystrophin production and their regulatory approval.
Original languageEnglish
JournalNeurology
Volume83
Issue number22
Early online date29 Oct 2014
DOIs
Publication statusPublished - 25 Nov 2014

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Dystrophin
Translational Medical Research
Immunohistochemistry
Western Blotting
Clinical Trials
Sarcolemma
Investigational Therapies
Duchenne Muscular Dystrophy
Clinical Protocols
Outcome Assessment (Health Care)
Biopsy
Muscles

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Anthony, K., Arechavala-Gomeza, V., Taylor, L. E., Vulin, A., Kaminoh, Y., Torelli, S., ... Muntoni, F. (2014). Dystrophin quantification: biological and translational research implications. Neurology, 83(22). https://doi.org/10.1212/WNL.0000000000001025
Anthony, Karen ; Arechavala-Gomeza, Virginia ; Taylor, Laura E ; Vulin, Adeline ; Kaminoh, Yuuki ; Torelli, Silvia ; Feng, Lucy ; Janghra, Narinder ; Bonne, Gisele ; Beuvin, Maud ; Barresi, Rita ; Henderson, Matt ; Laval, Steven ; Lourbakos, Afrodite ; Campion, Giles ; Straub, Volker ; Voit, Thomas ; Sewry, Caroline A ; Morgan, Jennifer E ; Flanigan, Kevin M ; Muntoni, Francesco. / Dystrophin quantification: biological and translational research implications. In: Neurology. 2014 ; Vol. 83, No. 22.
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Anthony, K, Arechavala-Gomeza, V, Taylor, LE, Vulin, A, Kaminoh, Y, Torelli, S, Feng, L, Janghra, N, Bonne, G, Beuvin, M, Barresi, R, Henderson, M, Laval, S, Lourbakos, A, Campion, G, Straub, V, Voit, T, Sewry, CA, Morgan, JE, Flanigan, KM & Muntoni, F 2014, 'Dystrophin quantification: biological and translational research implications', Neurology, vol. 83, no. 22. https://doi.org/10.1212/WNL.0000000000001025

Dystrophin quantification: biological and translational research implications. / Anthony, Karen; Arechavala-Gomeza, Virginia; Taylor, Laura E; Vulin, Adeline; Kaminoh, Yuuki; Torelli, Silvia; Feng, Lucy; Janghra, Narinder; Bonne, Gisele; Beuvin, Maud; Barresi, Rita; Henderson, Matt; Laval, Steven; Lourbakos, Afrodite; Campion, Giles; Straub, Volker; Voit, Thomas; Sewry, Caroline A; Morgan, Jennifer E; Flanigan, Kevin M; Muntoni, Francesco.

In: Neurology, Vol. 83, No. 22, 25.11.2014.

Research output: Contribution to JournalArticle

TY - JOUR

T1 - Dystrophin quantification: biological and translational research implications

AU - Anthony, Karen

AU - Arechavala-Gomeza, Virginia

AU - Taylor, Laura E

AU - Vulin, Adeline

AU - Kaminoh, Yuuki

AU - Torelli, Silvia

AU - Feng, Lucy

AU - Janghra, Narinder

AU - Bonne, Gisele

AU - Beuvin, Maud

AU - Barresi, Rita

AU - Henderson, Matt

AU - Laval, Steven

AU - Lourbakos, Afrodite

AU - Campion, Giles

AU - Straub, Volker

AU - Voit, Thomas

AU - Sewry, Caroline A

AU - Morgan, Jennifer E

AU - Flanigan, Kevin M

AU - Muntoni, Francesco

PY - 2014/11/25

Y1 - 2014/11/25

N2 - Objective: We formed a multi-institution collaboration in order to compare dystrophin quantification methods, reach a consensus on the most reliable method, and report its biological significance in the context of clinical trials. Methods: Five laboratories with expertise in dystrophin quantification performed a data-driven comparative analysis of a single reference set of normal and dystrophinopathy muscle biopsies using quantitative immunohistochemistry and Western blotting. We developed standardized protocols and assessed inter- and intralaboratory variability over a wide range of dystrophin expression levels. Results: Results from the different laboratories were highly concordant with minimal inter- and intralaboratory variability, particularly with quantitative immunohistochemistry. There was a good level of agreement between data generated by immunohistochemistry and Western blotting, although immunohistochemistry was more sensitive. Furthermore, mean dystrophin levels determined by alternative quantitative immunohistochemistry methods were highly comparable. Conclusions: Considering the biological function of dystrophin at the sarcolemma, our data indicate that the combined use of quantitative immunohistochemistry and Western blotting are reliable biochemical outcome measures for Duchenne muscular dystrophy clinical trials, and that standardized protocols can be comparable between competent laboratories. The methodology validated in our study will facilitate the development of experimental therapies focused on dystrophin production and their regulatory approval.

AB - Objective: We formed a multi-institution collaboration in order to compare dystrophin quantification methods, reach a consensus on the most reliable method, and report its biological significance in the context of clinical trials. Methods: Five laboratories with expertise in dystrophin quantification performed a data-driven comparative analysis of a single reference set of normal and dystrophinopathy muscle biopsies using quantitative immunohistochemistry and Western blotting. We developed standardized protocols and assessed inter- and intralaboratory variability over a wide range of dystrophin expression levels. Results: Results from the different laboratories were highly concordant with minimal inter- and intralaboratory variability, particularly with quantitative immunohistochemistry. There was a good level of agreement between data generated by immunohistochemistry and Western blotting, although immunohistochemistry was more sensitive. Furthermore, mean dystrophin levels determined by alternative quantitative immunohistochemistry methods were highly comparable. Conclusions: Considering the biological function of dystrophin at the sarcolemma, our data indicate that the combined use of quantitative immunohistochemistry and Western blotting are reliable biochemical outcome measures for Duchenne muscular dystrophy clinical trials, and that standardized protocols can be comparable between competent laboratories. The methodology validated in our study will facilitate the development of experimental therapies focused on dystrophin production and their regulatory approval.

UR - https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4248450/

U2 - 10.1212/WNL.0000000000001025

DO - 10.1212/WNL.0000000000001025

M3 - Article

VL - 83

JO - Neurology

JF - Neurology

SN - 0028-3878

IS - 22

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Anthony K, Arechavala-Gomeza V, Taylor LE, Vulin A, Kaminoh Y, Torelli S et al. Dystrophin quantification: biological and translational research implications. Neurology. 2014 Nov 25;83(22). https://doi.org/10.1212/WNL.0000000000001025