Validation of a Muscle-Specific Tissue Image Analysis Tool for Quantitative Assessment of Dystrophin Staining in Frozen Muscle Biopsies

Famke Aeffner; Crystal Faelan; Steven A Moore; Alexander Moody; Joshua C Black; Jay S Charleston; Diane E Frank; Johannes Dworzak; J Kris Piper; Manish Ranjitkar; Kristin Wilson; Suzanne Kanaly; Daniel G Rudmann; Holger Lange; G David Young; Anthony J Milici

doi:10.5858/arpa.2017-0536-OA

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Validation of a Muscle-Specific Tissue Image Analysis Tool for Quantitative Assessment of Dystrophin Staining in Frozen Muscle Biopsies

Journal article

Open access

Peer reviewed

Validation of a Muscle-Specific Tissue Image Analysis Tool for Quantitative Assessment of Dystrophin Staining in Frozen Muscle Biopsies

Famke Aeffner, Crystal Faelan, Steven A Moore, Alexander Moody, Joshua C Black, Jay S Charleston, Diane E Frank, Johannes Dworzak, J Kris Piper, Manish Ranjitkar, …

Archives of pathology & laboratory medicine (1976), Vol.143(2), pp.197-205

02/2019

DOI: 10.5858/arpa.2017-0536-OA

PMCID: PMC6813837

PMID: 30168727

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Abstract

Duchenne muscular dystrophy is a rare, progressive, and fatal neuromuscular disease caused by dystrophin protein loss. Common investigational treatment approaches aim at increasing dystrophin expression in diseased muscle. Some clinical trials include assessments of novel dystrophin production as a surrogate biomarker of efficacy, which may predict a clinical benefit from treatment. To establish an immunofluorescent scanning and digital image analysis workflow that provides an objective approach for staining intensity assessment of the immunofluorescence dystrophin labeling and determination of the percentage of biomarker-positive fibers in muscle cryosections. Optimal and repeatable digital image capture was achieved by a rigorously qualified fluorescent scanning process. After scanning qualification, the MuscleMap (Flagship Biosciences, Westminster, Colorado) algorithm was validated by comparing high-power microscopic field total and dystrophin-positive fiber counts obtained by trained pathologists to data derived by MuscleMap. Next, the algorithm was tested on whole-slide images of immunofluorescent-labeled muscle sections from Duchenne muscular dystrophy, Becker muscular dystrophy, and control patients. When used under the guidance of a trained pathologist, the digital image analysis tool met predefined validation criteria and demonstrated functional and statistical equivalence with manual assessment. This work is the first, to our knowledge, to qualify and validate immunofluorescent scanning and digital tissue image-analysis workflow, respectively, with the rigor required to support the clinical trial environments. MuscleMap enables analysis of all fibers within an entire muscle biopsy section and provides data on a fiber-by-fiber basis. This will allow future clinical trials to objectively investigate myofibers' dystrophin expression at a greater level of consistency and detail.

Frozen Sections

Humans

Image Interpretation, Computer-Assisted - methods

Middle Aged

Child, Preschool

Male

Dystrophin - analysis

Biopsy

Adolescent

Female

Muscle, Skeletal - pathology

Child

Muscular Dystrophy, Duchenne - diagnosis

Details

Title: Subtitle: Validation of a Muscle-Specific Tissue Image Analysis Tool for Quantitative Assessment of Dystrophin Staining in Frozen Muscle Biopsies
Creators: Famke Aeffner - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Crystal Faelan - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Steven A Moore - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Alexander Moody - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Joshua C Black - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Jay S Charleston - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Diane E Frank - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Johannes Dworzak - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
J Kris Piper - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Manish Ranjitkar - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Kristin Wilson - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Suzanne Kanaly - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Daniel G Rudmann - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Holger Lange - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
G David Young - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Anthony J Milici - From Flagship Biosciences Inc, Westminster, Colorado (Drs Aeffner, Faelan, Black, Wilson, Kanaly, Rudmann, Lange, Young, and Milici and Mr Moody); the Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City (Dr Moore); and Sarepta Therapeutics Inc, Cambridge, Massachusetts (Drs Charleston and Frank and Messrs Dworzak, Piper, and Ranjitkar). Dr Moore is now with Oregon Health and Science University, Portland, and Dr Rudmann is now with Charles River Laboratories, Ashland, Ohio
Resource Type: Journal article
Publication Details: Archives of pathology & laboratory medicine (1976), Vol.143(2), pp.197-205
Publisher: United States
DOI: 10.5858/arpa.2017-0536-OA
PMID: 30168727
PMCID: PMC6813837
ISSN: 0003-9985
eISSN: 1543-2165
Grant note: U54 NS053672 / NINDS NIH HHS
Language: English
Date published: 02/2019
Academic Unit: Pathology
Record Identifier: 9984047753902771

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