Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Nov 24;85(21):1886-93.
doi: 10.1212/WNL.0000000000002147. Epub 2015 Oct 23.

Histologic muscular history in steroid-treated and untreated patients with Duchenne dystrophy

Lorenzo Peverelli  1 , Silvia Testolin  1 , Luisa Villa  1 , Adele D'Amico  1 , Stefania Petrini  1 , Chiara Favero  1 , Francesca Magri  1 , Lucia Morandi  1 , Marina Mora  1 , Tiziana Mongini  1 , Enrico Bertini  1 , Monica Sciacco  1 , Giacomo P Comi  1 , Maurizio Moggio  2
Affiliations

Histologic muscular history in steroid-treated and untreated patients with Duchenne dystrophy

Lorenzo Peverelli et al. Neurology. .

Abstract

Objective: Duchenne muscular dystrophy (DMD) is a lethal disease. The outcome measures used in numerous therapeutic trials include skeletal muscle biopsy. We studied the natural history of DMD from the standpoint of muscle histology with the aim of providing a reproducible tool for use in evaluating and comparing any histologic changes occurring in patients with DMD undergoing treatment and hence be able to determine how therapy modulates the histologic evolution of the disease.

Methods: Three independent operators analyzed 56 muscle biopsies from 40 patients not treated with steroids, aged 1 to 10 years and 16 individuals treated with steroids, aged 7 to 10 years. We analyzed morphologic measures, normalized every measure for the average number of fibers observed for each year of age, and calculated intraclass correlation coefficients.

Results: The average proportion of connective tissue in patients not treated with steroids was 16.98% from ages 1 to 6 years and 30% from ages 7 to 10 years (p < 0.0001). The average proportion in patients treated with steroids was 24.90%. Muscle fiber area mirrored that of connective tissue in both groups.

Conclusions: Having provided a reproducible tool for evaluation and comparison of histologic changes occurring in patients undergoing clinical trials, it was observed that at ages 6 to 7 years, fibrotic tissue rapidly peaks to 29.85%; this is a crucial moment when muscle tissue loses its self-regeneration ability, veering toward fibrotic degeneration. These data should be considered when deciding the most suitable time to begin therapy.

PubMed Disclaimer

Figures

Figure 1
Figure 1. Connective tissue assessment (perimysial and endomysial)
(A and C) Hematoxylin & eosin staining. (B and D) Color subtraction of A and C using ImageJ 1.46r software to measure total fibrosis. Leica DC200 optical microscope, 25× magnification.
Figure 2
Figure 2. Connective tissue and MFA evaluation in untreated and treated patients
(A) Connective tissue in patients not treated with steroids (40 patients) shows a more consistent and rapid increase in fibrotic tissue between 6 and 7 years of age. At the age of 7, the proportion rises to 29.85% (SD 7.75) corresponding to an increase of more than 10% in 1 year. Connective tissue in patients treated with steroids (16 patients aged 7–10 years) ranged from 22.38% (SD 8.5) to 29.85% (SD 6.49) with increasing age. (B) MFA in the steroid-free group (40 patients). The values remained quite stable between 1 and 6 years of age (range from 79.14% [SD 0.05] to 85.04% [SD 0.06]), but a reduction was evident in patients older than 7 years (values between 69.84% [SD 0.1] and 74.47% [SD 0.05]). In the steroid-treated group (16 patients aged 7–10 years), MFA values ranged between 68.44% (SD 0.08) and 77.09% (SD 0.13).
Figure 3
Figure 3. Evaluation of morphologic measures in patients without and with steroids
(A) Necrotic fibers, regenerating fibers, internal nuclei, hypercontracted fibers, and cellular reactions in patients not treated with steroids (40 patients). (B) Necrotic fibers, regenerating fibers, internal nuclei, hypercontracted fibers, and cellular reactions in patients treated with steroids (16 patients).
See this image and copyright information in PMC

References

    1. Mendell JR, Rodino-Klapac LR, Sahenk Z, et al. Eteplirsen for the treatment of Duchenne muscular dystrophy. Ann Neurol 2013;74:637–647. - PubMed
    1. Hoffman EP, Reeves E, Damsker J, et al. Novel approaches to corticosteroid treatment in Duchenne muscular dystrophy. Phys Med Rehabil Clin N Am 2012;23:821–828. - PMC - PubMed
    1. Finkel RS, Flanigan KM, Wong B, et al. Phase 2a study of ataluren-mediated dystrophin production in patients with nonsense mutation Duchenne muscular dystrophy. PLoS One 2013;8 e81302. - PMC - PubMed
    1. ClinicalTrials.gov. Available at: www.Clinicaltrials.gov. Accessed February 20, 2015.
    1. Goemans NM, Tulinius M, van den Akker JT, et al. Systemic administration of PRO051 in Duchenne's muscular dystrophy. N Engl J Med 2011;364:1513–1522. - PubMed