Muscle-Derived Proteins as Serum Biomarkers for Monitoring Disease Progression in Three Forms of Muscular Dystrophy

Peter M Burch, Oksana Pogoryelova, Richard Goldstein, Donald Bennett, Michela Guglieri, Volker Straub, Kate Bushby, Hanns Lochmüller, Carl Morris, Peter M Burch, Oksana Pogoryelova, Richard Goldstein, Donald Bennett, Michela Guglieri, Volker Straub, Kate Bushby, Hanns Lochmüller, Carl Morris

Abstract

Background: Identifying translatable, non-invasive biomarkers of muscular dystrophy that better reflect the disease pathology than those currently available would aid the development of new therapies, the monitoring of disease progression and the response to therapy.

Objective: The goal of this study was to evaluate a panel of serum protein biomarkers with the potential to specifically detect skeletal muscle injury.

Method: Serum concentrations of skeletal troponin I (sTnI), myosin light chain 3 (Myl3), fatty acid binding protein 3 (FABP3) and muscle-type creatine kinase (CKM) proteins were measured in 74 Duchenne muscular dystrophy (DMD), 38 Becker muscular dystrophy (BMD) and 49 Limb-girdle muscular dystrophy type 2B (LGMD2B) patients and 32 healthy controls.

Results: All four proteins were significantly elevated in the serum of these three muscular dystrophy patient populations when compared to healthy controls, but, interestingly, displayed different profiles depending on the type of muscular dystrophy. Additionally, the effects of patient age, ambulatory status, cardiac function and treatment status on the serum concentrations of the proteins were investigated. Statistical analysis revealed correlations between the serum concentrations and certain clinical endpoints including forced vital capacity in DMD patients and the time to walk ten meters in LGMD2B patients. Serum concentrations of these proteins were also elevated in two preclinical models of muscular dystrophy, the mdx mouse and the golden-retriever muscular dystrophy dog.

Conclusions: These proteins, therefore, are potential muscular dystrophy biomarkers for monitoring disease progression and therapeutic response in both preclinical and clinical studies.

Keywords: Muscular dystrophy; biomarker; creatine kinase; fatty acid binding protein 3; myosin light chain 3; skeletal troponin I.

Figures

Fig.1
Fig.1
Scatter plots of the serum clinical chemistry and protein serum concentrations in muscular dystrophy patients. (A) Total serum CK, AST and ALT levels and (B) serum concentrations of sTnI, Myl3, FABP3 and CKM for the DMD, BMD, LGMD2B patient groups and healthy controls are shown. The line and error bars represent the mean and standard deviation of each group.  ***P <  0.001;  ****P <  0.0001.
Fig.2
Fig.2
Scatter plots of the protein serum concentrations in DMD patients based on clinical status. Serum concentrations of sTnI, Myl3, FABP3, CKM and total CK in (A) ambulant (N = 46) and non-ambulant (N = 29) DMD patients or (B) those classified as with (N = 25) and without (N = 50) cardiomyopathy are shown. The line and error bars represent the mean and standard deviation of each group.  ****P <  0.0001.
Fig.3
Fig.3
No significant change in patient’s serum biomarker concentrations taken approximately one year apart. Serum biomarker concentrations in serial blood samples from DMD patients (N = 26). Patients were grouped by age and individual serum biomarker concentrations from the initial (circles) and follow-up samples (squares) were plotted. P values (P) were calculated by two-way ANOVA. For age groups where no P values is given then P >  0.99 for that comparison.
Fig.4
Fig.4
Correlations of protein serum concentration and clinical measures in DMD patients. A graph of (A) FVC measurement and (B) NSAA score versus the serum concentrations of sTnI, Myl3, FABP3, CKM and total serum CK for each DMD patient. The Spearman’s rank-order correlation coefficient (r), P value (P) and number of patients in the sample set (N) is shown for each analysis.
Fig.5
Fig.5
Correlations of protein serum concentrations and clinical measures in LGMD2B patients. A graph of the (A) FVC measurement and (B) the time to walk 10 m test versus serum concentrations of sTnI, Myl3, FABP3, CKM and total serum CK for each LGMD2B patients is shown. The Spearman’s correlation coefficient (r), P value (P) and number of patients in the sample set (N) is shown for each analysis.
Fig.6
Fig.6
Scatter plots of protein serum concentrations and age in DMD, BMD and LGMD2B patients. A graph of the serum concentrations of sTnI, Myl3, FABP3, CKM and total serum CK are shown versus the age of the patient for the (A) DMD (N = 61), (B) BMD (N = 38) and(C) LGMD2B (N = 49) samples.

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