Duchenne Muscular Dystrophy: recent advances in protein biomarkers and the clinical application

Figures & data

Table 1. List of DMD biomarkers identified. The biomarkers are ranked based on the number of publications providing evidence for association of the biomarker with DMD. Biomarkers identified in both human and animal models are listed first, followed by biomarkers identified only in DMD patients and subsequently in only DMD animal models.

Biomarker candidate	Sample	Species	Proteomics method	Reference
Creatine kinase	Serum	Human, mdx, GRMD	Aptamer-, immuno-based	[34,47–52,80,98,101]
Carbonic anhydrase 3	Serum, plasma	Human, mdx‐4cv	Aptamer-, immuno-, mass spectrometry-based	[36,77,80,92,94,96,107,124]
Fatty acid binding protein 3	Serum	Human, mdx‐4cv, mdx-23, mdx-52	Aptamer-based, mass spectrometry-based	[34,80,92,93,96,107,142]
Myoglobin	Serum	Human, mdx-23, mdx-52	Aptamer-, immuno-, mass spectrometry-based	[34,48,92,94,96,107,144]
Cardiac muscle troponin I	Serum	Human, mdx-23	Aptamer-, immuno-based	[92–94,107,144]
Calcium/calmodulin- dependent protein kinases	Serum	Human, mdx-23	Aptamer-, immuno-based	[92–94,96,107]
Lactate dehydrogenase	Serum, plasma	Human, mdx-4cv	Aptamer-, immuno-, mass spectrometry based	[47,77,80,93,124]
Myosin light chain 1/3	Serum, plasma	Human, mdx-23, mdx-52, mdx-4cv	Immuno-, mass spectrometry based	[34,36,55,80,142]
Myostatin	Serum	Human, mdx-23	Aptamer-, mass spectrometry-based	[53,59,92,94,107]
Haptoglobin	Serum, plasma	Human, mouse mdx-4cv	Immuno-, mass spectrometry-based	[77,80,119,120]
Fibrinogens	Serum, plasma	Human, mouse mdx-23, mdx-52	Aptamer-, mass spectrometry-based	[34,58,94,96]
Malate dehydrogenase 2	Serum, plasma	Human, mdx-4cv	Immuno-, mass spectrometry-based	[36,80,84,124]
Enolase αβ	Serum, plasma	Human, mdv-4cv	Immuno-, mass spectrometry-based	[34,36,80]
Adiponectin	Serum	Human, mdx	Immuno-, mass spectrometry-based	[34,56,77]
Cytochrome-c	Serum	Human, mouse mdx-23, mdx‐4cv	Aptamer-, immuno-, mass spectrometry-based	[34,80,144]
Pyruvate kinase	Serum	Human, mdx-23, mdx-52, mdx-4cv	Mass spectrometry-based	[34,77,80]
Fructose bisphosphate aldolase	Serum	Human, mdx-23, mdx-52, mdx-4cv	Mass spectrometry-based	[34,77,80]
Titin	Serum	Human, mdx	Immuno-, mass spectrometry-based	[34,54,77]
Tumor necrosis factor receptor	Serum, plasma	Human, mouse mdx-23	Aptamer-, immuno-based	[59,118,121]
Interleukins (IL-6, IL-10, IL-13)	Serum, plasma	Human, Mouse mdx	Aptamer-, Immuno-based	[94,121]
Myomesin-3	Serum	Human, mdx-23, mdx-52	Mass spectrometry-based	[34,77]
Transforming growth factor β	Plasma	Human, mdx	Immuno-based	[55,121]
Tissue inhibitor TIMP-1 of metalloproteinase	Plasma	Human, mdx	Immuno-based	[55,112]
Filamin C	Serum	Human, mdx-23, mdx-52	Mass spectrometry-based	[34,77]
Fibronectin	Serum	Human,mdx-4cv	Mass spectrometry-based	[37,80]
A disintegrin and metalloproteinase with thrombospondin motifs 5	Serum	Human, mdx	Aptamer-, immuno-based	[144]
N-terminal fragment of α dystro-glycan	Serum	Human, mdx	Immuno-based	[57]
Phosphglycerate mutase	Serum	Human, mouse mdx-23	Aptamer-, immuno-based	[144]
Matrix metalloproteinase MMP-9	Serum, plasma	Human	Aptamer-, immuno-based	[95,112,113]
Interleukin 1 Receptor-Like 1 Protein	Serum	Human	Aptamer-, immuno-based	[95]
Calmodulin-like protein	Serum	Human	Mass spectrometry-based	[54]
Electron transfer flavoprotein A	Serum, plasma	Human	Immuno-based	[36]
Ankyrin repeat domain 2	Serum, plasma	Human	Immuno-based	[84]
Carbonic anhydrase 1	Serum	Human	Mass spectrometry-based	[54]
Fibulin 1	Serum	Human	Mass spectrometry-based	[54]
Gelsolin	Serum	Human	Mass spectrometry-based	[54]
Coagulation factor XIIIa	Serum	Mouse mdx-23, mdx-3cv	Mass spectrometry-based	[54,76]
Tumor necrosis factor-alpha	Plasma	Mouse mdx	Immuno-based	[60,121]
Interferon-γ	Plasma	Mouse, mdx	Immuno-based	[60,121]
Glutathione peroxidase	Plasma	Mouse mdx-23	Mass spectrometry-based	[58]
Lumican	Serum	Mouse mdx-23, mdx-52	Mass spectrometry-based	[34]
Leukemia inhibitory factor receptor	Serum	Mouse mdx-23, mdx-52	Mass spectrometry-based	[34]
Plasminogen	Serum	Mouse mdx-23, mdx-52	Mass spectrometry-based	[34]

Figure 1. Biomarker development for DMD requires (1) discovery, (2) analytical validation, (3) clinical validation, and (4) qualification of biomarker. The discovery phase comprises formulation of the context of use, assessment of feasibility in terms of eg. samples resources, selection of appropriate analytical method, etc., discovery of the biomarker and confirmation in additional samples collections to assess reproducibility in several cohorts. The analytical validation phase includes orthogonal validation of the biomarker using different analytical methods, development of biomarker quantification assays for targeted analysis and validation of the biomarker quantification assay, to evaluate biomarker sensitivity, specificity, dynamic range, and variation using the developed quantification assay and refine the context of use. Clinical validation requires testing the specificity and sensitivity of the biomarker in clinical setup, development of clinical assays, assess the context of use, robustness and variability, and its utility in clinical practice.

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