Proteomic profiling of mdx-4cv serum reveals highly elevated levels of the inflammation-induced plasma marker haptoglobin in muscular dystrophy

Murphy,Sandra; Dowling,Paul; Zweyer,Margit; Henry,Michael; Meleady,Paula; Mundegar,Rustam  R.; Swandulla,Dieter; Ohlendieck,Kay

doi:10.3892/ijmm.2017.2952

Proteomic profiling of mdx-4cv serum reveals highly elevated levels of the inflammation-induced plasma marker haptoglobin in muscular dystrophy

Authors:
- Sandra Murphy
- Paul Dowling
- Margit Zweyer
- Michael Henry
- Paula Meleady
- Rustam R. Mundegar
- Dieter Swandulla
- Kay Ohlendieck
View Affiliations

Affiliations: Department of Biology, Maynooth University, National University of Ireland, Maynooth, Co. Kildare, Ireland, Department of Physiology II, University of Bonn, D‑53115 Bonn, Germany, National Institute for Cellular Biotechnology, Dublin City University, Dublin 9, Ireland
Published online on: April 18, 2017 https://doi.org/10.3892/ijmm.2017.2952
Pages: 1357-1370
Copyright: © Murphy et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

X-linked muscular dystrophy is caused by primary abnormalities in the Dmd gene and is characterized by the almost complete loss of the membrane cytoskeletal protein dystrophin, which triggers sarcolemmal instability, abnormal calcium homeostasis, increased proteolysis and impaired excitation‑contraction coupling. In addition to progressive necrosis, crucial secondary pathologies are represented by myofibrosis and the invasion of immune cells in damaged muscle fibres. In order to determine whether these substantial changes within the skeletal musculature are reflected by an altered rate of protein release into the circulatory system or other plasma fluctuations, we used label‑free mass spectrometry to characterize serum from the mdx‑4cv model of Duchenne muscular dystrophy. Comparative proteomics revealed a large number of increased vs. decreased protein species in mdx‑4cv serum. A serum component with greatly elevated levels was identified as the inflammation‑inducible plasma marker haptoglobin. This acute phase response protein is usually secreted in relation to tissue damage and sterile inflammation. Both immunoblot analyses and enzyme‑linked immunosorbent assays confirmed the increased concentration of haptoglobin in crude mdx‑4cv serum. This suggests that haptoglobin, in conjunction with other altered serum proteins, represents a novel diagnostic, prognostic and/or therapy‑monitoring biomarker candidate to evaluate the inflammatory response in the mdx‑4cv animal model of dystrophinopathy.

View Figures

View References

1	Flanigan KM: Duchenne and Becker muscular dystrophies. Neurol Clin. 32:671–688. viii2014. View Article : Google Scholar : PubMed/NCBI
2	Mah JK, Korngut L, Dykeman J, Day L, Pringsheim T and Jette N: A systematic review and meta-analysis on the epidemiology of Duchenne and Becker muscular dystrophy. Neuromuscul Disord. 24:482–491. 2014. View Article : Google Scholar : PubMed/NCBI
3	Guiraud S, Aartsma-Rus A, Vieira NM, Davies KE, van Ommen GJ and Kunkel LM: The pathogenesis and therapy of muscular dystrophies. Annu Rev Genomics Hum Genet. 16:281–308. 2015. View Article : Google Scholar : PubMed/NCBI
4	Ohlendieck K: Towards an understanding of the dystrophin-glycoprotein complex: Linkage between the extracellular matrix and the membrane cytoskeleton in muscle fibers. Eur J Cell Biol. 69:1–10. 1996.PubMed/NCBI
5	Allen DG, Zhang BT and Whitehead NP: Stretch-induced membrane damage in muscle: Comparison of wild-type and mdx mice. Adv Exp Med Biol. 682:297–313. 2010. View Article : Google Scholar : PubMed/NCBI
6	Allen DG, Whitehead NP and Froehner SC: Absence of dystrophin disrupts skeletal muscle signaling: Roles of Ca²⁺, reactive oxygen species, and nitric oxide in the development of muscular dystrophy. Physiol Rev. 96:253–305. 2016. View Article : Google Scholar
7	Hopf FW, Turner PR and Steinhardt RA: Calcium misregulation and the pathogenesis of muscular dystrophy. Subcell Biochem. 45:429–464. 2007. View Article : Google Scholar
8	Shin J, Tajrishi MM, Ogura Y and Kumar A: Wasting mechanisms in muscular dystrophy. Int J Biochem Cell Biol. 45:2266–2279. 2013. View Article : Google Scholar : PubMed/NCBI
9	Mázala DA, Grange RW and Chin ER: The role of proteases in excitation-contraction coupling failure in muscular dystrophy. Am J Physiol Cell Physiol. 308:C33–C40. 2015. View Article : Google Scholar :
10	Holland A, Murphy S, Dowling P and Ohlendieck K: Pathoproteomic profiling of the skeletal muscle matrisome in dystrophinopathy associated myofibrosis. Proteomics. 16:345–366. 2016. View Article : Google Scholar
11	Serra F, Quarta M, Canato M, Toniolo L, De Arcangelis V, Trotta A, Spath L, Monaco L, Reggiani C and Naro F: Inflammation in muscular dystrophy and the beneficial effects of non-steroidal anti-inflammatory drugs. Muscle Nerve. 46:773–784. 2012. View Article : Google Scholar : PubMed/NCBI
12	De Paepe B and De Bleecker JL: Cytokines and chemokines as regulators of skeletal muscle inflammation: Presenting the case of Duchenne muscular dystrophy. Mediators Inflamm. 540370:2013. View Article : Google Scholar : PubMed/NCBI
13	Mojumdar K, Liang F, Giordano C, Lemaire C, Danialou G, Okazaki T, Bourdon J, Rafei M, Galipeau J, Divangahi M and Petrof BJ: Inflammatory monocytes promote progression of Duchenne muscular dystrophy and can be therapeutically targeted via CCR2. EMBO Mol Med. 6:1476–1492. 2014. View Article : Google Scholar : PubMed/NCBI
14	Villalta SA, Rosenberg AS and Bluestone JA: The immune system in Duchenne muscular dystrophy: Friend or foe. Rare Dis. 3:e10109662015. View Article : Google Scholar : PubMed/NCBI
15	Tidball JG: Mechanisms of muscle injury, repair, and regeneration. Compr Physiol. 1:2029–2062. 2011.PubMed/NCBI
16	Rosenberg AS, Puig M, Nagaraju K, Hoffman EP, Villalta SA, Rao VA, Wakefield LM and Woodcock J: Immune-mediated pathology in Duchenne muscular dystrophy. Sci Transl Med. 7:299rv42015. View Article : Google Scholar : PubMed/NCBI
17	Villalta SA, Rosenthal W, Martinez L, Kaur A, Sparwasser T, Tidball JG, Margeta M, Spencer MJ and Bluestone JA: Regulatory T cells suppress muscle inflammation and injury in muscular dystrophy. Sci Transl Med. 6:258ra1422014. View Article : Google Scholar : PubMed/NCBI
18	Proud CM: 50 years ago in the Journal of Pediatrics: The use of serum creatine phosphokinase and other serum enzymes in the diagnosis of progressive muscular dystrophy. J Pediatr. 163:16562013. View Article : Google Scholar : PubMed/NCBI
19	Percy ME, Andrews DF and Thompson MW: Duchenne muscular dystrophy carrier detection using logistic discrimination: Serum creatine kinase, hemopexin, pyruvate kinase, and lactate dehydrogenase in combination. Am J Med Genet. 13:27–38. 1982. View Article : Google Scholar : PubMed/NCBI
20	Carter ND, Heath R, Jeffery S, Jackson MJ, Newham DJ and Edwards RH: Carbonic anhydrase III in Duchenne muscular dystrophy. Clin Chim Acta. 133:201–208. 1983. View Article : Google Scholar : PubMed/NCBI
21	Fröhlich T, Reitter B, Scheffner D, Schirmer RH and Untucht-Grau R: Muscle adenylate kinase in Duchenne muscular dystrophy. Biochim Biophys Acta. 883:598–603. 1986. View Article : Google Scholar : PubMed/NCBI
22	Percy ME, Chang LS, Murphy EG, Oss I, Verellen-Dumoulin C and Thompson MW: Serum creatine kinase and pyruvate kinase in Duchenne muscular dystrophy carrier detection. Muscle Nerve. 2:329–339. 1979. View Article : Google Scholar : PubMed/NCBI
23	D'Amore PA, Brown RH Jr, Ku PT, Hoffman EP, Watanabe H, Arahata K, Ishihara T and Folkman J: Elevated basic fibroblast growth factor in the serum of patients with Duchenne muscular dystrophy. Ann Neurol. 35:362–365. 1994. View Article : Google Scholar : PubMed/NCBI
24	Bernasconi P, Torchiana E, Confalonieri P, Brugnoni R, Barresi R, Mora M, Cornelio F, Morandi L and Mantegazza R: Expression of transforming growth factor-beta 1 in dystrophic patient muscles correlates with fibrosis. Pathogenetic role of a fibrogenic cytokine. J Clin Invest. 96:1137–1144. 1995. View Article : Google Scholar : PubMed/NCBI
25	Sun G, Haginoya K, Chiba Y, Uematsu M, Hino-Fukuyo N, Tanaka S, Onuma A, Iinuma K and Tsuchiya S: Elevated plasma levels of tissue inhibitors of metalloproteinase-1 and their overexpression in muscle in human and mouse muscular dystrophy. J Neurol Sci. 297:19–28. 2010. View Article : Google Scholar : PubMed/NCBI
26	Nadarajah VD, van Putten M, Chaouch A, Garrood P, Straub V, Lochmüller H, Ginjaar HB, Aartsma-Rus AM, van Ommen GJ, den Dunnen JT and 't Hoen PA: Serum matrix metalloproteinase-9 (MMP-9) as a biomarker for monitoring disease progression in Duchenne muscular dystrophy (DMD). Neuromuscul Disord. 21:569–578. 2011. View Article : Google Scholar : PubMed/NCBI
27	Holland A, Carberry S and Ohlendieck K: Proteomics of the dystrophin-glycoprotein complex and dystrophinopathy. Curr Protein Pept Sci. 14:680–697. 2013. View Article : Google Scholar : PubMed/NCBI
28	Fuller HR, Graham LC, Llavero Hurtado M and Wishart TM: Understanding the molecular consequences of inherited muscular dystrophies: Advancements through proteomic experimentation. Expert Rev Proteomics. 13:659–671. 2016. View Article : Google Scholar : PubMed/NCBI
29	Hathout Y, Seol H, Han MH, Zhang A, Brown KJ and Hoffman EP: Clinical utility of serum biomarkers in Duchenne muscular dystrophy. Clin Proteomics. 13:92016. View Article : Google Scholar : PubMed/NCBI
30	Alagaratnam S, Mertens BJ, Dalebout JC, Deelder AM, van Ommen GJ, den Dunnen JT and 't Hoen PA: Serum protein profiling in mice: Identification of Factor XIIIa as a potential biomarker for muscular dystrophy. Proteomics. 8:1552–1563. 2008. View Article : Google Scholar : PubMed/NCBI
31	Duguez S, Duddy W, Johnston H, Lainé J, Le Bihan MC, Brown KJ, Bigot A, Hathout Y, Butler-Browne G and Partridge T: Dystrophin deficiency leads to disturbance of LAMP1-vesicle-associated protein secretion. Cell Mol Life Sci. 70:2159–2174. 2013. View Article : Google Scholar : PubMed/NCBI
32	Hathout Y, Marathi RL, Rayavarapu S, Zhang A, Brown KJ, Seol H, Gordish-Dressman H, Cirak S, Bello L, Nagaraju K, et al: Discovery of serum protein biomarkers in the mdx mouse model and cross-species comparison to Duchenne muscular dystrophy patients. Hum Mol Genet. 23:6458–6469. 2014. View Article : Google Scholar : PubMed/NCBI
33	Cynthia Martin F, Hiller M, Spitali P, Oonk S, Dalebout H, Palmblad M, Chaouch A, Guglieri M, Straub V, Lochmüller H, et al: Fibronectin is a serum biomarker for Duchenne muscular dystrophy. Proteomics Clin Appl. 8:269–278. 2014. View Article : Google Scholar : PubMed/NCBI
34	Ayoglu B, Chaouch A, Lochmüller H, Politano L, Bertini E, Spitali P, Hiller M, Niks EH, Gualandi F, Pontén F, et al: Affinity proteomics within rare diseases: A BIO-NMD study for blood biomarkers of muscular dystrophies. EMBO Mol Med. 6:918–936. 2014. View Article : Google Scholar : PubMed/NCBI
35	Rouillon J, Zocevic A, Leger T, Garcia C, Camadro JM, Udd B, Wong B, Servais L, Voit T and Svinartchouk F: Proteomics profiling of urine reveals specific titin fragments as biomarkers of Duchenne muscular dystrophy. Neuromuscul Disord. 24:563–573. 2014. View Article : Google Scholar : PubMed/NCBI
36	Coenen-Stass AM, McClorey G, Manzano R, Betts CA, Blain A, Saleh AF, Gait MJ, Lochmüller H, Wood MJ and Roberts TC: Identification of novel, therapy-responsive protein biomarkers in a mouse model of Duchenne muscular dystrophy by aptamer-based serum proteomics. Sci Rep. 5:170142015. View Article : Google Scholar : PubMed/NCBI
37	Rouillon J, Poupiot J, Zocevic A, Amor F, Léger T, Garcia C, Camadro JM, Wong B, Pinilla R, Cosette J, et al: Serum proteomic profiling reveals fragments of MYOM3 as potential biomarkers for monitoring the outcome of therapeutic interventions in muscular dystrophies. Hum Mol Genet. 24:4916–4932. 2015. View Article : Google Scholar : PubMed/NCBI
38	Hathout Y, Brody E, Clemens PR, Cripe L, DeLisle RK, Furlong P, Gordish-Dressman H, Hache L, Henricson E, Hoffman EP, et al: Large-scale serum protein biomarker discovery in Duchenne muscular dystrophy. Proc Natl Acad Sci USA. 112:7153–7158. 2015. View Article : Google Scholar : PubMed/NCBI
39	Oonk S, Spitali P, Hiller M, Switzar L, Dalebout H, Calissano M, Lochmüller H, Aartsma-Rus A, 't Hoen PA and van der Burgt YE: Comparative mass spectrometric and immunoassay-based proteome analysis in serum of Duchenne muscular dystrophy patients. Proteomics Clin Appl. 10:290–299. 2016. View Article : Google Scholar
40	Im WB, Phelps SF, Copen EH, Adams EG, Slightom JL and Chamberlain JS: Differential expression of dystrophin isoforms in strains of mdx mice with different mutations. Hum Mol Genet. 5:1149–1153. 1996. View Article : Google Scholar : PubMed/NCBI
41	Danko I, Chapman V and Wolff JA: The frequency of revertants in mdx mouse genetic models for Duchenne muscular dystrophy. Pediatr Res. 32:128–131. 1992. View Article : Google Scholar : PubMed/NCBI
42	Mitrpant C, Fletcher S, Iversen PL and Wilton SD: By-passing the nonsense mutation in the 4 CV mouse model of muscular dystrophy by induced exon skipping. J Gene Med. 11:46–56. 2009. View Article : Google Scholar
43	Wang Y, Kinzie E, Berger FG, Lim SK and Baumann H: Haptoglobin, an inflammation-inducible plasma protein. Redox Rep. 6:379–385. 2001. View Article : Google Scholar
44	Chapman VM, Miller DR, Armstrong D and Caskey CT: Recovery of induced mutations for X chromosome-linked muscular dystrophy in mice. Proc Natl Acad Sci USA. 86:1292–1296. 1989. View Article : Google Scholar : PubMed/NCBI
45	Partridge TA: The mdx mouse model as a surrogate for Duchenne muscular dystrophy. FEBS J. 280:4177–4186. 2013. View Article : Google Scholar : PubMed/NCBI
46	Carberry S, Zweyer M, Swandulla D and Ohlendieck K: Comparative proteomic analysis of the contractile-protein-depleted fraction from normal versus dystrophic skeletal muscle. Anal Biochem. 446:108–115. 2014. View Article : Google Scholar
47	Hortin GL and Sviridov D: The dynamic range problem in the analysis of the plasma proteome. J Proteomics. 73:629–636. 2010. View Article : Google Scholar
48	Anderson L and Anderson NG: The human plasma proteome: History, character, and diagnostic prospects. Mol Cell Proteomics. 1:845–867. 2002. View Article : Google Scholar : PubMed/NCBI
49	Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 72:248–254. 1976. View Article : Google Scholar : PubMed/NCBI
50	Holland A, Henry M, Meleady P, Winkler CK, Krautwald M, Brinkmeier H and Ohlendieck K: Comparative label-free mass spectrometric analysis of mildly versus severely affected mdx mouse skeletal muscles identifies annexin, lamin, and vimentin as universal dystrophic markers. Molecules. 20:11317–11344. 2015. View Article : Google Scholar : PubMed/NCBI
51	Holland A, Dowling P, Meleady P, Henry M, Zweyer M, Mundegar RR, Swandulla D and Ohlendieck K: Label-free mass spectrometric analysis of the mdx-4cv diaphragm identifies the matricellular protein periostin as a potential factor involved in dystrophinopathy-related fibrosis. Proteomics. 15:2318–2331. 2015. View Article : Google Scholar : PubMed/NCBI
52	Murphy S, Zweyer M, Mundegar RR, Henry M, Meleady P, Swandulla D and Ohlendieck K: Concurrent label-free mass spectrometric analysis of dystrophin isoform Dp427 and the myofibrosis marker collagen in crude extracts from mdx-4cv skeletal muscles. Proteomes. 3:298–327. 2015. View Article : Google Scholar : PubMed/NCBI
53	Di Luca A, Henry M, Meleady P and O'Connor R: Label-free LC-MS analysis of HER2⁺ breast cancer cell line response to HER2 inhibitor treatment. Daru. 23:402015. View Article : Google Scholar
54	Linge A, Maurya P, Friedrich K, Baretton GB, Kelly S, Henry M, Clynes M, Larkin A and Meleady P: Identification and functional validation of RAD23B as a potential protein in human breast cancer progression. J Proteome Res. 13:3212–3222. 2014. View Article : Google Scholar : PubMed/NCBI
55	Murphy S, Dowling P, Zweyer M, Mundegar RR, Henry M, Meleady P, Swandulla D and Ohlendieck K: Proteomic analysis of dystrophin deficiency and associated changes in the aged mdx-4cv heart model of dystrophinopathy-related cardiomyopathy. J Proteomics. 145:24–36. 2016. View Article : Google Scholar : PubMed/NCBI
56	Mi H, Muruganujan A and Thomas PD: PANTHER in 2013: Modeling the evolution of gene function, and other gene attributes, in the context of phylogenetic trees. Nucleic Acids Res. 41(D1): D377–D386. 2013. View Article : Google Scholar :
57	Staunton L, Zweyer M, Swandulla D and Ohlendieck K: Mass spectrometry-based proteomic analysis of middle-aged vs. aged vastus lateralis reveals increased levels of carbonic anhydrase isoform 3 in senescent human skeletal muscle. Int J Mol Med. 30:723–733. 2012.PubMed/NCBI
58	Holland A, Dowling P, Zweyer M, Swandulla D, Henry M, Clynes M and Ohlendieck K: Proteomic profiling of cardiomyopathic tissue from the aged mdx model of Duchenne muscular dystrophy reveals a drastic decrease in laminin, nidogen and annexin. Proteomics. 13:2312–2323. 2013. View Article : Google Scholar : PubMed/NCBI
59	Lewis C, Jockusch H and Ohlendieck K: Proteomic profiling of the dystrophin-deficient MDX heart reveals drastically altered levels of key metabolic and contractile proteins. J Biomed Biotechnol. 2010:6485012010. View Article : Google Scholar : PubMed/NCBI
60	Murphy S, Henry M, Meleady P, Zweyer M, Mundegar RR, Swandulla D and Ohlendieck K: Simultaneous pathoproteomic evaluation of the dystrophin-glycoprotein complex and secondary changes in the mdx-4cv mouse model of Duchenne muscular dystrophy. Biology (Basel). 4:397–423. 2015.
61	Murphy S, Zweyer M, Henry M, Meleady P, Mundegar RR, Swandulla D and Ohlendieck K: Label-free mass spectrometric analysis reveals complex changes in the brain proteome from the mdx-4cv mouse model of Duchenne muscular dystrophy. Clin Proteomics. 12:272015. View Article : Google Scholar : PubMed/NCBI
62	Dowling P, Holland A and Ohlendieck K: Mass spectrometry-based identification of muscle-associated and muscle-derived proteomic biomarkers of dystrophinopathies. J Neuromuscul Dis. 1:15–40. 2014.PubMed/NCBI
63	Gianazza E, Miller I, Palazzolo L, Parravicini C and Eberini I: With or without you - Proteomics with or without major plasma/serum proteins. J Proteomics. 140:62–80. 2016. View Article : Google Scholar : PubMed/NCBI
64	Omenn GS, States DJ, Adamski M, Blackwell TW, Menon R, Hermjakob H, Apweiler R, Haab BB, Simpson RJ, Eddes JS, et al: Overview of the HUPO Plasma Proteome Project: Results from the pilot phase with 35 collaborating laboratories and multiple analytical groups, generating a core dataset of 3020 proteins and a publicly-available database. Proteomics. 5:3226–3245. 2005. View Article : Google Scholar : PubMed/NCBI
65	Pietrowska M, Marczak L, Polanska J, Behrendt K, Nowicka E, Walaszczyk A, Chmura A, Deja R, Stobiecki M, Polanski A, et al: Mass spectrometry-based serum proteome pattern analysis in molecular diagnostics of early stage breast cancer. J Transl Med. 7:602009. View Article : Google Scholar : PubMed/NCBI
66	Smith MP, Wood SL, Zougman A, Ho JT, Peng J, Jackson D, Cairns DA, Lewington AJ, Selby PJ and Banks RE: A systematic analysis of the effects of increasing degrees of serum immunodepletion in terms of depth of coverage and other key aspects in top-down and bottom-up proteomic analyses. Proteomics. 11:2222–2235. 2011. View Article : Google Scholar : PubMed/NCBI
67	Dowling P, Hayes C, Ting KR, Hameed A, Meiller J, Mitsiades C, Anderson KC, Clynes M, Clarke C, Richardson P and O'Gorman P: Identification of proteins found to be significantly altered when comparing the serum proteome from Multiple Myeloma patients with varying degrees of bone disease. BMC Genomics. 15:9042014. View Article : Google Scholar : PubMed/NCBI
68	Araújo JE, Jorge S, Teixeira E, Costa F, Ramos A, Lodeiro C, Santos HM and Capelo JL: A cost-effective method to get insight into the peritoneal dialysate effluent proteome. J Proteomics. 145:207–213. 2016. View Article : Google Scholar : PubMed/NCBI
69	Gundry RL, Fu Q, Jelinek CA, Van Eyk JE and Cotter RJ: Investigation of an albumin-enriched fraction of human serum and its albuminome. Proteomics Clin Appl. 1:73–88. 2007. View Article : Google Scholar : PubMed/NCBI
70	Gundry RL, White MY, Nogee J, Tchernyshyov I and Van Eyk JE: Assessment of albumin removal from an immunoaffinity spin column: Critical implications for proteomic examination of the albuminome and albumin-depleted samples. Proteomics. 9:2021–2028. 2009. View Article : Google Scholar : PubMed/NCBI
71	Tirumalai RS, Chan KC, Prieto DA, Issaq HJ, Conrads TP and Veenstra TD: Characterization of the low molecular weight human serum proteome. Mol Cell Proteomics. 2:1096–1103. 2003. View Article : Google Scholar : PubMed/NCBI
72	Rayavarapu S, Coley W, Cakir E, Jahnke V, Takeda S, Aoki Y, Grodish-Dressman H, Jaiswal JK, Hoffman EP, Brown KJ, et al: Identification of disease specific pathways using in vivo SILAC proteomics in dystrophin deficient mdx mouse. Mol Cell Proteomics. 12:1061–1073. 2013. View Article : Google Scholar : PubMed/NCBI
73	Roberts TC, Johansson HJ, McClorey G, Godfrey C, Blomberg KE, Coursindel T, Gait MJ, Smith CI, Lehtiö J, El Andaloussi S and Wood MJ: Multi-level omics analysis in a murine model of dystrophin loss and therapeutic restoration. Hum Mol Genet. 24:6756–6768. 2015. View Article : Google Scholar : PubMed/NCBI
74	Turk R, Hsiao JJ, Smits MM, Ng BH, Pospisil TC, Jones KS, Campbell KP and Wright ME: Molecular signatures of membrane protein complexes underlying muscular dystrophy. Mol Cell Proteomics. 15:2169–2185. 2016. View Article : Google Scholar : PubMed/NCBI
75	Schrödl W, Büchler R, Wendler S, Reinhold P, Muckova P, Reindl J and Rhode H: Acute phase proteins as promising biomarkers: Perspectives and limitations for human and veterinary medicine. Proteomics Clin Appl. 10:1077–1092. 2016. View Article : Google Scholar : PubMed/NCBI
76	Ohlendieck K: Proteomic identification of biomarkers of skeletal muscle disorders. Biomarkers Med. 7:169–186. 2013. View Article : Google Scholar
77	Levy AP, Asleh R, Blum S, Levy NS, Miller-Lotan R, Kalet-Litman S, Anbinder Y, Lache O, Nakhoul FM, Asaf, et al: Haptoglobin: Basic and clinical aspects. Antioxid Redox Signal. 12:293–304. 2010. View Article : Google Scholar
78	John HA and Purdom IF: Elevated plasma levels of haptoglobin in Duchenne muscular dystrophy: Electrophoretic variants in patients with a severe form of the disease. Electrophoresis. 10:489–493. 1989. View Article : Google Scholar : PubMed/NCBI
79	Górecki DC: Dystrophin: The dead calm of a dogma. Rare Dis. 4:e11537772016. View Article : Google Scholar : PubMed/NCBI
80	Kharraz Y, Guerra J, Mann CJ, Serrano AL and Muñoz-Cánoves P: Macrophage plasticity and the role of inflammation in skeletal muscle repair. Mediators Inflamm. 2013:4914972013. View Article : Google Scholar : PubMed/NCBI
81	Sinadinos A, Young CN, Al-Khalidi R, Teti A, Kalinski P, Mohamad S, Floriot L, Henry T, Tozzi G, Jiang T, et al: 2R X7 purinoceptor: A therapeutic target for ameliorating the symptoms of duchenne muscular dystrophy. PLoS Med. 12:e10018882015. View Article : Google Scholar
82	Veenstra TD, Conrads TP, Hood BL, Avellino AM, Ellenbogen RG and Morrison RS: Biomarkers: Mining the biofluid proteome. Mol Cell Proteomics. 4:409–418. 2005. View Article : Google Scholar : PubMed/NCBI
83	Savino R, Paduano S, Preianò M and Terracciano R: The proteomics big challenge for biomarkers and new drug-targets discovery. Int J Mol Sci. 13:13926–13948. 2012. View Article : Google Scholar : PubMed/NCBI
84	Stastna M and Van Eyk JE: Secreted proteins as a fundamental source for biomarker discovery. Proteomics. 12:722–735. 2012. View Article : Google Scholar : PubMed/NCBI
85	Drucker E and Krapfenbauer K: Pitfalls and limitations in translation from biomarker discovery to clinical utility in predictive and personalised medicine. EPMA J. 4:72013. View Article : Google Scholar : PubMed/NCBI
86	Sun L, Hu S, Yu L, Guo C, Sun L, Yang Z, Qi J and Ran Y: Serum haptoglobin as a novel molecular biomarker predicting colorectal cancer hepatic metastasis. Int J Cancer. 138:2724–2731. 2016. View Article : Google Scholar : PubMed/NCBI