Restricted accessResearch articleFirst published online 2018-07
Cardiac-Specific Expression of ΔH2-R15 Mini-Dystrophin Normalized All Electrocardiogram Abnormalities and the End-Diastolic Volume in a 23-Month-Old Mouse Model of Duchenne Dilated Cardiomyopathy
Heart disease is a major health threat for Duchenne/Becker muscular dystrophy patients and carriers. Expression of a 6–8 kb mini-dystrophin gene in the heart holds promise to change the disease course dramatically. However, the mini-dystrophin gene cannot be easily studied with adeno-associated virus (AAV) gene delivery because the size of the minigene exceeds AAV packaging capacity. Cardiac protection of the ΔH2-R19 minigene was previously studied using the cardiac-specific transgenic approach. Although this minigene fully normalized skeletal muscle force, it only partially corrected electrocardiogram and heart hemodynamics in dystrophin-null mdx mice that had moderate cardiomyopathy. This study evaluated the ΔH2-R15 minigene using the same transgenic approach in mdx mice that had more severe cardiomyopathy. In contrast to the ΔH2-R19 minigene, the ΔH2-R15 minigene carries dystrophin spectrin-like repeats 16 to 19 (R16–19), a region that has been suggested to protect the heart in clinical studies. Cardiac expression of the ΔH2-R15 minigene normalized all aberrant electrocardiogram changes and improved hemodynamics. Importantly, it corrected the end-diastolic volume, an important diastolic parameter not rescued by ΔH2-R19 mini-dystrophin. It is concluded that that ΔH2-R15 mini-dystrophin is a superior candidate gene for heart protection. This finding has important implications in the design of the mini/micro-dystrophin gene for Duchenne cardiomyopathy therapy.
Get full access to this article
View all access options for this article.
References
1.
BaxterP. Treatment of the heart in Duchenne muscular dystrophy. Dev Med Child Neurol, 2006; 48:163.
American Academy of Pediatrics Section on Cardiology and Cardiac Surgery. Cardiovascular health supervision for individuals affected by Duchenne or Becker muscular dystrophy. Pediatrics, 2005; 116:1569–1573.
4.
DuanD. Challenges and opportunities in dystrophin-deficient cardiomyopathy gene therapy. Hum Mol Genet, 2006; 15:R253–261.
5.
LaiY, DuanD. Progress in gene therapy of dystrophic heart disease. Gene Ther, 2012; 19:678–685.
6.
YueY, BinalsheikhIM, LeachSB, et al.Prospect of gene therapy for cardiomyopathy in hereditary muscular dystrophy. Exp Opin Orphan Drugs, 2016; 4:169–183.
7.
GregorevicP, BlankinshipMJ, AllenJM, et al.Systemic delivery of genes to striated muscles using adeno-associated viral vectors. Nat Med, 2004; 10:828–834.
8.
WangZ, ZhuT, QiaoC, et al.Adeno-associated virus serotype 8 efficiently delivers genes to muscle and heart. Nat Biotechnol, 2005; 23:321–328.
9.
YueY, GhoshA, LongC, et al.A single intravenous injection of adeno-associated virus serotype-9 leads to whole body skeletal muscle transduction in dogs. Mol Ther, 2008; 16:1944–1952.
10.
YueY, PanX, HakimCH, et al.Safe and bodywide muscle transduction in young adult Duchenne muscular dystrophy dogs with adeno-associated virus. Hum Mol Genet, 2015; 24:5880–5890.
HarperSQ, HauserMA, DelloRussoC, et al.Modular flexibility of dystrophin: implications for gene therapy of Duchenne muscular dystrophy. Nat Med, 2002; 8:253–261.
13.
EnglandSB, NicholsonLV, JohnsonMA, et al.Very mild muscular dystrophy associated with the deletion of 46% of dystrophin. Nature, 1990; 343:180–182.
14.
LaiY, ThomasGD, YueY, et al.Dystrophins carrying spectrin-like repeats 16 and 17 anchor nNOS to the sarcolemma and enhance exercise performance in a mouse model of muscular dystrophy. J Clin Invest, 2009; 119:624–635.
15.
BostickB, YueY, LongC, et al.Cardiac expression of a mini-dystrophin that normalizes skeletal muscle force only partially restores heart function in aged mdx mice. Mol Ther, 2009; 17:253–261.
16.
NigroG, ComiLI, PolitanoL, et al.The incidence and evolution of cardiomyopathy in Duchenne muscular dystrophy. Int J Cardiol, 1990; 26:271–277.
17.
PapaAA, D'AmbrosioP, PetilloR, et al.Heart transplantation in patients with dystrophinopathic cardiomyopathy: review of the literature and personal series. Intractable Rare Dis Res, 2017; 6:95–101.
18.
BostickB, YueY, DuanD. Gender influences cardiac function in the mdx model of Duchenne cardiomyopathy. Muscle Nerve, 2010; 42:600–603.
19.
ArbustiniE, DiegoliM, MorbiniP, et al.Prevalence and characteristics of dystrophin defects in adult male patients with dilated cardiomyopathy. J Am Coll Cardiol, 2000; 35:1760–1768.
20.
PalmucciL, DoriguzziC, MonginiT, et al.Dilating cardiomyopathy as the expression of Xp21 Becker type muscular dystrophy. J Neurol Sci, 1992; 111:218–221.
21.
MelaciniP, FaninM, DanieliGA, et al.Cardiac involvement in Becker muscular dystrophy. J Am Coll Cardiol, 1993; 22:1927–1934.
22.
YoshidaK, IkedaS, NakamuraA, et al.Molecular analysis of the Duchenne muscular dystrophy gene in patients with Becker muscular dystrophy presenting with dilated cardiomyopathy. Muscle Nerve, 1993; 16:1161–1166.
23.
NigroG, PolitanoL, NigroV, et al.Mutation of dystrophin gene and cardiomyopathy. Neuromuscul Disord, 1994; 4:371–379.
24.
PiccoloG, AzanG, ToninP, et al.Dilated cardiomyopathy requiring cardiac transplantation as initial manifestation of Xp21 Becker type muscular dystrophy. Neuromuscul Disord, 1994; 4:143–146.
25.
SicilianoG, FaninM, AngeliniC, et al.Prevalent cardiac involvement in dystrophin Becker type mutation. Neuromuscul Disord, 1994; 4:381–386.
26.
MuntoniF, Di LenardaA, PorcuM, et al.Dystrophin gene abnormalities in two patients with idiopathic dilated cardiomyopathy. Heart, 1997; 78:608–612.
27.
MelisMA, CauM, DeiddaF, et al.Mutation of dystrophin gene in two families with X-linked dilated cardiomyopathy. Neuromuscul Disord, 1998; 8:244.
28.
TodorovaA, ConstantinovaD, KremenskyI. Dilated cardiomyopathy and new 16 bp deletion in exon 44 of the Dystrophin gene: the possible role of repeated motifs in mutation generation. Am J Med Genet A, 2003; 120A:5–7.
29.
NakamuraA, YoshidaK, FukushimaK, et al.Follow-up of three patients with a large in-frame deletion of exons 45–55 in the Duchenne muscular dystrophy (DMD) gene. J Clin Neurosci, 2008; 15:757–763.
30.
MiyazakiD, YoshidaK, FukushimaK, et al.Characterization of deletion breakpoints in patients with dystrophinopathy carrying a deletion of exons 45–55 of the Duchenne muscular dystrophy (DMD) gene. J Hum Genet, 2009; 54:127–130.
31.
YazakiM, YoshidaK, NakamuraA, et al.Clinical characteristics of aged Becker muscular dystrophy patients with onset after 30 years. Eur Neurol, 1999; 42:145–149.
32.
TasakiN, YoshidaK, HarutaSI, et al.X-linked dilated cardiomyopathy with a large hot-spot deletion in the dystrophin gene. Intern Med, 2001; 40:1215–1221.
33.
KasparRW, AllenHD, RayWC, et al.Analysis of dystrophin deletion mutations predicts age of cardiomyopathy onset in Becker muscular dystrophy. Circ Cardiovasc Genet, 2009; 2:544–551.
34.
BostickB, YueY, LongC, et al.Prevention of dystrophin-deficient cardiomyopathy in twenty-one-month-old carrier mice by mosaic dystrophin expression or complementary dystrophin/utrophin expression. Circ Res, 2008; 102:121–130.
35.
WasalaNB, ZhangK, WasalaLP, et al.The FVB background does not dramatically alter the dystrophic phenotype of mdx mice. PLoS Curr, 2015; 7:pii: ecurrents.md.28266819ca28266810ec28266815fefcac28266767ea28266819a28263461c.
36.
YueY, SkimmingJW, LiuM, et al.Full-length dystrophin expression in half of the heart cells ameliorates beta-isoproterenol-induced cardiomyopathy in mdx mice. Hum Mol Genet, 2004; 13:1669–1675.
37.
KodippiliK, VinceL, ShinJH, et al.Characterization of 65 epitope-specific dystrophin monoclonal antibodies in canine and murine models of Duchenne muscular dystrophy by immunostaining and western blot. PLoS One, 2014; 9:e88280.
38.
WasalaNB, BostickB, YueY, et al.Exclusive skeletal muscle correction does not modulate dystrophic heart disease in the aged mdx model of Duchenne cardiomyopathy. Hum Mol Genet, 2013; 22:2634–2641.
39.
LiD, YueY, LaiY, et al.Nitrosative stress elicited by nNOSmu delocalization inhibits muscle force in dystrophin-null mice. J Pathol, 2011; 223:88–98.
DuanD, Rafael-FortneyJA, BlainA, et al.Standard Operating Procedures (SOPs) for evaluating the heart in preclinical studies of Duchenne muscular dystrophy. J Cardiovasc Transl Res, 2016; 9:85–86.
42.
BostickB, ShinJ-H, YueY, et al.AAV-microdystrophin therapy improves cardiac performance in aged female mdx mice. Mol Ther, 2011; 19:1826–1832.
43.
MitchellGF, JeronA, KorenG. Measurement of heart rate and Q-T interval in the conscious mouse. Am J Physiol, 1998; 274:H747–751.
44.
NigroG, ComiLI, PolitanoL, et al.Cardiomyopathies associated with muscular dystrophies. In: EngelA, Franzini-ArmstrongC, eds. Myology: Basic and Clinical. New York: McGraw-Hill, Medical Publishing Division, 2004:1239–1256.
45.
YueY, WasalaNB, BostickB, et al.100-fold but not 50-fold dystrophin overexpression aggravates electrocardiographic defects in the mdx model of Duchenne muscular dystrophy. Mol Ther Methods Clin Dev, 2016; 3:16045.
46.
YinFC, SpurgeonHA, RakusanK, et al.Use of tibial length to quantify cardiac hypertrophy: application in the aging rat. Am J Physiol, 1982; 243:H941–947.
47.
RohiniA, AgrawalN, KoyaniCN, et al.Molecular targets and regulators of cardiac hypertrophy. Pharmacol Res, 2010; 61:269–280.
48.
BalakumarP, JagadeeshG. Multifarious molecular signaling cascades of cardiac hypertrophy: can the muddy waters be cleared?. Pharmacol Res, 2010; 62:365–383.
49.
BarrySP, DavidsonSM, TownsendPA. Molecular regulation of cardiac hypertrophy. Int J Biochem Cell Biol, 2008; 40:2023–2039.
50.
HeinekeJ, MolkentinJD. Regulation of cardiac hypertrophy by intracellular signalling pathways. Nat Rev Mol Cell Biol, 2006; 7:589–600.
YokotaT, WangY. p38 MAP kinases in the heart. Gene, 2016; 575:369–376.
53.
MarberMS, RoseB, WangY. The p38 mitogen-activated protein kinase pathway—a potential target for intervention in infarction, hypertrophy, and heart failure. J Mol Cell Cardiol, 2011; 51:485–490.
54.
LiangQ, MolkentinJD. Redefining the roles of p38 and JNK signaling in cardiac hypertrophy: dichotomy between cultured myocytes and animal models. J Mol Cell Cardiol, 2003; 35:1385–1394.
PillaiVB, SundaresanNR, GuptaMP. Regulation of Akt signaling by sirtuins: its implication in cardiac hypertrophy and aging. Circ Res, 2014; 114:368–378.
57.
SalazarNC, ChenJ, RockmanHA. Cardiac GPCRs: GPCR signaling in healthy and failing hearts. Biochim Biophys Acta, 2007; 1768:1006–1018.
58.
KangM, ChungKY, WalkerJW. G-protein coupled receptor signaling in myocardium: not for the faint of heart. Physiology (Bethesda), 2007; 22:174–184.
59.
EspositoG, RapacciuoloA, Naga PrasadSV, et al.Cardiac hypertrophy: role of G protein-coupled receptors. J Card Fail, 2002; 8:S409–414.
60.
MailletM, van BerloJH, MolkentinJD. Molecular basis of physiological heart growth: fundamental concepts and new players. Nat Rev Mol Cell Biol, 2013; 14:38–48.
61.
CrispA, YinH, GoyenvalleA, et al.Diaphragm rescue alone prevents heart dysfunction in dystrophic mice. Hum Mol Genet, 2011; 20:413–421.
62.
TownsendD, YasudaS, ChamberlainJ, et al.Cardiac consequences to skeletal muscle-centric therapeutics for Duchenne muscular dystrophy. Trends Cardiovasc Med, 2009; 19:50–55.
63.
LaiY, ZhaoJ, YueY, et al.alpha2 and alpha3 helices of dystrophin R16 and R17 frame a microdomain in the alpha1 helix of dystrophin R17 for neuronal NOS binding. Proc Natl Acad Sci U S A, 2013; 110:525–530.
64.
XuKY, HusoDL, DawsonTM, et al.Nitric oxide synthase in cardiac sarcoplasmic reticulum. Proc Natl Acad Sci U S A, 1999; 96:657–662.
65.
KanaiAJ, PearceLL, ClemensPR, et al.Identification of a neuronal nitric oxide synthase in isolated cardiac mitochondria using electrochemical detection. Proc Natl Acad Sci U S A, 2001; 98:14126–14131.
66.
JohnsonEK, ZhangL, AdamsME, et al.Proteomic analysis reveals new cardiac-specific dystrophin-associated proteins. PLoS One, 2012; 7:e43515.
67.
LaiY, ZhaoJ, YueY, et al.Partial restoration of cardiac function with ΔPDZ nNOS in aged mdx model of Duchenne cardiomyopathy. Hum Mol Genet, 2014; 23:3189–3199.
68.
MengH, LeddyJJ, FrankJ, et al.The association of cardiac dystrophin with myofibrils/Z-disc regions in cardiac muscle suggests a novel role in the contractile apparatus. J Biol Chem, 1996; 271:12364–12371.
69.
ZhangY, YueY, LiL, et al.Dual AAV therapy ameliorates exercise-induced muscle injury and functional ischemia in murine models of Duchenne muscular dystrophy. Hum Mol Genet, 2013; 22:3720–3729.
70.
PillersDA. Dystrophin and the retina. Mol Genet Metab, 1999; 68:304–309.
71.
RicottiV, MandyWP, ScotoM, et al.Neurodevelopmental, emotional, and behavioural problems in Duchenne muscular dystrophy in relation to underlying dystrophin gene mutations. Dev Med Child Neurol, 2016; 58:77–84.
72.
RajabA, StraubV, McCannLJ, et al.Fatal cardiac arrhythmia and long-QT syndrome in a new form of congenital generalized lipodystrophy with muscle rippling (CGL4) due to PTRF-CAVIN mutations. PLoS genetics, 2010; 6:e1000874.
73.
TaniguchiT, MaruyamaN, OgataT, et al.PTRF/Cavin-1 deficiency causes cardiac dysfunction accompanied by cardiomyocyte hypertrophy and cardiac fibrosis. PLoS One, 2016; 11:e0162513.
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
