Short hairpin RNA treatment improves gait in a mouse model of Charcot‑Marie‑Tooth disease type 1A

Doo,Hyun  Myung; Hong,Young Bin; Han,Jiyou; Moon,Hyo Won; Hwang,Hyun; Kwak,Geon; Nam,Soo   Hyun; Kim,Sang   Beom; Chung,Ki   Wha; Kim,Jong   Hyun; Choi,Byung‑Ok

doi:10.3892/mmr.2020.11579

Short hairpin RNA treatment improves gait in a mouse model of Charcot‑Marie‑Tooth disease type 1A

Authors:
- Hyun Myung Doo
- Young Bin Hong
- Jiyou Han
- Hyo Won Moon
- Hyun Hwang
- Geon Kwak
- Soo Hyun Nam
- Sang Beom Kim
- Ki Wha Chung
- Jong Hyun Kim
- Byung‑Ok Choi
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Affiliations: Department of Health Sciences and Technology, Samsung Advanced Institute for Health Science and Technology (SAIHST), Sungkyunkwan University, Seoul 06351, Republic of Korea, Department of Biochemistry, College of Medicine, Dong‑A University, Busan, South Gyeongsang 49201, Republic of Korea, Department of Biological Sciences, Laboratory of Stem Cell Research and Biotechnology, Hyupsung University, Hwasung‑si, Gyeonggi 18330, Republic of Korea, Department of Neurology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul 06351, Republic of Korea, Stem Cell and Regenerative Medicine Institute, Samsung Medical Center, Seoul 06351, Republic of Korea, Department of Neurology, Kyung Hee University Hospital at Gangdong, Kyung Hee University School of Medicine, Seoul 05278, Republic of Korea, Department of Biological Sciences, Kongju National University, Gongju, Chungcheong 32588, Republic of Korea
Published online on: October 11, 2020 https://doi.org/10.3892/mmr.2020.11579
Pages: 4947-4955

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Abstract

Charcot‑Marie‑Tooth disease (CMT) is the most common inherited neurological disorder of the peripheral nervous system. The major subtype, CMT type 1A (CMT1A), accounts for ~40% of CMT cases and is characterized by distal muscle atrophy and gait disturbances. Short hairpin (sh) RNA sequences are potentially advantageous therapeutic tools for distal muscle atrophy‑induced gait disturbance. Therefore, the current study focused on the effects of an optimal shRNA injection using the myostatin (mstn) gene inhibition system. shLenti‑Mstn A demonstrated significant suppression of endogenous mstn gene expression (>40%) via RT‑qPCR following direct injection into the gastrocnemius and rectus femoris of the hind limb in C22 mice. The results also reported that shLenti‑Mstn A treatment increased muscle mass and size of the hind limbs compared with mock‑treated mice via measurement of the mass of injected muscles and magnetic resonance imaging study. Furthermore, electrophysiological measurement using a Nicolet Viking Quest device revealed significantly improved compound muscle action potential (CMAP) in shLenti‑Mstn A‑treated mice compared with the mock group (P<0.05) whereas nerve conduction velocity (NCV) showed no difference between groups. The shLenti‑Mstn A treatment directly affected increased muscle regeneration, including mass and size, but not regeneration of peripheral nerve. Additionally, shLenti‑Mstn A treatment significantly enhanced mobility, including locomotor coordination (P<0.01) and grip strength of the hindlimbs (P<0.01). Furthermore, MotoRater analysis using real‑time recording with a high‑speed camera revealed that shLenti‑Mstn‑treated mice exhibited an improved walking pattern in terms of step length, base support and duty factor compared with the mock group. It was hypothesized that treatment with shLenti‑Mstn A may provide a novel therapeutic strategy for improving gait in patients with CMT1A.

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1	Skre H: Genetic and clinical aspects of Charcot-Marie-Tooth's disease. Clin Genet. 6:98–118. 1974. View Article : Google Scholar : PubMed/NCBI
2	Levitan IB and Kaczmarek LK: Intercellular Communication. The Neuron: Cell and Molecular Biology. 4th edition. Oxford University Press; New York, NY: pp. 153–328. 2015, View Article : Google Scholar
3	Rotthier A, Baets J, Timmerman V and Janssens K: Mechanisms of disease in hereditary sensory and autonomic neuropathies. Nat Rev Neurol. 8:73–85. 2012. View Article : Google Scholar : PubMed/NCBI
4	Rossor AM, Kalmar B, Greensmith L and Reilly MM: The distal hereditary motor neuropathies. J Neurol Neurosurg Psychiatr. 83:6–14. 2012. View Article : Google Scholar
5	Harding AE and Thomas PK: The clinical features of hereditary motor and sensory neuropathy types I and II. Brain. 103:259–280. 1980. View Article : Google Scholar : PubMed/NCBI
6	Kim JY, Woo SY, Hong YB, Choi H, Kim J, Choi H, Mook-Jung I, Ha N, Kyung J, Koo SK, et al: HDAC6 inhibitors rescued the defective axonal mitochondrial movement in motor neurons derived from the induced pluripotent stem cells of peripheral neuropathy patients with HSPB1 mutation. Stem Cells Int. 2016:94759812016. View Article : Google Scholar : PubMed/NCBI
7	Fledrich R, Stassart RM and Sereda MW: Murine therapeutic models for Charcot-Marie-Tooth (CMT) disease. Br Med Bull. 102:89–113. 2012. View Article : Google Scholar : PubMed/NCBI
8	Robertson AM, Perea J, McGuigan A, King RH, Muddle JR, Gabreels-Festen AA, Thomas PK and Huxley C: Comparison of a new pmp22 transgenic mouse line with other mouse models and human patients with CMT1A. J Anat. 200:377–390. 2002. View Article : Google Scholar : PubMed/NCBI
9	Meyer zu Horste G, Prukop T, Liebetanz D, Mobius W, Nave KA and Sereda MW: Antiprogesterone therapy uncouples axonal loss from demyelination in a transgenic rat model of CMT1A neuropathy. Ann Neurol. 61:61–72. 2007. View Article : Google Scholar : PubMed/NCBI
10	Sereda MW, Meyer zu Horste G, Suter U, Uzma N and Nave KA: Therapeutic administration of progesterone antagonist in a model of Charcot-Marie-Tooth disease (CMT-1A). Nat Med. 9:1533–1537. 2003. View Article : Google Scholar : PubMed/NCBI
11	Passage E, Norreel JC, Noack-Fraissignes P, Sanguedolce V, Pizant J, Thirion X, Robaglia-Schlupp A, Pellissier JF and Fontes M: Ascorbic acid treatment corrects the phenotype of a mouse model of Charcot-Marie-Tooth disease. Nat Med. 10:396–401. 2004. View Article : Google Scholar : PubMed/NCBI
12	Lee JS, Kwak G, Kim HJ, Park HT, Choi BO and Hong YB: MiR-381 attenuates peripheral neuropathic phenotype caused by overexpression of PMP22. Exp Neurobiol. 28:279–288. 2019. View Article : Google Scholar : PubMed/NCBI
13	Lee JS, Lee JY, Song DW, Bae HS, Doo HM, Yu HS, Lee KJ, Kim HK, Hwang H, Kwak G, et al: Targeted PMP22 TATA-box editing by CRISPR/Cas9 reduces demyelinating neuropathy of Charcot-Marie-Tooth disease type 1A in mice. Nucleic Acids Res. 48:130–140. 2020.PubMed/NCBI
14	Robaglia-Schlupp A, Pizant J, Norreel JC, Passage E, Saberan-Djoneidi D, Ansaldi JL, Vinay L, Figarella-Branger D, Levy N, Clarac F, et al: PMP22 overexpression causes dysmyelination in mice. Brain. 125:2213–2221. 2002. View Article : Google Scholar : PubMed/NCBI
15	McPherron AC, Lawler AM and Lee SJ: Regulation of skeletal muscle mass in mice by a new TGF-beta superfamily member. Nature. 387:83–90. 1997. View Article : Google Scholar : PubMed/NCBI
16	Lee SJ and McPherron AC: Regulation of myostatin activity and muscle growth. Proc Natl Acad Sci USA. 98:9306–9311. 2001. View Article : Google Scholar : PubMed/NCBI
17	Mariot V, Joubert R, Hourde C, Feasson L, Hanna M, Muntoni F, Maisonobe T, Servais L, Bogni C, Le Panse R, et al: Downregulation of myostatin pathway in neuromuscular diseases may explain challenges of anti-myostatin therapeutic approaches. Nat Commun. 8:18592017. View Article : Google Scholar : PubMed/NCBI
18	Wagner KR, McPherron AC, Winik N and Lee SJ: Loss of myostatin attenuates severity of muscular dystrophy in mdx mice. Ann Neurol. 52:832–836. 2002. View Article : Google Scholar : PubMed/NCBI
19	Holzbaur EL, Howland DS, Weber N, Wallace KE, She Y, Kwak S, Tchistiakova LA, Murphy E, Hinson J, Karim R, et al: Myostatin inhibition slows muscle atrophy in rodent models of amyotrophic lateral sclerosis. Neurobiol Dis. 23:697–707. 2006. View Article : Google Scholar : PubMed/NCBI
20	Hennebry A, Oldham J, Shavlakadze T, Grounds MD, Sheard P, Fiorotto ML, Falconer S, Smith HK, Berry C, Jeanplong F, et al: IGF1 stimulates greater muscle hypertrophy in the absence of myostatin in male mice. J Endocrinol. 234:187–200. 2017. View Article : Google Scholar : PubMed/NCBI
21	Dorman CW, Krug HE, Frizelle SP, Funkenbusch S and Mahowald ML: A comparison of DigiGait™ and TreadScan™ imaging systems: Assessment of pain using gait analysis in murine monoarthritis. J Pain Res. 7:25–35. 2013.PubMed/NCBI
22	Xu Y, Tian N, Bai Q, Chen Q, Sun XH and Wang Y: Gait assessment of pain and analgesics: Comparison of the DigiGait™ and CatWalk™ gait imaging systems. Neurosci Bull. 35:401–418. 2019. View Article : Google Scholar : PubMed/NCBI
23	Mancuso R, Olivan S, Osta R and Navarro X: Evolution of gait abnormalities in SOD1(G93A) transgenic mice. Brain Res. 1406:65–73. 2011. View Article : Google Scholar : PubMed/NCBI
24	DiGiusto DL, Krishnan A, Li L, Li H, Li S, Rao A, Mi S, Yam P, Stinson S, Kalos M, et al: RNA-based gene therapy for HIV with lentiviral vector-modified CD34(+) cells in patients undergoing transplantation for AIDS-related lymphoma. Sci Transl Med. 2:36ra432010. View Article : Google Scholar : PubMed/NCBI
25	Maples PB, Kumar P, Yu Y, Wang Z, Jay C, Pappen BO, Rao DD, Kuhn J, Nemunaitis J and Senzer N: FANG Vaccine: Autologous tumor cell vaccine genitically modified to express GM-CSF and block production of furin. BioProcess J. 8:4–14. 2009. View Article : Google Scholar
26	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-tie quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
27	Hasenberg A, Hasenberg M, Männ L, Neumann F, Borkenstein L, Stecher M, Kraus A, Engel DR, Klingberg A, Seddigh P, et al: Catchup: A mouse model for imaging-based tracking and modulation of neutrophil granulocytes. Nat Methods. 12:445–452. 2015. View Article : Google Scholar : PubMed/NCBI
28	Boivin GP, Hickman DL, Creamer-Hente MA, Pritchett-Corning KR and Bratcher NA: Review of CO₂ as a euthanasia agent for laboratory rats and mice. J Am Assoc Lab Anim Sci. 56:491–499. 2017.PubMed/NCBI
29	Marquardt N, Feja M, Hünigen H, Plendl J, Menken L, Fink H and Bert B: Euthanasia of laboratory mice: Are isoflurane and sevoflurane real alternatives to carbon dioxide? PLoS One. 13:e02037932018. View Article : Google Scholar : PubMed/NCBI
30	Makowska IJ and Weary DM: Rat aversion to induction with inhalant anaesthetics. Appl Anim Behav Sci. 119:229–235. 2009. View Article : Google Scholar
31	Leach MC, Bowell VA, Allan TF and Morton DB: Degrees of aversion shown by rats and mice to different concentrations of inhalational anaesthetics. Vet Rec. 150:808–815. 2002. View Article : Google Scholar : PubMed/NCBI
32	Verhamme C, King RH, ten Asbroek AL, Muddle JR, Nourallah M, Wolterman R, Baas F and van Schaik IN: Myelin and axon pathology in a long-term study of PMP22-overexpressing mice. J Neuropathol Exp Neurol. 70:386–398. 2011. View Article : Google Scholar : PubMed/NCBI
33	Komyathy K, Neal S, Feely S, Miller LJ, Lewis RA, Trigge G, Siskind CE, Shy ME and Ramchandren S: Anterior tibialis CMAP amplitude correlations with impairment in CMT1A. Muscle Nerve. 47:493–496. 2013. View Article : Google Scholar : PubMed/NCBI
34	Stojkovic T: Hereditary neuropathies: An update. Rev Neurol (Paris). 172:775–778. 2016. View Article : Google Scholar : PubMed/NCBI
35	Lee JS, Chang EH, Koo OJ, Jwa DH, Mo WM, Kwak G, Moon HW, Park HT, Hong YB and Choi BO: Pmp22 mutant allele-specific siRNA alleviates demyelinating neuropathic phenotype in vivo. Neurobiol Dis. 100:99–107. 2017. View Article : Google Scholar : PubMed/NCBI
36	Chal J and Pourquie O: Making muscle: Skeletal myogenesis in vivo and in vitro. Development. 144:2104–2122. 2017. View Article : Google Scholar : PubMed/NCBI
37	McPherron AC and Lee SJ: Double muscling in cattle due to mutations in the myostatin gene. Proc Natl Acad Sci USA. 94:12457–12461. 1997. View Article : Google Scholar : PubMed/NCBI
38	Grobet L, Martin LJ, Poncelet D, Pirottin D, Brouwers B, Riquet J, Schoeberlein A, Dunner S, Menissier F, Massabanda J, et al: A deletion in the bovine myostatin gene causes the double-muscled phenotype in cattle. Nat Genet. 17:71–74. 1997. View Article : Google Scholar : PubMed/NCBI
39	Kambadur R, Sharma M, Smith TP and Bass JJ: Mutations in myostatin (GDF8) in double-muscled Belgian blue and piedmontese cattle. Genome Res. 7:910–916. 1997. View Article : Google Scholar : PubMed/NCBI
40	Grobet L, Poncelet D, Royo LJ, Brouwers B, Pirottin D, Michaux C, Menissier F, Zanotti M, Dunner S and Georges M: Molecular definition of an allelic series of mutations disrupting the myostatin function and causing double-muscling in cattle. Mamm Genome. 9:210–213. 1998. View Article : Google Scholar : PubMed/NCBI
41	Lin J, Arnold HB, Della-Fera MA, Azain MJ, Hartzell DL and Baile CA: Myostatin knockout in mice increases myogenesis and decreases adipogenesis. Biochem Biophys Res Commun. 291:701–706. 2002. View Article : Google Scholar : PubMed/NCBI
42	Pearsall RS, Davies MV, Cannell M, Li J, Widrick J, Mulivor AW, Wallner S, Troy ME, Spaits M, Liharska K, et al: Follistatin-based ligand trap ACE-083 induces localized hypertrophy of skeletal muscle with functional improvement in models of neuromuscular disease. Sci Rep. 9:113922019. View Article : Google Scholar : PubMed/NCBI
43	Bonnefoy A and Armand S: Normal Gait. Orthopedic Management of Children With Cerebral Palsy: A Comprehensive Approach. Nova Science Publishers Inc.; Hauppauge, NY: pp. 5672015
44	Chester VL, Biden EN and Tingley M: Gait analysis. Biomed Instrum Technol. 39:64–74. 2005.PubMed/NCBI