Abstract
Cerebral palsy (CP) is a non-progressive motor disorder that affects posture and gait due to contracture development. The purpose of this study is to analyze a possible relation between muscle stiffness and gene expression levels in muscle tissue of children with CP. Next-generation sequencing (NGS) of gene transcripts was carried out in muscle biopsies from gastrocnemius muscle (n = 13 children with CP and n = 13 typical developed (TD) children). Passive stiffness of the ankle plantarflexors was measured. Structural changes of the basement membranes and the sarcomere length were measured. Twelve pre-defined gene target sub-categories of muscle function, structure and metabolism showed significant differences between muscle tissue of CP and TD children. Passive stiffness was significantly correlated to gene expression levels of HSPG2 (p = 0.02; R2 = 0.67), PRELP (p = 0.002; R2 = 0.84), RYR3 (p = 0.04; R2 = 0.66), C COL5A3 (p = 0.0007; R2 = 0.88), ASPH (p = 0.002; R2 = 0.82) and COL4A6 (p = 0.03; R2 = 0.97). Morphological differences in the basement membrane were observed between children with CP and TD children. The sarcomere length was significantly increased in children with CP when compared with TD (p = 0.04). These findings show that gene targets in the categories: calcium handling, basement membrane and collagens, were significantly correlated to passive muscle stiffness. A Reactome pathway analysis showed that pathways involved in DNA repair, ECM proteoglycans and ion homeostasis were amongst the most upregulated pathways in CP, while pathways involved in collagen fibril crosslinking, collagen fibril assembly and collagen turnover were amongst the most downregulated pathways when compared with TD children. These results underline that contracture formation and motor impairment in CP is an interplay between multiple factors.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alamowitch S, Plaisier E, Favrole P, Prost C, Chen Z, Van Agtmael T et al (2009) Cerebrovascular disease related to COL4A1 mutations in HANAC syndrome. Neurology 73(22):1873–1882. https://doi.org/10.1212/WNL.0b013e3181c3fd12
Arikawa-Hirasawa E, Le AH, Nishino I, Nonaka I, Ho NC, Francomano CA et al (2002) Structural and functional mutations of the perlecan gene cause Schwartz-Jampel syndrome, with myotonic myopathy and chondrodysplasia. Am J Hum Genet 70(5):1368–1375
Barber L, Barrett R, Lichtwark G (2012) Medial gastrocnemius muscle fascicle active torque-length and Achilles tendon properties in young adults with spastic cerebral palsy. J Biomech 45(15):2526–2530. https://doi.org/10.1016/j.jbiomech.2012.07.018
Bartels EM, Korbo L, Harrison AP (2020) Novel insights into cerebral palsy. J Muscle Res Cell Motil. https://doi.org/10.1007/s10974-020-09577-4
Baschal EE, Wethey CI, Swindle K, Baschal RM, Gowan K, Tang NL et al (2015). Exome sequencing identifies a Rare HSPG2 variant associated with familial idiopathic scoliosis. G3 (Bethesda) 5(2), 167–174. https://doi.org/10.1534/g3.114.015669
Bax M, Goldstein M, Rosenbaum P, Leviton A, Paneth N, Dan B et al (2005) Proposed definition and classification of cerebral palsy, April 2005. Dev Med Child Neurol 47(8):571–576
Beard NA, Wei L, Dulhunty AF (2009) Control of muscle ryanodine receptor calcium release channels by proteins in the sarcoplasmic reticulum lumen. Clin Exp Pharmacol Physiol 36(3):340–345. https://doi.org/10.1111/j.1440-1681.2008.05094.x
Bengtsson E, Neame PJ, Heinegard D, Sommarin Y (1995) The primary structure of a basic leucine-rich repeat protein, PRELP, found in connective tissues. J Biol Chem 270(43):25639–25644. https://doi.org/10.1074/jbc.270.43.25639
Bonnemann CG (2011) The collagen VI-related myopathies: muscle meets its matrix. Nat Rev Neurol 7(7):379–390. https://doi.org/10.1038/nrneurol.2011.81
Booth CM, Cortina-Borja MJ, Theologis TN (2001) Collagen accumulation in muscles of children with cerebral palsy and correlation with severity of spasticity. Dev Med Child Neurol 43(5):314–320
Borg L, Sporring J, Dam EB, Dahl VA, Dyrby TB, Feidenhans’l R et al (2019) Muscle fibre morphology and microarchitecture in cerebral palsy patients obtained by 3D synchrotron X-ray computed tomography. Comput Biol Med 107:265–269. https://doi.org/10.1016/j.compbiomed.2019.02.008
Breedveld G, de Coo IF, Lequin MH, Arts WF, Heutink P, Gould DB et al (2006) Novel mutations in three families confirm a major role of COL4A1 in hereditary porencephaly. J Med Genet 43(6):490–495. https://doi.org/10.1136/jmg.2005.035584
Cohn RD, Campbell KP (2000) Molecular basis of muscular dystrophies. Muscle Nerve 23(10):1456–1471
Collins M, Mokone GG, September AV, van der Merwe L, Schwellnus MP (2009) The COL5A1 genotype is associated with range of motion measurements. Scand J Med Sci Sports 19(6):803–810. https://doi.org/10.1111/j.1600-0838.2009.00915.x
Cosgrove D, Liu S (2017) Collagen IV diseases: a focus on the glomerular basement membrane in Alport syndrome. Matrix Biol 57–58:45–54. https://doi.org/10.1016/j.matbio.2016.08.005
Croft D, Mundo AF, Haw R, Milacic M, Weiser J, Wu G et al (2014) The Reactome pathway knowledgebase. Nucleic Acids Res 42(Database issue), D472–477. https://doi.org/10.1093/nar/gkt1102
Croft D, O'Kelly G, Wu G, Haw R, Gillespie M, Matthews L et al (2011) Reactome: a database of reactions, pathways and biological processes. Nucleic Acids Res 39(Database issue), D691–697.https://doi.org/10.1093/nar/gkq1018
DiFranco M, Kramerova I, Vergara JL, Spencer MJ (2016) Attenuated Ca(2+) release in a mouse model of limb girdle muscular dystrophy 2A. Skelet Muscle 6:11. https://doi.org/10.1186/s13395-016-0081-y
Divet A, Paesante S, Bleunven C, Anderson A, Treves S, Zorzato F (2005) Novel sarco(endo)plasmic reticulum proteins and calcium homeostasis in striated muscles. J Muscle Res Cell Motil 26(1):7–12. https://doi.org/10.1007/s10974-005-9001-1
Durbeej M, Campbell KP (2002) Muscular dystrophies involving the dystrophin-glycoprotein complex: an overview of current mouse models. Curr Opin Genet Dev 12(3):349–361
Fabregat A, Sidiropoulos K, Garapati P, Gillespie M, Hausmann K, Haw R et al (2016) The Reactome pathway Knowledgebase. Nucleic Acids Res 44(D1):D481-487. https://doi.org/10.1093/nar/gkv1351
Fabregat A, Sidiropoulos K, Viteri G, Forner O, Marin-Garcia P, Arnau V et al (2017) Reactome pathway analysis: a high-performance in-memory approach. BMC Bioinformatics 18(1):142. https://doi.org/10.1186/s12859-017-1559-2
Gehlert S, Bloch W, Suhr F (2015) Ca2+-dependent regulations and signaling in skeletal muscle: from electro-mechanical coupling to adaptation. Int J Mol Sci 16(1):1066–1095. https://doi.org/10.3390/ijms16011066
Germany L, Ehlinger V, Klapouszczak D, Delobel M, Hollody K, Sellier E et al (2013) Trends in prevalence and characteristics of post-neonatal cerebral palsy cases: a European registry-based study. Res Dev Disabil 34(5):1669–1677. https://doi.org/10.1016/j.ridd.2013.02.016
Gillett JG, Lichtwark GA, Boyd RN, and Barber LA (2018) Functional Capacity in Adults With Cerebral Palsy: Lower Limb Muscle Strength Matters. Arch Phys Med Rehabil 99(5), 900–906 e901. https://doi.org/10.1016/j.apmr.2018.01.020
Gong S, Su BB, Tovar H, Mao C, Gonzalez V, Liu Y et al (2018) Polymorphisms within RYR3 gene are associated with risk and age at onset of hypertension, diabetes, and Alzheimer’s disease. Am J Hypertens 31(7):818–826. https://doi.org/10.1093/ajh/hpy046
Gough M, Shortland AP (2012) Could muscle deformity in children with spastic cerebral palsy be related to an impairment of muscle growth and altered adaptation? Dev Med Child Neurol 54(6):495–499. https://doi.org/10.1111/j.1469-8749.2012.04229.x
Ha TT, Sadleir LG, Mandelstam SA, Paterson SJ, Scheffer IE, Gecz J et al (2016) A mutation in COL4A2 causes autosomal dominant porencephaly with cataracts. Am J Med Genet A 170A(4):1059–1063. https://doi.org/10.1002/ajmg.a.37527
Hagglund G, Wagner P (2011) Spasticity of the gastrosoleus muscle is related to the development of reduced passive dorsiflexion of the ankle in children with cerebral palsy: a registry analysis of 2,796 examinations in 355 children. Acta Orthop 82(6):744–748. https://doi.org/10.3109/17453674.2011.618917
Himmelmann K, Uvebrant P (2014) The panorama of cerebral palsy in Sweden. XI. Changing patterns in the birth-year period 2003–2006. Acta Paediatr 103(6), 618–624.https://doi.org/10.1111/apa.12614
Hussain AW, Onambele GL, Williams AG, Morse CI (2014) Muscle size, activation, and coactivation in adults with cerebral palsy. Muscle Nerve 49(1):76–83. https://doi.org/10.1002/mus.23866
Jones LR, Zhang L, Sanborn K, Jorgensen AO, Kelley J (1995) Purification, primary structure, and immunological characterization of the 26-kDa calsequestrin binding protein (junctin) from cardiac junctional sarcoplasmic reticulum. J Biol Chem 270(51):30787–30796. https://doi.org/10.1074/jbc.270.51.30787
Ko IH, Kim JH, Lee BH (2014) Relationship between lower limb muscle structure and function in cerebral palsy. J Phys Ther Sci 26(1):63–66. https://doi.org/10.1589/jpts.26.63
Kushnir A, Wajsberg B, Marks AR (2018) Ryanodine receptor dysfunction in human disorders. Biochim Biophys Acta. https://doi.org/10.1016/j.bbamcr.2018.07.011
Labelle-Dumais C, Dilworth DJ, Harrington EP, de Leau M, Lyons D, Kabaeva Z et al (2011) COL4A1 mutations cause ocular dysgenesis, neuronal localization defects, and myopathy in mice and Walker-Warburg syndrome in humans. PLoS Genet 7(5):e1002062. https://doi.org/10.1371/journal.pgen.1002062
Lewis M (2003) PRELP, collagen, and a theory of Hutchinson-Gilford progeria. Ageing Res Rev 2(1):95–105. https://doi.org/10.1016/s1568-1637(02)00044-2
Li C, Li HJ, Chen Q, Zhang XM, Li CL (2018) Expression of ASPH gene in invasive breast cancer and its clinical significance in promoter methylation. Sichuan Da Xue Xue Bao Yi Xue Ban 49(1):54–58
Lieber RL, Friden J (2002) Spasticity causes a fundamental rearrangement of muscle-joint interaction. Muscle Nerve 25(2):265–270
Lieber RL, Friden J (2019) Muscle contracture and passive mechanics in cerebral palsy. J Appl Physiol (1985) 126(5), 1492–1501. https://doi.org/10.1152/japplphysiol.00278.2018
Malaiya R, McNee AE, Fry NR, Eve LC, Gough M, Shortland AP (2007) The morphology of the medial gastrocnemius in typically developing children and children with spastic hemiplegic cerebral palsy. J Electromyogr Kinesiol 17(6):657–663. https://doi.org/10.1016/j.jelekin.2007.02.009
Malfait F, De Paepe A (2014) The Ehlers-Danlos syndrome. Adv Exp Med Biol 802:129–143. https://doi.org/10.1007/978-94-007-7893-1_9
Mao M, Alavi MV, Labelle-Dumais C, Gould DB (2015) Type IV collagens and basement membrane diseases: cell biology and pathogenic mechanisms. Curr Top Membr 76:61–116. https://doi.org/10.1016/bs.ctm.2015.09.002
Mathewson MA, Chambers HG, Girard PJ, Tenenhaus M, Schwartz AK, Lieber RL (2014) Stiff muscle fibers in calf muscles of patients with cerebral palsy lead to high passive muscle stiffness. J Orthop Res 32(12):1667–1674. https://doi.org/10.1002/jor.22719
Mathewson MA, Lieber RL (2015) Pathophysiology of muscle contractures in cerebral palsy. Phys Med Rehabil Clin N Am 26(1):57–67. https://doi.org/10.1016/j.pmr.2014.09.005
Mathewson MA, Ward SR, Chambers HG, Lieber RL (2015) High resolution muscle measurements provide insights into equinus contractures in patients with cerebral palsy. J Orthop Res 33(1):33–39. https://doi.org/10.1002/jor.22728
Missiaen L, De Smedt H, Droogmans G, Casteels R (1992) Ca2+ release induced by inositol 1,4,5-trisphosphate is a steady-state phenomenon controlled by luminal Ca2+ in permeabilized cells. Nature 357(6379):599–602. https://doi.org/10.1038/357599a0
Pingel J, Andersen JD, Christiansen SL, Borsting C, Morling N, Lorentzen J et al (2019) Sequence variants in muscle tissue-related genes may determine the severity of muscle contractures in cerebral palsy. Am J Med Genet B Neuropsychiatr Genet 180(1):12–24. https://doi.org/10.1002/ajmg.b.32693
Pingel J, Bartels EM, Nielsen JB (2016) New perspectives on the development of muscle contractures following central motor lesions. J Physiol. https://doi.org/10.1113/jp272767
Robertson CM, Ricci MF, O'Grady K, Oskoui M, Goez H, Yager JY et al (2017) Prevalence estimate of cerebral palsy in Northern Alberta: Births, 2008–2010 Can J Neurol Sci 1–9 https://doi.org/10.1017/cjn.2017.33
Rosenbaum P, Paneth N, Leviton A, Goldstein M, Bax M, Damiano D et al (2007) A report: the definition and classification of cerebral palsy April 2006. Dev Med Child Neurol Suppl 109:8–14
Rosenbaum PL, Palisano RJ, Bartlett DJ, Galuppi BE, Russell DJ (2008) Development of the gross motor function classification system for cerebral palsy. Dev Med Child Neurol 50(4):249–253. https://doi.org/10.1111/j.1469-8749.2008.02045.x
Sigurdardottir S, Thorkelsson T, Halldorsdottir M, Thorarensen O, Vik T (2009) Trends in prevalence and characteristics of cerebral palsy among Icelandic children born 1990 to 2003. Dev Med Child Neurol 51(5):356–363
Smith LR, Chambers HG, Subramaniam S, Lieber RL (2012) Transcriptional abnormalities of hamstring muscle contractures in children with cerebral palsy. PLoS ONE 7(8):e40686. https://doi.org/10.1371/journal.pone.0040686
Smith LR, Lee KS, Ward SR, Chambers HG, Lieber RL (2011) Hamstring contractures in children with spastic cerebral palsy result from a stiffer extracellular matrix and increased in vivo sarcomere length. J Physiol 589(Pt 10):2625–2639. https://doi.org/10.1113/jphysiol.2010.203364
Smith LR, Ponten E, Hedstrom Y, Ward SR, Chambers HG, Subramaniam S et al (2009) Novel transcriptional profile in wrist muscles from cerebral palsy patients. BMC Med Genomics 2:44. https://doi.org/10.1186/1755-8794-2-44
Spence HJ, Chen YJ, Winder SJ (2002) Muscular dystrophies, the cytoskeleton and cell adhesion. BioEssays 24(6):542–552. https://doi.org/10.1002/bies.10098
Terry EE, Zhang X, Hoffmann C, Hughes LD, Lewis SA, Li J et al (2018) Transcriptional profiling reveals extraordinary diversity among skeletal muscle tissues Elife 7 https://doi.org/10.7554/eLife.34613
van der Knaap MS, Smit LM, Barkhof F, Pijnenburg YA, Zweegman S, Niessen HW et al (2006) Neonatal porencephaly and adult stroke related to mutations in collagen IV A1. Ann Neurol 59(3):504–511. https://doi.org/10.1002/ana.20715
Von Walden F, Gantelius S, Liu C, Borgstrom H, Bjork L, Gremark O et al (2018) Muscle contractures in patients with cerebral palsy and acquired brain injury are associated with extracellular matrix expansion, pro-inflammatory gene expression, and reduced rRNA synthesis. Muscle Nerve 58(2):277–285. https://doi.org/10.1002/mus.26130
Vos P, Zietse R, Geel M, van Brooks AS, Cransberg K (2018) Diagnosing Alport syndrome: lessons from the pediatric ward Nephron 1–8 https://doi.org/10.1159/000492438
Willerslev-Olsen M, Lorentzen J, Sinkjaer T, Nielsen JB (2013) Passive muscle properties are altered in children with cerebral palsy before the age of 3 years and are difficult to distinguish clinically from spasticity. Dev Med Child Neurol. https://doi.org/10.1111/dmcn.12124
Williams PE, Goldspink G (1978) Changes in sarcomere length and physiological properties in immobilized muscle. J Anat 127(Pt 3):459–468
Xu Z, Ichikawa N, Kosaki K, Yamada Y, Sasaki T, Sakai LY et al (2010) Perlecan deficiency causes muscle hypertrophy, a decrease in myostatin expression, and changes in muscle fiber composition. Matrix Biol 29(6):461–470. https://doi.org/10.1016/j.matbio.2010.06.001
Yurchenco PD (2011). Basement membranes: cell scaffoldings and signaling platforms. Cold Spring Harb Perspect Biol 3(2). https://doi.org/10.1101/cshperspect.a004911
Yurchenco PD, Patton BL (2009) Developmental and pathogenic mechanisms of basement membrane assembly. Curr Pharm Des 15(12):1277–1294
Zhang L, Kelley J, Schmeisser G, Kobayashi YM, and Jones LR (1997). Complex formation between junctin, triadin, calsequestrin, and the ryanodine receptor. Proteins of the cardiac junctional sarcoplasmic reticulum membrane. J Biol Chem 272(37), 23389–23397.https://doi.org/10.1074/jbc.272.37.23389
Funding
This project was funded by the Danish research Council (DFF-1333-00197), and the Elsass Foundation.
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interests
The authors declare that they have no conflict of interest.
Informed consent
All the parents and/or legally authorized representatives of the participants gave informed consent.
Ethical approval
The protocol was approved by the Regional Ethics Committee for Copenhagen (H-4-2014-047). All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Pingel, J., Kampmann, ML., Andersen, J.D. et al. Gene expressions in cerebral palsy subjects reveal structural and functional changes in the gastrocnemius muscle that are closely associated with passive muscle stiffness. Cell Tissue Res 384, 513–526 (2021). https://doi.org/10.1007/s00441-020-03399-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00441-020-03399-z