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Omentum acts as a regulatory organ controlling skeletal muscle repair of mdx mice diaphragm

  • Douglas Florindo Pinheiro
  • Rafael F. da Silva
  • Thiago Morais Barbosa
  • Jaciara F. G. Gama
  • Aline C. Gomes
  • Thereza Quirico-Santos
  • Jussara Lagrota-CandidoEmail author
Regular Article
  • 125 Downloads

Abstract

Duchenne muscular dystrophy is a lethal X-linked muscle wasting disease due to mutations of the dystrophin gene leading to distinct susceptibility to degeneration and fibrosis among skeletal muscles. This study aims at verifying whether intense mdx diaphragm remodeling could be attributed to influences from the omentum, a lymphohematopoietic tissue rich in progenitor cells and trophic factors. Mdx omentum produces growth factors HGF and FGF and increased amounts of VEGF with pleiotropic actions upon muscular progenitors and myoblast differentiation. Histology revealed that the absence of the omentum reduced inflammation and collagen deposition in the diaphragm. The diaphragm from omentectomized mdx mice presents impaired repair with a predominance of collagen type I deposition, decreased muscle regeneration and a reduction in collagen type IV and indication of altered basal lamina integrity in the diaphragm. Omentectomy further reduced inflammatory infiltration and NFκ-B activation but a change in the pattern of muscle inflammation with low numbers of the F4/80+CD206+ M-2 macrophage subset. Although omentectomized mice had high levels of Pax7, myogenin and TNF-α, the percentage of myofibers undergoing regeneration was low thus suggesting that a lack of the omentum halts the muscle differentiation program. Such results support that omentum exerts a regulatory function inducing an inflammatory process that favors regeneration and inhibits fibrosis selectively in the diaphragm muscle thus being a potential site for therapeutic interventions in DMD.

Keywords

Omentum mdx Macrophages Fibrosis Diaphragm Muscle regeneration 

Notes

Acknowledgments

We are grateful to Nina Cortez and Diogo G. Garcia for technical assistance and Giselle M. Faria with the statistical analysis.

Funding information

This study was financed in part by FAPERJ (Fundação de Amparo a Pesquisa do Rio de Janeiro), PROPPI (UFF) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001.

Supplementary material

441_2019_3012_Fig6_ESM.png (2 mb)
Figure S1

Gastrocnemius muscle from mdx at 12 weeks old is not affected by omentectomy. Sirius Red stain of gastrocnemius muscle and respective quantification of area occupied by collagen and regenerating myofibres from SHAM and OTX mdx mice at 12 (a) and 24 weeks old (b). (PNG 2026 kb)

441_2019_3012_MOESM1_ESM.tif (8.8 mb)
High-resolution Image (TIF 8991 kb)
441_2019_3012_Fig7_ESM.png (94 kb)
Figure S2

Measurements of TNF-α and myogenin by real-time PCR in OTX mdx gastrocnemius at 12 and 24 weeks old. Test samples were normalized against the mRNA levels detected for the beta-actin gene and expressed as fold change compared to SHAM. (*p < 0.05) (PNG 93 kb)

441_2019_3012_MOESM2_ESM.tif (588 kb)
High-resolution Image (TIF 587 kb)

References

  1. Allen DG, Gervasio OL, Yeung EW, Whitehead NP (2010) Calcium and the damage pathways in muscular dystrophy. Can J Physiol Pharmacol 88:83–91.  https://doi.org/10.1139/Y09-058 CrossRefGoogle Scholar
  2. Ansel KM, Harris RBS, Cyster JG (2002) CXCL13 is required for B1 cell homing, natural antibody production, and body cavity immunity. Immunity 16:67–76CrossRefGoogle Scholar
  3. Arnold L, Henry A, Poron F et al (2007) Inflammatory monocytes recruited after skeletal muscle injury switch into antiinflammatory macrophages to support myogenesis. J Exp Med 204:1057–1069.  https://doi.org/10.1084/jem.20070075 CrossRefGoogle Scholar
  4. Bakkar N, Guttridge DC (2010) NF-κB signaling: a tale of two pathways in skeletal myogenesis. Physiol Rev 90:495–511.  https://doi.org/10.1152/physrev.00040.2009 CrossRefGoogle Scholar
  5. Bakkar N, Wang J, Ladner KJ et al (2008) IKK/NF-κB regulates skeletal myogenesis via a signaling switch to inhibit differentiation and promote mitochondrial biogenesis. J Cell Biol 180:787–802.  https://doi.org/10.1083/jcb.200707179 CrossRefGoogle Scholar
  6. Balachandran K, Konduri S, Sucosky P et al (2006) An ex vivo study of the biological properties of porcine aortic valves in response to circumferential cyclic stretch. Ann Biomed Eng 34:1655–1665.  https://doi.org/10.1007/s10439-006-9167-8 CrossRefGoogle Scholar
  7. Bani C, Lagrota-Candido J, Pinheiro DF et al (2008) Pattern of metalloprotease activity and myofiber regeneration in skeletal muscles of mdx mice. Muscle Nerve 37:583–592.  https://doi.org/10.1002/mus.20970 CrossRefGoogle Scholar
  8. Berberich S, Dähne S, Schippers A et al (2008) Differential molecular and anatomical basis for B cell migration into the peritoneal cavity and omental milky spots. J Immunol 180:2196–2203CrossRefGoogle Scholar
  9. Bryan BA, Walshe TE, Mitchell DC et al (2008) Coordinated vascular endothelial growth factor expression and signaling during skeletal myogenic differentiation. Mol Biol Cell 19:994–1006.  https://doi.org/10.1091/mbc.E07-09-0856 CrossRefGoogle Scholar
  10. Canicio J, Ruiz-Lozano P, Carrasco M et al (2001) Nuclear factor κB-inducing kinase and IκB kinase-α signal skeletal muscle cell differentiation. J Biol Chem 276:20228–20233.  https://doi.org/10.1074/jbc.M100718200 CrossRefGoogle Scholar
  11. Cavalla F, Reyes M, Vernal R et al (2013) High levels of CXC ligand 12/stromal cell-derived factor 1 in apical lesions of endodontic origin associated with mast cell infiltration. J Endod 39:1234–1239.  https://doi.org/10.1016/j.joen.2013.06.020 CrossRefGoogle Scholar
  12. Chamberlain JS, Metzger J, Reyes M et al (2007) Dystrophin-deficient mdx mice display a reduced life span and are susceptible to spontaneous rhabdomyosarcoma. FASEB J 21:2195–2204.  https://doi.org/10.1096/fj.06-7353com CrossRefGoogle Scholar
  13. Chen S-E, Jin B, Li Y-P (2007) TNF-alpha regulates myogenesis and muscle regeneration by activating p38 MAPK. Am J Physiol Cell Physiol 292:C1660–C1671.  https://doi.org/10.1152/ajpcell.00486.2006 CrossRefGoogle Scholar
  14. Collins RA, Grounds MD (2001) The role of tumor necrosis factor-alpha (TNF-alpha) in skeletal muscle regeneration. Studies in TNF-alpha(-/-) and TNF-alpha(-/-)/LT-alpha(-/-) mice. J Histochem Cytochem 49:989–1001.  https://doi.org/10.1177/002215540104900807 CrossRefGoogle Scholar
  15. Cuttle L, Nataatmadja M, Fraser JF et al (2005) Collagen in the scarless fetal skin wound: detection with picrosirius-polarization. Wound Repair Regen 13:198–204.  https://doi.org/10.1111/j.1067-1927.2005.130211.x CrossRefGoogle Scholar
  16. De Siena R, Balducci L, Blasi A et al (2010) Omentum-derived stromal cells improve myocardial regeneration in pig post-infarcted heart through a potent paracrine mechanism. Exp Cell Res 316:1804–1815.  https://doi.org/10.1016/j.yexcr.2010.02.009 CrossRefGoogle Scholar
  17. Demoule A, Divangahi M, Danialou G et al (2005) Expression and regulation of CC class chemokines in the dystrophic (mdx) diaphragm. Am J Respir Cell Mol Biol 33:178–185.  https://doi.org/10.1165/rcmb.2004-0347OC CrossRefGoogle Scholar
  18. Dinulovic I, Furrer R, Handschin C (2017) Plasticity of the muscle stem cell microenvironment. Adv Exp Med Biol 1041:141–169.  https://doi.org/10.1007/978-3-319-69194-7_8 CrossRefGoogle Scholar
  19. Dorchies OM, Reutenauer-Patte J, Dahmane E et al (2013) The anticancer drug tamoxifen counteracts the pathology in a mouse model of Duchenne muscular dystrophy. Am J Pathol 182:485–504.  https://doi.org/10.1016/j.ajpath.2012.10.018 CrossRefGoogle Scholar
  20. Goldsmith HS (2007) Omental transposition in treatment of Alzheimer disease. J Am Coll Surg 205:800–804.  https://doi.org/10.1016/j.jamcollsurg.2007.06.294 CrossRefGoogle Scholar
  21. Goldsmith HS (2009) Treatment of acute spinal cord injury by omental transposition: a new approach. J Am Coll Surg 208:289–292.  https://doi.org/10.1016/j.jamcollsurg.2008.10.021 CrossRefGoogle Scholar
  22. Hirai K, Takemori N, Namiki M (1994) Erythropoiesis in mouse omental milky spots induced by erythropoietin: light and electron microscopic study. Int J Exp Pathol 75:375–383Google Scholar
  23. Ito A, Yamamoto M, Ikeda K et al (2015) Effects of type IV collagen on myogenic characteristics of IGF-I gene-engineered myoblasts. J Biosci Bioeng 119:596–603.  https://doi.org/10.1016/j.jbiosc.2014.10.008 CrossRefGoogle Scholar
  24. Johnson EK, Li B, Yoon JH et al (2013) Identification of new dystroglycan complexes in skeletal muscle. PLoS One 8:e73224.  https://doi.org/10.1371/journal.pone.0073224 CrossRefGoogle Scholar
  25. Kieny P, Chollet S, Delalande P et al (2013) Evolution of life expectancy of patients with Duchenne muscular dystrophy at AFM Yolaine de Kepper centre between 1981 and 2011. Ann Phys Rehabil Med 56:443–454.  https://doi.org/10.1016/j.rehab.2013.06.002 CrossRefGoogle Scholar
  26. Kosmac K, Peck BD, Walton RG, et al. (2018) Immunohistochemical identification of human skeletal muscle macrophages. Biotechnol Protoc 8(12).  https://doi.org/10.21769/BioProtoc.2883
  27. Kowalski K, Kołodziejczyk A, Sikorska M et al (2016) Stem cells migration during skeletal muscle regeneration - the role of Sdf-1/Cxcr4 and Sdf-1/Cxcr7 axis. Cell Adhes Migr 11:384–398.  https://doi.org/10.1080/19336918.2016.1227911 CrossRefGoogle Scholar
  28. Lees JG, Ching YW, Adams DH et al (2013) Tropomyosin regulates cell migration during skin wound healing. J Invest Dermatol 133:1330–1339.  https://doi.org/10.1038/jid.2012.489 CrossRefGoogle Scholar
  29. Liebermann-Meffert D (2000) The greater omentum. Anatomy, embryology, and surgical applications. Surg Clin North Am 80:275–293 xiiCrossRefGoogle Scholar
  30. Litbarg NO, Gudehithlu KP, Sethupathi P et al (2007) Activated omentum becomes rich in factors that promote healing and tissue regeneration. Cell Tissue Res 328:487–497.  https://doi.org/10.1007/s00441-006-0356-4 CrossRefGoogle Scholar
  31. Loufrani L, Dubroca C, You D et al (2004) Absence of dystrophin in mice reduces NO-dependent vascular function and vascular density: total recovery after a treatment with the aminoglycoside gentamicin. Arterioscler Thromb Vasc Biol 24:671–676.  https://doi.org/10.1161/01.ATV.0000118683.99628.42 CrossRefGoogle Scholar
  32. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  33. McGreevy JW, Hakim CH, McIntosh MA, Duan D (2015) Animal models of Duchenne muscular dystrophy: from basic mechanisms to gene therapy. Dis Model Mech 8:195–213.  https://doi.org/10.1242/dmm.018424 CrossRefGoogle Scholar
  34. Mendell JR, Shilling C, Leslie ND et al (2012) Evidence-based path to newborn screening for Duchenne muscular dystrophy. Ann Neurol 71:304–313.  https://doi.org/10.1002/ana.23528 CrossRefGoogle Scholar
  35. Messina S, Mazzeo A, Bitto A et al (2007) VEGF overexpression via adeno-associated virus gene transfer promotes skeletal muscle regeneration and enhances muscle function in mdx mice. FASEB J 21:3737–3746.  https://doi.org/10.1096/fj.07-8459com CrossRefGoogle Scholar
  36. Meza-Perez S, Randall TD (2017) Immunological Functions of the Omentum. Trends Immunol 38:526–536.  https://doi.org/10.1016/j.it.2017.03.002 CrossRefGoogle Scholar
  37. Michailova KN (2001) Postinflammatory changes of the diaphragmatic stomata. Ann Anat 183:309–317.  https://doi.org/10.1016/S0940-9602(01)80168-0 CrossRefGoogle Scholar
  38. Moens P, Baatsen PHWW, Maréchal G (1993) Increased susceptibility of EDL muscles from mdx mice to damage induced by contractions with stretch. J Muscle Res Cell Motil 14:446–451.  https://doi.org/10.1007/BF00121296 CrossRefGoogle Scholar
  39. Nawaz A, Aminuddin A, Kado T et al (2017) CD206 + M2-like macrophages regulate systemic glucose metabolism by inhibiting proliferation of adipocyte progenitors. Nat Commun 8:286.  https://doi.org/10.1038/s41467-017-00231-1 CrossRefGoogle Scholar
  40. Odemis V, Boosmann K, Dieterlen MT, Engele J (2007) The chemokine SDF1 controls multiple steps of myogenesis through atypical PKCzeta. J Cell Sci 120:4050–4059.  https://doi.org/10.1242/jcs.010009 CrossRefGoogle Scholar
  41. Pigozzo SR, Da Re L, Romualdi C, et al (2013) Revertant fibers in the mdx murine model of Duchenne muscular dystrophy: an age- and muscle-related reappraisal. PLoS One 8.  https://doi.org/10.1371/journal.pone.0072147
  42. Pinheiro DF, da Silva RF, Carvalho LP et al (2012) Persistent activation of omentum influences the pattern of muscular lesion in the mdx diaphragm. Cell Tissue Res 350:77–88.  https://doi.org/10.1007/s00441-012-1443-3 CrossRefGoogle Scholar
  43. Pinho M de FB, Hurtado SP, El-Cheikh MC, Borojevic R (2005) Haemopoietic progenitors in the adult mouse omentum: permanent production of B lymphocytes and monocytes. Cell Tissue Res 319:91–102.  https://doi.org/10.1007/s00441-004-0998-z CrossRefGoogle Scholar
  44. Pituch-Noworolska A, Majka M, Janowska-Wieczorek A et al (2003) Circulating CXCR4-positive stem/progenitor cells compete for SDF-1-positive niches in bone marrow, muscle and neural tissues: an alternative hypothesis to stem cell plasticity. Folia Histochem Cytobiol 41:13–21Google Scholar
  45. Proto JD, Tang Y, Lu A et al (2015) NF-κB inhibition reveals a novel role for HGF during skeletal muscle repair. Cell Death Dis 6:e1730.  https://doi.org/10.1038/cddis.2015.66 CrossRefGoogle Scholar
  46. Rangel-Moreno J, Moyron-Quiroz JE, Carragher DM et al (2009) Milky spots in the omentum develop in the absence of lymphoid tissue inducer cells and independently support B and T cell responses to peritoneal antigens. Immunity 30:731–743.  https://doi.org/10.1016/j.immuni.2009.03.014 CrossRefGoogle Scholar
  47. Roggendorf W, Opitz H, Schuppan D (1988) Altered expression of collagen type VI in brain vessels of patients with chronic hypertension. A comparison with the distribution of collagen IV and procollagen III. Acta Neuropathol 77:55–60CrossRefGoogle Scholar
  48. Sanes JR (2003) The basement membrane/basal lamina of skeletal muscle. J Biol Chem 278:12601–12604.  https://doi.org/10.1074/jbc.R200027200 CrossRefGoogle Scholar
  49. Selsby JT, Morine KJ, Pendrak K et al (2012) Rescue of dystrophic skeletal muscle by PGC-1α involves a fast to slow fiber type shift in the mdx mouse. PLoS ONE 7:e30063.  https://doi.org/10.1371/journal.pone.0030063 CrossRefGoogle Scholar
  50. Shaik-Dasthagirisaheb YB, Varvara G, Murmura G et al (2013) Vascular endothelial growth factor (VEGF), mast cells and inflammation. Int J Immunopathol Pharmacol 26:327–335.  https://doi.org/10.1177/039463201302600206
  51. Sharpe KM, Premsukh MD, Townsend D (2013) Alterations of dystrophin associated glycoproteins in the heart lacking dystrophin or dystrophin and utrophin. J Muscle Res Cell Motil 34:395–405.  https://doi.org/10.1007/s10974-013-9362-9 CrossRefGoogle Scholar
  52. Siemionow M, Cwykiel J, Heydemann A et al (2018) Dystrophin Expressing Chimeric (DEC) Human Cells Provide a Potential Therapy for Duchenne Muscular Dystrophy. Stem Cell Rev 14:370–384.  https://doi.org/10.1007/s12015-018-9807-z CrossRefGoogle Scholar
  53. Sienkiewicz D, Kulak W, Okurowska-Zawada B et al (2015) Duchenne muscular dystrophy: current cell therapies. Ther Adv Neurol Disord 8:166–177.  https://doi.org/10.1177/1756285615586123 CrossRefGoogle Scholar
  54. Singh AK, Patel J, Litbarg NO et al (2008) Stromal cells cultured from omentum express pluripotent markers, produce high amounts of VEGF, and engraft to injured sites. Cell Tissue Res 332:81–88.  https://doi.org/10.1007/s00441-007-0560-x CrossRefGoogle Scholar
  55. Singh AK, Pancholi N, Patel J et al (2009) Omentum facilitates liver regeneration. World J Gastroenterol 15:1057–1064Google Scholar
  56. Smith LR, Hammers DW, Sweeney HL, Barton ER (2016) Increased collagen cross-linking is a signature of dystrophin-deficient muscle. Muscle Nerve 54:71–78.  https://doi.org/10.1002/mus.24998 CrossRefGoogle Scholar
  57. Tatsumi R, Anderson JE, Nevoret CJ et al (1998) HGF/SF is present in normal adult skeletal muscle and is capable of activating satellite cells. Dev Biol 194:114–128.  https://doi.org/10.1006/dbio.1997.8803 CrossRefGoogle Scholar
  58. Tidball JG, Wehling-Henricks M (2007) Macrophages promote muscle membrane repair and muscle fibre growth and regeneration during modified muscle loading in mice in vivo. J Physiol 578:327–336.  https://doi.org/10.1113/jphysiol.2006.118265 CrossRefGoogle Scholar
  59. Van Ruiten HJA, Marini Bettolo C, Cheetham T et al (2016) Why are some patients with Duchenne muscular dystrophy dying young: an analysis of causes of death in North East England. Eur J Paediatr Neurol 20:904–909.  https://doi.org/10.1016/j.ejpn.2016.07.020 CrossRefGoogle Scholar
  60. Villalta SA, Rosenberg AS, Bluestone JA (2015) The immune system in Duchenne muscular dystrophy: friend or foe. Rare Dis 3.  https://doi.org/10.1080/21675511.2015.1010966
  61. Warren GL, Hulderman T, Jensen N et al (2002) Physiological role of tumor necrosis factor alpha in traumatic muscle injury. FASEB J 16:1630–1632.  https://doi.org/10.1096/fj.02-0187fje CrossRefGoogle Scholar
  62. Weir AP, Morgan JE, Davies KE (2004) A-utrophin up-regulation in mdx skeletal muscle is independent of regeneration. Neuromuscul Disord 14:19–23CrossRefGoogle Scholar
  63. Witt R, Weigand A, Boos AM et al (2017) Mesenchymal stem cells and myoblast differentiation under HGF and IGF-1 stimulation for 3D skeletal muscle tissue engineering. BMC Cell Biol 18:15.  https://doi.org/10.1186/s12860-017-0131-2 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Douglas Florindo Pinheiro
    • 1
  • Rafael F. da Silva
    • 1
  • Thiago Morais Barbosa
    • 1
  • Jaciara F. G. Gama
    • 1
  • Aline C. Gomes
    • 2
  • Thereza Quirico-Santos
    • 2
  • Jussara Lagrota-Candido
    • 1
    Email author
  1. 1.Departamento de Imunobiologia, Instituto de BiologiaUniversidade Federal FluminenseNiteróiBrazil
  2. 2.Departamento de Biologia Celular e Molecular, Instituto de BiologiaUniversidade Federal FluminenseNiteróiBrazil

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