Skip to main content

Advertisement

SpringerLink
Log in

RGC-32 regulates reactive astrocytosis and extracellular matrix deposition in experimental autoimmune encephalomyelitis

  • Original Article
  • Published:
Immunologic Research Aims and scope Submit manuscript

Abstract

Extracellular matrix (ECM) deposition in active demyelinating multiple sclerosis (MS) lesions may impede axonal regeneration and can modify immune reactions. Response gene to complement (RGC)-32 plays an important role in the mediation of TGF-β downstream effects, but its role in gliosis has not been investigated. To gain more insight into the role played by RGC-32 in gliosis, we investigated its involvement in TGF-β-induced ECM expression and the upregulation of the reactive astrocyte markers α-smooth muscle actin (α-SMA) and nestin. In cultured neonatal rat astrocytes, collagens I, IV, and V, fibronectin, α-SMA, and nestin were significantly induced by TGF-β stimulation, and RGC-32 silencing resulted in a significant reduction in their expression. Using astrocytes isolated from RGC-32 knock-out (KO) mice, we found that the expression of TGF-β-induced collagens I, IV, and V, fibronectin, and α-SMA was significantly reduced in RGC-32 KO mice when compared with wild-type (WT) mice. SIS3 inhibition of Smad3 phosphorylation was also associated with a significant reduction in RGC-32 nuclear translocation and TGF-β-induced collagen I expression. In addition, during experimental autoimmune encephalomyelitis (EAE), RGC-32 KO mouse astrocytes displayed an elongated, bipolar phenotype, resembling immature astrocytes and glial progenitors whereas those from WT mice had a reactive, hypertrophied phenotype. Taken together, our data demonstrate that RGC-32 plays an important role in mediating TGF-β-induced reactive astrogliosis in EAE. Therefore, RGC-32 may represent a new target for therapeutic intervention in MS.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12
Fig. 13

Similar content being viewed by others

Abbreviations

ACTA2:

alpha smooth muscle actin

COL1A1 :

collagen type I alpha 1

COL4A1 :

collagen type IV alpha 1

COL5A1 :

collagen type V alpha 1

EAE :

experimental autoimmune encephalomyelitis

ECM :

extracellular matrix

EMT :

epithelial to mesenchymal transition

FN :

fibronectin

GFAP :

glial fibrillary acidic protein

KO :

knock-out

MOG :

myelin oligodendrocyte glycoprotein

MS :

multiple sclerosis

NAGM :

normal-appearing gray matter

NAWM :

normal-appearing white matter

PVS :

perivascular space

RGC-32 :

response gene to complement 32

ROCK :

rho-associated coiled-coil-containing protein kinase

WT :

wild-type

References

  1. Badea TC, Niculescu FI, Soane L, Shin ML, Rus H. Molecular cloning and characterization of RGC-32, a novel gene induced by complement activation in oligodendrocytes. J Biol Chem. 1998;273:26977–81.

    Article  PubMed  CAS  Google Scholar 

  2. Badea T, Niculescu F, Soane L, Fosbrink M, Sorana H, Rus V, et al. RGC-32 increases p34CDC2 kinase activity and entry of aortic smooth muscle cells into S-phase. J Biol Chem. 2002;277:502–8.

    Article  PubMed  CAS  Google Scholar 

  3. Vlaicu SI, Tatomir A, Boodhoo D, Ito T, Fosbrink M, Cudrici C, et al. RGC-32 is expressed in the human atherosclerotic arterial wall: role in C5b-9-induced cell proliferation and migration. Exp Mol Pathol. 2016;101:221–30.

    Article  PubMed  CAS  Google Scholar 

  4. Fosbrink M, Cudrici C, Tegla CA, Soloviova K, Ito T, Vlaicu S, et al. Response gene to complement 32 is required for C5b-9 induced cell cycle activation in endothelial cells. Exp Mol Pathol. 2009;86:87–94.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  5. Rus H, Cudrici C, Niculescu F, Shin ML. Complement activation in autoimmune demyelination: dual role in neuroinflammation and neuroprotection. J Neuroimmunol. 2006;180:9–16.

    Article  PubMed  CAS  Google Scholar 

  6. Tegla CA, Cudrici CD, Azimzadeh P, Singh AK, Trippe R 3rd, Khan A, et al. Dual role of response gene to complement-32 in multiple sclerosis. Exp Mol Pathol. 2013;94:17–28.

    Article  PubMed  CAS  Google Scholar 

  7. Rus V, Nguyen V, Tatomir A, Lees JR, Mekala AP, Boodhoo D, et al. RGC-32 promotes Th17 cell differentiation and enhances experimental autoimmune encephalomyelitis. J Immunol. 2017;198:3869–77.

    Article  PubMed  CAS  Google Scholar 

  8. Vlaicu SI, Cudrici C, Ito T, Fosbrink M, Tegla CA, Rus V, et al. Role of response gene to complement 32 in diseases. Arch Immunol Ther Exp. 2008;56:115–22.

    Article  CAS  Google Scholar 

  9. Huang W-Y, Li Z-G, Rus H, Wang X, Jose PA, Chen S-Y. RGC-32 mediates transforming growth factor-β-induced epithelial-mesenchymal transition in human renal proximal tubular cells. J Biol Chem. 2009;284:9426–32.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Guo X, Jose PA, Chen S-Y. Response gene to complement 32 interacts with Smad3 to promote epithelial–mesenchymal transition of human renal tubular cells. Am J Physiol Cell Physiol. 2011;300:C1415–C21.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  11. Sofroniew MV. Astrogliosis. Cold Spring Harb Perspect Biol. 2015;7:a020420.

    Article  PubMed Central  CAS  Google Scholar 

  12. Voskuhl RR, Peterson RS, Song B, Ao Y, Morales LB, Tiwari-Woodruff S, et al. Reactive astrocytes form scar-like perivascular barriers to leukocytes during adaptive immune inflammation of the CNS. J Neurosci. 2009;29:11511–22.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  13. Anderson MA, Ao Y, Sofroniew MV. Heterogeneity of reactive astrocytes. Neurosci Lett. 2014;565:23–9.

    Article  PubMed  CAS  Google Scholar 

  14. Zamanian JL, Xu L, Foo LC, Nouri N, Zhou L, Giffard RG, et al. Genomic analysis of reactive astrogliosis. J Neurosci. 2012;32:6391–410.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  15. Cudrici C, Ito T, Zafranskaia E, Weerth S, Rus V, Chen H, et al. Complement C5 regulates the expression of insulin-like growth factor binding proteins in chronic experimental allergic encephalomyelitis. J Neuroimmunol. 2008;203:94–103.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  16. Tegla CA, Azimzadeh P, Andrian-Albescu M, Martin A, Cudrici CD, Trippe R 3rd, et al. SIRT1 is decreased during relapses in patients with multiple sclerosis. Exp Mol Pathol. 2014;96:139–48.

    Article  PubMed  CAS  Google Scholar 

  17. Hoffman WH, Cudrici CD, Zafranskaia E, Rus H. Complement activation in diabetic ketoacidosis brains. Exp Mol Pathol. 2006;80:283–8.

    Article  PubMed  CAS  Google Scholar 

  18. Rus HG, Kim LM, Niculescu FI, Shin ML. Induction of C3 expression in astrocytes is regulated by cytokines and Newcastle disease virus. J Immunol. 1992;148:928–33.

    PubMed  CAS  Google Scholar 

  19. Tegla CA, Cudrici CD, Nguyen V, Danoff J, Kruszewski AM, Boodhoo D, et al. RGC-32 is a novel regulator of the T-lymphocyte cell cycle. Exp Mol Pathol. 2015;98:328–37.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  20. Rus HG, Niculescu F, Shin ML. Sublytic complement attack induces cell cycle in oligodendrocytes. J Immunol. 1996;156:4892–900.

    PubMed  CAS  Google Scholar 

  21. Mohan H, Krumbholz M, Sharma R, Eisele S, Junker A, Sixt M, et al. Extracellular matrix in multiple sclerosis lesions: fibrillar collagens, biglycan and decorin are upregulated and associated with infiltrating immune cells. Brain Pathol. 2010;20:966–75.

    PubMed  CAS  Google Scholar 

  22. Moreels M, Vandenabeele F, Dumont D, Robben J, Lambrichts I. Alpha-smooth muscle actin (alpha-SMA) and nestin expression in reactive astrocytes in multiple sclerosis lesions: potential regulatory role of transforming growth factor-beta 1 (TGF-beta1). Neuropathol Appl Neurobiol. 2008;34:532–46.

    Article  PubMed  CAS  Google Scholar 

  23. Derynck R, Zhang YE. Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature. 2003;425:577–84.

    Article  PubMed  CAS  Google Scholar 

  24. Jinnin M, Ihn H, Tamaki K. Characterization of SIS3, a novel specific inhibitor of Smad3, and its effect on transforming growth factor-beta1-induced extracellular matrix expression. Mol Pharmacol. 2006;69:597–607.

    Article  PubMed  CAS  Google Scholar 

  25. Chen S, Crawford M, Day RM, Briones VR, Leader JE, Jose PA, et al. RhoA modulates Smad signaling during transforming growth factor-beta-induced smooth muscle differentiation. J Biol Chem. 2006;281:1765–70.

    Article  PubMed  CAS  Google Scholar 

  26. Kuivaniemi H, Tromp G, Prockop DJ. Mutations in fibrillar collagens (types I, II, III, and XI), fibril-associated collagen (type IX), and network-forming collagen (type X) cause a spectrum of diseases of bone, cartilage, and blood vessels. Hum Mutat. 1997;9:300–15.

    Article  PubMed  CAS  Google Scholar 

  27. Marjamaa J, Tulamo R, Abo-Ramadan U, Hakovirta H, Frosen J, Rahkonen O, et al. Mice with a deletion in the first intron of the Col1a1 gene develop dissection and rupture of aorta in the absence of aneurysms: high-resolution magnetic resonance imaging, at 4.7 T, of the aorta and cerebral arteries. Magn Reson Med. 2006;55:592–7.

    Article  PubMed  CAS  Google Scholar 

  28. van Horssen J, Dijkstra CD, de Vries HE. The extracellular matrix in multiple sclerosis pathology. J Neurochem. 2007;103:1293–301.

    Article  PubMed  CAS  Google Scholar 

  29. Hara M, Kobayakawa K, Ohkawa Y, Kumamaru H, Yokota K, Saito T, et al. Interaction of reactive astrocytes with type I collagen induces astrocytic scar formation through the integrin–N-cadherin pathway after spinal cord injury. Nat Med. 2017;23:818–28.

    Article  PubMed  CAS  Google Scholar 

  30. Liesi P, Kauppila T. Induction of type IV collagen and other basement-membrane-associated proteins after spinal cord injury of the adult rat may participate in formation of the glial scar. Exp Neurol. 2002;173:31–45.

    Article  PubMed  CAS  Google Scholar 

  31. Zhu D, Tapadia MD, Palispis W, Luu M, Wang W, Gupta R. Attenuation of robust glial scar formation facilitates functional recovery in animal models of chronic nerve compression injury. J Bone Joint Surg Am. 2017;99:e132.

    Article  PubMed  Google Scholar 

  32. Xia M, Zhu Y. Fibronectin enhances spinal cord astrocyte proliferation by elevating P2Y1 receptor expression. J Neurosci Res. 2014;92:1078–90.

    Article  PubMed  CAS  Google Scholar 

  33. van Horssen J, Vos CM, Admiraal L, van Haastert ES, Montagne L, van der Valk P, et al. Matrix metalloproteinase-19 is highly expressed in active multiple sclerosis lesions. Neuropathol Appl Neurobiol. 2006;32:585–93.

    Article  PubMed  CAS  Google Scholar 

  34. Massagué J, Wotton D. Transcriptional control by the TGF-β/Smad signaling system. EMBO J. 2000;19:1745–54.

    Article  PubMed  PubMed Central  Google Scholar 

  35. Zhao P, Gao D, Wang Q, Song B, Shao Q, Sun J, et al. Response gene to complement 32 (RGC-32) expression on M2-polarized and tumor-associated macrophages is M-CSF-dependent and enhanced by tumor-derived IL-4. Cell Mol Immunol. 2015;12:692–9.

    Article  PubMed  CAS  Google Scholar 

  36. Cregg JM, DePaul MA, Filous AR, Lang BT, Tran A, Silver J. Functional regeneration beyond the glial scar. Exp Neurol. 2014;253:197–207.

    Article  PubMed  Google Scholar 

  37. Harlow DE, Macklin WB. Inhibitors of myelination: ECM changes, CSPGs and PTPs. Exp Neurol. 2014;251:39–46.

    Article  PubMed  CAS  Google Scholar 

  38. Kawano H, Kimura-Kuroda J, Komuta Y, Yoshioka N, Li HP, Kawamura K, et al. Role of the lesion scar in the response to damage and repair of the central nervous system. Cell Tissue Res. 2012;349:169–80.

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

We thank Dr. Deborah McClellan for editing this manuscript.

Funding

This work was supported in part by Veterans Administration Merit Award I01BX001458 (to H.R.).

Author information

Authors and Affiliations

  1. Department of Neurology, University of Maryland School of Medicine, 655 W Baltimore St, BRB 12-033, Baltimore, MD, 21201, USA

    Alexandru Tatomir, Cosmin A. Tegla, Alvaro Martin, Dallas Boodhoo, Adam J. Sugarman, Armugam Mekala, Freidrich Anselmo, Anamaria Talpos-Caia & Horea Rus

  2. Research Service, Veterans Administration Maryland Health Care System, Baltimore, MD, USA

    Cosmin A. Tegla, Violeta Rus & Horea Rus

  3. Department of Medicine, Division of Rheumatology and Clinical Immunology, University of Maryland School of Medicine, Baltimore, MD, USA

    Vinh Nguyen & Violeta Rus

  4. Department of Rheumatology, “Iuliu Hatieganu” University of Medicine and Pharmacy, Cluj-Napoca, Romania

    Anamaria Talpos-Caia

  5. National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD, USA

    Cornelia Cudrici

  6. Retinal Circuit Development and Genetics Unit, N-NRL, National Eye Institute, Bethesda, MD, USA

    Tudor C. Badea

  7. Veterans Administration Multiple Sclerosis Center of Excellence–East, Baltimore, MD, USA

    Horea Rus

Authors
  1. Alexandru Tatomir
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Cosmin A. Tegla
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alvaro Martin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Dallas Boodhoo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Vinh Nguyen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Adam J. Sugarman
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Armugam Mekala
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Freidrich Anselmo
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Anamaria Talpos-Caia
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Cornelia Cudrici
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Tudor C. Badea
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Violeta Rus
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Horea Rus
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Horea Rus.

Ethics declarations

Conflict of interest

H.R. has received a grant from TEVA Neuroscience (CNS-2014-174). All other authors declare that they have no conflict of interest.

Electronic supplementary material

Supplemental Table 1

(DOC 45 kb)

Supplemental Fig. 1

Quantification of ECM deposits in MS and control brains Collagen I–V deposits in MS and normal brains were quantified by two independent investigators. Statistically significant higher staining intensity was found in MS brains when compared with normal brains. Results are expressed as mean ± SEM. **p < 0.01; ***p < 0.001; ****p < 0.0001 (PNG 70 kb)

High Resolution Image

(TIF 133 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tatomir, A., Tegla, C.A., Martin, A. et al. RGC-32 regulates reactive astrocytosis and extracellular matrix deposition in experimental autoimmune encephalomyelitis. Immunol Res 66, 445–461 (2018). https://doi.org/10.1007/s12026-018-9011-x

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12026-018-9011-x

Keywords

Search

Navigation