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

  • Alexandru Tatomir
  • Cosmin A. Tegla
  • Alvaro Martin
  • Dallas Boodhoo
  • Vinh Nguyen
  • Adam J. Sugarman
  • Armugam Mekala
  • Freidrich Anselmo
  • Anamaria Talpos-Caia
  • Cornelia Cudrici
  • Tudor C. Badea
  • Violeta Rus
  • Horea Rus
Original Article

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.

Keywords

RGC-32 Astrocyte Multiple sclerosis Experimental autoimmune encephalomyelitis Extracellular matrix 

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

Notes

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.).

Compliance with ethical standards

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.

Supplementary material

12026_2018_9011_MOESM1_ESM.doc (45 kb)
Supplemental Table 1 (DOC 45 kb)
12026_2018_9011_Fig14_ESM.png (70 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)

12026_2018_9011_MOESM2_ESM.tif (134 kb)
High Resolution Image (TIF 133 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Alexandru Tatomir
    • 1
  • Cosmin A. Tegla
    • 1
    • 2
  • Alvaro Martin
    • 1
  • Dallas Boodhoo
    • 1
  • Vinh Nguyen
    • 3
  • Adam J. Sugarman
    • 1
  • Armugam Mekala
    • 1
  • Freidrich Anselmo
    • 1
  • Anamaria Talpos-Caia
    • 1
    • 4
  • Cornelia Cudrici
    • 5
  • Tudor C. Badea
    • 6
  • Violeta Rus
    • 2
    • 3
  • Horea Rus
    • 1
    • 2
    • 7
  1. 1.Department of NeurologyUniversity of Maryland School of MedicineBaltimoreUSA
  2. 2.Research ServiceVeterans Administration Maryland Health Care SystemBaltimoreUSA
  3. 3.Department of Medicine, Division of Rheumatology and Clinical ImmunologyUniversity of Maryland School of MedicineBaltimoreUSA
  4. 4.Department of Rheumatology“Iuliu Hatieganu” University of Medicine and PharmacyCluj-NapocaRomania
  5. 5.National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of HealthBethesdaUSA
  6. 6.Retinal Circuit Development and Genetics Unit, N-NRLNational Eye InstituteBethesdaUSA
  7. 7.Veterans Administration Multiple Sclerosis Center of Excellence–EastBaltimoreUSA

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