Overexpression of Glutamate Decarboxylase in Mesenchymal Stem Cells Enhances Their Immunosuppressive Properties and Increases GABA and Nitric Oxide Levels.
The neurotransmitter GABA has been recently identified as a potent immunosuppressive agent that targets both innate and adaptive immune systems and prevents disease progression of several autoimmunity models. Mesenchymal stem cells (MSCs) are self-renewing progenitor cells that differentiate into various cell types under specific conditions, including neurons. In addition, MSC possess strong immunosuppressive capabilities. Upon cytokine priming, undifferentiated MSC suppress T-cell proliferation via cell-to-cell contact mechanisms and the secretion of soluble factors like nitric oxide, prostaglandin E2 and IDO. Although MSC and MSC-derived neuron-like cells express some GABAergic markers in vitro, the role for GABAergic signaling in MSC-mediated immunosuppression remains completely unexplored. Here, we demonstrate that pro-inflammatory cytokines selectively regulate GAD-67 expression in murine bone marrow-MSC. However, expression of GAD-65 is required for maximal GABA release by MSC. Gain of function experiments using GAD-67 and GAD-65 co-expression demonstrates that GAD increases immunosuppressive function in the absence of pro-inflammatory licensing. Moreover, GAD expression in MSC evokes an increase in both GABA and NO levels in the supernatants of co-cultured MSC with activated splenocytes. Notably, the increase in NO levels by GAD expression was not observed in cultures of isolated MSC expressing GAD, suggesting crosstalk between these two pathways in the setting of immunosuppression. These results indicate that GAD expression increases MSC-mediated immunosuppression via secretion of immunosuppressive agents. Our findings may help reconsider GABAergic activation in MSC for immunological disorders.
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Mariana
M
Urrutia
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Sebastián
S
Fernández
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Marisol
M
González
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Rodrigo
R
Vilches
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Pablo
P
Rojas
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Manuel
M
Vásquez
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Mónica
M
Kurte
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Ana María
AM
Vega-Letter
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Flavio
F
Carrión
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Fernando
F
Figueroa
Departamento de Biología, Universidad de Santiago de Chile, Santiago, Chile.
Patricio
P
Rojas
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Carlos
C
Irarrázabal
Centro de Investigación Biomédica, Facultad de Medicina, Universidad de Los Andes, Santiago, Chile.
Rodrigo A
http://orcid.org/0000-0001-8724-091X
RA
Fuentealba
Proc Natl Acad Sci U S A. 2003 Apr 1;100(7):4293-8
12634427
Ciba Found Symp. 1988;136:42-60
3068016
Nat Genet. 2003 Jul;34(3):263-4
12796781
J Mol Neurosci. 2012 Jul;47(3):619-30
22215379
Nat Cell Biol. 2003 Sep;5(9):834-9
12942087
Biol Res. 2012;45(3):269-77
23283436
Exp Hematol. 2002 Jan;30(1):42-8
11823036
Anal Biochem. 1993 Oct;214(1):11-6
7504409
J Neurochem. 1996 May;66(5):2082-90
8780039
Stem Cells. 2011 Oct;29(10):1572-9
21837654
Stem Cells Dev. 2015 Sep 15;24(18):2171-80
26153898
Blood. 2004 Jun 15;103(12):4619-21
15001472
Blood. 2007 Jan 1;109(1):228-34
16985180
Stem Cells. 2009 Aug;27(8):1954-62
19544427
Proc Natl Acad Sci U S A. 1997 Jun 10;94(12):6496-9
9177246
Blood. 2005 Feb 15;105(4):1815-22
15494428
Stem Cell Res Ther. 2010 Mar 15;1(1):2
20504283
PLoS One. 2011;6(9):e25338
21966503
Annu Rev Immunol. 2013;31:285-316
23298209
Mol Ther. 2012 Jan;20(1):187-95
21934657
Nat Struct Mol Biol. 2007 Apr;14(4):280-6
17384644
Neuron. 2007 Dec 6;56(5):851-65
18054861
PLoS One. 2010 Apr 26;5(4):e10088
20436665
J Cell Biol. 2002 Sep 30;158(7):1229-38
12356867
Nat Biotechnol. 2014 Mar;32(3):252-60
24561556
PLoS One. 2012;7(8):e42959
22927941
Neuroscience. 2014 Jul 25;273:24-38
24814730
J Cell Biol. 2010 Sep 6;190(5):911-25
20805323
J Neuroimmunol. 2013 Feb 15;255(1-2):45-53
23194644
Eur J Neurosci. 2008 Jun;27(11):2783-802
18588525
Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):14060-5
9391152
J Inflamm (Lond). 2005 Jul 26;2:8
16045800
J Neuroimmunol. 2007 Aug;188(1-2):64-8
17599468
Neuroscience. 1999;93(4):1475-82
10501472
J Immunol. 2004 Jun 1;172(11):6545-9
15153468
J Pathol. 2009 Jan;217(2):318-24
19023885
Lancet. 2004 May 1;363(9419):1439-41
15121408
Eur J Immunol. 2008 Jun;38(6):1745-55
18493986
J Neurochem. 2003 Aug;86(4):879-87
12887686
EMBO J. 1991 May;10(5):1275-84
2022191
J Physiol. 2004 Jan 1;554(Pt 1):100-10
14678495
FASEB J. 2014 Sep;28(9):3856-66
24830384
Nat Med. 2007 Jul;13(7):862-7
17589520
Scand J Immunol. 2003 Jan;57(1):11-20
12542793
Clin Pharmacol Ther. 2010 Jan;87(1):23-6
20019698
Immunol Rev. 2005 Oct;207:19-30
16181324
Stem Cells. 2008 Jan;26(1):212-22
17932417
Cell Stem Cell. 2009 Jul 2;5(1):54-63
19570514
J Biol Chem. 1981 Oct 10;256(19):9816-9
7196914
J Neuroimmunol. 2008 Dec 15;205(1-2):44-50
18954912
J Biol Chem. 1995 Feb 3;270(5):2241-6
7836456
J Neurosci Res. 2000 Aug 15;61(4):364-70
10931522
Transplantation. 2013 Oct 15;96(7):616-23
23851932
Free Radic Biol Med. 2003 Apr 1;34(7):856-61
12654474
Mediators Inflamm. 2010;2010:865601
20628526
Mol Immunol. 2006 Mar;43(9):1432-42
16213022
Stem Cells. 2005 Mar;23(3):392-402
15749934
Nat Rev Neurosci. 2002 Sep;3(9):715-27
12209120
Stem Cells. 2009 Apr;27(4):909-19
19353519
Stem Cells Dev. 2010 May;19(5):657-68
19886804
J Immunol. 1998 Jun 15;160(12):5729-34
9637481
PLoS One. 2010 Dec 07;5(12):e14247
21151872
Cytotherapy. 2006;8(4):315-7
16923606
Lancet. 2008 May 10;371(9624):1579-86
18468541
Cell Mol Immunol. 2012 Nov;9(6):473-81
23085948
Cell Stem Cell. 2008 Feb 7;2(2):141-50
18371435
Blood. 2007 Feb 15;109(4):1422-32
17038530
Proc Natl Acad Sci U S A. 2011 Jul 12;108(28):11692-7
21709230
Gene Ther. 2003 May;10(10):928-31
12732877
Blood. 2003 Nov 15;102(10):3837-44
12881305
Amino Acids. 2013 Jul;45(1):87-94
22160261
Blood. 2004 Mar 1;103(5):1662-8
14592819
Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2115-9
1549570
Nat Biotechnol. 2005 Nov;23(11):1399-405
16273073
PLoS One. 2014 Feb 13;9(2):e88667
24551133
Blood. 2011 May 5;117(18):4826-35
21389316
Curr Top Microbiol Immunol. 2007;316:97-117
17969445
J Immunol. 2004 Oct 15;173(8):5298-304
15470076
Proc Natl Acad Sci U S A. 2010 Feb 9;107(6):2580-5
20133656
J Immunol. 2002 Jan 15;168(2):689-95
11777962
PLoS One. 2012;7(10):e47559
23133515
J Neurosci. 1997 Jan 15;17 (2):625-34
8987785
J Biol Chem. 1999 Dec 24;274(52):37200-9
10601283
Blood. 2003 May 1;101(9):3722-9
12506037
Exp Hematol. 1976 Sep;4(5):267-74
976387
J Cell Sci. 2006 Jun 1;119(Pt 11):2204-13
16684817
J Neurosci. 2004 Oct 13;24(41):8940-9
15483113
J Neuroimmunol. 1999 Apr 1;96(1):21-8
10227421
Autoimmun Rev. 2015 Nov;14(11):1048-56
26226414
J Nucl Med. 2006 Mar;47(3):486-91
16513618
PLoS One. 2015 Feb 03;10(2):e0117130
25647668
Nat Med. 2012 Apr 15;18(5):759-65
22504485
Leukemia. 2011 Sep;25(9):1408-14
21617697
Diabetes Metab Syndr Obes. 2015 Feb 03;8:79-87
25678807
Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11451-5
9326630
J Neurochem. 1992 Feb;58(2):618-27
1729406
Nat Med. 2009 Jan;15(1):42-9
19098906
Biochem Biophys Res Commun. 1996 Dec 24;229(3):891-5
8954991
Glia. 2011 Nov;59(11):1600-11
21748804
Stem Cells Int. 2015;2015:140170
25838828
Brain Res. 2009 Nov 10;1297:23-31
19699726
Autoimmunity. 2011 Sep;44(6):465-70
21604972
Stem Cells. 2016 Feb;34(2):456-69
26528946
Blood. 2002 May 15;99(10):3838-43
11986244
PLoS One. 2010 Jul 29;5(7):e11884
20686698
J Physiol. 1993 May;464:423-39
8229811
PLoS One. 2012;7(9):e45590
23029122
Cell Res. 2008 Aug;18(8):846-57
18607390
Nature. 2013 Apr 11;496(7444):238-42
23535595
Pharmacol Res. 2008 May;57(5):374-82
18467116
Neuropharmacology. 2011 Feb-Mar;60(2-3):513-9
21093461