Induction of Hematopoietic Microchimerism by Gene-Modified BMT Elicits Antigen-Specific B and T Cell Unresponsiveness toward Gene Therapy Products.
BACKGROUND
Gene therapy is a promising treatment option for hemophilia and other protein deficiencies. However, immune responses against the transgene product represent an obstacle to safe and effective gene therapy, urging for the implementation of tolerization strategies. Induction of a hematopoietic chimerismviabone marrow transplantation (BMT) is a potent means for inducing immunological tolerance in solid organ transplantation.


OBJECTIVE
We reasoned, here, that the same viral vector could be used, first, to transduce BM cells for inducing chimerism-associated transgene-specific immune tolerance and, second, for correcting protein deficiencies by vector-mediated systemic production of the deficient coagulation factor.


METHODS
Evaluation of strategies to induce B and T cell tolerance was performed usingex vivogene transfer with lentiviral (LV) vectors encoding coagulation factor IX (FIX) or the SIINFEKL epitope of ovalbumin. Following induction of microchimerismviaBMT, animals were challenged within vivogene transfer with LV vectors.


RESULTS
The experimental approach prevented humoral immune response against FIX, resulting in persistence of therapeutic levels of circulating FIX, after LV-mediated gene transferin vivo. In an ovalbumin model, we also demonstrated that this approach effectively tolerized the CD8+T cell compartment in an antigen-specific manner.


CONCLUSIONS
These results provide the proof-of-concept that inducing a microchimerism by gene-modified BMT is a powerful tool to provide transgene-specific B and T cell tolerance in a gene therapy setting.
Normandie University, UNIROUEN, Pathophysiology and Biotherapy of Inflammatory and Autoimmune Diseases, INSERM, CHU Rouen , Rouen , France.
Jérémie
J
Martinet
Normandie University, UNIROUEN, Pathophysiology and Biotherapy of Inflammatory and Autoimmune Diseases, INSERM, CHU Rouen , Rouen , France.
Gwladys
G
Bourdenet
U974, INSERM, University Pierre and Marie Curie, Paris, France; Genethon, Evry, France.
Amine
A
Meliani
Normandie University, UNIROUEN, Pathophysiology and Biotherapy of Inflammatory and Autoimmune Diseases, INSERM, CHU Rouen , Rouen , France.
Laetitia
L
Jean
Normandie University, UNIROUEN, Pathophysiology and Biotherapy of Inflammatory and Autoimmune Diseases, INSERM, CHU Rouen , Rouen , France.
Sahil
S
Adriouch
U955 Team 21, Inserm , Créteil , France.
Jose L
JL
Cohen
U974, INSERM, University Pierre and Marie Curie, Paris, France; Genethon, Evry, France.
Federico
F
Mingozzi
Normandie University, UNIROUEN, Pathophysiology and Biotherapy of Inflammatory and Autoimmune Diseases, INSERM, CHU Rouen , Rouen , France.
Olivier
O
Boyer
animal model; gene therapy; hemophilia B; microchimerism; tolerance
Blood. 2014 Jul 17;124(3):344-53
24914142
Thromb Haemost. 2011 Apr;105(4):676-87
21264447
Transl Res. 2013 Apr;161(4):230-40
23360745
Gene Ther. 2013 May;20(5):478-86
22833118
J Thromb Haemost. 2007 Jan;5(1):16-24
17002653
Lancet. 2008 Oct 11;372(9646):1342-53
18929906
Blood. 2007 Apr 1;109(7):2797-805
17170119
N Engl J Med. 2008 Jan 24;358(4):353-61
18216355
EMBO Mol Med. 2013 Nov;5(11):1698-709
24106230
Gene Ther. 2005 Oct;12(19):1453-64
15889137
Blood. 2006 Nov 15;108(10 ):3321-8
16868252
J Clin Invest. 2003 May;111(9):1347-56
12727926
Br J Haematol. 2012 Oct;159(2):123-34
22924753
Blood. 2007 Dec 15;110(13):4144-52
17726165
Haemophilia. 2008 Jul;14 Suppl 3:36-42
18510520
Arch Virol. 2012 Feb;157(2):217-23
22042211
EMBO Mol Med. 2013 Nov;5(11):1684-97
24106222
J Immunol. 2002 Aug 15;169(4):1930-5
12165518
Blood. 2005 Jun 15;105(12):4865-70
15769892
J Transl Med. 2014 Jan 25;12:25
24460861
J Immunol. 2006 Mar 15;176(6):3410-6
16517709
N Engl J Med. 2010 Oct 7;363(15):1429-37
20925545
Mol Ther. 2001 Sep;4(3):201-10
11545610
Mol Ther. 2006 Feb;13(2):260-9
16324888
Blood. 2007 Oct 15;110(8):2855-63
17569821
Mol Ther. 2016 Feb;24(1):87-95
26265250
Front Microbiol. 2011 Sep 29;2:199
22046170
Gene Ther. 2006 Jan;13(2):117-26
16163377
Gene Ther. 2010 Mar;17(3):295-304
19907498
J Immunol. 2002 Apr 1;168(7):3145-9
11907065
Blood. 1999 Nov 1;94(9):3234-41
10556212
Blood. 2012 Nov 29;120(23):4482-7
22829631
Blood. 2015 Mar 5;125(10):1553-61
25568350
Mol Ther. 2015 Apr;23(4):617-26
25655313
Mol Ther. 2005 Dec;12(6):1034-42
16226058
Hum Gene Ther. 2012 Jul;23(7):754-68
22463386
Nat Genet. 2000 Jun;25(2):217-22
10835641
Blood. 2003 Oct 1;102(7):2412-9
12791653