Integration profile of retroviral vector in gene therapy treated patients is cell-specific according to gene expression and chromatin conformation of target cell.
The analysis of genomic distribution of retroviral vectors is a powerful tool to monitor 'vector-on-host' effects in gene therapy (GT) trials but also provides crucial information about 'host-on-vector' influences based on the target cell genetic and epigenetic state. We had the unique occasion to compare the insertional profile of the same therapeutic moloney murine leukemia virus (MLV) vector in the context of the adenosine deaminase-severe combined immunodeficiency (ADA-SCID) genetic background in two GT trials based on infusions of transduced mature lymphocytes (peripheral blood lymphocytes, PBL) or a single infusion of haematopoietic stem/progenitor cells (HSC). We found that vector insertions are cell-specific according to the differential expression profile of target cells, favouring, in PBL-GT, genes involved in immune system and T-cell functions/pathways as well as T-cell DNase hypersensitive sites, differently from HSC-GT. Chromatin conformations and histone modifications influenced integration preferences but we discovered that only H3K27me3 was cell-specifically disfavoured, thus representing a key epigenetic determinant of cell-type dependent insertion distribution. Our study shows that MLV vector insertional profile is cell-specific according to the genetic/chromatin state of the target cell both in vitro and in vivo in patients several years after GT.
San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milano, Italy.
Luca
L
Biasco
Alessandro
A
Ambrosi
Danilo
D
Pellin
Cynthia
C
Bartholomae
Immacolata
I
Brigida
Maria Grazia
MG
Roncarolo
Clelia
C
Di Serio
Christof
C
von Kalle
Manfred
M
Schmidt
Alessandro
A
Aiuti
D000243:Adenosine Deaminase / Q000172:deficiency; D002843:Chromatin / Q000378:metabolism; D020869:Gene Expression Profiling; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000821:virology; D006657:Histones / Q000378:metabolism; D006801:Humans; D008214:Lymphocytes / Q000821:virology; D012190:Retroviridae / Q000235:genetics* / Q000502:physiology; D016511:Severe Combined Immunodeficiency / Q000628:therapy; D016662:Virus Integration
D002843:Chromatin; D006657:Histones; D000243:Adenosine Deaminase
Proc Natl Acad Sci U S A. 2006 Jan 31;103(5):1457-62
16432223
J Clin Invest. 2008 Sep;118(9):3132-42
18688285
N Engl J Med. 2009 Jan 29;360(5):447-58
19179314
Mol Ther. 2009 May;17(5):851-6
19293778
Curr Gene Ther. 2008 Dec;8(6):419-29
19075625
Blood. 2007 Mar 1;109(5):1897-907
17119121
PLoS Pathog. 2006 Jun;2(6):e60
16789841
AIDS. 2009 Jul 31;23(12):1461-71
19550285
J Clin Invest. 2008 Sep;118(9):3143-50
18688286
Immunity. 2009 Jan 16;30(1):155-67
19144320
Science. 2002 Jun 28;296(5577):2410-3
12089448
Cell. 2007 May 18;129(4):823-37
17512414
N Engl J Med. 2009 Jan 29;360(5):518-21
19179320
Blood. 2007 Sep 15;110(6):1770-8
17507662
AIDS. 2008 Jul 31;22(12):1411-5
18614863
Blood. 2008 Apr 15;111(8):4209-19
18218852
Science. 2003 Jun 13;300(5626):1749-51
12805549
Nat Med. 2006 Apr;12(4):401-9
16582916
Blood. 2009 Oct 22;114(17):3546-56
19652199
J Clin Invest. 2007 Aug;117(8):2083-6
17671645
PLoS One. 2009;4(2):e4571
19238208
Genome Res. 2007 Aug;17(8):1186-94
17545577
Blood. 2010 Jun 3;115(22):4356-66
20228274
Nat Methods. 2007 Dec;4(12):1051-7
18049469
Blood. 2008 Sep 15;112(6):2278-86
18566328
Blood. 2008 Oct 1;112(7):2722-9
18625888
Nat Med. 2002 May;8(5):423-5
11984564
N Engl J Med. 2003 Jan 16;348(3):255-6
12529469
Nat Biotechnol. 2006 Jun;24(6):687-96
16732270
J Clin Invest. 2007 Aug;117(8):2225-32
17671652
Science. 2009 Nov 6;326(5954):805-6
19892968
Cell Stem Cell. 2009 Jan 9;4(1):80-93
19128795
Science. 2003 Oct 17;302(5644):415-9
14564000
Mol Ther. 2009 Sep;17(9):1537-47
19532134
PLoS Genet. 2007 Aug;3(8):e136
17708682
J Clin Invest. 2009 Apr;119(4):964-75
19307726
PLoS Biol. 2004 Aug;2(8):E234
15314653
Science. 1995 Oct 20;270(5235):470-5
7570000
J Clin Invest. 2007 Aug;117(8):2233-40
17671653
Cell. 2008 Jan 25;132(2):311-22
18243105