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K Pike-Overzet,
M Rodijk,
Y-Y Ng,
M R M Baert,
C Lagresle-Peyrou,
A Schambach,
F Zhang,
R C Hoeben, S Hacein-Bey-Abina,
A C Lankester,
R G M Bredius,
G J A Driessen,
A J Thrasher,
C Baum,
M Cavazzana-Calvo,
J J M van Dongen,
F J T Staal
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ABSTRACT: Severe combined immunodeficiency (SCID) patients with an inactivating mutation in recombination activation gene 1 (RAG1) lack B and T cells due to the inability to rearrange immunoglobulin (Ig) and T-cell receptor (TCR) genes. Gene therapy is a valid treatment option for RAG-SCID patients, especially for patients lacking a suitable bone marrow donor, but developing such therapy has proven challenging. As a preclinical model for RAG-SCID, we used Rag1-/- mice and lentiviral self-inactivating (SIN) vectors harboring different internal elements to deliver native or codon-optimized human RAG1 sequences. Treatment resulted in the appearance of B and T cells in peripheral blood and developing B and T cells were detected in central lymphoid organs. Serum Ig levels and Ig and TCR Vβ gene segment usage was comparable to wild-type (WT) controls, indicating that RAG-mediated rearrangement took place. Remarkably, relatively low frequencies of B cells produced WT levels of serum immunoglobulins. Upon stimulation of the TCR, corrected spleen cells proliferated and produced cytokines. In vivo challenge resulted in production of antigen-specific antibodies. No leukemia development as consequence of insertional mutagenesis was observed. The functional reconstitution of the B- as well as the T-cell compartment provides proof-of-principle for therapeutic RAG1 gene transfer in Rag1-/- mice using lentiviral SIN vectors.
Leukemia: official journal of the Leukemia Society of America, Leukemia Research Fund, U.K 05/2011; 25(9):1471-83. · 8.30 Impact Factor
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F J T Staal,
C Baum,
C Cowan,
E Dzierzak, S Hacein-Bey-Abina,
S Karlsson,
T Lapidot,
I Lemischka,
S Mendez-Ferrer,
H Mikkers,
K Moore,
E Moreno,
C L Mummery,
C Robin,
T Suda,
M Van Pel,
G Vanden Brink,
J J Zwaginga,
W E Fibbe
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ABSTRACT: The hematopoietic stem cell (HSC) is the prototype organ-regenerating stem cell (SC), and by far the most studied type of SC in the body. Currently, HSC-based therapy is the only routinely used SC therapy; however, advances in the field of embryonic SCs and induced pluripotent SCs may change this situation. Interest into in vitro generation of HSCs, including signals for HSC expansion and differentiation from these more primitive SCs, as well as advances in other organ-specific SCs, in particular the intestine, provide promising new applications for SC therapies. Here, we review the basic principles of different SC systems, and on the basis of the experience with HSC-based SC therapy, provide recommendations for clinical application of emerging SC technologies.
Leukemia: official journal of the Leukemia Society of America, Leukemia Research Fund, U.K 04/2011; 25(7):1095-102. · 8.30 Impact Factor
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ABSTRACT: The concept of gene therapy emerged as a way of correcting monogenic inherited diseases by introducing a normal copy of the mutated gene into at least some of the patients' cells. Although this concept has turned out to be quite complicated to implement, it is in the field of primary immunodeficiencies (PIDs) that proof of feasibility has been undoubtedly achieved. There is now a strong rationale in support of gene therapy for at least some PIDs, as discussed in this article.
Hematology/oncology clinics of North America 02/2011; 25(1):89-100. · 2.05 Impact Factor
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A. Deichmann,
M. H. Brugman,
C. C. Bartholomae,
K. Schwarzwaelder,
M. M. Verstegen,
S. J. Howe,
A. Arens,
M. G. Ott,
D. Hoelzer,
R. Seger, [......],
F. Mavilio,
B. Cassani,
A. Aiuti,
C. E. Dunbar,
C. Baum,
H. B. Gaspar,
A. J. Thrasher,
C. von Kalle,
M. Schmidt,
G. Wagemaker
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ABSTRACT: Vector-associated side effects in clinical gene therapy have provided insights into the molecular mechanisms of hematopoietic regulation in vivo. Surprisingly, many retrovirus insertion sites (RIS) present in engrafted cells have been found to cluster nonrandomly in close association with specific genes. Our data demonstrate that these genes directly influence the in vivo fate of hematopoietic cell clones. Analysis of insertions thus far has been limited to individual clinical studies. Here, we studied >7,000 insertions retrieved from various studies. More than 40% of all insertions found in engrafted gene-modified cells were clustered in the same genomic areas covering only 0.36% of the genome. Gene classification analyses displayed significant overrepresentation of genes associated with hematopoietic functions and relevance for cell growth and survival in vivo. The similarity of insertion distributions indicates that vector insertions in repopulating cells cluster in predictable patterns. Thus, insertion analyses of preclinical in vitro and murine in vivo studies as well as vector insertion repertoires in clinical trials yielded concerted results and mark a small number of interesting genomic loci and genes that warrants further investigation of the biological consequences of vector insertions.
Mol Ther. 01/2011;
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ABSTRACT: The concept of gene therapy emerged as a way of correcting monogenic inherited diseases by introducing a normal copy of the mutated gene into at least some of the patients' cells. Although this concept has turned out to be quite complicated to implement, it is in the field of primary immunodeficiencies (PIDs) that proof of feasibility has been undoubtedly achieved. There is now a strong rationale in support of gene therapy for at least some PIDs, as discussed in this article.
Immunology and allergy clinics of North America 05/2010; 30(2):237-48. · 3.18 Impact Factor
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ABSTRACT: X-linked severe-combined immunodeficiency (SCID-X1) has been treated by therapeutic gene transfer using gammaretroviral vectors, but insertional activation of proto-oncogenes contributed to leukemia in some patients. Here we report a longitudinal study of gene-corrected progenitor cell populations from 8 patients using 454 pyrosequencing to map vector integration sites, and extensive resampling to allow quantification of clonal abundance. The number of transduced cells infused into patients initially predicted the subsequent diversity of circulating cells. A capture-recapture analysis was used to estimate the size of the gene-corrected cell pool, revealing that less than 1/100th of the infused cells had long-term repopulating activity. Integration sites were clustered even at early time points, often near genes involved in growth control, and several patients harbored expanded cell clones with vectors integrated near the cancer-implicated genes CCND2 and HMGA2, but remain healthy. Integration site tracking also documented that chemotherapy for adverse events resulted in successful control. The longitudinal analysis emphasizes that key features of transduced cell populations--including diversity, integration site clustering, and expansion of some clones--were established early after transplantation. The approaches to sequencing and bioinformatics analysis reported here should be widely useful in assessing the outcome of gene therapy trials.
Blood 03/2010; 115(22):4356-66. · 9.90 Impact Factor
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ABSTRACT: Following gene therapy of SCID-X1 using murine leukemia virus (MLV) derived vector, two patients developed leukemia owing to an activating vector integration near the LMO2 gene. We found that these integrations reside within FRA11E, a common fragile site known to correlate with chromosomal breakpoints in tumors. Further analysis showed that fragile sites attract a nonrandom number of MLV integrations, shedding light on its integration mechanism and risk-to-benefit ratio in gene therapy.
Gene Therapy 08/2006; 13(13):1057-9. · 3.71 Impact Factor
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S Caillat-Zucman,
F Le Deist,
E Haddad,
M Gannagé,
L Dal Cortivo,
N Jabado, S Hacein-Bey-Abina,
S Blanche,
J-L Casanova,
A Fischer,
M Cavazzana-Calvo
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ABSTRACT: Hematological inherited diseases can be cured by hematopoietic stem cell transplantation (HSCT) from an human leukocyte antigen (HLA)-identical sibling donor (MSD), but the outcome of unrelated donors (URD) or haploidentical donors (HMD) has been a cause of concern. In all, 94 children affected with inherited diseases underwent HSCT at a single center using MSD (group A, n=31), URD (group B, n=23) or HMD (group C, n=40). There was no difference in the rate of engraftment or in the incidence of grades III-IV acute graft-versus-host disease (GVHD) between the groups. Survival rate was 80.6% in group A, 62.5% in group B and 47.5% in group C (P=0.023). In group B, survival rate was 73.7% in the subgroup with zero or one class I mismatch, and 25% in the subgroup with two or more class I mismatches (P=0.04). In group C, survival rate was 83.3% in the 9/10-identical subgroup, 64.3% in the seven or 8/10 subgroup, and 25% in the five or 6/10 subgroup (P=0.0007). Thus, engraftment, incidence of GVHD and survival are similar in recipients of grafts from MSD, URD with 0-1 class I-mismatch, or HMD with at least 7/10 HLA matches. The low success of HSCT using more disparate donors suggests reserving them for patients with very poor prognosis.
Bone Marrow Transplantation 07/2004; 33(11):1089-95. · 3.75 Impact Factor
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S Hacein-Bey-Abina,
C Von Kalle,
M Schmidt,
M P McCormack,
N Wulffraat,
P Leboulch,
A Lim,
C S Osborne,
R Pawliuk,
E Morillon, [......],
E Macintyre,
F Sigaux,
J Soulier,
L E Leiva,
M Wissler,
C Prinz,
T H Rabbitts,
F Le Deist,
A Fischer,
M Cavazzana-Calvo
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ABSTRACT: We have previously shown correction of X-linked severe combined immunodeficiency [SCID-X1, also known as gamma chain (gamma(c)) deficiency] in 9 out of 10 patients by retrovirus-mediated gamma(c) gene transfer into autologous CD34 bone marrow cells. However, almost 3 years after gene therapy, uncontrolled exponential clonal proliferation of mature T cells (with gammadelta+ or alphabeta+ T cell receptors) has occurred in the two youngest patients. Both patients' clones showed retrovirus vector integration in proximity to the LMO2 proto-oncogene promoter, leading to aberrant transcription and expression of LMO2. Thus, retrovirus vector insertion can trigger deregulated premalignant cell proliferation with unexpected frequency, most likely driven by retrovirus enhancer activity on the LMO2 gene promoter.
Science 11/2003; 302(5644):415-9. · 31.20 Impact Factor
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ABSTRACT: Recent clinical trials in patients with a severe combined immunodeficiency disease demonstrate that gene therapy is a powerful tool in the treatment of genetic blood defects. Recent identification of the genes involved in the pathogenesis of inherited lymphohemopoietic disorders led to animal models of gene transfer. Extensive preclinical studies have overcome some of the obstacles involved in the transduction of hemopoietic cells. These promising results led to the approval of several clinical trials that are currently underway. This review focuses on the clinical outcome in patients with genetic blood defects treated by gene transfer and examines the progress achieved to date and the problems that have been encountered. Despite the obstacles, improved clinical results for several of these diseases are expected within the next 5 years.
Current Opinion in Hematology 12/2001; 8(6):360-7. · 4.52 Impact Factor
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ABSTRACT: Hematopoietic stem cells (HSC) have been widely used for autologous and allodeneic transplantation during decades, although little was known about their migration, survival, self-renewal and differentiation process. Their sorting by the CD34(+) marker they express at the cell surface in human has been challenged by the recent discovery of HSC in the CD34(-) compartment that may precede CD34(+) HSC in the differentiation process. Until recently, stem cells present in the bone marrow were thought to be specific for hematopoiesis. Some experiments including clinical trials showing the formation of various tissues, muscle, neural cells and hepatocytes for instance, after transplantation of medullar cells, have challenged this dogma. In fact, the proofs of such a transdifferentiation process by HSC are still missing and the observations may result from the differentiation of other mulipotent stem cells present in the bone marrow, such as mesenchymal stem cells and more primitive multipotent adult progenitor cells (MAPC) and side population (SP) cells.
Annales de biologie clinique 62(2):131-8. · 0.34 Impact Factor
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S Hacein-Bey-Abina,
C von Kalle,
M Schmidt,
F Le Deist,
N.M. Wulffraat,
Elisabeth McIntyre,
Isabelle Radford,
J.-L. Villeval,
Christophe Fraser,
M Cavazzana-Calvo,
A Fischer
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S Hacein-Bey-Abina,
A. Garrigue,
GP. Wang,
J Soulier,
A Lim,
E Morillon,
E Clappier,
L. Caccavelli,
E Delabesse,
K Beldjord, [......],
B H Belohradsky,
U Wintergerst,
M.C. Velez,
L.E. Leiva,
R. Sorensen,
N.M. Wulffraat,
S Blanche,
FD. Bushman,
A Fischer,
M Cavazzana-Calvo
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S Hacein-Bey-Abina,
C von Kalle,
M Schmidt,
M P McCormack,
N.M. Wulffraat,
P Leboulch,
A Lim,
C S Osborne,
R Pawliuk,
E Morillon, [......],
E Macintyre,
F Sigaux,
J Soulier,
L.E. Leiva,
M Wissler,
C. Prinz,
T H Rabbitts,
F Le Deist,
A Fischer,
M Cavazzana-Calvo
