scholarly journals Ex vivo γ-retroviral gene therapy of dogs with X-linked severe combined immunodeficiency and the development of a thymic T cell lymphoma

2011 ◽  
Vol 142 (1-2) ◽  
pp. 36-48 ◽  
Author(s):  
Douglas R. Kennedy ◽  
Brian J. Hartnett ◽  
Jeffrey S. Kennedy ◽  
William Vernau ◽  
Peter F. Moore ◽  
...  
Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 195-195 ◽  
Author(s):  
S. Hacein-Bey-Abina ◽  
M. Schmidt ◽  
F. Le Deist ◽  
A. Garrigue ◽  
A. Borkhardt ◽  
...  

Abstract We have previously reported that ex vivo retroviraly-mediated gc gene transfer into CD34 (+) bone marrow precursor cells led to the correction of the immunodeficiency in 9 out of 10 patients with X-linked severe combined immunodeficiency. Follow-up now reaches more that 6 years for the first 2 treated patients. Patients’immune function has been restored. The distribution of both TCR Vb family usage and TCR Vb CDR3 length still reveals a broadly diversified T cell repertoire. Moreover 6 years after treatment the thymus is still seeded by transduced progenitor cells as attested by the presence of TRECS in peripheral blood RTE. Among these patients, three (P4, P5 and P10) developed at 30 to 34 months after gene therapy a monoclonal T cell proliferation requiring a chemotherapy. P4 received also an allogenic HSCT from a MUD but died 26 months after the occurence of the lymphoproliferation. For P5 and P10, chemotherapy has led to an overall control of the clonal proliferation. These two patients are doing well and P5 is off treatment with a good immunological recovery. Genetic analysis of the blastic cells showed that in the two first cases the vector had integrated within or upstream of the LMO2 locus causing an insertional activation of LMO2 transcription. The last case revealed the involvement of several targeted sites, but their exact contribution to the lymphoproliferation is still under investigation. The repeated involvment of LMO2 as a site of vector integration in the proliferating T-cells points to an insertional activation of this gene as at least one of the causes of the oncogenic process. However, the long latency observed in all cases (> 30 months) suggests that additional “hits” have been required for overt desease. Synergy with gc expression and thereby induced proliferative signals (explaining occurrence in SCID-X1 patients only) is the most obvious hypothesis which we are trying to analyse in a mouse model. A deep analysis of retroviral integration patterns has been performed on patients’PBMCS by LAM-PCR to estimate the frequency of potentially harmful integration events and to assess the risk factors associated with the LTR’s strong enhancer effect of the MLV-based retroviral vector. 708 unique integration sites (IS) have been obtained from all analysed patients post-gene therapy and among them, 577 could be mapped unequivocally to the human genome. *Most of these insertions (63%) are located in the vicinity of 10kb or within the coding sequence of a known gene*. A significant peak of insertion frequency is related closely to the transcription strart site *among the 577 IS, 43 are common integration sites. Among the latter, we found out a high selection of genes involved in human oncogenic process.


Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 410-410 ◽  
Author(s):  
Javier Chinen ◽  
Jennifer M. Puck ◽  
Joie Davis ◽  
Gilda F. Linton ◽  
Narda L. Whiting-Theobald ◽  
...  

Abstract X-linked severe combined immunodeficiency (XSCID) results from mutations in IL2RG, which encodes the common gamma chain (γc) shared by receptors for IL-2, 4, 7, 9, 15 and 21. XSCID is best treated with bone marrow transplantation (BMT) from an HLA-matched sibling. Patients lacking a matched sibling can benefit from a T-cell depleted haploidentical BMT, but some do not achieve adequate immune reconstitution. Ex vivo autologous hematopoietic stem cell (HSC) gene therapy may be an alternative to haploidentical BMT. In a French trial, 9 of 10 XSCID infants had immune reconstitution following ex vivo transduction of autologous HSC with a retroviral vector encoding γc. Selective development and expansion of T, NK and B cells from progenitors expressing γc was important to the success of this therapy. However, the 2 youngest patients, treated at 1 and 3 months of age, later developed T cell leukemias associated with retrovirus insertions that activated the LMO2 transcription factor. Young age at treatment might have had a role in the development of these adverse events. We have developed an XSCID gene transfer protocol as salvage treatment for older patients who have failed haploidentical BMT. An 11 year-old XSCID patient with no detectable engraftment from prior haploidentical BMTs had lymphocytopenia, growth failure, infections, chronic diarrhea and skin rashes. After G-CSF mobilization and harvest by apheresis, his purified autologous peripheral blood CD34+ cells were transduced daily for 4 days with GALV-MFGS-γc retrovirus in the presence of growth factors and Retronectin®. Eighty million cells/kg (80% CD34+; 40% γc transgene positive) were reinfused. At 1, 2 and 3 months after treatment, provirus marking by PCR of unseparated blood leukocytes was 1.4%, 2.3% and <0.01%, respectively. At 4.5 months, marking reappeared in lineages dependent on IL2RG expression: 0.5% in T cells, 0.1% in NK cells and 0.05% in B cells. This lineage-specific marking persisted at the same level at 6 months. LAM PCR showed polyclonal marking. T-lymphocytes have not yet increased above 300/μl. However, from 2 months after gene therapy the patient experienced a sustained improvement in well-being with resolution of lifelong diarrhea and rashes. No infections have occurred except one episode of otitis externa 3 months post therapy that resolved promptly to oral antibiotics. At six months submandibular lymph nodes were palpable for the first time in his life. Thymus size on chest CT images increased from 1.8 cm3 to 3.2 cm3. Additional follow up is necessary to know if gene marking and clinical improvement persist and if significant expansion of corrected lymphocytes occurs. Our preliminary results suggest that ex vivo retrovirus-mediated gene therapy, targeting CD34+ cells from peripheral blood, may benefit older children with XSCID who have failed haploidentical BMT.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Koen Debackere ◽  
Lukas Marcelis ◽  
Sofie Demeyer ◽  
Marlies Vanden Bempt ◽  
Nicole Mentens ◽  
...  

AbstractPeripheral T-cell lymphoma (PTCL) is a heterogeneous group of non-Hodgkin lymphomas with poor prognosis. Up to 30% of PTCL lack distinctive features and are classified as PTCL, not otherwise specified (PTCL-NOS). To further improve our understanding of the genetic landscape and biology of PTCL-NOS, we perform RNA-sequencing of 18 cases and validate results in an independent cohort of 37 PTCL cases. We identify FYN-TRAF3IP2, KHDRBS1-LCK and SIN3A-FOXO1 as new in-frame fusion transcripts, with FYN-TRAF3IP2 as a recurrent fusion detected in 8 of 55 cases. Using ex vivo and in vivo experiments, we demonstrate that FYN-TRAF3IP2 and KHDRBS1-LCK activate signaling pathways downstream of the T cell receptor (TCR) complex and confer therapeutic vulnerability to clinically available drugs.


2018 ◽  
Vol 10 (1) ◽  
Author(s):  
Erik L. Clarke ◽  
A. Jesse Connell ◽  
Emmanuelle Six ◽  
Nadia A. Kadry ◽  
Arwa A. Abbas ◽  
...  

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Sanjay de Mel ◽  
Masturah B. M. Rashid ◽  
Xi Yun Zhang ◽  
Jasmine Goh ◽  
Chun Tsu Lee ◽  
...  

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4651-4651
Author(s):  
Johnathan Ledet ◽  
Joshua May ◽  
Seth Billelo ◽  
Ellen Friday ◽  
Mary Nordberg ◽  
...  

Abstract Cutaneous T-cell lymphoma (CTCL) represents a spectrum of diseases characterized by the accumulation of clonal lymphocytes in the skin. It has been well established that primary cutaneous lymphomas of T-cell lineage have shown ALK protein expression. Furthermore, we have previously shown that ALK-ALCL cell lines highly express the coxsackie-adenovirus receptor (CAR), primary receptor for recombinant wild type-fiber adenovirus-derived vectors for gene therapy. Since skin-directed therapies are the preferred first-line treatment the presence of CAR in CTCL may be helpful in developing therapeutic modalities such gene or targeted molecular therapy. By using ALK-immunostaining, we analyzed retrospectively a series of human tissue primarily diagnosed as CTCL [5 mycosis fungoides (MF), 4 (ALCL)] or peripheral T cell lymphoma (1 PTCL) in order to establish the level of CAR expression in primary CTCL tissue. Fluorescence in situ hybridization (FISH) analyses were also performed on these cases using a locus specific DNA probe for the ALK/p2 locus. We used ALK identification because at the time when the primary diagnosis was placed, the samples were not tested for ALK expression. Patients ranged in age from 32–73 years old with a median age of 58. Normal skin biopsies from 8 patients were used as controls. The results of our analysis are illustrated in Table 1. TABLE 1. DIAGNOSIS SPECIMEN CAR INTENSITY/% ALK INTENSITY/% FISH CD30 ALCL RT ANKLE MASS 0 / 0 N/A N/A POS ALCL RT FLANK MASS 1+/ 80 1+ / 80 32% B-ANEUPLOID ALCL SKIN, LOWER RIGHT EXTREMITY 0 / 0 2+/ 10 NL POS PTCL SKIN, RT ARM 3+ / 95 3 +/ 70 MF SKIN, RT AXILLA 1 + / 5 3+ / 5 ALK POS MF SKIN 1+ / 5 3+ / 5 NL 21% ANEUPLOID MF SKIN, ARM 0 / 0 3+ / 90 NL (18% ANEUPLOID) POS (RARE) MF SKIN, LEFT BACK 2+ / 40 3+ / 95 NL MF SKIN, RT FOREARM 2+ / 30 1 + / 30 In this study, we show that the immunostaining of CAR in combination with classic ALK-immuno or ALK-FISH is feasible and helps in identifying CTCL cases candidate for skin targeted adenoviral-mediated gene therapy.


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