Showing papers by "Laurent Poirot published in 2015"
TL;DR: The applicability of TALEN-mediated genome editing to a scalable process enables the manufacturing of third-party CAR T-cell immunotherapies against arbitrary targets and can therefore be used in an "off-the-shelf" manner akin to other biologic immunopharmaceuticals.
Abstract: Adoptive immunotherapy using autologous T cells endowed with chimeric antigen receptors (CAR) has emerged as a powerful means of treating cancer. However, a limitation of this approach is that autologous CAR T cells must be generated on a custom-made basis. Here we show that electroporation of transcription activator-like effector nuclease (TALEN) mRNA allows highly efficient multiplex gene editing in primary human T cells. We use this TALEN-mediated editing approach to develop a process for the large-scale manufacturing of T cells deficient in expression of both their αβ T-cell receptor (TCR) and CD52, a protein targeted by alemtuzumab, a chemotherapeutic agent. Functionally, T cells manufactured with this process do not mediate graft-versus-host reactions and are rendered resistant to destruction by alemtuzumab. These characteristics enable the administration of alemtuzumab concurrently or prior to engineered T cells, supporting their engraftment. Furthermore, endowing the TALEN-engineered cells with a CD19 CAR led to efficient destruction of CD19(+) tumor targets even in the presence of the chemotherapeutic agent. These results demonstrate the applicability of TALEN-mediated genome editing to a scalable process, which enables the manufacturing of third-party CAR T-cell immunotherapies against arbitrary targets. As such, CAR T-cell immunotherapies can therefore be used in an "off-the-shelf" manner akin to other biologic immunopharmaceuticals
515 citations
TL;DR: This work proposes to address three requirements of allogeneic CAR T-cell infusion through the development of multidrug-resistant T cell receptor αβ-deficient CAR T cells and demonstrates that these engineered T cells displayed efficient antitumor activity and proliferated in the presence of purine and pyrimidine nucleoside analogues.
83 citations
Patent•
11 Mar 2015
TL;DR: In this paper, a method for the preparation and use of engineered T-cells, particularly for immunotherapy, is presented. But the method is not suitable for allogeneic transplantations, especially because it reduces both the risk of rejection by the host's immune system and risk of developing graft versus host disease.
Abstract: The present invention pertains to engineered T-cells, method for their preparation and their use as medicament, particularly for immunotherapy. The engineered T-cells of the invention are characterized in that the expression of beta 2-microglobulin (B2M) and/or class I I major histocompatibility complex transactivator (CIITA) is inhibited, e.g., by using rare-cutting endonucleases able to selectively inactivating by DNA cleavage the gene encoding B2M and/or CIITA, or by using nucleic acid molecules which inhibit the expression of B2M and/or CIITA. In order to further render the T-cell non-alloreactive, at least one gene encoding a component of the T-cell receptor is inactivated, e.g., by using a rare-cutting endonucleases able to selectively inactivating by DNA cleavage the gene encoding said TCR component. In addition, expression of immunosuppressive polypeptide can be performed on those modified T-cells in order to prolong the survival of these modified T cells in host organism. Such modified T-cell is particularly suitable for allogeneic transplantations, especially because it reduces both the risk of rejection by the host's immune system and the risk of developing graft versus host disease. The invention opens the way to standard and affordable adoptive immunotherapy strategies using T-Cells for treating cancer, infections and auto-immune diseases.
81 citations
Patent•
13 Feb 2015
TL;DR: In this paper, the authors proposed a method of developing genetically engineered immune cells for immunotherapy, which can be endowed with Chimeric Antigen Receptors targeting an antigen marker that is common to both the pathological cells and the immune cells (ex: CD38, CSl or CD70).
Abstract: Methods of developing genetically engineered immune cells for immunotherapy, which can be endowed with Chimeric Antigen Receptors targeting an antigen marker that is common to both the pathological cells and said immune cells (ex: CD38, CSl or CD70) by the fact that the genes encoding said markers are inactivated in said immune cells by a rare cutting endonuclease such as TALEN, Cas9 or argonaute
75 citations
TL;DR: By implementing new non-conventional RVDs (ncRVDs) that possess novel intrinsic targeting specificity features, it is demonstrated in living cells the possibility to efficiently promote TALEN-mediated processing of a target in the HBB locus and alleviate undesired off-site cleavage.
Abstract: A key feature when designing DNA targeting tools and especially nucleases is specificity. The ability to control and tune this important parameter represents an invaluable advance to the development of such molecular scissors. Here, we identified and characterized new non-conventional RVDs (ncRVDs) that possess novel intrinsic targeting specificity features. We further report a strategy to control TALEN targeting based on the exclusion capacities of ncRVDs (discrimination between different nucleotides). By implementing such ncRVDs, we demonstrated in living cells the possibility to efficiently promote TALEN-mediated processing of a target in the HBB locus and alleviate undesired off-site cleavage. We anticipate that this method can greatly benefit to designer nucleases, especially for therapeutic applications and synthetic biology.
39 citations
Patent•
20 Feb 2015
TL;DR: In this article, the authors present a method for the preparation and use of engineered T-cells for immunotherapy, particularly for cancer and bacterial or viral infections, which opens the way to standard and affordable adoptive immunotherapy strategies, especially for treating or preventing cancer, and bacterial and viral infections.
Abstract: The present invention pertains to engineered T-cells, method for their preparation and their use as medicament, particularly for immunotherapy. The engineered T-cells of the invention are designed to express both a Chimeric Antigen Receptor (CAR) directed against at least one antigen expressed at the surface of a malignant or infected cell, and a secreted inhibitor of regulatory T-cells (Treg). Preferably, such secreted inhibitor is a peptide inhibitor of forkhead/winged helix transcription factor 3 (FoxP3), a specific factor involved into the differentiation of T-cells into regulatory T-cells. The engineered T-cells of the invention direct theirimmune activity towards specific malignant or infected cells, while at the same time will prevent neighbouring regulatory T-cells from modulating the immune response. The invention opens the way to standard and affordable adoptive immunotherapy strategies, especially for treating or preventing cancer, and bacterial or viral infections.
38 citations
TL;DR: High numbers of erythroid cells derived from gene-edited PKDiPSCs showed correction of the energetic imbalance, providing an approach to correct metabolic erythyroid diseases and demonstrating the practicality of this approach to generate the large cell numbers required for comprehensive biochemical and metabolic ERYthroid analyses.
Abstract: Pyruvate kinase deficiency (PKD) is a rare erythroid metabolic disease caused by mutations in the PKLR gene. Erythrocytes from PKD patients show an energetic imbalance causing chronic non-spherocytic hemolytic anemia, as pyruvate kinase defects impair ATP production in erythrocytes. We generated PKD induced pluripotent stem cells (PKDiPSCs) from peripheral blood mononuclear cells (PB-MNCs) of PKD patients by non-integrative Sendai viral vectors. PKDiPSCs were gene edited to integrate a partial codon-optimized R-type pyruvate kinase cDNA in the second intron of the PKLR gene by TALEN-mediated homologous recombination (HR). Notably, we found allele specificity of HR led by the presence of a single-nucleotide polymorphism. High numbers of erythroid cells derived from gene-edited PKDiPSCs showed correction of the energetic imbalance, providing an approach to correct metabolic erythroid diseases and demonstrating the practicality of this approach to generate the large cell numbers required for comprehensive biochemical and metabolic erythroid analyses.
31 citations
Patent•
07 Oct 2015
TL;DR: In this paper, a method to modulate the level of activation of an engineered immune cell (such as a Chimeric Antigen Receptor T-cell) for immunotherapy is presented.
Abstract: The present invention relates to a method to modulate the level of activation of an engineered immune cell (such as a Chimeric Antigen Receptor T-cell) for immunotherapy. The present invention also relates to cells obtained by the present method, preferably comprising said modulable/tunable chimeric antigen receptors for use in therapeutic or prophylactic treatment.
26 citations
Patent•
17 Dec 2015
TL;DR: In this article, negative T-cell signal inducing chimeric antigen receptor (N-CAR or i-CAR) and to T-cells comprising such N-CAR as well as a positive T-Cell signal inducing CAR (P-CAR).
Abstract: The invention relates to negative T-cell signal inducing chimeric antigen receptor (N-CAR or i-CAR) and to T-cells comprising such N-CAR as well as a positive T-cell signal inducing CAR (P-CAR) as well as their use in therapy.
25 citations
Patent•
09 Nov 2015
TL;DR: In this article, an inhibitory chimeric antigen receptor (N-CAR) was proposed, where an extracellular domain comprising an antigen binding domain, a transmembrane domain, and an intracellular domain comprises an Immunoreceptor Tyrosine-based Switch Motif ITSM.
Abstract: The invention relates to an inhibitory chimeric antigen receptor (N-CAR) comprising an extracellular domain comprising an antigen binding domain, a transmembrane domain, and, an intracellular domain wherein the intracellular domain comprises an Immunoreceptor Tyrosine-based Switch Motif ITSM, wherein said ITSM is a sequence of amino acid TX 1 YX 2 X 3 X 4 , wherein X 1 is an amino acid X 2 is an amino acid X 3 is an amino acid and X 4 is V or I.
7 citations
TL;DR: In this paper, autologous T-cells engineered to express chimeric antigen receptors (CARs) that target specific tumor antigens are of high potential in treating different kinds of cancer.
Abstract: 3069 Background: Autologous T-cells engineered to express chimeric antigen receptors (CARs) that target specific tumor antigens are of high potential in treating different kinds of cancer. However,...
Patent•
09 Nov 2015
TL;DR: In this article, a sequence of acides amines is defined: TX 1 YX 2 X 3 X 4, X 1 designant un acide amine, X 2 designant u-n, X 3 designant U n, and X 4 designant V ou I.
Abstract: L'invention concerne un recepteur antigenique chimerique inhibiteur (N-CAR) comprenant un domaine extracellulaire comprenant un domaine de liaison a l'antigene, un domaine transmembranaire, un domaine intracellulaire, le domaine intracellulaire comprenant un motif de commutation d'immunorecepteur a base de tyrosine (ITSM), ledit ITSM etant une sequence d'acides amines : TX 1 YX 2 X 3 X 4 , X 1 designant un acide amine, X 2 designant un acide amine, X 3 designant un acide amine, et X 4 designant V ou I.
Patent•
20 Feb 2015
TL;DR: The presente invention concerne des cellules T genetiquement modifiees, un procede pour leur preparation, and leur utilisation en tant que medicament, en particulier pour une immunotherapie as discussed by the authors.
Abstract: La presente invention concerne des cellules T genetiquement modifiees, un procede pour leur preparation, et leur utilisation en tant que medicament, en particulier pour une immunotherapie. Les cellules T genetiquement modifiees de l'invention sont concues pour exprimer tant un Recepteur d'Antigene Chimere (CAR) dirige contre au moins un antigene exprime sur la surface d'une cellule maligne ou infectee, qu'un inhibiteur secrete de cellules T regulatrices (Treg). De preference, un tel inhibiteur secrete est un inhibiteur peptidique du facteur de transcription tete de fourche/helice en forme d'aile (forkhead/winged helix 3) (FoxP3), un facteur specifique implique dans la differenciation des cellules T en cellules T regulatrices. Les cellules T genetiquement modifiees de l'invention dirigent leur activite immune vers des cellules malignes ou infectees specifiques, tout en empechant simultanement une modulation de la reponse immune par des cellules T regulatrices voisines. L'invention ouvre la voie a des strategies immunotherapeutiques adoptives standard et accessibles, en particulier pour traiter ou prevenir un cancer, et des infections bacteriennes ou virales.
Patent•
11 Mar 2015
TL;DR: In this article, the authors present a procedure for leur preparation and leur utilisation en tant que medicament, en particulier for l'immunotherapie.
Abstract: La presente invention se rapporte a des lymphocytes T obtenus par genie genetique, a un procede pour leur preparation et leur utilisation en tant que medicament, en particulier pour l'immunotherapie. Les lymphocytes T de l'invention obtenus par genie genetique sont caracterises en ce que l'expression de la microglobuline beta 2 (B2M) et/ou du transactivateur du complexe majeur d'histocompatibilite de classe II (CIITA) est inhibee, par exemple a l'aide d'endonucleases de clivage rares, capables d'inactiver selectivement, par clivage d'ADN, le gene codant pour la microglobuline beta 2 (B2M) et/ou CIITA, ou a l'aide de molecules d'acide nucleique qui inhibent l'expression de la B2M et/ou de CIITA. De plus, afin de rendre les lymphocytes T non alloreactifs, au moins un gene codant pour un composant du recepteur des lymphocytes T est inactive, par exemple a l'aide d'endonucleases de clivage rares, capables d'inactiver selectivement, par clivage d'ADN, le gene codant pour ledit composant TCR. De plus, l'expression du polypeptide immunosuppresseur peut etre effectuee sur ces lymphocytes T modifies afin de prolonger la survie de ces lymphocytes T modifies dans un organisme hote. Un tel lymphocyte T modifie est particulierement approprie aux transplantations allogeniques, en particulier du fait qu'il permet de reduire a la fois le risque de rejet par le systeme immunitaire de l'hote et le risque de developpement de la maladie du greffon contre l'hote. La presente invention ouvre la voie a des strategies d'immunotherapie adoptive standard et abordables utilisant des lymphocytes T pour le traitement du cancer, d'infections et de maladies auto-immunes.
Patent•
13 Feb 2015
TL;DR: In this article, a procedure de developpement de cellules immunitaires genetiquement modifiees destinees a l'immunotherapie is described.
Abstract: L'invention concerne des procedes de developpement de cellules immunitaires genetiquement modifiees destinees a l'immunotherapie, qui peuvent etre dotees de recepteurs d'antigenes chimeriques ciblant un marqueur antigenique commun aux cellules pathologiques et auxdites cellules immunitaires (ex : CD38, CSl ou CD70) du fait que les genes codant pour lesdits marqueurs sont inactives dans lesdites cellules immunitaires par une endonuclease a activite de clivage rare, telle que TALEN, Cas9 ou Argonaute