Method of producing T cell receptor-deficient T cells expressing a chimeric receptor
AssigneeTrustees of Dartmouth College
InventorCharles L. Sentman
Life of this patent
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Abstract
The invention is directed to modified T cells, methods of making and using isolated, modified T cells, and methods of using these isolated, modified T cells to address diseases and disorders. In one embodiment, this invention broadly relates to TCR-deficient T cells, isolated populations thereof, and compositions comprising the same. In another embodiment of the invention, these TCR-deficient T cells are designed to express a functional non-TCR receptor. The invention also pertains to methods of making said TCR-deficient T cells, and methods of reducing or ameliorating, or preventing or treating, diseases and disorders using said TCR-deficient T cells, populations thereof, or compositions comprising the same.
Claims30
A method of producing one or more, modified primary human T cells, which modified primary human T cells are derived from primary human T cells isolated from a human donor that (i) are modified to reduce expression of the endogenous T cell receptor (TCR), and (ii) are further modified to express at least one functional exogenous non-TCR that comprises a chimeric receptor comprising a ligand binding domain and a signaling domain, said method including the following:
- (a) introducing into one or more primary human T cells isolated from a human donor at least one small-hairpin RNA (shRNA) which each target a nucleic acid that encodes a component of a functional TCR that is endogenously expressed by an unmodified primary human T cell isolated from said human donor, wherein expression of said at least one shRNA reduces the expression of the endogenous TCR by blocking or inhibiting the expression of at least one targeted component of said endogenous TCR, and
- (b) further introducing into said one or more primary human T cells isolated from a human donor at least one nucleic acid that results in the expression of a functional exogenous non-TCR receptor that comprises a chimeric receptor comprising a ligand binding domain and a signaling domain;
- wherein the one or more cells obtained by steps (a) and (b) are suitable for use in human therapy, and further wherein the cells obtained by steps (a) and (b) elicit no or a reduced graft-versus-host disease (GVHD) response in a histoincompatible human recipient as compared to the GVHD response elicited by a primary human T cell or primary human T cells isolated from the same human donor either of which are only modified as in (b).
A method of producing one or more modified primary human T cells, which modified primary human T cells are derived from primary human T cells isolated from a human donor that (i) are modified to functionally impair or to reduce expression of the endogenous T cell receptor (TCR), and (ii) are further modified to express at least one functional exogenous non-TCR that comprises a chimeric receptor comprising a ligand binding domain and a signaling domain, said method including the following:
- (a) introducing into one or more primary human T cells isolated from a human donor at least one nucleic acid that encodes a dominant-negative inhibitor protein that is capable of interrupting expression or function of the endogenous TCR, wherein said nucleic acid comprises a polynucleotide that encodes a variant of a polypeptide component of a functional TCR complex endogenously expressed by an unmodified primary T human cell isolated from said human donor, wherein said variant polypeptide TCR component is modified, compared to the unmodified polypeptide TCR component, by one or more of the following:
- (1) the variant polypeptide TCR component lacks key signaling motifs required for function of the unmodified polypeptide TCR component;
- (2) the variant polypeptide TCR component is modified such that it does not associate properly with other endogenous TCR components;
- or (3) the variant polypeptide TCR component is modified such that it is still capable of associating properly with other endogenous TCR components, but the resultant TCR complex containing the variant polypeptide TCR component is incapable of binding to ligands;
- and
- (b) further introducing into said one or more primary human T cells isolated from a human donor at least one nucleic acid that results in the expression of a functional exogenous non-TCR receptor that comprises a chimeric receptor comprising a ligand binding domain and a signaling domain;
- wherein the one or more cells obtained by steps (a) and (b) are suitable for use in human therapy, and further wherein the cells obtained by steps (a) and (b) elicit no or a reduced graft-versus-host disease (GVHD) response in a histoincompatible human lrecipient as compared to the GVHD response elicited by a primary human T cell or primary human T cells isolated from the same human donor either of which are only modified as in (b).
Dependencies are inferred from the claim text.