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AssigneesNovartis AG, The Trustees of the University of Pennsylvania
InventorsSteven M. Albelda, Edmund K. Moon
Estimating the term…
Compositions and methods for treating diseases associated with expression of a cancer associated antigen are disclosed. The invention also relates to chimeric antigen receptor (CAR) specific to a cancer associated antigen as described herein, SHP inhibitory molecules, vectors encoding the same, and recombinant immune effector cells comprising the CARs and SHP inhibitory molecules. Methods of administering a genetically modified immune effector cell expressing a CAR that comprises an antigen binding domain that binds to a cancer associated antigen and a SHP inhibitory polypeptide are also disclosed.
A nucleic acid composition comprising
The nucleic acid composition of any of the preceding claims, wherein the binding of the SHP inhibitor polypeptide to the ITIM domain is reduced by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, or 99% compared to a wild-type SHP.
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide (e.g., SHP-1 polypeptide or SHP-2 polypeptide) is less than 240, 220, 180, 160, 140, 120, 100, 80, 60, or 40 amino acids in length.
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide has reduced phosphatase activity, compared to wild-type SHP, to one or more SHP substrates (e.g., substrates comprising phosphorylated tyrosine).
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide has a deletion of at least part or all of the phosphatase domain.
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide lacks its phosphatase domain.
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide, when expressed in an immune effector cell (e.g., a T cell), does not result (e.g., substantially result, e.g., results in less than 10%, 9%, 8%, 7%, 6%, 5% or less change) in one of more of the following:
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide, when expressed in an immune effector cell (e.g., a T cell), results in one or more of:
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide, when expressed in an immune effector cell (e.g., a T cell) that also expresses a CAR polypeptide, results in increased cytokine secretion and/or increases the percentage of cytokine-expressing cells, wherein the cytokine is optionally IL-2, compared to an otherwise similar cell lacking the SHP inhibitor polypeptide or an otherwise similar cell comprising a wild-type SHP polypeptide or a wild type SH2-N terminal fragment thereof (e.g., an SHP polypeptide according to amino acids 1-100 of SEQ ID NO:
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide, when expressed in an immune effector cell (e.g., a T cell) that also expresses a CAR polypeptide, results in increased lysis, e.g., in vitro, of cancer cells that express PD-L1 and an antigen recognized by the CAR polypeptide, compared to an otherwise similar cell that lacks the SHP inhibitor polypeptide or an otherwise similar cell comprising a wild type SHP polypeptide or a wild type SH2-N terminal fragment thereof (e.g., an SHP polypeptide according to amino acids 1-100 of SEQ ID NO:
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide, when expressed in an immune effector cell (e.g., a T cell) that also expresses a CAR polypeptide (e.g., an immune effector cell that expresses PD-1), results in decreased tumor volume (e.g., of a tumor having cells expressing PD-L1 and an antigen recognized by the CAR polypeptide), e.g., in a mouse model, compared to an otherwise similar animal treated with otherwise similar immune effector cells that that lack the SHP inhibitor polypeptide or an otherwise similar cell comprising a wild type SHP polypeptide, or a wild type SH2-N terminal fragment thereof according to amino acids 1-100 of SEQ ID NO:
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide, when expressed in an immune effector cell (e.g., a T cell) that also expresses a CAR polypeptide (e.g., an immune effector cell that expresses PD-1), results in increased T lymphocyte infiltration into a tumor, e.g., in a mouse model, compared to an otherwise similar animal treated with otherwise similar immune effector cells that lack the SHP inhibitor polypeptide or an otherwise similar cell comprising a wild type SHP polypeptide, or a wild type SH2-N terminal fragment thereof according to amino acids 1-100 of SEQ ID NO:
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide, when expressed in an immune effector cell (e.g., a T cell) that also expresses a CAR polypeptide, results in increased phosphorylation of ZAP70, e.g., in the presence of PD-L1-expressing tumor cells, compared to an otherwise similar immune effector cell that lacks the SHP inhibitor polypeptide or an otherwise similar cell comprising a wild type SHP polypeptide, or a wild type SH2-N terminal fragment thereof according to amino acids 1-100 of SEQ ID NO:
The nucleic acid composition of any of the preceding claims, wherein the SHP inhibitor polypeptide, when expressed in an immune effector cell (e.g., a T cell) that also expresses a CAR polypeptide, results in increased expression of IFNγ or IL-2 (or increased percentage of IFNγ positive or IL-2 positive cells), e.g., in the presence of PD-L1-expressing tumor cells, compared to an otherwise similar immune effector cell that lacks the SHP inhibitor polypeptide or an otherwise similar cell comprising a wild type SHP polypeptide, or a wild type SH2-N terminal fragment thereof according to amino acids 1-100 of SEQ ID NO:
The nucleic acid composition of any of the preceding claims, comprising
The nucleic acid composition of any of the preceding claims, wherein the CAR polypeptide and SHP inhibitor polypeptide are encoded by a single nucleic acid molecule in the same frame and as a single polypeptide chain.
The nucleic acid composition of any of the preceding claims, wherein said SHP inhibitor polypeptide is attached to the N-terminus of said CAR polypeptide or the C-terminus of said CAR polypeptide.
The nucleic acid composition of any of the preceding claims, wherein said SHP inhibitor polypeptide and CAR polypeptide are separated by one or more peptide cleavage sites, optionally wherein the peptide cleavage site is an auto-cleavage site or a substrate for an intracellular protease, optionally wherein the peptide cleavage site is a T2A or P2A site.
The nucleic acid composition of any of the preceding claims, wherein the nucleic acid molecule encoding the CAR polypeptide and the nucleic acid molecule encoding the SHP inhibitor polypeptide are separated by a nucleic acid sequence encoding T2A or P2A.
The nucleic acid composition of any of the preceding claims, wherein said composition consists of a single isolated nucleic acid.
The nucleic acid composition of any of the preceding claims, wherein the encoded CAR polypeptide comprises an antigen binding domain, a transmembrane domain, and an intracellular signaling domain.
The nucleic acid composition of any of the preceding claims, which is DNA or RNA.
A polypeptide comprising a CAR polypeptide and a SHP inhibitor polypeptide, e.g., with a peptide cleavage site disposed therebetween, wherein the SHP inhibitor polypeptide comprises:
An immune effector cell (e.g., a population of immune effector cells) comprising a CAR polypeptide and a SHP inhibitor polypeptide as recited in any of the preceding claims.
An immune effector cell (e.g., a population of immune effector cells) comprising
A composition comprising:
Dependencies are inferred from the claim text.
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From animals; from humans: receptors; Cell surface antigens; Cell surface determinants
Against receptors, cell surface antigens or cell surface determinants: from tumour cells
Carbohydrates; Sugars; Derivatives thereof: compounds having three or more nucleosides or nucleotides
Compounds having three or more nucleosides or nucleotides: double-stranded nucleic acids or oligonucleotides
Term end not estimated
The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Feb. 27, 2018, is named N2067-7118WO_SL.txt and is 1,566,610 bytes in size.
The present invention relates generally to compositions and uses of immune effector cells (e.g., T cells, NK cells) engineered to express a Chimeric Antigen Receptor (CAR) to treat a disease associated with expression of a tumor antigen.
Adoptive cell transfer (ACT) therapy with autologous T-cells, especially with T-cells transduced with Chimeric Antigen Receptors (CARs), has shown promise in cancer clinical trials. Although CAR technology has demonstrated tremendous success in eliminating hematologic tumors, the need exists for decreasing the effect of immunosuppressive factors that exist with the microenvironment of solid tumors that reduce the activity of CAR T cells.
FIG. 1 shows a diagram of examples of inhibitory receptors (IRs) involved in immunosuppression of CAR T cells.
FIG. 2 shows a diagram of TCR signaling, highlighting the role of SHP1.
FIG. 3 shows graphs of tumor cell killing (top) and IFNg secretion (bottom) of anti-mesothelin CAR TIL cells recovered after CAR T cells were injected into NSG flank tumors; recovered TIL cells were treated or not treated with SSG. “cryo mesoCAR” represents T cells that were not injected but cryopreserved, “mesoCAR TIL” represents T cells that were injected, then isolated from flank tumors at the experiment endpoint.
Compositions and uses that improve an activity (e.g., one or more of function, persistence, cancer killing effect, or tumor infiltration) of an immune effector cell, e.g., a population of immune effector cells (e.g., T cells, NK cells) are disclosed. In some embodiments, the immune effector cell expresses a Chimeric Antigen Receptor molecule (e.g., a CAR polypeptide) that binds to a tumor antigen. In some embodiments, the immune effector cell comprises, or is contacted with an inhibitor of a Src homology region 2 domain-containing phosphatase (SHP). In one embodiment, the inhibitor is an inhibitor of SHP-1. In another embodiment, the inhibitor is an inhibitor of SHP-2. In one embodiment, the SHP inhibitor interferes with SHP signaling (e.g., interferes with SHP-1 signaling or SHP-2 signaling, or both), also referred to herein as an SHP inhibitor molecule (e.g., an SHP inhibitor polypeptide). In general, the invention features, at least in part, immune cells, e.g., T-cells, containing a CAR molecule and an SHP inhibitor molecule, e.g., an SHP inhibitor polypeptide. The invention is based, at least in part, on the discovery that immune effector cells comprising one or more SHP inhibitor polypeptides result in one or more of: increased killing of tumor cells, increased cytokine release, and increased tumor infiltration in vitro and in vivo.
Without wishing to be bound by theory, SHP1 (and SHP2) regulates T cell receptor signaling, and is activated by inhibitory receptors (IRs). IR signaling down-regulates T cell function, lowering the efficacy of CAR T cell therapies in targeting and killing tumor cells. SHP inhibition is expected to interfere with the signaling of immunosuppressive factors, such as IRs, or checkpoint molecules. In certain embodiments, the IRs are present in the microenvironment of a tumor, e.g., a solid tumor, thus resulting in decreased effectiveness of a therapy, e.g., a CAR therapy, in the tumor microenvironment. SHP inhibitor molecules, e.g., polypeptides that inhibit SHP1 and/or SHP2, and, when co-expressed with a CAR in an immune effector cell, result in one or more of: increase killing of tumor cells, increase cytokine release, and increase tumor infiltration in vitro and in vivo. The SHP inhibitor molecules disclosed herein are compatible with a wide array of CARs, also described herein.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains.