EP-4739340-A1 - IMMUNE CELL EXPRESSING CHIMERIC ANTIGEN RECEPTOR AND TRANSGENIC T CELL RECEPTOR

Abstract

The present invention provides T cells that express both a CAR and a tTCR (CARTCR T cells) and compositions comprising said CARTCR T cells together with T cells that express only said CAR ("CAR T cells") and/or with T cells that express only said tTCR ("tTCR T cells") for treatment of a disease. Methods for generation of these compositions are also provided.

Inventors

• LOCK, Dominik
• TEPPERT, Karin
• JOHNSTON, IAN
• BRANDES, Caroline
• ENGELS, Boris
• ASSENMACHER, MARIO

Assignees

• Miltenyi Biotec B.V. & Co. KG

Dates

Publication Date

20260513

Application Date

20240524

Claims (1)

1. Claims 1) An in-vitro method for the generation of a composition, wherein the composition comprises A) engineered immune cells co-expressing a CAR and a transgenic TCR, B) engineered immune cells expressing said CAR, and C) engineered immune cells expressing said transgenic TCR, the method comprising the step: transducing simultaneously immune cells a) with a first viral vector comprising a first nucleic acid sequence comprising encoding a chimeric antigen receptor (CAR) comprising 1) an antigen binding domain specific for a first epitope of an antigen ii) a transmembrane domain iii) an intracellular signaling domain, and b) with a second viral vector comprising a second nucleic acid sequence encoding a transgenic T cell receptor (tTCR) having specificity for a second epitope of an antigen, wherein said antigen of said first epitope and said antigen of said second epitope are the same antigen or are different antigens, thereby obtaining said composition. 2) The in-vitro method of claim 1, wherein said immune cells are T cells. 3) The in-vitro method of claim 1 or 2, wherein the multiplicity of infection (MOI) ratio of said first viral vector and said second viral vector used for the transduction step is between 1 : 10 and 10:1. 4) The in-vitro method of claim 3, wherein said MOI ratio is about 1 : 10. 5) A composition comprising A) I) engineered immune cells comprising a) a chimeric antigen receptor (CAR) comprising i) an antigen binding domain specific for a first epitope of an antigen ii) a transmembrane domain iii) an intracellular signaling domain, and b) a transgenic T cell receptor (tTCR) having specificity for a second epitope of an antigen, and II) engineered immune cells comprising said chimeric antigen receptor (CAR) comprising i) an antigen binding domain specific for said first epitope of an antigen ii) a transmembrane domain iii) an intracellular signaling domain, and III) engineered immune cells comprising said transgenic T cell receptor (tTCR) having specificity for said second epitope of an antigen, wherein said antigen of said first epitope and said antigen of said second epitope are the same antigen or are different antigens; or B) I) engineered immune cells comprising a) a chimeric antigen receptor (CAR) comprising i) an antigen binding domain specific for a first epitope of an antigen ii) a transmembrane domain iii) an intracellular signaling domain, and b) a transgenic T cell receptor (tTCR) having specificity for a second epitope of an antigen, and II) engineered immune cells comprising said chimeric antigen receptor (CAR) comprising i) an antigen binding domain specific for said first epitope of an antigen ii) a transmembrane domain iii) an intracellular signaling domain, wherein said antigen of said first epitope and said antigen of said second epitope are the same antigen or are different antigens; or C) I) engineered immune cells comprising a)a chimeric antigen receptor (CAR) comprising i) an antigen binding domain specific for a first epitope of an antigen ii) a transmembrane domain iii) an intracellular signaling domain, and b) a transgenic T cell receptor (tTCR) having specificity for a second epitope of an antigen, and II) engineered immune cells comprising said transgenic T cell receptor (TCR) having specificity for said second epitope of an antigen, wherein said antigen of said first epitope and said antigen of said second epitope are the same antigen or are different antigens. 6) The composition of claim 5, wherein said engineered immune cells are T cells. 7) The composition of claim 5 or 6, wherein said CAR is specific for an epitope of the antigen CD33 and wherein said tTCR has specificity for an epitope of the antigen ANPM1. 8) The composition of claim 7, wherein said antigen binding domain specific for said epitope of the antigen CD33 comprises SEQ ID NO:2, and wherein said tTCR having specificity for the epitope of the antigen ANPM1 comprises: (a) a polypeptide comprising a CDR3 of a TCR alpha chain polypeptide that specifically binds to the peptide CLAVEEVSL (SEQ ID NO: 1) and (b) a polypeptide comprising a CDR3 of a TCR beta chain polypeptide that specifically binds to the peptide CLAVEEVSL (SEQ ID NO: 1), wherein the CDR3 of (a) is within a TCR a chain variable region having at least 90% sequence identity to SEQ ID NO:7, wherein the CDR3 has an amino acid sequence of SEQ ID NO: 5; and wherein (a) comprises a TCR alpha chain constant region, wherein the TCR alpha chain variable region CDR1 has an amino acid sequence of SEQ ID NO: 9 and the TCR a chain variable region CDR2 has an amino acid sequence of SEQ ID NO: 10; and wherein the CDR3 of (b) is within a TCR beta chain variable region having at least 90% sequence identity to SEQ ID NO: 8, wherein the CDR3 has an amino acid sequence of SEQ ID NO: 6; and wherein (b) comprises a TCR beta chain constant region, and wherein the TCR beta chain variable region CDR1 has an amino acid sequence of SEQ ID NO: 11 and the TCR P chain variable region CDR2 has an amino acid sequence of SEQ ID NO: 12. 9) The composition of claim 5 to 8, wherein said composition is composition A, and wherein said composition A is obtained by the method of any one of claims 1 to 4. 10) A pharmaceutical composition comprising the composition of claims 5 to 9, and optionally a pharmaceutical acceptable carrier. 11) An engineered immune cell comprising a) a chimeric antigen receptor (CAR) comprising i) an antigen binding domain specific for a first epitope of an antigen ii) a transmembrane domain iii) an intracellular signaling domain, b) a transgenic T cell receptor (tTCR) having specificity for a second epitope of an antigen, wherein said antigen of said first epitope and said antigen of said second epitope are different antigens, and wherein said CAR and said tTCR are expressed constitutively in said immune cell, wherein said CAR is specific for an epitope of the antigen CD33 and wherein said tTCR has specificity for an epitope of the antigen ANPM1. 12) The engineered immune cell of claim 11, wherein said antigen binding domain specific for said epitope of the antigen CD33 comprises SEQ ID NO:2, and wherein said tTCR having specificity for the epitope of the antigen ANPM1 comprises: (a) a polypeptide comprising a CDR3 of a TCR alpha chain polypeptide that specifically binds to the peptide CLAVEEVSL (SEQ ID NO: 1) and (b) a polypeptide comprising a CDR3 of a TCR beta chain polypeptide that specifically binds to the peptide CLAVEEVSL (SEQ ID NO: 1), wherein the CDR3 of (a) is within a TCR a chain variable region having at least 90% sequence identity to SEQ ID NO:7, wherein the CDR3 has an amino acid sequence of SEQ ID NO: 5; and wherein (a) comprises a TCR alpha chain constant region, wherein the TCR alpha chain variable region CDR1 has an amino acid sequence of SEQ ID NO: 9 and the TCR a chain variable region CDR2 has an amino acid sequence of SEQ ID NO: 10; and wherein the CDR3 of (b) is within a TCR beta chain variable region having at least 90% sequence identity to SEQ ID NO: 8, wherein the CDR3 has an amino acid sequence of SEQ ID NO: 6; and wherein (b) comprises a TCR beta chain constant region, and wherein the TCR beta chain variable region CDR1 has an amino acid sequence of SEQ ID NO: 11 and the TCR P chain variable region CDR2 has an amino acid sequence of SEQ ID NO: 12. 13) The engineered immune cell of claim 11 or 12, wherein said immune cell is a T cell. 14) A pharmaceutical composition comprising the engineered immune cell of any one of claims 11 to 13, and optionally a pharmaceutical acceptable carrier.

Description

Title Immune cell expressing chimeric antigen receptor and transgenic T cell receptor Field of the invention The present invention generally relates to the field of adoptive T cell therapies, e.g. use of T cell receptor and/or chimeric antigen binding receptor (CAR) expressed on the surface of immune cells for treatment of a disease such as cancer, in particular to immune cells that coexpress TCR and CAR and/or to the generation of a composition of immune cells that comprise TCR-expressing immune cells and/or, CAR-expressing T cells together with TCR and CAR co-expressing immune cells. Background of the invention The use of T cell receptor (TCR)- or chimeric antigen receptor (CAR)-expressing T cells redirected to specifically recognize and eliminate malignant cells, greatly increased the scope and potential of adoptive immunotherapy and is being assessed for new standard of care in certain human malignancies or other diseases. TCR-engineered T cell recognize their intracellularly processed peptide fragments in a human leukocyte antigen (HLA)-dependent manner. CARs are recombinant receptors that typically target surface molecules HLA-independently. Generally, CARs comprise an extracellular antigen recognition moiety, often a single-chain variable fragment (scFv) derived from antibodies or a Fab fragment, linked to an extracellular spacer, a transmembrane domain and intracellular co-stimulatory and signaling domains. Therapies using TCR- or CAR-modified T cells, although sometimes efficacious, have a high potential for improvements. Especially novel strategies are required to ameliorate T cell function. In Uslu et al. 2016 the authors combined a T-cell receptor and a chimeric antigen receptor, specific for different common melanoma antigens, gplOO (PMEL) and MCSP (HMW-MAA), to generate functional CD8+ T cells expressing two additional receptors (TETARs) by electroporation of receptor-encoding mRNA, thereby generating CD8+ T cells that express transiently a transgenic TCR and a CAR. In Simon et al. 2019 the authors generated T cells expressing two additional receptors (TETARs) by lentiviral transduction of a gplOO-specific T cell receptor (TCR) and subsequent electroporation of mRNA encoding a second-generation CSPG4-specific chimeric antigen receptor (CAR). Again, the CAR is expressed transiently only. Miyao et al. 2018 disclose a transgenic TCR together with artificial T cell-activating adapter molecules (ATAM), generated by inserting the intracellular domain (ICD) of activating T-cell signaling moieties into CD3z. ATAMs with the ICD of either CD28 or 4-1BB were generated, assembled into the TCR complex as a part of CD3z, and enhanced downstream signaling from the supramolecular activation cluster. In Omer et al. 2018 the authors evaluated first and second generation GD2.CARs containing costimulatory endodomains derived from 4-1BB or CD28 in T cells specific for varicella zoster virus and EBV, i.e. these T cells have native TCRs. Omer et al. 2022 discloses a T cell co-expressing a tTCR targeting an immunodominant epitope of surviving and a costimulatory CAR (CoCAR) that lacked the cytotoxic zeta chain of conventional CARs, but still provide T cell co-stimulation upon CD 19 ligation. W02019004831 discloses transgenic TCRs with specificity for ANPM1. WO2015150526A2 discloses engineered immune cells expressing CARs with specificity for CD33. There is a need in the art for improved or alternative adoptive immune cell such as T cell therapies including TCR and/or CAR immune cells such as T cells, especially with regard to the potential risk of tumor relapse due to HLA downregulation or surface antigen loss. Brief description of the invention Surprisingly, it was found that an immune cell, preferentially a T cell that expresses both a chimeric antigen receptor (CAR) specific for (targeting) a first epitope of an antigen and a transgenic T cell receptor (tTCR) having specificity for a second epitope of an antigen can be used beneficially for use in treatment of a disease such as cancer in a subject suffering from said disease. It is even more surprising that a composition comprising immune cells, preferentially T cells that express both a CAR and a tTCR (“CARTCR T cells”) together with immune cells, preferentially T cells that express only said CAR (“CAR T cells”) and/or with immune cells, preferentially T cells that express only said tTCR (“tTCR T cells”) is superior for said treatment of a disease as compared to the single CARTCR T cell or a composition comprising immune cells, preferentially T cells that express only said CAR together with immune cells, preferentially T cells that express only said tTCR, i.e. a composition without said CARTCR T cell. Combinations of specificity of CARs and tTCRs that may be usable for the immune cells and compositions as disclosed herein are, but without intention to be limited to these kind of combinations (specificity for CAR/specificity for tTCR): CD33/ANPM1, BCMA/BOB1, C