COMPOSITIONS AND METHODS FOR TCR REPROGRAMMING USING FUSION PROTEINS

1 .- 20 . (canceled)

21 . A pharmaceutical composition comprising

(I) a T cell from a human subject, wherein the T cell comprises a recombinant nucleic acid molecule encoding a T cell receptor (TCR) fusion protein (TFP) comprising

(a) a TCR subunit comprising

(i) an extracellular domain,

(ii) a TCR transmembrane domain, and

(iii) a TCR intracellular domain; and

(b) a murine, human or humanized scFv or single domain antibody comprising an antigen binding domain; and

(II) a pharmaceutically acceptable carrier;

wherein the TCR subunit and the antigen binding domain are operatively linked; wherein the TCR subunit does not comprise at least one of the following: a functional CD3 zeta intracellular domain, a costimulatory domain, and a heterologous stimulatory domain;

wherein the TFP functionally interacts with an endogenous TCR when expressed in a T cell; and

wherein the T cell exhibits increased cytotoxicity to a human cell expressing an antigen that specifically interacts with the antigen binding domain compared to a T cell not containing the TFP.

22 . The pharmaceutical composition of claim 21 , wherein the antigen binding domain is connected to the TCR extracellular domain by a linker.

23 . The pharmaceutical composition of claim 22 , wherein the linker comprises (G4S) n , wherein G is glycine, S is serine, and n is an integer from 1 to 4.

24 . The pharmaceutical composition of claim 21 , wherein the antigen binding domain comprises an anti-CD19 binding domain or an anti-BCMA binding domain.

25 . The pharmaceutical composition of claim 21 , wherein the antigen binding domain comprises

(i) a light chain (LC) CDR1, LC CDR2 and LC CDR3 sequence of SEQ ID NO: 25, SEQ ID NO: 27 and SEQ ID NO: 29, respectively;

(ii) a heavy chain (HC) CDR1, HC CDR2 and HC CDR3 sequence of SEQ ID NO: 31, SEQ ID NO: 33 and SEQ ID NO: 35, respectively; or

(iii) a combination thereof.

26 . The pharmaceutical composition of claim 21 , wherein the extracellular domain, the TCR transmembrane domain and the TCR intracellular domain are derived from a single TCR subunit.

27 . The pharmaceutical composition of claim 26 , wherein the single TCR subunit is CD3 gamma, CD3 delta or CD3 epsilon.

28 . The pharmaceutical composition of claim 26 , wherein the single TCR subunit is TCR alpha, TCR beta, TCR delta or TCR gamma.

29 . The pharmaceutical composition of claim 26 , wherein the extracellular domain comprises a full-length TCR extracellular domain of the single TCR subunit.

30 . The pharmaceutical composition of claim 21 , wherein the TCR subunit does not comprise a functional CD3 zeta intracellular domain.

31 . The pharmaceutical composition of claim 21 , wherein the TCR subunit does not comprise a costimulatory domain.

32 . The pharmaceutical composition of claim 21 , wherein the TCR subunit does not comprise a heterologous stimulatory domain.

33 . The pharmaceutical composition of claim 21 , wherein the TCR subunit does not comprise at least two of the following: a functional CD3 zeta intracellular domain, a costimulatory domain, and a heterologous stimulatory domain.

34 . The pharmaceutical composition of claim 21 , wherein the TCR subunit does not comprise a functional CD3 zeta intracellular domain, a costimulatory domain and a heterologous stimulatory domain.

35 . The pharmaceutical composition of claim 21 , wherein in the presence of a human cell expressing an antigen that specifically interacts with the antigen binding domain the T cell has greater than or more efficient cytotoxic activity than a T cell comprising a nucleic acid encoding a chimeric antigen receptor (CAR) comprising (a) the antigen binding domain operatively linked to (b) a CD28 extracellular domain (c) a CD28 transmembrane domain (d) a CD28 intracellular domain and (e) a CD3 zeta intracellular domain.

36 . The pharmaceutical composition of claim 21 , wherein production of a pro-inflammatory cytokine by the T cell is lower compared to production of the pro-inflammatory cytokine by a T cell comprising a nucleic acid encoding a CAR comprising the antigen binding domain operatively linked to a CD28 extracellular domain, a CD28 transmembrane domain, a CD28 intracellular domain, and a CD3 zeta intracellular domain in the presence of a human cell expressing an antigen that specifically interacts with the antigen binding domain.

37 . The pharmaceutical composition of claim 36 , wherein the pro-inflammatory cytokine is TNFα, GM-CSF or IL-2.

38 . The pharmaceutical composition of claim 21 , wherein production of IFNγ or IL-2 by the T cell is increased compared to a T cell not containing the TFP in the presence of a human cell expressing an antigen that specifically interacts with the antigen binding domain.

39 . The pharmaceutical composition of claim 26 , wherein the single TCR subunit comprises a sequence with at least 90% sequence identity to SEQ ID NO: 58.

40 . The pharmaceutical composition of claim 26 , wherein the single TCR subunit comprises a sequence with at least 90% sequence identity to SEQ ID NO: 60.

41 . The pharmaceutical composition of claim 26 , wherein the single TCR subunit comprises a sequence with at least 90% sequence identity to SEQ ID NO: 63.

42 . The pharmaceutical composition of claim 26 , wherein the single TCR subunit is CD3 delta.

43 . The pharmaceutical composition of claim 26 , wherein the single TCR subunit is TCR alpha.

44 . The pharmaceutical composition of claim 26 , wherein the single TCR subunit is TCR beta.

45 . A method of treating cancer in a subject in need thereof comprising administering a T cell to the subject, wherein the T cell comprises a recombinant nucleic acid molecule encoding a T cell receptor (TCR) fusion protein (TFP) comprising

(a) a TCR subunit comprising

(i) a TCR extracellular domain,

(ii) a TCR transmembrane domain, and

(iii) a TCR intracellular domain; and

(b) an antibody domain comprising an antigen binding domain;

wherein the TCR subunit and the antigen binding domain are operatively linked;

wherein the TCR extracellular domain is a full length TCR extracellular domain;

wherein the TFP functionally interacts with an endogenous TCR when expressed in a T cell.

46 . The method of claim 45 , wherein the TCR subunit does not comprise at least one of: a functional CD3 zeta intracellular domain, a costimulatory domain or a heterologous stimulatory domain.

47 . A method of treating cancer in a subject in need thereof comprising administering a T cell to the subject, wherein the T cell comprises a recombinant nucleic acid molecule encoding a T cell receptor (TCR) fusion protein (TFP) comprising

(a) a TCR subunit comprising

(i) an extracellular domain,

(ii) a TCR transmembrane domain, and

(iii) a TCR intracellular domain; and

(b) an antibody domain comprising an antigen binding domain;

wherein the TCR subunit and the antigen binding domain are operatively linked;

wherein the TCR subunit does not comprise at least one of: a functional CD3 zeta intracellular domain, a costimulatory domain or a heterologous stimulatory domain;

wherein the TCR transmembrane domain and the TCR intracellular domain are derived from a single TCR subunit; and

wherein the TFP functionally interacts with an endogenous TCR when expressed in a T cell.

48 . The method of claim 47 , wherein the single TCR subunit is selected from the group consisting of CD3 epsilon, CD3 gamma, CD3 delta, TCR alpha, TCR beta, TCR gamma and TCR delta.

49 . The method of claim 47 , wherein the extracellular domain, the TCR transmembrane domain and the TCR intracellular domain are derived from a single TCR subunit.

50 . The method of claim 47 , wherein the extracellular domain is a full length TCR extracellular domain.