Engineered Expression of Cell Surface and Secreted Sialidase by CAR T Cells for Increased Efficacy in Solid Tumors

Abstract

The present disclosure provides modified immune cells or precursors thereof (e.g. modified T cells) comprising a chimeric cell surface sialidase or a variant sialidase precursor protein. Compositions and methods of treatment are also provided.

What is claimed: 
     
         1 . A modified immune cell or precursor cell thereof, comprising:
 (a) a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising a sialidase (neuraminidase) or an enzymatically functional portion thereof, and   (b) a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface.   
     
     
         2 . A modified immune cell or precursor cell thereof, comprising:
 (a) a chimeric cell surface sialidase consisting of the amino acid sequence set forth in any one of SEQ ID NOs: 10, 37, or 38; and/or   (b) a variant sialidase precursor protein comprising a heterologous secretory sequence operably linked to a sialidase (neuraminidase) or an enzymatically functional portion thereof, wherein the variant sialidase precursor protein lacks a transmembrane domain, and wherein the sialidase or enzymatically functional portion thereof is capable of being secreted from an immune or precursor cell thereof when the variant sialidase precursor protein is expressed in the cell; and/or   (c) a variant sialidase precursor protein comprising a heterologous secretory sequence operably linked to a sialidase (neuraminidase) or an enzymatically functional portion thereof, wherein the sialidase comprises the amino acid sequence set forth in SEQ ID NO: 34; and/or   (d) a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising a sialidase (neuraminidase) or an enzymatically functional portion thereof, and a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface; and a chimeric antigen receptor (CAR) and/or a T cell receptor (TCR); and/or   (e) a variant sialidase precursor protein comprising a heterologous secretory sequence operably linked to a sialidase (neuraminidase) or an enzymatically functional portion thereof, wherein the variant sialidase precursor protein lacks a transmembrane domain, and wherein the sialidase or enzymatically functional portion thereof is capable of being secreted from an immune or precursor cell thereof when the variant sialidase precursor protein is expressed in the cell; and a chimeric antigen receptor (CAR) and/or a T cell receptor (TCR); and/or   (f) a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising Neu2 or an enzymatically functional portion thereof, and a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface; and a chimeric antigen receptor (CAR); and/or   (g) a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising Neu2 or an enzymatically functional portion thereof, and a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface; and a chimeric antigen receptor (CAR) having specificity for TnMUC1, CD19, or PSMA; and/or   (h) a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising Neu2 or an enzymatically functional portion thereof, a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface, a hinge domain, and an intracellular region comprising a costimulatory signaling domain and an intracellular signaling domain; and a chimeric antigen receptor (CAR); and/or   (i) a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising Neu2 or an enzymatically functional portion thereof, a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface, a hinge domain, and an intracellular region comprising a costimulatory signaling domain and an intracellular signaling domain; and a chimeric antigen receptor (CAR) having specificity for TnMUC1, CD19, or PSMA; and/or   (j) a variant sialidase precursor protein comprising a heterologous secretory sequence operably linked to a sialidase (neuraminidase) or an enzymatically functional portion thereof, wherein the variant sialidase precursor protein lacks a transmembrane domain, and wherein the sialidase or enzymatically functional portion thereof is capable of being secreted from an immune or precursor cell thereof when the variant sialidase precursor protein is expressed in the cell; and a chimeric antigen receptor (CAR) having specificity for TnMUC1, CD19, or PSMA.   
     
     
         3 . The modified cell of  claim 1 , wherein:
 (a) the sialidase is a human or humanized sialidase; and/or   (b) the sialidase is a human or humanized sialidase and wherein the human sialidase is selected from the group consisting of Neu1, Neu2, Neu3, and Neu4; and/or   (c) the sialidase is a human or humanized sialidase and wherein the human sialidase is Neu2; and/or   (d) the sialidase comprises the amino acid sequence set forth in SEQ ID NO: 13; and/or   (e) the sialidase comprises the amino acid sequence set forth in SEQ ID NO: 10; and/or   (f) the sialidase consists of the amino acid sequence set forth in SEQ ID NO: 10; and/or   (g) the sialidase comprises the amino acid sequence set forth in any one of SEQ ID NOs: 10, 37, or 38; and/or   (h) the sialidase consists of the amino acid sequence set forth in any one of SEQ ID NOs: 10, 37, or 38; and/or   (i) the sialidase comprises the amino acid sequence set forth in SEQ ID NO: 34; and/or   (j) the sialidase consists of the amino acid sequence set forth in SEQ ID NO: 34; and/or   (k) the transmembrane domain is selected from the group consisting of an artificial hydrophobic sequence, and a transmembrane domain of a type I transmembrane protein, an alpha, beta, or zeta chain of a T cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, OX40 (CD134), 4-1BB (CD137), and CD154; and/or   (l) the transmembrane domain comprises a transmembrane domain of CD8; and/or   (m) the transmembrane domain comprises a transmembrane domain of CD8 alpha; and/or   (n) the transmembrane domain comprises the amino acid sequence set forth in SEQ ID NO: 16.   
     
     
         4 . The modified cell of  claim 2 , wherein:
 (a) the sialidase is a human or humanized sialidase; and/or   (b) the sialidase is a human or humanized sialidase and wherein the human sialidase is selected from the group consisting of Neu1, Neu2, Neu3, and Neu4; and/or   (c) the sialidase is a human or humanized sialidase and wherein the human sialidase is Neu2; and/or   (d) the sialidase comprises the amino acid sequence set forth in SEQ ID NO: 13; and/or   (e) the sialidase comprises the amino acid sequence set forth in SEQ ID NO: 10; and/or   (f) the sialidase consists of the amino acid sequence set forth in SEQ ID NO: 10; and/or   (g) the sialidase comprises the amino acid sequence set forth in any one of SEQ ID NOs: 10, 37, or 38; and/or   (h) the sialidase consists of the amino acid sequence set forth in any one of SEQ ID NOs: 10, 37, or 38; and/or   (i) the sialidase comprises the amino acid sequence set forth in SEQ ID NO: 34; and/or   (j) the sialidase consists of the amino acid sequence set forth in SEQ ID NO: 34; and/or   (k) the variant sialidase precursor protein comprises the amino acid sequence set forth in SEQ ID NO: 13; and/or   (l) the variant sialidase precursor protein consists of the amino acid sequence set forth in SEQ ID NO: 13   (m) the transmembrane domain is selected from the group consisting of an artificial hydrophobic sequence, and a transmembrane domain of a type I transmembrane protein, an alpha, beta, or zeta chain of a T cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, OX40 (CD134), 4-1BB (CD137), and CD154; and/or   (n) the transmembrane domain comprises a transmembrane domain of CD8; and/or   (o) the transmembrane domain comprises a transmembrane domain of CD8 alpha; and/or   (p) the transmembrane domain comprises the amino acid sequence set forth in SEQ ID NO: 16.   
     
     
         5 . The modified cell of  claim 1 , wherein the chimeric cell surface sialidase further comprises:
 (a) a hinge domain; and/or   (b) an intracellular region comprising a costimulatory signaling domain and an intracellular signaling domain.   
     
     
         6 . The modified cell of  claim 5 , wherein:
 (a) the hinge domain is selected from the group consisting of an Fc fragment of an antibody, a hinge region of an antibody, a CH2 region of an antibody, a CH3 region of an antibody, an artificial hinge domain, a hinge comprising an amino acid sequence of CD8, or any combination thereof; and/or   (b) the hinge domain is a hinge comprising an amino acid sequence of CD8; and/or   (c) the hinge domain is a hinge comprising an amino acid sequence of CD8 alpha; and/or   (d) the hinge domain comprises the amino acid sequence set forth in SEQ ID NO: 15; and/or   (e) the costimulatory signaling domain comprises a costimulatory domain of a protein selected from the group consisting of proteins in the TNFR superfamily, CD28, 4-1BB (CD137), OX40 (CD134), PD-1, CD7, LIGHT, CD83L, DAP10, DAP12, CD27, CD2, CD5, ICAM-1, LFA-1, Lck, TNFR-I, TNFI-II, Fas, CD30, CD40, ICOS, NKG2C, and B7-H3 (CD276), or a variant thereof; and/or   (f) the costimulatory signaling domain comprises a costimulatory domain of 4-1BB; and/or   (g) the costimulatory signaling domain comprises the amino acid sequence set forth in SEQ ID NO: 17; and/or   (h) the costimulatory signaling domain comprises a costimulatory domain of CD2; and/or   (i) the intracellular signaling domain comprises an intracellular domain selected from the group consisting of cytoplasmic signaling domains of a human CD3 zeta chain (CD3z), FcγRIII, FcsRI, a cytoplasmic tail of an Fc receptor, an immunoreceptor tyrosine-based activation motif (ITAM) bearing cytoplasmic receptor, TCR zeta, FcR gamma, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, and CD66d, or a variant thereof; and/or   (j) the intracellular signaling domain comprises an intracellular domain of CD3z; and/or   (k) the intracellular signaling domain comprises the amino acid sequence set forth in SEQ ID NO: 18.   
     
     
         7 . The modified immune cell of  claim 1 , further comprising a chimeric antigen receptor (CAR) and/or a T cell receptor (TCR). 
     
     
         8 . The modified cell of  claim 2 , wherein:
 (a) the CAR comprises an antigen binding domain, a transmembrane domain, and an intracellular region; and/or   (b) the CAR comprises an antigen binding domain selected from the group consisting of an antibody, an scFv, and a Fab; and/or   (c) the CAR comprises an antigen binding domain comprising specificity for a tumor associated antigen (TAA); and/or   (d) the CAR comprises an antigen binding domain comprising specificity for TnMUC1; and/or   (e) the CAR comprises an antigen binding domain comprising specificity for CD19; and/or   (f) the CAR comprises an antigen binding domain comprising specificity for PSMA; and/or   (g) the CAR further comprises a hinge domain.   
     
     
         9 . The modified cell of  claim 7 , wherein the TCR:
 (a) is specific for a tumor associated antigen (TAA); and/or   (b) is selected from the group consisting of a wild-type TCR, a high affinity TCR, and a chimeric TCR; and/or   (c) comprises a TCR alpha chain and a TCR beta chain.   
     
     
         10 . The modified cell of  claim 1 , wherein the modified cell is:
 (a) a modified immune cell; and/or   (b) a modified T cell; and/or   (c) an autologous cell; and/or   (d) an autologous cell obtained from a human subject.   
     
     
         11 . A pharmaceutical composition comprising a therapeutically effective amount of the modified cell of  claim 1 . 
     
     
         12 . The composition of  claim 11 , further comprising a therapeutically effective population of innate immune cells. 
     
     
         13 . The composition of  claim 12 , wherein:
 (a) the innate immune cells are NK cells; and/or   (b) the innate immune cells are NK cells and wherein the NK cells are autologous NK cells; and/or   (c) the innate immune cells are NK cells and wherein the NK cells are autologous NK cells obtained from a human subject.   
     
     
         14 . The pharmaceutical composition of  claim 11 , comprising:
 (a) a therapeutically effective amount of a modified immune cell or precursor cell thereof, wherein the modified cell comprises a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising Neu2 or an enzymatically functional portion thereof, and a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface, and a chimeric antigen receptor (CAR); and a therapeutically effective amount of an NK cell; and/or   (b) a therapeutically effective amount of a modified immune cell or precursor cell thereof, wherein the modified cell comprises a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising Neu2 or an enzymatically functional portion thereof, a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface, a hinge domain, and an intracellular region comprising a costimulatory signaling domain and an intracellular signaling domain, and a chimeric antigen receptor (CAR); and a therapeutically effective amount of an NK cell; and/or   (c) a therapeutically effective amount of a modified immune cell or precursor cell thereof, wherein the modified cell comprises a variant sialidase precursor protein comprising a heterologous secretory sequence operably linked to a sialidase (neuraminidase) or an enzymatically functional portion thereof, wherein the variant sialidase precursor protein lacks a transmembrane domain, and wherein the sialidase or enzymatically functional portion thereof is capable of being secreted from an immune or precursor cell thereof when the variant sialidase precursor protein is expressed in the cell, and a chimeric antigen receptor (CAR); and a therapeutically effective amount of an NK cell.   
     
     
         15 . A chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising a sialidase (neuraminidase) or an enzymatically functional portion thereof, and a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface. 
     
     
         16 . The cell surface sialidase of  claim 15 , wherein:
 (a) the sialidase is a human or humanized sialidase; and/or   (b) the sialidase is a human or humanized sialidase and wherein the human sialidase is selected from the group consisting of Neu1, Neu2, Neu3, and Neu4; and/or   (c) the sialidase is a human or humanized sialidase and wherein the human sialidase is Neu2; and/or   (d) the sialidase comprises the amino acid sequence set forth in SEQ ID NO: 13; and/or   (e) the cell surface sialidase comprises the amino acid sequence set forth in SEQ ID NO: 10; and/or   (f) the cell surface sialidase comprises the amino acid sequence set forth any one of SEQ ID NOs: 10, 37, or 38; and/or   (g) the transmembrane domain is selected from the group consisting of an artificial hydrophobic sequence, and a transmembrane domain of a type I transmembrane protein, an alpha, beta, or zeta chain of a T cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, OX40 (CD134), 4-1BB (CD137), and CD154; and/or   (h) the transmembrane domain comprises a transmembrane domain of CD8; and/or   (i) the transmembrane domain comprises a transmembrane domain of CD8 alpha; and/or   (j) the transmembrane domain comprises the amino acid sequence set forth in SEQ ID NO: 16.   
     
     
         17 . The cell surface sialidase of  claim 15 , further comprising:
 (a) a hinge domain; and/or   (b) an intracellular region comprising a costimulatory signaling domain and an intracellular signaling domain.   
     
     
         18 . The cell surface sialidase of  claim 17 , wherein:
 (a) the hinge domain is selected from the group consisting of an Fc fragment of an antibody, a hinge region of an antibody, a CH2 region of an antibody, a CH3 region of an antibody, an artificial hinge domain, a hinge comprising an amino acid sequence of CD8, or any combination thereof; and/or   (b) the hinge domain is a hinge comprising an amino acid sequence of CD8; and/or   (c) the hinge domain is a hinge comprising an amino acid sequence of CD8 alpha; and/or   (d) the hinge domain comprises the amino acid sequence set forth in SEQ ID NO: 15; and/or   (e) the costimulatory signaling domain comprises a costimulatory domain of a protein selected from the group consisting of proteins in the TNFR superfamily, CD28, 4-1BB (CD137), OX40 (CD134), PD-1, CD7, LIGHT, CD83L, DAP10, DAP12, CD27, CD2, CD5, ICAM-1, LFA-1, Lck, TNFR-I, TNFI-II, Fas, CD30, CD40, ICOS, NKG2C, and B7-H3 (CD276), or a variant thereof; and/or   (f) the costimulatory signaling domain comprises a costimulatory domain of 4-1BB; and/or   (g) the costimulatory signaling domain comprises the amino acid sequence set forth in SEQ ID NO: 17; and/or   (h) the intracellular signaling domain comprises an intracellular domain selected from the group consisting of cytoplasmic signaling domains of a human CD3 zeta chain (CD3z), FcγRIII, FcsRI, a cytoplasmic tail of an Fc receptor, an immunoreceptor tyrosine-based activation motif (ITAM) bearing cytoplasmic receptor, TCR zeta, FcR gamma, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, and CD66d, or a variant thereof; and/or   (i) the intracellular signaling domain comprises an intracellular domain of CD3z; and/or   (j) the intracellular signaling domain comprises the amino acid sequence set forth in SEQ ID NO: 18.   
     
     
         19 . A variant sialidase precursor protein, comprising:
 (a) a heterologous secretory sequence operably linked to a sialidase (neuraminidase) or an enzymatically functional portion thereof, wherein the variant sialidase precursor protein lacks a transmembrane domain, and wherein the sialidase or enzymatically functional portion thereof is capable of being secreted from an immune or precursor cell thereof when the variant sialidase precursor protein is expressed in the cell; and/or   (b) a heterologous secretory sequence operably linked to a sialidase (neuraminidase) or an enzymatically functional portion thereof, wherein the sialidase comprises the amino acid sequence set forth in SEQ ID NO: 34.   
     
     
         20 . The variant sialidase of  claim 19 , wherein:
 (a) the sialidase is a human sialidase; and/or   (b) the sialidase is a human sialidase and wherein the human sialidase is selected from the group consisting of Neu1, Neu2, Neu3, and Neu4; and/or   (c) the variant sialidase is a variant Neu2; and/or   (d) the variant sialidase is a variant Neu2 and wherein the variant Neu2 comprises the amino acid sequence set forth in SEQ ID NO: 13.   
     
     
         21 . A nucleic acid comprising:
 (a) a first nucleic acid encoding a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising a sialidase (neuraminidase) or an enzymatically functional portion thereof, and a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface; and/or   (b) a first nucleic acid sequence encoding a variant sialidase precursor protein comprising SEQ ID NO: 30; and/or   (c) first nucleic acid sequence encoding a variant sialidase precursor protein comprising SEQ ID NO: 30; and/or   (d) a first nucleic acid sequence encoding a chimeric cell surface sialidase comprising SEQ ID NO: 1 or 40; and/or   (e) a first nucleic acid sequence encoding a variant sialidase precursor protein comprising a heterologous secretory sequence operably linked to a sialidase (neuraminidase) or an enzymatically functional portion thereof, wherein the variant sialidase precursor protein lacks a transmembrane domain, and wherein the sialidase or enzymatically functional portion thereof is capable of being secreted from an immune or precursor cell thereof when the variant sialidase precursor protein is expressed in the cell; and/or   (f) a first nucleic acid sequence encoding a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising a sialidase (neuraminidase) or an enzymatically functional portion thereof, and a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface, and a second nucleic acid sequence encoding a chimeric antigen receptor (CAR) and/or a T cell receptor (TCR); and/or   (g) a first nucleic acid sequence encoding a variant sialidase precursor protein comprising a heterologous secretory sequence operably linked to a sialidase (neuraminidase) or an enzymatically functional portion thereof, wherein the variant sialidase precursor protein lacks a transmembrane domain, and wherein the sialidase or enzymatically functional portion thereof is capable of being secreted from an immune or precursor cell thereof when the variant sialidase precursor protein is expressed in the cell, and a second nucleic acid sequence encoding a chimeric antigen receptor (CAR) and/or a T cell receptor (TCR).   
     
     
         22 . The nucleic acid of  claim 21 , wherein:
 (a) the sialidase is a human or humanized sialidase; and/or   (b) the sialidase is a human or humanized sialidase and wherein the human sialidase is selected from the group consisting of Neu1, Neu2, Neu3, and Neu4; and/or   (c) the sialidase is a human or humanized sialidase and wherein the sialidase is Neu2; and/or   (d) the sialidase is encoded by a nucleic acid sequence comprising SEQ ID NO: 4; and/or   (e) the transmembrane domain is selected from the group consisting of an artificial hydrophobic sequence, and a transmembrane domain of a type I transmembrane protein, an alpha, beta, or zeta chain of a T cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, OX40 (CD134), 4-1BB (CD137), and CD154; and/or   (f) the transmembrane domain comprises a transmembrane domain of CD8; and/or   (g) the transmembrane domain comprises a transmembrane domain of CD8 alpha; and/or   (h) the transmembrane domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 7; and/or   (i) the nucleic acid further comprises a leader sequence; and/or   (j) the nucleic acid further comprises a leader sequence and wherein the leader sequence is a CD8 alpha leader sequence; and/or   (k) the nucleic acid further comprises a leader sequence and wherein the leader sequence is encoded by a nucleic acid sequence comprising SEQ ID NO: 2; and/or   (l) the nucleic acid further comprises a hinge domain; and/or   (m) the nucleic acid further comprises a hinge domain and wherein the hinge domain is selected from the group consisting of an Fc fragment of an antibody, a hinge region of an antibody, a CH2 region of an antibody, a CH3 region of an antibody, an artificial hinge domain, a hinge comprising an amino acid sequence of CD8, or any combination thereof; and/or   (n) the nucleic acid further comprises a hinge domain and wherein the hinge domain is a hinge comprising an amino acid sequence of CD8; and/or   (o) the nucleic acid further comprises a hinge domain and wherein the hinge domain is a hinge comprising an amino acid sequence of CD8 alpha; and/or   (p) the nucleic acid further comprises a hinge domain and wherein the hinge domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 6; and/or   (q) the nucleic acid further comprises an intracellular region comprising a costimulatory signaling domain and an intracellular signaling domain, and optionally wherein:
 (i) the costimulatory signaling domain comprises a costimulatory domain of a protein selected from the group consisting of proteins in the TNFR superfamily, CD28, 4-1BB (CD137), OX40 (CD134), PD-1, CD7, LIGHT, CD83L, DAP10, DAP12, CD27, CD2, CD5, ICAM-1, LFA-1, Lck, TNFR-I, TNFI-II, Fas, CD30, CD40, ICOS, NKG2C, and B7-H3 (CD276), or a variant thereof; and/or 
 (ii) the costimulatory signaling domain comprises a costimulatory domain of 4-1BB; and/or 
 (iii) the costimulatory signaling domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 8; and/or 
 (iv) the intracellular signaling domain comprises an intracellular domain selected from the group consisting of cytoplasmic signaling domains of a human CD3 zeta chain (CD3z), FcγRIII, FcsRI, a cytoplasmic tail of an Fc receptor, an immunoreceptor tyrosine-based activation motif (ITAM) bearing cytoplasmic receptor, TCR zeta, FcR gamma, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, and CD66d, or a variant thereof; and/or 
 (v) the intracellular signaling domain comprises an intracellular domain of CD3z; and/or 
 (vi) the intracellular signaling domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 9. 
   
     
     
         23 . The nucleic acid of  claim 21 , further comprising a second nucleic acid sequence encoding an exogenous T cell receptor (TCR) and/or chimeric antigen receptor (CAR). 
     
     
         24 . The nucleic acid of  claim 23 , wherein:
 (a) the CAR comprises an antigen binding domain, a transmembrane domain, and an intracellular region, and optionally wherein:
 (i) the transmembrane domain is selected from the group consisting of an artificial hydrophobic sequence and transmembrane domain of a type I transmembrane protein, an alpha, beta, or zeta chain of a T cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, and CD154; and/or 
 (ii) the intracellular region comprises a costimulatory signaling domain and an intracellular signaling domain; and/or 
 (iii) the costimulatory signaling domain comprises a costimulatory domain of a protein selected from the group consisting of proteins in the TNFR superfamily, CD28, 4-1BB (CD137), OX40 (CD134), PD-1, CD7, LIGHT, CD83L, DAP10, DAP12, CD27, CD2, CD5, ICAM-1, LFA-1, Lck, TNFR-I, TNFI-II, Fas, CD30, CD40, ICOS, NKG2C, and B7-H3 (CD276), or a variant thereof; and/or 
 (iv) the costimulatory signaling domain comprises a costimulatory domain of CD2; and/or 
 (v) the costimulatory signaling domain comprises a costimulatory domain of 4-1BB; and/or 
 (vi) the intracellular signaling domain comprises an intracellular domain selected from the group consisting of cytoplasmic signaling domains of a human CD3 zeta chain (CD3z), FcγRIII, FcsRI, a cytoplasmic tail of an Fc receptor, an immunoreceptor tyrosine-based activation motif (ITAM) bearing cytoplasmic receptor, TCR zeta, FcR gamma, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, and CD66d, or a variant thereof; and/or 
 (vii) the intracellular signaling domain comprises an intracellular domain of CD3z; and/or 
   (b) the CAR comprises an antigen binding domain selected from the group consisting of an antibody, an scFv, and a Fab; and/or   (c) the CAR comprises an antigen binding domain comprising specificity for a tumor associated antigen (TAA); and/or   (d) the CAR comprises an antigen binding domain comprising specificity for TnMUC1; and/or   (e) the CAR comprises an antigen binding domain comprising specificity for CD19; and/or   (f) the CAR comprises an antigen binding domain comprising specificity for PSMA; and/or   (g) the CAR further comprises a hinge domain, and optionally wherein the hinge domain is a hinge domain selected from the group consisting of an Fc fragment of an antibody, a hinge region of an antibody, a CH2 region of an antibody, a CH3 region of an antibody, an artificial hinge domain, a hinge comprising an amino acid sequence of CD8, or any combination thereof; and/or   (h) the TCR is specific for a tumor associated antigen (TAA); and/or   (i) the TCR is selected from the group consisting of a wild-type TCR, a high affinity TCR, and a chimeric TCR; and/or   (j) the TCR comprises a TCR alpha chain coding sequence and a TCR beta chain coding sequence; and/or   (k) the TCR alpha chain coding sequence and the TCR beta chain coding sequence are separated by a first linker, and optionally wherein:
 (i) the first linker comprises a nucleic acid sequence encoding an internal ribosome entry site (IRES), a furin cleavage site, a self-cleaving peptide, or any combination thereof; and/or 
 (ii) the first linker comprises a furin cleavage site and a self-cleaving peptide, and optionally wherein the self-cleaving peptide is a 2A peptide selected from the group consisting of porcine teschovirus-1 2A (P2A), Thoseaasigna virus 2A (T2A), equine rhinitis A virus 2A (E2A), and foot-and-mouth disease virus 2A (F2A); and/or 
   (l) the first nucleic acid sequence and the second nucleic acid sequence are separated by a second linker, and optionally wherein:
 (i) the second linker comprises a nucleic acid sequence encoding an internal ribosome entry site (IRES); and/or 
 (ii) the second linker comprises a cleavage site and/or a self-cleaving peptide, and optionally wherein the cleavage site is a furin cleavage site and/or wherein the self-cleaving peptide is a 2A peptide, and optionally wherein the 2A peptide (A) is selected from the group consisting of porcine teschovirus-1 2A (P2A), Thoseaasigna virus 2A (T2A), equine rhinitis A virus 2A (E2A), and foot-and-mouth disease virus 2A (F2A); (B) is P2A; and/or (C) is T2A; and/or 
   (m) the nucleic acid comprises from 5′ to 3′: the first nucleic acid sequence, the second linker, and the second nucleic acid sequence; and/or   (n) the nucleic acid comprises from 5′ to 3′: the second nucleic acid sequence, the second linker, and the first nucleic acid sequence.   
     
     
         25 . An expression vector comprising the nucleic acid of  claim 21 . 
     
     
         26 . The expression vector of  claim 25 , wherein:
 (a) the expression vector is a viral vector selected from the group consisting of a retroviral vector, a lentiviral vector, an adenoviral vector, and an adeno-associated viral vector; and/or   (b) the expression vector is a lentiviral vector, and optionally wherein the lentiviral vector is a self-inactivating lentiviral vector.   
     
     
         27 . A method of treating cancer in a subject in need thereof, the method comprising:
 (a) administering the modified cell of  claim 1  or the pharmaceutical composition of  claim 11  to the subject; and/or   (b) administering a modified T cell comprising a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising a sialidase (neuraminidase) or an enzymatically functional portion thereof, and a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface, and a chimeric antigen receptor (CAR); and/or   (c) administering a modified T cell comprising a variant sialidase precursor protein comprising a heterologous secretory sequence operably linked to a sialidase (neuraminidase) or an enzymatically functional portion thereof, wherein the variant sialidase precursor protein lacks a transmembrane domain, and wherein the sialidase or enzymatically functional portion thereof is capable of being secreted from an immune or precursor cell thereof when the variant sialidase precursor protein is expressed in the cell, and a chimeric antigen receptor (CAR); and/or   (d) administering to the subject a therapeutically effective amount of a modified T cell comprising a chimeric cell surface sialidase (neuraminidase) enzyme comprising an extracellular portion comprising a sialidase (neuraminidase) or an enzymatically functional portion thereof, and a heterologous transmembrane domain capable of tethering the extracellular portion to a cell surface, and a chimeric antigen receptor (CAR); and administering to the subject a therapeutically effective amount of a NK cell; and/or   (e) administering to the subject a therapeutically effective amount of a modified T cell comprising a variant sialidase precursor protein comprising a heterologous secretory sequence operably linked to a sialidase (neuraminidase) or an enzymatically functional portion thereof, wherein the variant sialidase precursor protein lacks a transmembrane domain, and wherein the sialidase or enzymatically functional portion thereof is capable of being secreted from an immune or precursor cell thereof when the variant sialidase precursor protein is expressed in the cell, and a chimeric antigen receptor (CAR); and administering to the subject a therapeutically effective amount of a NK cell.   
     
     
         28 . The method of  claim 27 , wherein:
 (a) the CAR comprises specificity for TnMUC1; and/or   (b) the CAR comprises specificity for CD19; and/or   (c) the CAR comprises specificity for PSMA; and/or   (d) the method further comprises administering to the subject a population of innate immune cells, and optionally wherein:
 (i) the innate immune cells are NK cells; and/or 
 (ii) the innate immune cells are NK cells and wherein the NK cells are autologous NK cells obtained from a human subject; and/or 
   (e) the modified T cell and the NK cell are administered simultaneously; and/or   (f) the modified T cell and the NK cell are administered separately; and/or   (g) the NK cell is autologous; and/or   (h) the modified T cell is autologous; and/or   (i) the subject is human.