Immunoeffector cells derived from induced pluripotent stem cells genetically engineered with membrane bound il12 and uses thereof

An induced pluripotent stem cell (iPSC), a natural killer (NK) cell or a T cell comprising:

• (i) a first exogenous polynucleotide encoding a chimeric antigen receptor (CAR);
• (ii) a second exogenous polynucleotide encoding:
• a membrane-bound interleukin 12 (IL-12) comprising a first polypeptide comprising an IL-12 alpha subunit p35 or a polypeptide at least 90% similar thereto, a second polypeptide comprising an IL-12 beta subunit p40 or a polypeptide at least 90% similar thereto, and a transmembrane domain fused to the terminus of the first and/or second IL-12 subunit polypeptide;
• or a membrane bound IL-12 fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide;
• (iii) a third exogenous polynucleotide encoding a human leukocyte antigen E (HLA-E) and/or human leukocyte antigen G (HLA-G);
• (iv) optionally a fourth exogenous polynucleotide encoding a safety switch;
• and
• (v) optionally a fifth exogeneous polynucleotide encoding a cytokine;
• wherein one or more of the exogenous polynucleotides are integrated at loci of CIITA and B2M genes to thereby delete or reduce expression of CIITA and B2M.

An iPSC, a natural killer (NK) cell or a T cell, comprising:

• (i) a first exogenous polynucleotide encoding a chimeric antigen receptor (CAR);
• (ii) a second exogenous polynucleotide encoding:
• i. a membrane-bound interleukin 12 (IL-12) having the amino acid sequence of SEQ ID NO:
• 96;
• ii. a membrane bound IL-12 fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide having the amino acid sequence of SEQ ID NO:
• 98;
• iii. a membrane bound IL-12 fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide having the amino acid sequence of SEQ ID NO:
• 108;
• iv. a membrane bound IL-12 fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide having the amino acid sequence of SEQ ID NO:
• 110;
• v. a membrane bound IL-12 fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide having the amino acid sequence of SEQ ID NO:
• 112;
• vi. a membrane bound IL-12 fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide having the amino acid sequence of SEQ ID NO:
• 114;
• vii. a membrane bound IL-12 fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide having the amino acid sequence of SEQ ID NO:
• 116;
• viii. a membrane bound IL-12 fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide having the amino acid sequence of SEQ ID NO:
• 118;
• ix. a membrane bound IL-12 fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide having the amino acid sequence of SEQ ID NO:
• 120;
• x. a membrane bound IL-12 fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide having the amino acid sequence of SEQ ID NO:
• 122;
• or
• xi. a membrane bound IL-12 fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide having the amino acid sequence of SEQ ID NO:
• 124;
• (iii) optionally, a third exogenous polynucleotide encoding a human leukocyte antigen E (HLA-E) having the amino acid sequence of SEQ ID NO:
• 66 and/or an exogenous polynucleotide encoding a human leukocyte antigen G (HLA-G) having the amino acid sequence of SEQ ID NO:
• 69;
• and
• (iv) optionally, a fourth exogeneous polynucleotide encoding an IL-15 protein according to SEQ ID NO:
• 72;
• wherein one or more of the exogenous polynucleotides are integrated at loci of CIITA and B2M genes to thereby delete or reduce expression of CIITA and/or B2M.