Methods and Compositions Comprising Orthogonal Cytokine Responsive Immune Cells
AssigneesThe Trustees of the University of Pennsylvania, The Board of Trustees of the Leland Stanford Junior University
InventorsMichael C. Milone, Qian Zhang, Kenan Christopher Garcia
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Abstract
The present disclosure provides methods of producing a modified immune cell responsive to orthogonal cytokine signaling and a modified immune cell produced by said method. The present disclosure further provides a modified immune cell responsive to orthogonal cytokine signaling and methods for treating cancer comprising the modified immune cell.
Claims53
A method of producing a modified immune cell responsive to orthogonal cytokine signaling, the method comprising:
- (a) genetically engineering an immune effector cell responsive to interleukin-2 (IL-2) and interleukin-15 (IL-15) to express a T cell receptor (TCR) or a chimeric antigen receptor (CAR) from an exogenous nucleic acid inserted at a locus within endogenous IL-2 gene of the immune cell such that the modified immune cell is an IL-2−/− immune cell;
- and
- (b) genetically engineering the immune effector cell to express an orthogonal IL-2 receptor beta (oIL2Rβ);
- wherein step (a) and step (b) are performed in any order.
A modified immune cell responsive to orthogonal cytokine signaling, wherein the modified immune cell is derived from an immune effector cell responsive to interleukin-2 (IL-2) and interleukin-15 (IL-15);
- and wherein the modified immune cell:
- (a) expresses a T cell receptor (TCR) or a chimeric antigen receptor (CAR) from an exogenous nucleic acid inserted at a locus within endogenous IL-2 gene of the immune cell, wherein the exogenous nucleic acid comprises a polynucleotide sequence encoding the TCR or the CAR, such that the modified immune cell is an IL2−/− immune cell;
- and
- (b) expresses an orthogonal IL-2 receptor beta (oIL2Rβ).
A method of producing a modified immune cell responsive to orthogonal cytokine signaling, the method comprising:
- (a) genetically engineering an immune effector cell responsive to interleukin-2 (IL-2) and interleukin-15 (IL-15) to express a T cell receptor (TCR) or a chimeric antigen receptor (CAR) from an exogenous nucleic acid inserted at a locus within endogenous IL-2 gene of the immune cell such that the modified immune cell is an IL-2−/− immune cell;
- and
- (b) genetically engineering endogenous IL-2 receptor beta (IL2Rβ) gene of the immune effector cell to express an orthogonal IL-2 receptor beta (oIL2Rβ) such that the modified immune cell is an endogenous IL2Rβ−/− immune cell and an oIL2Rβ+/+ immune cell;
- wherein step (a) and step (b) are performed in any order;
- and
- further wherein step (a) comprises CRISPR/Cas-mediated homology directed repair (HDR) and step (b) comprises prime editing.
A modified immune cell responsive to orthogonal cytokine signaling, wherein the modified immune cell is derived from an immune effector cell responsive to interleukin-2 (IL-2) and interleukin-15 (IL-15);
- and wherein the modified immune cell:
- (a) expresses a T cell receptor (TCR) or a chimeric antigen receptor (CAR) from an exogenous nucleic acid inserted at a locus within endogenous IL-2 gene of the immune cell, wherein the exogenous nucleic acid comprises a polynucleotide sequence encoding the TCR or the CAR, such that the modified immune cell is an IL2−/− immune cell;
- and
- (b) expresses an orthogonal IL-2 receptor beta (oIL2Rβ);
- wherein the endogenous IL2Rβ gene is edited such that it encodes the oIL2Rβ.
A method of producing a modified immune cell responsive to orthogonal cytokine signaling, the method comprising genetically engineering at least one endogenous IL-2 receptor beta (IL2Rβ) gene of the immune effector cell to express an orthogonal IL-2 receptor beta (oIL2Rβ), wherein the modified immune cell is derived from an immune effector cell responsive to interleukin-2 (IL-2) and interleukin-15 (IL-15);
- further wherein the genetic engineering comprises prime editing, and wherein the prime editing comprises a prime editing guide RNA (pegRNA) comprising or consisting of SEQ ID NO:
- 1.
Dependencies are inferred from the claim text.