Tumor infiltrating cells engineered to express a pro-inflammatory polypeptide
The present disclosure provides methods of preparing tumor infiltrating cells engineered to express a pro-inflammatory polypeptide. The pro-inflammatory polypeptide is expressed from the tumor infiltrating cell to counter a generally immunosuppressive state in and around tumors resulting from an imbalance between the number and activation state of immune effector cells versus those of suppressor cells. Delivering the proinflammatory polypeptide via expression from the TICs, as distinct from systemic administration, reduces side effects from increased inflammation at sides remote from a tumor to be treated.
1. An isolated tumor-infiltrating cell genetically engineered to provide increased expression of a pro-inflammatory protein other than a chimeric antigen receptor (CAR), wherein the tumor-infiltrating cell is derived from a cell isolated from a solid tumor biopsy and 1s genetically engineered for expression of an exogenous nucleic acid encoding the pro-inflammatory protein; and wherein the tumor-infiltrating cell further comprises a nucleic acid encoding a matrix degrading enzyme, wherein the matrix degrading enzyme 1s chosen from a matrix metalloproteinase and plasminogen activator, wherein the cell is selected from a T cell or a natural killer (NK) cell, and wherein the pro-inflammatory protein encoded by the exogenous nucleic acid comprises
a. an IL-21 cytokine;
b. a chemokine chosen from RANTES, IP-10, CXCL9, and CXCL10; or
c. a fusion protein comprising any of the foregoing cytokines and chemokines.
2. The isolated tumor-infiltrating cell of claim 1 , further comprising a nucleic acid encoding a CAR.
3. The isolated tumor-infiltrating cell of claim 1 , wherein the cell is a T cell or wherein the cell expresses at least one marker from CD3, CD4, and CD8.
4. A composition comprising a mixture of genetically engineered tumor-infiltrating cell populations, each population clonally derived from a single genetically engineered tumor-infiltrating cell of claim 1 .
5. A composition comprising a mixture comprising clonal genetically engineered tumor-infiltrating cell populations, wherein at least 95% of the cells in the mixture are from one of the clonal populations, each clonal population being derived from a single genetically engineered tumor-infiltrating cell of claim 1 .
6. The isolated tumor-infiltrating cell of claim 1 , wherein the cell is a NK cell or wherein the cell expresses at least one marker chosen from CD56 and CD16.
7. A method of treating a patient having a cancer, the method comprising administering to the patient a genetically engineered tumor-infiltrating cell of claim 1 .
8. The method of claim 7 , wherein the cancer is a melanoma, a breast cancer, or a lung cancer.
9. The method of claim 7 , wherein the genetically engineered tumor infiltrating cells comprises an exogenous nucleic acid operably linked to an inducible promoter, and wherein the method further comprises administering to the patient an agent capable of inducing expression of the exogenous nucleic acid, further wherein the exogenous nucleic acid comprises a tetracycline-inducible promoter and the agent administered to the patient is tetracycline.