Universal anti-tag chimeric antigen receptor-expressing t cells and methods of treating cancer
The present invention provides a universal, yet adaptable, anti-tag chimeric antigen receptor (AT-CAR) system which provides T cells with the ability and specificity to recognize and kill target cells, such as tumor cells, that have been marked by tagged antibodies. As an example, αFITC-CAR-expressing T cells have been developed that specifically recognize various human cancer cells when those cells are bound by cancer-reactive FITC-labeled antibodies. The activation of αFITC-CAR-expressing T cells is shown to induce efficient target lysis, T cell proliferation, and cytokine/chemokine production. The system can be used to treating subjects having cancer.
1. A method of treating cancer in a subject, comprising:
(a) administering a formulation of tagged proteins to a subject in need of treatment, wherein the tagged proteins bind a cancer cell in the subject, and
(b) administering a therapeutically-effective population of anti-tag chimeric receptor (AT-CAR)-expressing effector cells to the subject, wherein the AT-CAR comprises a tag-binding domain, a transmembrane domain, and a T cell activation domain, where the T cell activation domain consists of one or more of the cytoplasmic region of OX40, the cytoplasmic region of HVEM, and FcRε, where the T cell activation domain does not comprise CD3ζ, and wherein the AT-CAR-expressing effector cells bind the tagged proteins and induce cancer cell death, thereby treating cancer in a subject.
2. A method of treating cancer in a subject, comprising:
(a) administering one or more formulations of tagged proteins to a subject in need of treatment, wherein the tagged proteins bind a cancer cell in the subject, and
(b) administering one or more therapeutically-effective populations of AT-CAR-expressing effector cells to the subject, wherein the AT-CAR comprises a tag-binding domain, a transmembrane domain, and a T cell activation domain, where the T cell activation domain consists of one or more of the cytoplasmic region of OX40, the cytoplasmic region of HVEM, and FcRε, where the T cell activation domain does not comprise CD3ζ, and wherein the AT-CAR-expressing effector cells bind the tagged proteins and induce cancer cell death, thereby treating cancer in a subject.
3. A method of treating cancer in a subject, comprising:
(a) administering at least two formulations of tagged proteins to a subject in need of treatment, wherein the tagged proteins bind a cancer cell in the subject, and
(b) administering at least two therapeutically-effective populations of AT-CAR-expressing effector cells to the subject, wherein the AT-CAR comprises a tag-binding domain, a transmembrane domain, and a T cell activation domain, where the T cell activation domain consists of one or more of the cytoplasmic region of OX40, the cytoplasmic region of HVEM, and FcRε, where the T cell activation domain does not comprise CD3ζ, and wherein the AT-CAR-expressing effector cells bind the tagged proteins and induce cancer cell death, thereby treating cancer in a subject.
4. The method of claim 1 , wherein the tagged proteins are tagged with a tag selected from the group consisting of fluorescein isothiocyanate (FITC), streptavidin, biotin, dinitrophenol, peridinin chlorophyll protein complex, green fluorescent protein, phycoerythrin (PE), horse radish peroxidase, palmitoylation, nitrosylation, alkalanine phosphatase, glucose oxidase, and maltose binding protein.
5. The method of claim 1 , wherein the protein of the tagged proteins is an antibody or an antigen-binding fragment thereof.
6. The method of claim 5 , wherein the antibody or antigen-binding fragment thereof is cetuximab, nimotuzumab, panitumumab, retuximab, omalizumab, tositumomab, trastuzumab, gemtuzumab, or alemtuzumab, or an antigen-binding fragment of any one thereof.
7. The method of claim 1 , wherein the tag-binding domain is an antibody or an antigen-binding fragment thereof.
8. The method of claim 1 , wherein the tag-binding domain specifically binds FITC, biotin, PE, or streptavidin.
9. The method of claim 7 , wherein the antigen-binding fragment is a single chain variable fragment (scFv).
10. The method of claim 7 , wherein the antigen-binding fragment is a single chain variable fragment (scFv) that specifically binds FITC, biotin, PE, or streptavidin.
11. The method of claim 7 , wherein the transmembrane domain is the hinge and transmembrane regions of the human CD8α chain.
12. The method of claim 1 , wherein the effector cells of the population of AT-CAR-expressing effector cells comprises CD3 + CD8 + T cells, CD3 + CD4 + T cells, gamma delta T cells, cytokine induced killer (CIK) cells, natural killer (NK) cells, natural killer T (NKT) cells, tumor infiltrating lymphocytes (TILs), macrophages, or microglia.
13. The method of claim 1 , wherein the formulation of tagged proteins is administered to the subject prior to administration of the therapeutically-effective population of AT-CAR-expressing effector cells.
14. The method of claim 1 , wherein the formulation of tagged proteins are administered to the subject concurrently with administration of the therapeutically-effective population of AT-CAR-expressing effector cells.
15. The method of claim 1 , wherein the formulation of tagged proteins are administered to the subject after administration of the therapeutically-effective population of AT-CAR-expressing effector cells.
16. The method of claim 1 , wherein the formulation of tagged proteins and the therapeutically-effective population of AT-CAR-expressing effector cells are administered to the subject in any order.
17. The method of claim 1 , wherein AT-CAR-expressing effector cell binding to the tagged proteins, which are bound to a cancer cell, induces cytolytic activation of the effector cells.
18. The method of claim 1 , wherein the subject is a human.