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 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 wherein the tagged proteins are antibodies or antigen-binding fragments thereof, and
(b) administering one or more therapeutically-effective populations of anti-tag chimeric receptor (AT-CAR)-expressing T 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 T 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 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 wherein the tagged proteins are antibodies or antigen-binding fragments thereof, and
(b) administering at least two therapeutically-effective populations of AT-CAR-expressing T 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 T cells bind the tagged proteins and induce cancer cell death, thereby treating cancer in a subject.
3. The method of claim 1 , wherein the tags of the formulations of tagged proteins are the same or different and the tags are 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.
4. The method of claim 1 , wherein the antibodies or antigen-binding fragments thereof of the formulations are the same or different.
5. The method of claim 4 , wherein the antibodies or antigen-binding fragments thereof are selected from the group consisting of cetuximab, nimotuzumab, panitumumab, retuximab, omalizumab, tositumomab, trastuzumab, gemtuzumab, alemtuzumab or an antigen-binding fragment of any one thereof.
6. The method of claim 1 , wherein the AT-CAR of the AT-CAR-expressing T cells are the same or different.
7. The method of claim 6 , wherein the tag-binding domain is an antibody or an antigen-binding fragment thereof.
8. The method of claim 6 , 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 6 , wherein the transmembrane domain is the hinge and transmembrane regions of the human CD8a chain.
12. The method of claim 1 , wherein the T cells of the populations of AT-CAR-expressing T cells are the same or different and wherein the T cells are selected from the group consisting of T cells of any HLA-background from peripheral blood mononuclear cells (PBMC), T cells isolated from a tumor explant of the subject, and intratumoral T cells of the subject.
13. The method of claim 1 , wherein the T cells of the populations of AT-CAR-expressing T cells consist of HLA-A2+peripheral blood mononuclear cells (PBMC).
14. The method of claim 1 , wherein the formulations of tagged proteins are administered to the subject prior to or after administration of the therapeutically-effective populations of AT-CAR-expressing T cells.
15. The method of claim 1 , wherein the formulations of tagged proteins are administered to the subject concurrently with administration of the therapeutically-effective populations of AT-CAR-expressing T cells.
16. The method of claim 1 , wherein the formulations of tagged proteins and the therapeutically-effective populations of AT-CAR-expressing T cells are administered to the subject in any order.
17. The method of claim 1 , wherein AT-CAR-expressing T cell binding to the tagged proteins induces cytolytic activation of the T cells.
18. The method of claim 1 , wherein the subject is a human.