Chimeric antigen receptor and its use
In a first aspect, the present invention relates to a recombinant polypeptide containing a domain comprising at least two antibody units whereby the first antibody unit is an anti-CD30 single chain antibody unit while the second antibody unit is a antibody unit being specific for an antigen present on the surface of a pre-determined target cell. In particular, the present invention relates to a recombinant polypeptide containing at least the following domains starting from the N-terminus to the C-terminus: a first domain containing an anti-CD30 single chain antibody unit, in particular, HRS3 scFv of SEQ ID No. 2 or homologs thereof having at least 70% identity with SEQ ID No. 2 binding specifically to CD30, and an antibody unit said antibody unit being specific for an antigen present on the surface of a predetermined target cell, in particular, being specific for a tumor-associated antigen; optionally a spacer domain; a trans-membrane domain; and a cytoplasmatic signalling domain. In a further aspect, the present invention relates to a nucleic acid molecule encoding the polypeptide according to the present invention, as well as vectors and cells containing the same. Moreover, lymphocytes are provided, in particular T-cells like CD8 + or a CD4 + T-cell expressing on its surface chimeric antigen receptors containing an anti-CD30 single chain antibody unit and an antibody unit whereby said antibody unit being specific for an antigen present on the surface of a predetermined target cell. Immune cells modified with the polypeptide show improved functions, in particular in the treatment of cancer, in particular CD30 − cancer. That is, the cells are for use in adapted cell therapy for treating cancer in a subject in need thereof.
1. A method of treating CD30 negative cancer comprising the step of administering to a subject in need thereof a nucleic acid molecule comprising a nucleic acid sequence encoding a recombinant polypeptide with at least the following domains starting from the N-terminus to the C-terminus:
first domain containing i) an anti-CD30 single chain antibody unit and ii) an antibody unit specific for an antigen present on the surface of a predetermined target cell, wherein the target cell is a CD30 negative cancer cell;
optionally a spacer domain
a transmembrane domain; and
a cytoplasmatic signalling domain,
wherein the anti-CD30 single chain antibody unit in the first domain is HRS3 scFv of SEQ ID NO. 2 or a homolog thereof having at least 95% identity with SEQ ID NO. 2 binding specifically to CD30, and
wherein the nucleic acid sequence is present in a cell, cell line, host cell or a viral vector or other vector.
2. The method of treating cancer according to claim 1 wherein the cell, cell line, or host cell expresses the recombinant polypeptide.
3. The method of treating cancer according to claim 1 , wherein the cancer cells of said cancer express the antigen.
4. The method for treating cancer according to claim 1 , wherein the antibody unit used for the antigen present on the surface of the predetermined target cells being CD30 negative binds to a tumor-associated antigen selected from the group consisting of carcinoembryonic antigen (CEA), CA19-9, CA72-4 (TAG-72), PSCA, Muc-1, HMW-MAA, p97 melanotransferrin, fetal actelycholin receptor, ErbB2 (Her2/neu), multi-drug-resistance protein (MDR), CD19, CD20, and TOSO.
5. The method of treating cancer according to claim 1 , wherein said recombinant polypeptide further comprises a leader sequence being located N-termini to the first domain containing anti-CD30 single chain antibody unit and the antibody unit specific for the antigen present on the surface of a predetermined target cell being CD30 negative.
6. The method of treating cancer according to claim 1 , wherein the anti-CD30 single chain antibody unit is the HRS3 scFv peptide of SEQ ID NO. 2.
7. The method of treating cancer according to claim 1 , wherein the spacer domain is present and is an IgG1 CH2 CH3 domain of SEQ ID No. 6 or homologs thereof having at least 95% identity herewith.
8. The method of treating cancer according to claim 1 , wherein the transmembrane domain is obtained from CD28.
9. The method of treating cancer according to claim 1 , wherein the intracellular domain contains a CD3zeta or a Fc-epsilon receptor I-gamma signalling chain or a costimulatory wall.
10. The method of treating cancer according to claim 1 , wherein the cell or the host cell is provided, and wherein the cell or host cell is a modified peripheral blood cell.
11. The method of treating cancer according to claim 1 , wherein the spacer domain is a mutated IgG1 CH2 CH3 domain according to SEQ ID No. 6.
12. The method of treating cancer according to claim 1 , wherein said antigen present on the surface of a predetermined target cell is a tumor-associated antigen.
13. The method of treating cancer according to claim 1 , wherein the intracellular domain is the IgE Fc epsilon receptor-I gamma-signalling unit of SEQ ID No. 12 or a homolog thereof having at least 95% identity.