US20200085868A1 - Method For Treating Cancer - Google Patents

Method For Treating Cancer Download PDF

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US20200085868A1
US20200085868A1 US16/390,308 US201916390308A US2020085868A1 US 20200085868 A1 US20200085868 A1 US 20200085868A1 US 201916390308 A US201916390308 A US 201916390308A US 2020085868 A1 US2020085868 A1 US 2020085868A1
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chimeric antigen
antigen receptor
cell line
group
human
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Inventor
Der-Yang CHO
Shao-Chih CHIU
Chia-Ing JAN
Chih-Ming Pan
Shi-Wei Huang
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China Medical University Hospital
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China Medical University Hospital
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Assigned to CHINA MEDICAL UNIVERSITY HOSPITAL reassignment CHINA MEDICAL UNIVERSITY HOSPITAL ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: CHIU, SHAO-CHIH, CHO, DER-YANG, HUANG, Shi-wei, JAN, CHIA-ING, PAN, CHIH-MING
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Definitions

  • sequence listing submitted via EFS in compliance with 37 CFR ⁇ 1.52(e)(5), is incorporated herein by reference.
  • the sequence listing text file submitted via EFS contains the file “CP-4343-US_SequenceListing”, created on Mar. 19, 2019, which is 15,771 bytes in size.
  • the present disclosure relates to a method for treating cancer. More particularly, the present disclosure relates to a method for treating cancer with a chemotherapy drug and chimeric antigen receptor expressing cells.
  • Cancer also known as malignancy, is a state of abnormal proliferation of cells, and these proliferating cells may invade other parts of the body as a disease caused by a malfunction in the control of cell division and proliferation.
  • Cancer is one of the top ten causes of death for the Chinese people and has been the top ten causes of death for twenty-seven consecutive years.
  • Cancer immunotherapy is another method for treating cancer except the above methods.
  • the immune system of the patient is activated in the cancer immunotherapy by using tumor cells or tumor antigens to induce specific cellular and humoral immune responses for enhancing the anti-cancer ability of the patient, preventing the growth, spread, and recurrence of tumors, and achieving the purpose of removing or controlling tumors.
  • the chimeric antigen receptor immune cell technology is one of the cell therapy developing very rapidly in recent years.
  • the chimeric antigen receptor immune cell transfecting a chimeric protein, which couples the antigen binding portion having capable of recognizing a certain tumor antigen of the antibody to the intracellular portion of the CD3- ⁇ chain or Fc ⁇ Rl ⁇ in vitro, into the immune cell by a transduction method to express the chimeric antigen receptor.
  • the chimeric antigen receptor immune cell technology has a significant therapeutic effect in the treatment of acute leukemia and non-Hodgkin's lymphoma, and it is considered to be one of the most promising treatment for cancer.
  • the cell therapy of the chimeric antigen receptor immune cell currently has the following disadvantages: lack of unique tumor-associated antigens, low efficiency of homing of immune cells to tumor sites, and inability to overcome the immunosuppressive microenvironment of solid tumors. Accordingly, the efficacy of the chimeric antigen receptor immune cell in solid tumors is greatly limited.
  • a method for treating a cancer includes steps as follows.
  • a chemotherapy drug is administered to a subject in need for a treatment of cancer.
  • a composition containing a plurality of chimeric antigen receptor expressing cells is administered to the subject, wherein the chimeric antigen receptor expressing cells expresses a chimeric antigen receptor specific to human leukocyte antigen G (HLA-G).
  • HLA-G human leukocyte antigen G
  • FIGS. 1A, 1B, 1C, 1D, 1E, 1F, 1G and 1H are analytical results of immunofluorescence staining assay showing a HLA-G expression level of tumor cells treated with a chemotherapy drug.
  • FIGS. 2A, 2B, 2C, 2D and 2E are analytical results of flow cytometry showing a HLA-G expression of tumor cells treated with a chemotherapy drug.
  • FIG. 3 is a graph showing an expression level of chimeric antigen receptors in a chimeric antigen receptor expressing cell according to Example 1 of the present disclosure.
  • FIGS. 4A, 4B, 4C, 4D, 4E, 4F, 4G, 4H and 4I show analytical results of tumor cell death induced by chimeric antigen receptor expressing cells according to Example 1 of the present disclosure.
  • FIG. 5 is a graph showing an expression level of chimeric antigen receptors in a chimeric antigen receptor expressing cell according to Example 2 of the present disclosure.
  • FIGS. 6A, 6B, 6C, 6D, 6E, 6F, 6G, 6H and 6I show analytical results of tumor cell death induced by chimeric antigen receptor expressing cells according to Example 2 of the present disclosure.
  • FIG. 7 is a graph showing an expression level of a chimeric antigen receptor in a chimeric antigen receptor expressing cell according to Example 3 of the present disclosure.
  • FIGS. 8A, 8B, 8C, 8D, 8E, 8F, 8G, 8H and 8I show analytical results of tumor cell death induced by chimeric antigen receptor expressing cells according to Example 3 of the present disclosure.
  • FIG. 9 is a schematic view showing the theoretical structure and mechanism of a chimeric antigen receptor in the plasma membrane of a chimeric antigen receptor expressing cell of the present disclosure.
  • a method for treating cancer includes administering a chemotherapy drug to a subject in need for a treatment of cancer, and administering a composition containing a plurality of chimeric antigen receptor expressing cells to the subject, wherein the chimeric antigen receptor expressing cells expresses a chimeric antigen receptor specific to human leukocyte antigen G (HLA-G).
  • the chemotherapeutic drug can induce the plasma membrane of the tumor cells to express a large amount of the human leukocyte antigen G, thereby enhancing a toxicity of the chimeric antigen receptor cell to the tumor cell.
  • HLA-G human leukocyte antigen G
  • MHC major histocompatibility complex
  • HLA-G is expressed on fetal derived placental cells, and is active in the negative regulation of immune response. HLA-G may play a role in immune tolerance in pregnancy.
  • the cancer can include breast cancer, polymorphic glioblastoma, pancreatic cancer and ovarian cancer
  • the chemotherapy drug can include doxorubicin (Dox), temozolomide (TMZ), gemcitabine (Gem) and carboplatin (CB).
  • the chimeric antigen receptor can include, in order from an N-terminus to a C-terminus, an anti-HLA-G antibody including an amino acid sequence of SEQ ID NO: 1, an HLA-G receptor including an amino acid sequence of SEQ ID NO: 2, and a costimulatory domain including an amino acid sequence of SEQ ID NO: 3.
  • an anti-HLA-G antibody including an amino acid sequence of SEQ ID NO: 1
  • an HLA-G receptor including an amino acid sequence of SEQ ID NO: 2
  • a costimulatory domain including an amino acid sequence of SEQ ID NO: 3.
  • a suicide protein including an amino acid sequence of SEQ ID NO: 4 is linked to the C-terminus of the costimulatory domain
  • a 2A peptide including an amino acid sequence of SEQ ID NO: 10 links the HLA-G receptor and the costimulatory domain.
  • the anti-HLA-G antibody including the amino acid sequence of SEQ ID NO: 1 includes a heavy chain (HC) immunoglobulin variable domain sequence and a light chain (LC) immunoglobulin variable domain sequence.
  • the HC immunoglobulin variable domain sequence includes a CDRH1 including an amino acid sequence of SEQ ID NO: 5, a CDRH2 including an amino acid sequence of SEQ ID NO: 6, and a CDRH3 including an amino acid sequence of SEQ ID NO: 7.
  • the LC immunoglobulin variable domain sequence includes a CDRL2 including an amino acid sequence of SEQ ID NO: 8, and a CDRL3 including an amino acid sequence of SEQ ID NO: 9.
  • the costimulatory domain including an amino acid sequence of SEQ ID NO: 3 is DNAX activating protein 12 (DAP12).
  • the suicide protein including an amino acid sequence of SEQ ID NO: 4 is iCas9 protein.
  • the chimeric antigen receptor expression plasmid can include a promoter and a nucleic acid encoding the chimeric antigen receptor.
  • the nucleic acid encoding the chimeric antigen receptor can include, in order from a 5′ end to a 3′ end, an anti-HLA-G antibody coding fragment, an HLA-G receptor coding fragment, and a costimulatory domain coding fragment.
  • a suicide gene is linked to the 3′ end of the costimulatory domain coding fragment, and a 2A peptide coding fragment links the HLA-G receptor coding fragment and the costimulatory domain coding fragment.
  • the chimeric antigen receptor expression plasmid is Lenti-EF1a-CAR-100517-S1A plasmid, wherein the insert thereof includes a promoter, an anti-HLA-G antibody coding fragment, an HLA-G receptor coding fragment, and a costimulatory domain coding fragment.
  • the promoter is the EF-1 alpha promoter including a nucleic acid sequence of SEQ ID NO: 16.
  • the anti-HLA-G antibody coding fragment includes the nucleic acid sequence of SEQ ID NO: 11.
  • the HLA-G receptor coding fragment includes the nucleic acid sequence of SEQ ID NO: 12.
  • the costimulatory domain coding fragment includes the nucleic acid sequence of SEQ ID NO: 13.
  • the insert of the Lenti-EF1a-CAR-100517-S1A plasmid further includes a signal peptide coding fragment including a nucleic acid sequence of SEQ ID NO: 17, the suicide gene including the nucleic acid sequence of SEQ ID NO: 14, and the 2A peptide coding fragment including the nucleic acid sequence of SEQ ID NO: 15.
  • the signal peptide coding fragment is linked to the 5′ end of the anti-HLA-G antibody coding fragment
  • the suicide gene is linked to the 3′ end of the costimulatory domain coding fragment
  • the 2A peptide coding fragment links the HLA-G receptor coding fragment and the costimulatory domain coding fragment.
  • the insert is constructed on Creative Biolabs vector (Creative Biolabs, N.Y., USA) to obtain the Lenti-EF1a-CAR-100517-S1A plasmid.
  • the Creative Biolabs vector is a lentivirus vector system, so that the constructed chimeric antigen receptor expression plasmid can be transfected into expression cells to produce lentiviruses, and the chimeric antigen receptor can be subsequently transduced into the immune cells using lentiviruses.
  • the chimeric antigen receptor expressing cell can include an immune cell and a chimeric antigen receptor expression plasmid expresses the chimeric antigen receptor specific to HLA-G.
  • the chimeric antigen receptor expressing cell of the present disclosure is obtained by transducing the chimeric antigen receptor of the present disclosure into the immune cell using lentiviruses.
  • the immune cell can be a T lymphocyte or a natural killer (NK) cell. More preferably, the NK cell can be a NK-92 cell line or a primary NK cell.
  • the constructed Lenti-EF1a-CAR-100517-S1A plasmid is transfected into a 293T cell line using lipofectamine 3000 (Invitrogen) to prepare the lentiviruses carrying the chimeric antigen receptor of the present disclosure.
  • lipofectamine 3000 Invitrogen
  • the supernatant containing the prepared lentiviruses and Opti-MEM (Invitrogen) containing 8 ⁇ g/ml of polybrene (Sigma-Aldrich) are used to culture the primary T lymphocytes for 3 days to transduce the chimeric antigen receptor of the present disclosure into the primary T lymphocytes.
  • the supernatant containing the prepared lentiviruses and the Opti-MEM (Invitrogen) containing 50 ⁇ g/ml of protamine sulfate (Sigma-Aldrich) are used to culture the primary NK cells or the NK-92 cell line for 7 days to transduce the chimeric antigen receptor of the present disclosure into the primary NK cell or the NK-92 cell line.
  • the obtained chimeric antigen receptor expressing cell has an effect of inducing tumor cell death in mammals, so that the chimeric antigen receptor expressing cell can be used for inhibiting a proliferation of tumor cells in a subject in need for a treatment of a tumor.
  • the tumor cell can be a breast cancer cell, a polymorphic glioblastoma cell, a pancreatic cancer cell or an ovarian cancer cell.
  • the tumor cells are treated with the chemotherapy drug, and then detecting the HLA-G expression level of the tumor cells.
  • the tumor cells used are human breast cancer cell line MDA-MB-231, human malignant brain tumor cell line DBTRG-05MG (hereinafter referred to as DBTRG), human pancreatic cancer cell line AsPC1, and human ovarian cancer cell line SKOV3.
  • the tumor cell lines used are all purchased from the American Type Culture Collection (ATCC).
  • the human breast cancer cell line MDA-MB-231 is a triple-negative breast cancer cell line, that is, the hormone receptor (ER, PR) and HER-2 receptor thereof are negative, and the human breast cancer cell line MDA-MB-231 is cultured in RPMI culture medium containing 10% fetal bovine serum (FBS).
  • the human malignant brain tumor cell line DBTRG is cultured in DMEM culture medium containing 10% FBS.
  • the human pancreatic cancer cell line AsPC1 is cultured in RPMI culture medium containing 10% FBS.
  • the human ovarian cancer cell line SKOV3 is cultured in McCoy's 5A culture medium containing 10% FBS.
  • the human breast cancer cell line MDA-MB-231, the human malignant brain tumor cell line DBTRG, the human pancreatic cancer cell line AsPC1 and the human ovarian cancer cell line SKOV3 are seeded in a 6-well plate at a density of 2 ⁇ 10 5 cells/well. The cells are subsequently incubated for 24 hours. Each type of the tumor cells is divided into two groups. In a control, the tumor cells are untreated. In an experiment group, the tumor cells are treated with the chemotherapy drug for 48 hours.
  • the chemotherapy drug used for treating the human breast cancer cell line MDA-MB-231 is doxorubicin (200 nM), the chemotherapy drug used for treating the human malignant brain tumor cell line DBTRG is temozolomide (80 ⁇ g/mL), the chemotherapy drug used for treating the human pancreatic cancer cell line AsPC1 is gemcitabine (20 ⁇ M), the chemotherapy drug used for treating the human ovarian cancer cell line SKOV3 is carboplatin (20 ⁇ M). Then, the HLA-G expression level of the tumor cells are detected by immunofluorescence staining assay and flow cytometry.
  • FIGS. 1A, 1B, 10, 1D, 1E, 1F, 1G and 1H are analytical results of immunofluorescence staining assay showing the HLA-G expression level of tumor cells treated with the chemotherapy drug.
  • FIG. 1A is analytical result of immunofluorescence staining assay showing the HLA-G expression level in the control group of the human breast cancer cell line MDA-MB-231
  • FIG. 1B is analytical result of immunofluorescence staining assay showing the HLA-G expression level of the human breast cancer cell line MDA-MB-231 treated with doxorubicin (Dox).
  • Dox doxorubicin
  • FIG. 1C is analytical result of immunofluorescence staining assay showing the HLA-G expression level in the control group of the human malignant brain tumor cell line DBTRG
  • FIG. 1D is analytical result of immunofluorescence staining assay showing the HLA-G expression level of the human malignant brain tumor cell line DBTRG treated with temozolomide (TMZ)
  • FIG. 1E is analytical result of immunofluorescence staining assay showing the HLA-G expression level in the control group of the human pancreatic cancer cell line AsPC1
  • FIG. 1F is analytical result of immunofluorescence staining assay showing the HLA-G expression level of the human pancreatic cancer cell line AsPC1 treated with gemcitabine (Gem).
  • FIG. 1G is analytical result of immunofluorescence staining assay showing the HLA-G expression level in the control group of the human ovarian cancer cell line SKOV3
  • FIG. 1H is analytical result of immunofluorescence staining assay showing the HLA-G expression level of the human ovarian cancer cell line SKOV3 treated with carboplatin (CB).
  • treatment of doxorubicin can increase the HLA-G expression level on the plasma membrane of the human breast cancer cell line MDA-MB-231.
  • treatment of temozolomide can increase the HLA-G expression level on the plasma membrane of the human malignant brain tumor cell line DBTRG.
  • treatment of gemcitabine can increase the HLA-G expression level on the plasma membrane of the human pancreatic cancer cell line AsPC1.
  • treatment of carboplatin can increase the HLA-G expression level on the plasma membrane of the human ovarian cancer cell line SKOV3.
  • FIGS. 2A, 2B, 2C, 2D and 2E are analytical results of flow cytometry showing a HLA-G expression of tumor cells treated with a chemotherapy drug.
  • FIG. 2A is analytical result of flow cytometry of the human breast cancer cell line MDA-MB-231.
  • FIG. 2B is analytical result of flow cytometry of the human malignant brain tumor cell line DBTRG.
  • FIG. 2C is analytical result of flow cytometry of the human pancreatic cancer cell line AsPC1.
  • FIG. 2D is analytical result of flow cytometry of the human ovarian cancer cell line SKOV3.
  • FIG. 2E is a statistical chart of FIGS. 2A, 2B, 2C and 2D .
  • the mean fluorescence intensity (MFI) of the control group of the human breast cancer cell line MDA-MB-231 is only 12.27%, while the MFI of the experiment group of the human breast cancer cell line MDA-MB-231 can reach 64.45%, which is statistically significant (p ⁇ 0.001).
  • MFI of the control group of the human malignant brain tumor cell line DBTRG is only 14.01%, while the MFI of the experiment group of the human malignant brain tumor cell line DBTRG can reach 22.33%, which is statistically significant (p ⁇ 0.001).
  • FIG. 2A the mean fluorescence intensity (MFI) of the control group of the human breast cancer cell line MDA-MB-231 is only 12.27%, while the MFI of the experiment group of the human breast cancer cell line MDA-MB-231 can reach 64.45%, which is statistically significant (p ⁇ 0.001).
  • MFI of the control group of the human malignant brain tumor cell line DBTRG is only 14.01%, while the MFI of the experiment group of the human malignant brain tumor cell line DBTRG
  • the MFI of the control group of the human pancreatic cancer cell line AsPC1 is only 13.18%, while the MFI of the experiment group of the human pancreatic cancer cell line AsPC1 can reach 41.44%, which is statistically significant (p ⁇ 0.01).
  • the MFI of the control group of the human ovarian cancer cell line SKOV3 is only 14.69%, while the MFI of the experiment group of the human ovarian cancer cell line SKOV3 can reach 38.58%, which is statistically significant (p 21 0.01).
  • the method for treating cancer of the present disclosure further administers a composition containing a plurality of chimeric antigen receptor expressing cells to the subject in need for a treatment of cancer, in which the chimeric antigen receptor expressing cells expresses the chimeric antigen receptor specific to HLA-G, in order to enhance the effect of killing tumor cells.
  • the treatment of the chemotherapy drug and the composition containing the chimeric antigen receptor expressing cells can be in a sequence or simultaneous.
  • Example 1 an Example 1, an Example 2 and an Example 3 will be further provided to illustrate the accompanied efficacies of the method for treating cancer of the present disclosure on inducing tumor cell death.
  • the present disclosure is not limited thereto.
  • the Lenti-EF1a-CAR-100517-S1A plasmid is transduced into the NK-92 cell line to obtain the chimeric antigen receptor expressing cell of Example 1 of the present disclosure, and the expression level of the chimeric antigen receptor of the obtained chimeric antigen receptor expressing cell of Example 1 is analyzed by flow cytometry.
  • FIG. 3 is a graph showing the expression level of chimeric antigen receptors in the chimeric antigen receptor expressing cell according to Example 1 of the present disclosure.
  • FIG. 3 is a graph showing the expression level of chimeric antigen receptors in the chimeric antigen receptor expressing cell according to Example 1 of the present disclosure.
  • FIG. 3 shows the expression level of the chimeric antigen receptor of the parental NK-92 cell line without transducing the chimeric antigen receptor of the present disclosure, and the expression level of the chimeric antigen receptor of the chimeric antigen receptor expressing cell of Example 1 on day 3 and day 7 after transduction the chimeric antigen receptor.
  • the MFI of the parental NK-92 cell line is only 9.98%, while the MFI of the chimeric antigen receptor expressing cell of Example 1 on day 3 and day 7 after transduction can reach 20.11% and 65.07%, respectively.
  • the results indicate that the chimeric antigen receptor expressing cell of Example 1 can stably express the chimeric antigen receptor of the present disclosure.
  • the human breast cancer cell line MDA-MB-231, the human malignant brain tumor cell line DBTRG, the human pancreatic cancer cell line AsPC1 and the human ovarian cancer cell line SKOV3 are seeded in a 12-well plate at a density of 1 ⁇ 10 5 cells/well. The cells are subsequently incubated for 24 hours. Each type of the tumor cells is divided into six groups. In a control, the tumor cells are untreated. In a group 1, the tumor cells are treated with the chemotherapy drug. In a group 2, the tumor cells are treated with the parental NK-92 cell line. In a group 3, the tumor cells are treated with the parental NK-92 cell line and the chemotherapy drug.
  • the number of the parental NK-92 cell line treated is 1 ⁇ 10 5 cells.
  • the tumor cells are treated with the chimeric antigen receptor expressing cell of Example 1.
  • the tumor cells are treated with the chimeric antigen receptor expressing cell of Example 1 and the chemotherapy drug.
  • the number of the chimeric antigen receptor expressing cell of Example 1 treated is 1 ⁇ 10 5 cells.
  • the chemotherapy drug used for treating the human breast cancer cell line MDA-MB-231 is doxorubicin (200 nM), the chemotherapy drug used for treating the human malignant brain tumor cell line DBTRG is temozolomide (80 ⁇ g/mL), the chemotherapy drug used for treating the human pancreatic cancer cell line AsPC1 is gemcitabine (20 ⁇ M), the chemotherapy drug used for treating the human ovarian cancer cell line SKOV3 is carboplatin (20 ⁇ M).
  • the treated cells are stained with Annexin V-FITC and propidium iodide (PI), and the apoptosis and the death of the tumor cells are detected by the flow cytometry.
  • the sum of the percentage of cells stained with Annexin V-FITC and/or PI (that is the percentage of cells in the first quadrant, the second quadrant, and the fourth quadrant of the bivariate flow cytometry scatter plot) are calculated to obtain the cytotoxicity.
  • the results of the cytotoxicity are counted after the three independent trials in each group.
  • FIGS. 4A, 4B, 4C, 4D, 4E, 4F, 4G, 4H and 4I show analytical results of tumor cell death induced by the chimeric antigen receptor expressing cells according to Example 1 of the present disclosure.
  • FIG. 4A is a graph showing the analytical results of the death of the human breast cancer cell line MDA-MB-231 induced by the chimeric antigen receptor expressing cell of Example 1
  • FIG. 4B is a statistical chart of FIG. 4A after the three independent trials.
  • FIG. 4C is a graph showing the analytical results of the death of the human malignant brain tumor cell line DBTRG induced by the chimeric antigen receptor expressing cell of Example 1
  • FIG. 4D is a statistical chart of FIG. 4C after the three independent trials.
  • FIG. 4A is a graph showing the analytical results of the death of the human breast cancer cell line MDA-MB-231 induced by the chimeric antigen receptor expressing cell of Example 1
  • FIG. 4B is a statistical chart of FIG. 4
  • FIG. 4E is a graph showing the analytical results of the death of the human pancreatic cancer cell line AsPC1 induced by the chimeric antigen receptor expressing cell of Example 1, and FIG. 4F is a statistical chart of FIG. 4E after the three independent trials.
  • FIG. 4G is a graph showing the analytical results of the death of the human ovarian cancer cell line SKOV3 induced by the chimeric antigen receptor expressing cell of Example 1, and FIG. 4H is a statistical chart of FIG. 4G after the three independent trials.
  • FIG. 4I is a statistical chart of FIGS.
  • P represents the parental NK-92 cell line
  • H represents the chimeric antigen receptor expressing cell of Example 1
  • D represents doxorubicin
  • T represents temozolomide
  • G represents gemcitabine
  • C represents carboplatin.
  • the death rate of the human breast cancer cell line MDA-MB-231 is only about 10%.
  • the death rate of the human breast cancer cell line MDA-MB-231 is increased, but there is no statistically significant difference compared to the control.
  • the death rate of the human breast cancer cell line MDA-MB-231 can increase to 40%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 2.
  • the death rate of the human breast cancer cell line MDA-MB-231 is about 60%, and there is a statistically significant difference (p ⁇ 0.001) compared to the group 2. Furthermore, in the group 5 treated with the doxorubicin and the chimeric antigen receptor expressing cell of Example 1, the death rate of the human breast cancer cell line MDA-MB-231 can reach 80%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 4 and a statistically significant difference (p ⁇ 0.01) compared to the group 3, respectively.
  • the death rate of the human malignant brain tumor cell line DBTRG is less than 10%.
  • the death rate of the human malignant brain tumor cell line DBTRG is increased, but there is no statistically significant difference compared to the control.
  • the death rate of the human malignant brain tumor cell line DBTRG can increase to 40%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 2.
  • the death rate of the human malignant brain tumor cell line DBTRG is more than 60%, and there is a statistically significant difference (p ⁇ 0.001) compared to the group 2. Furthermore, in the group 5 treated with the temozolomide and the chimeric antigen receptor expressing cell of Example 1, the death rate of the human malignant brain tumor cell line DBTRG can reach more than 80%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 4 and a statistically significant difference (p ⁇ 0.001) compared to the group 3, respectively.
  • the death rate of the human pancreatic cancer cell line AsPC1 is less than 10%.
  • the death rate of the human pancreatic cancer cell line AsPC1 is increased, but there is no statistically significant difference compared to the control.
  • the death rate of the human pancreatic cancer cell line AsPC1 can increase to 30%, but there is no statistically significant difference compared to the control.
  • the death rate of the human pancreatic cancer cell line AsPC1 is approximately 40%, and there is a statistically significant difference (p ⁇ 0.01) compared to the group 2. Furthermore, in the group 5 treated with the gemcitabine and the chimeric antigen receptor expressing cell of Example 1, the death rate of the human pancreatic cancer cell line AsPC1 can reach 60%, and there is a statistically significant difference (p ⁇ 0.001) compared to the group 4 and a statistically significant difference (p ⁇ 0.001) compared to the group 3, respectively.
  • the death rate of the human ovarian cancer cell line SKOV3 is less than 10%.
  • the death rate of the human ovarian cancer cell line SKOV3 is increased, but there is no statistically significant difference compared to the control.
  • the death rate of the human ovarian cancer cell line SKOV3 can increase to 30%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 2.
  • the death rate of the human ovarian cancer cell line SKOV3 is approximately 40%, and there is a statistically significant difference (p ⁇ 0.01) compared to the group 2. Furthermore, in the group 5 treated with the carboplatin and the chimeric antigen receptor expressing cell of Example 1, the death rate of the human ovarian cancer cell line SKOV3 can reach 60%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 4 and a statistically significant difference (p ⁇ 0.01) compared to the group 3, respectively.
  • the results indicate that the chimeric antigen receptor expressing cell of Example 1 can be used to treat with the human breast cancer cell line MDA-MB-231, the human malignant brain tumor cell line DBTRG, the human pancreatic cancer cell line AsPC1 and the human ovarian cancer cell line SKOV3 for excellent cell killing. Therefore, the chimeric antigen receptor expressing cell of the present disclosure can be used for inhibiting the proliferation of the tumor cells in the subject in need for the treatment of the tumor.
  • the tumor cell can be the breast cancer cell, the polymorphic glioblastoma cell, the pancreatic cancer cell or the ovarian cancer cell.
  • the simultaneous treatment of the chemotherapy drug and the chimeric antigen receptor expressing cell of Example 1 can significantly increase the toxic effect on inducing death of the human breast cancer cell line MDA-MB-231, the human malignant brain tumor cell line DBTRG, the human pancreatic cancer cell line AsPC1 and the human ovarian cancer cell line SKOV3.
  • the results indicate that the method for treating cancer of the present disclosure can effectively inhibit the growth of the tumor cells and treat cancer.
  • the Lenti-EF1a-CAR-100517-S1A plasmid is transduced into the primary NK cell to obtain the chimeric antigen receptor expressing cell of Example 2 of the present disclosure, and the expression level of the chimeric antigen receptor of the obtained chimeric antigen receptor expressing cell of Example 2 is analyzed by the flow cytometry.
  • FIG. 5 is a graph showing an expression level of chimeric antigen receptors in a chimeric antigen receptor expressing cell according to Example 2 of the present disclosure.
  • FIG. 5 is a graph showing an expression level of chimeric antigen receptors in a chimeric antigen receptor expressing cell according to Example 2 of the present disclosure.
  • FIG. 5 shows the expression level of the chimeric antigen receptor of the parental primary NK cell without transducing the chimeric antigen receptor of the present disclosure, and the expression level of the chimeric antigen receptor of the chimeric antigen receptor expressing cell of Example 2 on day 3 and day 7 after transduction the chimeric antigen receptor.
  • the MFI of the parental primary NK cell is 22.09%, while the MFI of the chimeric antigen receptor expressing cell of Example 2 on day 3 and day 7 after transduction can reach 29.02% and 50.21%, respectively.
  • the results indicate that the chimeric antigen receptor expressing cell of Example 2 can stably express the chimeric antigen receptor of the present disclosure.
  • the human breast cancer cell line MDA-MB-231, the human malignant brain tumor cell line DBTRG, the human pancreatic cancer cell line AsPC1 and the human ovarian cancer cell line SKOV3 are seeded in a 12-well plate at a density of 1 ⁇ 10 5 cells/well. The cells are subsequently incubated for 24 hours. Each type of the tumor cells is divided into six groups. In a control, the tumor cells are untreated. In a group 1, the tumor cells are treated with the chemotherapy drug. In a group 2, the tumor cells are treated with the parental primary NK cell. In a group 3, the tumor cells are treated with the parental primary NK cell and the chemotherapy drug. In the groups 2 and 3, the number of the parental primary NK cell treated is 1 ⁇ 10 5 cells.
  • the tumor cells are treated with the chimeric antigen receptor expressing cell of Example 2.
  • the tumor cells are treated with the chimeric antigen receptor expressing cell of Example 2 and the chemotherapy drug.
  • the number of the chimeric antigen receptor expressing cell of Example 2 treated is 1 ⁇ 10 5 cells.
  • the chemotherapy drug used for treating the human breast cancer cell line MDA-MB-231 is doxorubicin (200 nM), the chemotherapy drug used for treating the human malignant brain tumor cell line DBTRG is temozolomide (80 ⁇ g/mL), the chemotherapy drug used for treating the human pancreatic cancer cell line AsPC1 is gemcitabine (20 ⁇ M), the chemotherapy drug used for treating the human ovarian cancer cell line SKOV3 is carboplatin (20 ⁇ M).
  • the treated cells are stained with Annexin V-FITC and propidium iodide (PI), and the apoptosis and the death of the tumor cells are detected by the flow cytometry.
  • the sum of the percentage of cells stained with Annexin V-FITC and/or PI (that is the percentage of cells in the first quadrant, the second quadrant, and the fourth quadrant of the bivariate flow cytometry scatter plot) are calculated to obtain the cytotoxicity.
  • the results of the cytotoxicity are counted after the three independent trials in each group.
  • FIGS. 6A, 6B, 6C, 6D, 6E, 6F, 6G, 6H and 6I show analytical results of tumor cell death induced by the chimeric antigen receptor expressing cells according to Example 2 of the present disclosure.
  • FIG. 6A is a graph showing the analytical results of the death of the human breast cancer cell line MDA-MB-231 induced by the chimeric antigen receptor expressing cell of Example 2
  • FIG. 6B is a statistical chart of FIG. 6A after the three independent trials.
  • FIG. 6C is a graph showing the analytical results of the death of the human malignant brain tumor cell line DBTRG induced by the chimeric antigen receptor expressing cell of Example 2
  • FIG. 6D is a statistical chart of FIG. 6C after the three independent trials.
  • FIG. 6A, 6B, 6C, 6D, 6F, 6G, 6H and 6I show analytical results of tumor cell death induced by the chimeric antigen receptor expressing cells according to Example 2 of the present disclosure.
  • FIG. 6E is a graph showing the analytical results of the death of the human pancreatic cancer cell line AsPC1 induced by the chimeric antigen receptor expressing cell of Example 2
  • FIG. 6F is a statistical chart of FIG. 6E after the three independent trials.
  • FIG. 6G is a graph showing the analytical results of the death of the human ovarian cancer cell line SKOV3 induced by the chimeric antigen receptor expressing cell of Example 2
  • FIG. 6H is a statistical chart of FIG. 6G after the three independent trials.
  • FIG. 6I is a statistical chart of FIGS.
  • P represents the parental primary NK cell
  • H represents the chimeric antigen receptor expressing cell of Example 2
  • D represents doxorubicin
  • T represents temozolomide
  • G represents gemcitabine
  • C represents carboplatin.
  • the death rate of the human breast cancer cell line MDA-MB-231 is only about 10%.
  • the death rate of the human breast cancer cell line MDA-MB-231 is increased, but there is no statistically significant difference compared to the control.
  • the death rate of the human breast cancer cell line MDA-MB-231 can increase to 30%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 2.
  • the death rate of the human breast cancer cell line MDA-MB-231 is more than 50%, and there is a statistically significant difference (p ⁇ 0.01) compared to the group 2. Furthermore, in the group 5 treated with the doxorubicin and the chimeric antigen receptor expressing cell of Example 2, the death rate of the human breast cancer cell line MDA-MB-231 can reach 80%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 4 and a statistically significant difference (p ⁇ 0.01) compared to the group 3, respectively.
  • the death rate of the human malignant brain tumor cell line DBTRG is less than 10%.
  • the death rate of the human malignant brain tumor cell line DBTRG is increased, but there is no statistically significant difference compared to the control.
  • the death rate of the human malignant brain tumor cell line DBTRG can increase to about 30%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 2.
  • the death rate of the human malignant brain tumor cell line DBTRG is more than 20%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 2. Furthermore, in the group 5 treated with the temozolomide and the chimeric antigen receptor expressing cell of Example 2, the death rate of the human malignant brain tumor cell line DBTRG can reach about 60%, and there is a statistically significant difference (p ⁇ 0.01) compared to the group 4 and a statistically significant difference (p ⁇ 0.05) compared to the group 3, respectively.
  • the death rate of the human pancreatic cancer cell line AsPC1 is less than 10%.
  • the death rate of the human pancreatic cancer cell line AsPC1 is increased, but there is no statistically significant difference compared to the control.
  • the death rate of the human pancreatic cancer cell line AsPC1 can increase to 30%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 2.
  • the death rate of the human pancreatic cancer cell line AsPC1 is approximately 20%, and there is a statistically significant difference (p ⁇ 0.01) compared to the group 2. Furthermore, in the group 5 treated with the gemcitabine and the chimeric antigen receptor expressing cell of Example 2, the death rate of the human pancreatic cancer cell line AsPC1 can reach 50%, and there is a statistically significant difference (p ⁇ 0.01) compared to the group 4 and a statistically significant difference (p ⁇ 0.05) compared to the group 3, respectively.
  • the death rate of the human ovarian cancer cell line SKOV3 is less than 10%.
  • the death rate of the human ovarian cancer cell line SKOV3 is comparable to that of the control.
  • the death rate of the human ovarian cancer cell line SKOV3 can increase to more than 20%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 2.
  • the death rate of the human ovarian cancer cell line SKOV3 is approximately 20%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 2. Furthermore, in the group 5 treated with the carboplatin and the chimeric antigen receptor expressing cell of Example 2, the death rate of the human ovarian cancer cell line SKOV3 can reach 50%, and there is a statistically significant difference (p 21 0.01) compared to the group 4 and a statistically significant difference (p ⁇ 0.05) compared to the group 3, respectively.
  • the results indicate that the chimeric antigen receptor expressing cell of Example 2 can be used to treat with the breast cancer cell, the polymorphic glioblastoma cell, the pancreatic cancer cell or the ovarian cancer cell for excellent cell killing. Therefore, the chimeric antigen receptor expressing cell of the present disclosure can be used for inhibiting the proliferation of the tumor cells in the subject in need for the treatment of the tumor. Further, the simultaneous treatment of the chemotherapy drug and the chimeric antigen receptor expressing cell of Example 2 can significantly increase the toxic effect on inducing death of the human breast cancer cell line MDA-MB-231, the human malignant brain tumor cell line DBTRG, the human pancreatic cancer cell line AsPC1 and the human ovarian cancer cell line SKOV3. The results indicate that the method for treating cancer of the present disclosure can effectively inhibit the growth of the tumor cells and treat cancer.
  • the chimeric antigen receptor of the present disclosure is transduced into the primary T lymphocyte to obtain the chimeric antigen receptor expressing cell of Example 3 of the present disclosure, and the expression level of the chimeric antigen receptor of the obtained chimeric antigen receptor expressing cell of Example 3 is analyzed by the flow cytometry.
  • FIG. 7 is a graph showing an expression level of a chimeric antigen receptor in a chimeric antigen receptor expressing cell according to Example 3 of the present disclosure.
  • FIG. 7 shows the expression level of the chimeric antigen receptor of the parental primary T lymphocyte without transducing the chimeric antigen receptor of the present disclosure, and the expression level of the chimeric antigen receptor of the chimeric antigen receptor expressing cell of Example 3 on day 3 and day 7 after transduction the chimeric antigen receptor.
  • the MFI of the parental primary T lymphocyte only is 9.36%, while the MFI of the chimeric antigen receptor expressing cell of Example 3 on day 3 and day 7 after transduction can reach 34.1% and 88.64%, respectively.
  • the results indicate that the chimeric antigen receptor expressing cell of Example 3 can stably express the chimeric antigen receptor of the present disclosure.
  • Example 3 of the present disclosure The effects of the chimeric antigen receptor expressing cell of Example 3 of the present disclosure on inducing the death of the breast cancer cells, the glioblastoma multiforme cells, the pancreatic cancer cells, and the ovarian cancer cells are further demonstrated in following experiments.
  • the human breast cancer cell line MDA-MB-231, the human malignant brain tumor cell line DBTRG, the human pancreatic cancer cell line AsPC1 and the human ovarian cancer cell line SKOV 3 are seeded in a 12-well plate at a density of 1 ⁇ 10 5 cells/well. The cells are subsequently incubated for 24 hours. Each type of the tumor cells is divided into six groups. In a control, the tumor cells are untreated. In a group 1, the tumor cells are treated with the chemotherapy drug. In a group 2, the tumor cells are treated with the parental primary T lymphocyte. In a group 3, the tumor cells are treated with the parental T lymphocyte and the chemotherapy drug. In the groups 2 and 3, the number of the parental primary T lymphocyte treated is 1 ⁇ 10 5 cells.
  • the tumor cells are treated with the chimeric antigen receptor expressing cell of Example 3.
  • the tumor cells are treated with the chimeric antigen receptor expressing cell of Example 3 and the chemotherapy drug.
  • the number of the chimeric antigen receptor expressing cell of Example 3 treated is 1 ⁇ 10 5 cells.
  • the chemotherapy drug used for treating the human breast cancer cell line MDA-MB-231 is doxorubicin (200 nM), the chemotherapy drug used for treating the human malignant brain tumor cell line DBTRG is temozolomide (80 ⁇ g/mL), the chemotherapy drug used for treating the human pancreatic cancer cell line AsPC1 is gemcitabine (20 ⁇ M), the chemotherapy drug used for treating the human ovarian cancer cell line SKOV3 is carboplatin (20 ⁇ M).
  • the treated cells are stained with Annexin V-FITC and propidium iodide (PI), and the apoptosis and the death of the tumor cells are detected by the flow cytometry.
  • the sum of the percentage of cells stained with Annexin V-FITC and/or PI (that is the percentage of cells in the first quadrant, the second quadrant, and the fourth quadrant of the bivariate flow cytometry scatter plot) are calculated to obtain the cytotoxicity.
  • the results of the cytotoxicity are counted after the three independent trials in each group.
  • FIGS. 8A, 8B, 8C, 8D, 8E, 8F, 8G, 8H and 8I show analytical results of tumor cell death induced by the chimeric antigen receptor expressing cells according to Example 3 of the present disclosure.
  • FIG. 8A is a graph showing the analytical results of the death of the human breast cancer cell line MDA-MB-231 induced by the chimeric antigen receptor expressing cell of Example 3, and
  • FIG. 8B is a statistical chart of FIG. 8A after the three independent trials.
  • FIG. 8C is a graph showing the analytical results of the death of the human malignant brain tumor cell line DBTRG induced by the chimeric antigen receptor expressing cell of Example 3
  • FIG. 8D is a statistical chart of FIG. 8C after the three independent trials.
  • FIG. 8A, 8B, 8C, 8D, 8F, 8G, 8H and 8I show analytical results of tumor cell death induced by the chimeric antigen receptor expressing cells according to Example 3 of the present disclosure.
  • FIG. 8E is a graph showing the analytical results of the death of the human pancreatic cancer cell line AsPC1 induced by the chimeric antigen receptor expressing cell of Example 3, and FIG. 8F is a statistical chart of FIG. 8E after the three independent trials.
  • FIG. 8G is a graph showing the analytical results of the death of the human ovarian cancer cell line SKOV3 induced by the chimeric antigen receptor expressing cell of Example 3, and
  • FIG. 8H is a statistical chart of FIG. 8G after the three independent trials.
  • FIG. 8I is a statistical chart of FIGS.
  • P represents the parental primary T lymphocyte
  • H represents the chimeric antigen receptor expressing cell of Example 3
  • D represents doxorubicin
  • T represents temozolomide
  • G represents gemcitabine
  • C represents carboplatin.
  • the death rate of the human breast cancer cell line MDA-MB-231 is only about 10%.
  • the death rate of the human breast cancer cell line MDA-MB-231 is increased, but there is no statistically significant difference compared to the control.
  • the death rate of the human breast cancer cell line MDA-MB-231 can increase to 20%, and there is a statistically significant difference (p ⁇ 0.01) compared to the group 2.
  • the death rate of the human breast cancer cell line MDA-MB-231 is more than 30%, and there is a statistically significant difference (p ⁇ 0.001) compared to the group 2. Furthermore, in the group 5 treated with the doxorubicin and the chimeric antigen receptor expressing cell of Example 3, the death rate of the human breast cancer cell line MDA-MB-231 can reach about 50%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 4 and a statistically significant difference (p ⁇ 0.001) compared to the group 3, respectively.
  • the death rate of the human malignant brain tumor cell line DBTRG is less than 20%.
  • the death rate of the human malignant brain tumor cell line DBTRG is increased, but there is no statistically significant difference compared to the control.
  • the death rate of the human malignant brain tumor cell line DBTRG can increase to about 30%, there is no statistically significant difference compared to the control.
  • the death rate of the human malignant brain tumor cell line DBTRG is more than 50%, and there is a statistically significant difference (p ⁇ 0.001) compared to the group 2. Furthermore, in the group 5 treated with the temozolomide and the chimeric antigen receptor expressing cell of Example 3, the death rate of the human malignant brain tumor cell line DBTRG can reach about 80%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 4 and a statistically significant difference (p ⁇ 0.001) compared to the group 3, respectively.
  • the death rate of the human pancreatic cancer cell line AsPC1 is less than 20%.
  • the death rate of the human pancreatic cancer cell line AsPC1 is comparable to that of the control.
  • the death rate of the human pancreatic cancer cell line AsPC1 can increase to 30%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 2.
  • the death rate of the human pancreatic cancer cell line AsPC1 is increased to more than 50%, and there is a statistically significant difference (p ⁇ 0.001) compared to the group 2. Furthermore, in the group 5 treated with the gemcitabine and the chimeric antigen receptor expressing cell of Example 3, the death rate of the human pancreatic cancer cell line AsPC1 can reach 60%, and there is a statistically significant difference (p ⁇ 0.01) compared to the group 4 and a statistically significant difference (p ⁇ 0.01) compared to the group 3, respectively.
  • the death rate of the human ovarian cancer cell line SKOV3 is less than 10%.
  • the death rate of the human ovarian cancer cell line SKOV3 is increased, but there is no statistically significant difference compared to the control.
  • the death rate of the human ovarian cancer cell line SKOV3 can increase to about 30%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 2.
  • the death rate of the human ovarian cancer cell line SKOV3 is approximately 60%, and there is a statistically significant difference (p ⁇ 0.001) compared to the group 2. Furthermore, in the group 5 treated with the carboplatin and the chimeric antigen receptor expressing cell of Example 3, the death rate of the human ovarian cancer cell line SKOV3 can reach more than 60%, and there is a statistically significant difference (p ⁇ 0.05) compared to the group 4 and a statistically significant difference (p ⁇ 0.001) compared to the group 3, respectively.
  • the results indicate that the chimeric antigen receptor expressing cell of Example 3 can be used to treat with the breast cancer cell, the polymorphic glioblastoma cell, the pancreatic cancer cell or the ovarian cancer cell for excellent cell killing. Therefore, the chimeric antigen receptor expressing cell of the present disclosure can be used for inhibiting the proliferation of the tumor cells in the subject in need for the treatment of the tumor. Further, the simultaneous treatment of the chemotherapy drug and the chimeric antigen receptor expressing cell of Example 3 can significantly increase the toxic effect on inducing death of the human breast cancer cell line MDA-MB-231, the human malignant brain tumor cell line DBTRG, the human pancreatic cancer cell line AsPC1 and the human ovarian cancer cell line SKOV3. The results indicate that the method for treating cancer of the present disclosure can effectively inhibit the growth of the tumor cells and treat cancer.
  • FIG. 9 is a schematic view showing the theoretical structure and mechanism of the chimeric antigen receptor in the plasma membrane of the chimeric antigen receptor expressing cell of the present disclosure.
  • the chimeric antigen receptor expressing cell of the present disclosure is a genetically engineered NK cell or T cell which expresses the chimeric antigen receptor of the present disclosure, and the chimeric antigen receptor of the present disclosure is a tumor-targeting receptor complex included the anti-HLA-G antibody (scFv), the HLA-G receptor (KIR) and the costimulatory domain (DAP12).
  • the chimeric antigen receptor of the present disclosure can further include the suicide protein iCas9.
  • the chimeric antigen receptor expressing cell of the present disclosure can specifically recognize the HLA-G on the tumor plasma membrane.
  • the HLA-G expression on the plasma membrane of the tumor cell can be positively regulated.
  • the chimeric antigen receptor expressing cell of the present disclosure binds to the HLA-G, which is specifically recognized on the surface of the tumor cell, signal transduction is triggered, and a signal cascade is generated to cause activation and proliferation of the chimeric antigen receptor expressing cell of the present disclosure. In turn, it also triggers exocytosis of lytic granules and killing of the target tumor cells.
  • the treatment of the chemotherapy drug can increase the HLA-G expression level on the plasma membrane of tumor cells.
  • the chimeric antigen receptor expressed by the chimeric antigen receptor expressing cell of the present disclosure has excellent specific binding ability to the tumor cells, especially specific binding to HLA-G expressed on the plasma membrane of tumor cells, and can specifically target the tumor cells to avoid the off-target effect, thereby effectively killing the tumor cells. Accordingly, the method for treating cancer of the present disclosure can effectively inhibit the proliferation of the tumor cells in the subject in need for the treatment of the tumor and thereby treat cancer.

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022083590A1 (zh) * 2020-10-19 2022-04-28 南京卡提医学科技有限公司 包含dap12和共刺激信号分子信号域的嵌合受体及其使用方法
US11325977B2 (en) 2018-08-31 2022-05-10 Invectys SA Chimeric antigen receptors against multiple HLA-G isoforms

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI717880B (zh) * 2019-10-24 2021-02-01 中國醫藥大學附設醫院 Hla-g特異性嵌合抗原受體、核酸、hla-g特異性嵌合抗原受體表達質體、表達hla-g特異性嵌合抗原受體細胞、其用途以及用於治療癌症之醫藥組合物
CN114369168A (zh) * 2020-10-19 2022-04-19 南京卡提医学科技有限公司 包含dap12和共刺激信号分子信号域的嵌合受体及其使用方法
TWI823402B (zh) * 2021-07-21 2023-11-21 長聖國際生技股份有限公司 多特異性單域抗體嵌合抗原受體及t細胞銜接體、核酸、其表達細胞、其用途以及治療癌症之醫藥組合物
WO2023230729A1 (en) * 2022-06-03 2023-12-07 Mcmaster University Synthetic antigen receptors for covalent immune recruiters and methods and uses thereof

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114621969A (zh) * 2014-09-17 2022-06-14 诺华股份有限公司 用于过继免疫疗法的具有嵌合受体的靶向细胞毒性细胞
CA2981166A1 (en) * 2015-03-27 2016-10-06 University Of Southern California Hla-g as a novel target for car t-cell immunotherapy
JP2020501512A (ja) * 2016-09-23 2020-01-23 ユニバーシティ オブ サザン カリフォルニア キメラ抗原受容体およびその組成物ならびにそれらの使用の方法

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
Jan et al (J. Immunother. Cancer, 2021, 9: e003050.doi:10.1136/jitc-2021-003050, pages 1-16) (Year: 2021) *
Jan et al (Proc. AACR Special Conf. on Tumor Immunology and Immunotherapy, Nov. 17, 2019, Cancer Immunol. Res. 2020, 8(3 Suppl): Abstract nr A61) (Year: 2020) *
Paridaens et al (Abstract of J. Clinical Oncol. 2000, 18.4.724, 5 pages) (Year: 2000) *
Ramakrishna et al (Expert Opinion on Biological Therapy, 2020, 20(5): 503-516) (Year: 2020) *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11325977B2 (en) 2018-08-31 2022-05-10 Invectys SA Chimeric antigen receptors against multiple HLA-G isoforms
US11505608B2 (en) 2018-08-31 2022-11-22 Invectys SA Chimeric antigen receptors against multiple HLA-G isoforms
WO2022083590A1 (zh) * 2020-10-19 2022-04-28 南京卡提医学科技有限公司 包含dap12和共刺激信号分子信号域的嵌合受体及其使用方法

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