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PD1-knockout chimeric antigen receptor T cell targeting DR5, preparation method and application thereof

A chimeric antigen receptor, DR5 technology, applied in the field of medical biology

Inactive Publication Date: 2019-06-04
SHENZHEN BINDEBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research on chimeric antigen receptor T cells targeting DR5 that can prevent tumor cells from escaping immune surveillance

Method used

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  • PD1-knockout chimeric antigen receptor T cell targeting DR5, preparation method and application thereof
  • PD1-knockout chimeric antigen receptor T cell targeting DR5, preparation method and application thereof
  • PD1-knockout chimeric antigen receptor T cell targeting DR5, preparation method and application thereof

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Embodiment 1

[0091] A method for preparing a chimeric antigen receptor T cell targeting DR5 that knocks out PD1, includes the following steps:

[0092] (1) Preparation of the gene sequence of the chimeric antigen receptor CAR-DR5 targeting DR5

[0093] The signal peptide, the single-chain antibody targeting DR5, the CD8α hinge region, the CD8 transmembrane region, the 4-1B B signal region and the CD3ζ signal region encoding genes were prepared separately, and the signal peptide encoding genes are as shown in SEQ ID NO: 8 As shown, the coding gene of the single-chain antibody targeting DR5 is shown in SEQ ID NO: 5, the coding gene of the CD8α hinge region is shown in SEQ ID NO: 10, and the coding gene of the CD8 transmembrane region is shown in The coding gene of the 4-1BB signal region is shown in SEQ ID NO: 12, and the coding gene of the CD3ζ signal region is shown in SEQ ID NO: 16.

[0094] The above-mentioned signal peptide, single-chain antibody targeting DR5, CD8α hinge region, CD8 transmem...

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Abstract

The invention provides a preparation method of a PD1-knockout chimeric antigen receptor T cell targeting DR5. The method comprises: linking an encoding gene of the chimeric antigen receptor CAR-DR5 and targeting DR5 to a vector, and constructing the recombinant lentivirus, performing transfection on CD3 positive T lymphocytes, and performing knockout of the PD1 gene of the transfected T cells to obtain the PD1-knockout chimeric antigen receptor T cell targeting DR5. The knockout of the PD1 gene is carried out by the chimeric antigen receptor T cell, which is in favor of amplification of T cells in the patient, and avoids the escape of immunosurveillance of the tumor cells, and the T cell has the performance of killing tumor cells with high efficiency and specificity, which maintains the viability and lethality of the cells for a long time and does not cause damage to normal cells.

Description

Technical field [0001] The present invention relates to the field of medical biology, in particular to a chimeric antigen receptor T cell targeting DR5 that knocks out PD1, and a preparation method and application thereof. Background technique [0002] Cancer has become the culprit threatening human life, and the incidence is on the rise. People started from the perspective of inducing tumor cell apoptosis or overcoming tumor cell tolerance to apoptosis, and discovered tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). ), is a member of the tumor necrosis factor superfamily that can induce apoptosis. Death receptor 5 (DR5) is a member of the tumor necrosis factor receptor superfamily (TN FRSF10B). Its cytoplasmic region contains a death domain, which is distributed in tumor cells, lymphatic tissues and activated T cells. TRAIL binds to DR5 on the cell surface and can induce cell apoptosis. DR5 is widely expressed in many solid tumor cells such as liver cancer, whi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/867C12N7/01C12N15/113A61P35/00
Inventor 张宏玲龙丽梅
Owner SHENZHEN BINDEBIOTECH CO LTD
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