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PD1-knockout chimeric antigen receptor T cell targeting CD317, preparation method and application thereof

A chimeric antigen receptor and targeting technology, applied in the field of medical biology, can solve problems such as limitations and no progress

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

However, the current application of CAR-T technology is still limited to hematological tumors, and has not yet made progress in various solid tumors such as liver cancer.
At present, there is no research on chimeric antigen receptor T cells targeting CD317 that knocks out PD1 and can prevent tumor cells from escaping immune surveillance

Method used

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  • PD1-knockout chimeric antigen receptor T cell targeting CD317, preparation method and application thereof
  • PD1-knockout chimeric antigen receptor T cell targeting CD317, preparation method and application thereof
  • PD1-knockout chimeric antigen receptor T cell targeting CD317, preparation method and application thereof

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Experimental program
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Embodiment 1

[0092] A method for preparing CD317-targeted chimeric antigen receptor T cells knocked out of PD1, comprising the following steps:

[0093] (1) Prepare the gene sequence of chimeric antigen receptor CAR-CD317

[0094] Prepare the coding genes of signal peptide, single-chain antibody targeting CD317, CD8α hinge region, CD8 transmembrane region, 4-1BB signal region and CD3ζ signal region respectively, the coding gene of the signal peptide is shown in SEQ ID NO: 8 , the coding gene of the single-chain antibody targeting CD317 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 SEQ ID NO: As shown in ID NO: 12, the coding gene of the 4-1BB signal region is shown in SEQ ID NO: 14, and the coding gene of the CD3ζ signal region is shown in SEQ ID NO: 16.

[0095] The genes encoding the above-mentioned signal peptide, single-chain antibody targeting CD317, CD8α hinge region, CD8 ...

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Abstract

The invention provides a preparation method of a PD1-knockout chimeric antigen receptor T cell targeting CD317. The method comprises: linking an encoding gene of the chimeric antigen receptor CAR-CD317 and targeting CD317 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 CD317. 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 CD317 positive 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 invention relates to the field of medical biology, in particular to a chimeric antigen receptor T cell targeting CD317 knocked out of PD1 and its preparation method and application. Background technique [0002] Malignant tumor (cancer) has become the culprit threatening human life, and the incidence rate is on the rise. CD317 is a lipid raft-related protein, which is mainly expressed in various solid tumor cells such as liver cancer and colon cancer, but rarely expressed in ordinary cells. The expression of CD317 is often the response of cells to the gamma-interferon pathway, so it is closely related to the occurrence of tumors. [0003] CAR-T (Chimeric Antigen Receptor T Cell) technology is a new type of cell therapy, which is to reinfuse CAR-modified T cells into the human body, activate the autoimmune system, and kill tumor cells. One of the effective ways to treat malignant tumors. However, the current application of CAR-T technology is sti...

Claims

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Application Information

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