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PD1-knockout chimeric antigen receptor T cell targeting ROR1, preparation method and application thereof

A chimeric antigen receptor, targeting technology, applied in the field of medical biology, can solve problems such as limitations

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 CART technology is still limited to hematological tumors, and there is no relevant research on most solid tumors such as lung cancer
There is no research on chimeric antigen receptor T cells targeting ROR1 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 ROR1, preparation method and application thereof
  • PD1-knockout chimeric antigen receptor T cell targeting ROR1, preparation method and application thereof
  • PD1-knockout chimeric antigen receptor T cell targeting ROR1, preparation method and application thereof

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Embodiment 1

[0092] A method for preparing chimeric antigen receptor T cells targeting ROR1 that knocks out PD1, comprising the following steps:

[0093] (1) Prepare the gene sequence of chimeric antigen receptor CAR-ROR1 targeting ROR1

[0094] The coding genes of the signal peptide, the single-chain antibody targeting ROR1, the CD8α hinge region, the CD8 transmembrane region, the 4-1BB signal region and the CD3ζ signal region are respectively prepared, and the coding genes of the signal peptide are shown in SEQ ID NO: 8 , the coding gene of the single-chain antibody targeting ROR1 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 signal peptide, ROR1-targeting single-ch...

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Abstract

The invention provides a preparation method of PD1-knockout chimeric antigen receptor T cell targeting ROR1. The method comprises: linking an encoding gene of the chimeric antigen receptor CAR-ROR1 toa 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 ROR1. The knockout of the PD1 gene by the chimeric antigen receptor T cell is in favor of amplification of T cells in the patient, and avoids the escapeof immunosurveillance of the tumor cells, 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 PD1-knockout ROR1-targeting chimeric antigen receptor T cell and its preparation method and application. Background technique [0002] Malignant tumors are the most serious diseases that endanger human health. Every year, about 3 million people in China are diagnosed with new tumors, and 2 million people die of cancer. Recent studies have shown that the tyrosine protein kinase transmembrane receptor (Receptortyrosine kinase like orphan receptor 1, ROR1) is expressed in a variety of B cell malignancies, and subsequently in including mantle cell lymphoma (MCL), acute lymphoblastic ROR1 has been identified on the surface of a number of cancers including blastic leukemia (ALL) and subsets of lung, breast, colon, pancreatic, renal and ovarian cancers. In addition, ROR1 is highly expressed on cancer stem cells, such as ovarian cancer stem cells, and mediates their migration an...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/867C12N7/01C12N15/113A61P35/00
Inventor 张宏玲龙丽梅钟春颖
Owner SHENZHEN BINDEBIOTECH CO LTD
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