PD1-knockout chimeric antigen receptor T cell targeting HER2, preparation method and application thereof

A chimeric antigen receptor and targeting technology, applied in the field of medical biology, can solve the problems of HER2-positive solid tumors without related research and 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 HE R2-positive solid tumors.
At present, there is no research on chimeric antigen receptor T cells targeting HER2 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 HER2, preparation method and application thereof
  • PD1-knockout chimeric antigen receptor T cell targeting HER2, preparation method and application thereof
  • PD1-knockout chimeric antigen receptor T cell targeting HER2, preparation method and application thereof

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Experimental program
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Embodiment 1

[0092] A method for preparing chimeric antigen receptor T cells targeting HER2 that knocks out PD1, comprising the following steps:

[0093] (1) Prepare the gene sequence of chimeric antigen receptor CAR-HER2 targeting HER2

[0094] The coding genes of the signal peptide, the single-chain antibody targeting HER2, 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 HER2 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, HER2-targeting single-ch...

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Abstract

The invention provides a preparation method of a PD1-knockout chimeric antigen receptor T cell targeting HER2. The method comprises: linking an encoding gene of the chimeric antigen receptor CAR-HER2to 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 chimeric antigen receptor T cell targeting HER2. 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 avoidsthe 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 thecells 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 HER2 knocked out of PD1 and its preparation method and application. Background technique [0002] The global incidence of breast cancer has been on the rise since the late 1970s, and breast cancer has become a major public health problem in the current society. Clinically, human epidermal growth factor receptor-2 (HER2) is an important prognostic factor for breast cancer, which is closely related to the spread and metastasis of breast cancer. HER2 is a member of the human epithelial growth factor receptor family. The HER2 gene product is normally expressed in secretory gland epithelial cells. When its gene copy increases abnormally, it will drive the malignant transformation and expansion of the cell. According to statistics, about 20-30% of breast cancer patients are HER2 positive. Compared with HER2-negative breast cancer, HER2-posit...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N7/01C12N15/113C12N5/10A61P35/00
Inventor 曾滢钟春颖
Owner SHENZHEN BINDEBIOTECH CO LTD
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