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Chimeric antigen receptor targeting NKG2D, chimeric antigen receptor T-cell targeting NKG2D and preparation method thereof and application thereof

A chimeric antigen receptor and targeting technology, which is applied in the field of biomedicine, can solve the problems of inability to exert high-killing tumor cells for a long time and the lack of research on CAR-T cells, so as to maintain tumor lethality and promote expansion , stable targeting effect

Pending Publication Date: 2020-07-03
SHENZHEN BINDEBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although NKG2D ligands provide a more precise target for tumor immunotherapy, there are few studies on NKG2D-targeting CAR-T cells, and in practical applications, they cannot play a long-term and durable role in killing tumor cells. ability

Method used

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  • Chimeric antigen receptor targeting NKG2D, chimeric antigen receptor T-cell targeting NKG2D and preparation method thereof and application thereof
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  • Chimeric antigen receptor targeting NKG2D, chimeric antigen receptor T-cell targeting NKG2D and preparation method thereof and application thereof

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Embodiment 1

[0059] The preparation of chimeric antigen receptor T cells targeting NKG2D specifically includes the following steps:

[0060] (1) Prepare the gene sequence of chimeric antigen receptor CAR-NKG2D targeting NKG2D

[0061] Prepare signal peptide, NKG2D-targeting single-chain antibody, CD8α hinge region, CD8 transmembrane region, 4-1BB signal region, and CD3ζ signal region encoding genes, and use PCR to synthesize the above-mentioned signal peptide, NKG2D-targeting single-chain The coding genes of the antibody, CD8α hinge region, CD8 transmembrane region, 4-1BB signal region and CD3ζ signal region are connected together from the 5' end to the 3' end in turn to obtain the coding gene of CAR-NKG2D, the coding gene of the CAR-NKG2D The gene is shown in SEQ ID NO:4. Wherein, the coding gene sequence of the signal peptide is shown in SEQ ID NO: 7, and the coding gene sequence of the NKG2D-targeting single-chain antibody is shown in SEQ ID NO: 2.

[0062] (2) Construction of pWPXLd-...

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Abstract

The invention provides a chimeric antigen receptor (CAR) targeting NKG2D. Amino acid sequences of the CAR targeting NKG2D comprises the amino acid sequences of a single-chain antibody targeting NKG2D,an extracellular hinge region, a transmembrane region and an intracellular signal region which are sequentially connected from an amino terminal to a carboxyl terminal, respectively. The single-chainantibody targeting NKG2D comprises the amino acid sequences shown in SEQ ID NO: 1. The invention also provides a chimeric antigen receptor T-cell (CAR-T) targeting NKG2D and a preparation method thereof, as well as application of the CAR targeting NKG2D and the CAR-T targeting NKG2D in the prevention, diagnosis and treatment of liver cancer, cervical cancer, pancreatic cancer and other malignanttumors.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a chimeric antigen receptor targeting NKG2D, a chimeric antigen receptor T cell, and a preparation method and application thereof. Background technique [0002] CAR-T (Chimeric Antigen Receptor T Cell) technology is a new type of immune cell therapy, which is to reinfuse CAR-modified T cells into the human body, activate the body's own immune system, and continuously kill targeted tumor cells , is considered to be one of the most effective treatments for malignant tumors. [0003] NKG2D is a lectin-like type II transmembrane glycoprotein, which is expressed on the surface of NK cells, CD8+ T cells, activated macrophages, and tumor-infiltrating γδ T cells, and the expression of NKG2D ligands is a key factor in the expression of NKG2D ligands. An indicator of "stressed states," such as when cells are infected by viruses and malignantly transformed, but rarely on the surface of healthy c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/10C12N15/867C12N15/62A61K39/00A61P35/00
CPCC07K16/2851C07K14/7051C12N15/86C12N5/0636A61K39/0011A61P35/00C07K2319/02C07K2319/03C07K2319/33C07K2317/622C12N2740/15043C12N2510/00A61K2039/844A61K2039/892A61K2039/852
Inventor 刘茂玄许晨光黎琴子
Owner SHENZHEN BINDEBIOTECH CO LTD
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