Fusion protein of cytokine combined chimeric antigen receptor and application of fusion protein

A chimeric antigen receptor and fusion protein technology, applied in the field of biomedicine, can solve the problems of lack of safe and deliberate targets, complex tumor immune microenvironment, poor treatment effect, etc. Tumor effect, good anti-tumor effect

Active Publication Date: 2021-09-17
SUN YAT SEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Based on the breakthrough of CAR T in the treatment of blood tumors, CAR T therapy has been further extended to the treatment of solid tumors, and has been explored in various solid tumors including breast cancer, liver cancer, colorectal cancer and lung cancer. Unlike hematological tumors, it lacks safe and specific targets, and solid tumors have a complex tumor immune microenvironment, which ultimately leads to poor therapeutic effects

Method used

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  • Fusion protein of cytokine combined chimeric antigen receptor and application of fusion protein
  • Fusion protein of cytokine combined chimeric antigen receptor and application of fusion protein
  • Fusion protein of cytokine combined chimeric antigen receptor and application of fusion protein

Examples

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Embodiment

[0070] 1 Construction of T2-mCAR molecules co-expressing IL-7 and CCL3

[0071] The mouse CAR gene constructed based on the retroviral vector backbone can be efficiently transduced into mouse T cells. Based on the original mouse second-generation CAR molecule T2-m28z, we constructed a co-expression mouse protein IL -7 and CCL3 mCAR fusion protein. Mouse IL-7 and CCL3 proteins are connected to the 3' end of the CAR gene without a stop codon by using a cleavable fusion protein adapter sequence (2A peptide, P2A). Since P2A can cut itself independently of proteases, The two structural domains are broken to achieve the result of co-expression of the three molecules. So far, we have constructed the murine CAR structure of T2-m28z IL7CCL3 ( 7P3 ), its amino acid sequence is shown in SEQ ID NO: 12, and its structure is shown in figure 1 shown.

[0072] 2 Preparation of CAR T cells co-expressing IL-7 and CCL3

[0073] The present invention prepares mCAR T cells according to the ex...

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Abstract

The invention relates to the field of biological medicine, in particular to fusion protein of a cytokine combined chimeric antigen receptor and an application of the fusion protein. The fusion protein comprises a chimeric antigen receptor, 2A peptide, IL-7, 2A peptide and CCL3 which are sequentially connected in series, the chimeric antigen receptor comprises an scFv region, a hinge region, a transmembrane domain and an intracellular signal transduction region. According to the fusion protein of the cytokine combined chimeric antigen receptor and the application of the fusion protein, two cytokines IL-7 and CCL-3 are added during construction of a CAR vector, and the two cytokines can enhance the anti-apoptosis effect of CAR T cells and reduce the immunosuppression effect of the CAR T cells, so that the CAR T cells are helped to enhance the anti-tumor effect.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a fusion protein of cytokine combined with chimeric antigen receptor and its application. Background technique [0002] Malignant tumors have become one of the leading causes of death worldwide. With the continuous development of science and technology, through the early diagnosis and treatment of cancer, the cure rate of cancer has been greatly improved and the quality of life of patients has been improved. However, most malignant tumors still have problems such as poor prognosis and easy recurrence. Among all tumor treatments, immunotherapy has received constant attention and expectations from researchers. The biggest feature of immunotherapy is to use the immune system itself to fight cancer, including antibody therapy, cell therapy and immunomodulator therapy. Among them, cell therapy, which has attracted much attention, refers to the reinfusion of cell preparations into the pat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/85C12N5/10A61K39/00A61K38/20A61K38/19A61P35/00
CPCC07K14/5418C07K14/7051C07K14/523C12N5/0636C12N15/85A61K39/001111A61K38/2046A61K38/195A61P35/00C12N2510/02C07K2319/02C07K2319/03A61K2039/5158A61K2300/00
Inventor 黄朝峰陈焕鹏
Owner SUN YAT SEN UNIV
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