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Chimeric antigen receptor adipose-derived stem cell and preparation method thereof

A chimeric antigen receptor, adipose stem cell technology, applied in the field of tumor treatment, can solve the problems of immune rejection, low virus infection efficiency, etc., and achieve the effect of improving the effect, easy virus infection, and low immunogenicity

Inactive Publication Date: 2016-05-11
BIOTOWNTEK CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a chimeric antigen receptor fat stem cell and its preparation method, aiming at solving the problem of "immune storm", virus Low infection efficiency and immune rejection of allografts

Method used

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  • Chimeric antigen receptor adipose-derived stem cell and preparation method thereof
  • Chimeric antigen receptor adipose-derived stem cell and preparation method thereof
  • Chimeric antigen receptor adipose-derived stem cell and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] CAR-Adipose Stem Cells Expressing TRAIL to Treat Tumors

[0052] In this example, for different solid tumors, the third-generation CAR, adipose-derived stem cells, and the expression of secreted TRAIL are first used to treat tumors, such as image 3 shown. First of all, it is necessary to synthesize the third-generation chimeric antigen receptor, that is, to synthesize the tandem sequence: soluble signal peptide sequence-scFv antigen-binding sequence-CD8 (hinge sequence and transmembrane sequence)-CD28-endo-4-1BB-endo-CD3ζ sequence, Among them, the two ends of the synthesized sequence need to synthesize EcoRI and BamHI restriction sites. In addition, it is also necessary to synthesize the TRAIL gene (also known as TNFSF10) that can be secreted into the extracellular space. See Seq-TRAIL for its sequence. Among them, the two ends of the TRAIL sequence also need to add EcoRI and BamHI restriction sites respectively. Then the above two sequences were respectively constr...

Embodiment 2

[0058] CAR-Trunc-FGFR1 Adipose Stem Cells Expressing TRAIL to Treat Tumors

[0059] In this example, for different solid tumors, the improved CAR-Trunc-FGFR1 for adipose stem cells, adipose stem cells and expression of secreted TRAIL can be combined to treat tumors, such as Figure 4 shown. Similarly, CAR-Trunc-FGFR1 for adipose-derived stem cells needs to be synthesized first, and the tandem sequence of each domain is: soluble signal peptide sequence-scFv antigen-binding sequence-FGFR1-trans-endo sequence, where the two ends of the synthesized sequence need to be synthesized EcoRI and BamHI restriction sites. In addition, it is also necessary to synthesize the TRAIL gene (also known as TNFSF10) that can be secreted into the extracellular space, and the two ends of its sequence also need to add EcoRI and BamHI restriction sites.

[0060] Then the above two sequences were respectively constructed on the PLVX lentiviral vector. The construction process is called: use EcoRI an...

Embodiment 3

[0066] CAR-Adipose Stem Cells Expressing TNF-α to Treat Tumors

[0067] In this example, for different solid tumors, the third-generation CAR, adipose stem cells, and the expression of tumor-killing factor TNF-α are first combined to treat tumors, such as Figure 5 shown. First, the third-generation chimeric antigen receptor needs to be synthesized, that is, the tandem sequence: soluble signal peptide sequence-scFv antigen-binding sequence-CD8 (hinge and transmembrane)-CD28-endo-4-1BB-endo-CD3ζ sequence, wherein, Both ends of the synthesized sequence need to synthesize EcoRI and BamHI restriction sites. In addition, it is also necessary to synthesize TNF-α that can be secreted extracellularly, and its sequence is shown in Seq-TNF-α.

[0068] Wherein, the two ends of the TNF-α sequence also need to add EcoRI and BamHI restriction sites respectively. Then the above two sequences were respectively constructed on the PLVX lentiviral vector. The construction process is called: ...

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Abstract

The invention discloses a chimeric antigen receptor adipose-derived stem cell and a preparation method thereof. The chimeric antigen receptor adipose-derived stem cell is composed of a chimeric antigen receptor, an adipose-derived stem cell body and a tumor killing factor. The high-targeting characteristic that the antigen receptor is specifically combined with a specific tumor type, the characteristics that a fatty liver cell is low in immunogenicity, prone to be infected with viruses and capable of being amplified in a large scale within a short time and the tumor resisting function of the tumor killing factor are combined, and therefore the problems of immune storms, low chimeric antigen expression efficiency and allotransplantation immunological rejection during existing CART tumor treatment can be solved, and the tumor treating effect is improved. The improved adipose-derived stem cell has the efficient targeting performance of being specifically transferred to a specific tumor position, the risk that an immune system is excessively activated, and consequently immunological rejection is caused is avoided, the tumor killing factor carried by the adipose-derived stem cell can efficiently kill a tumor at the position of the tumor, and then the aim of treating the tumor is achieved.

Description

technical field [0001] The invention relates to the field of tumor treatment, in particular to a chimeric antigen receptor fat stem cell and a preparation method thereof. Background technique [0002] Tumor is one of the major diseases that seriously endanger human health. In my country, tumors are a great threat to people's lives and health, greatly reducing the quality of life of cancer patients, and tumors have also caused a huge burden on families and society. Therefore, it is an important research topic in the world medical community to find an effective tumor treatment method and completely conquer the tumor. [0003] Tumor immunotherapy is one of the most promising research directions in the current field of tumor treatment, and it is a new method of tumor treatment that has emerged in recent years and has significant curative effects. Preliminary clinical trial results show that its treatment efficiency is very high, and major pharmaceutical companies have sought t...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/867
CPCC07K14/525C07K14/70575C07K2319/33
Inventor 陈光风史秀娟刘小青
Owner BIOTOWNTEK CO LTD
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