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Application of adm competitive antagonist ama in glioma immunotherapy

A technology of glioma stem cells and inhibitors, which is applied in the field of research on compounds that inhibit glioma growth and inhibit glioma growth, can solve problems such as the unknown role of proliferation and tumor growth, and achieve important clinical application value and potential business value effect

Active Publication Date: 2021-05-14
THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the role of ADM-CRLR signaling in GBM in GSC cell proliferation and tumor growth is still unknown, and there is no report on AMA acting on glioma through ADM-CRLR signaling

Method used

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  • Application of adm competitive antagonist ama in glioma immunotherapy
  • Application of adm competitive antagonist ama in glioma immunotherapy
  • Application of adm competitive antagonist ama in glioma immunotherapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Cell culture

[0043] Human U937 monocytes were purchased from the American Type Culture Collection (ATCC). U937 adds 10% fetal calf serum to the RPMI-1640 medium (Gibco company) containing the M-CSF of 10ng / ml, the IL-4, IL-10, IL-13, TGF-β (PeproTech company) of 20ng / ml (Gibco Company) were cultured for 7 days to induce TAM. GSCs were cultured with Neurobasal medium (Lifetechnology Company), which contained 20 μl / ml B27 (Life Technology Company) and 20 ng / ml EGF, bFGF growth factors (PeproTech Company). All cells were stored at 37°C, 5% CO 2 In an incubator (95% relative humidity), the cells are subcultured for 2-3 days, and digested with papain and trypsin for 2-5 minutes before subculture.

[0044] 2. Construction of intracranial orthotopic xenograft tumor model co-injected with GSC and TAM

[0045] (1) Experimental grouping: control group, AMA treatment group;

[0046] (2) Take 4-6 weeks old female NCG mice (Shanghai Nanfang Model Biotechnology Co., Ltd., bre...

Embodiment 2

[0047] Example 2 Analysis of the effect of AMA interfering with the ADM-CRLR pathway on tumorigenesis in tumor-bearing mice

[0048] On day 9, intraperitoneal injection of AMA (0.25 mg / kg, Beijing Boaosen Co., Ltd.) was performed once every two days. Tumor-bearing mice were subjected to live animal imaging tests on days 8, 11, and 17. The method was as follows: 200 μl of Luciferin substrate (BioVision Company) was injected intraperitoneally into each mouse. After 10 minutes, the tumor formation situation and the Luciferase signal intensity of each group of mice were detected with a living animal imager (PerkinElmer Company), and the experimental results were as follows: figure 2 shown.

[0049] Conclusion: TAM secretes ADM to promote glioma tumor growth. Antagonist AMA blocks the binding of ADM secreted by TAMs to CRLR and inhibits glioma tumor growth. Such as figure 2 As shown, A is the imaging result of a mouse live animal, and B is the corresponding quantitative anal...

Embodiment 3

[0050] Example 3 Cell Spheroidization Experiment

[0051] 1) Experimental grouping: control group, TAM supernatant treatment group, AMA single treatment group, TAM supernatant and AMA combined treatment group;

[0052] 2) The number of TAM cells is 1×10 7 , cultured with 6ml stem cell starvation medium containing 5μl / ml B27, 2ng / ml EGF, and bFGF growth factors, and collected the supernatant after 2 days;

[0053] 3) Collect GSC, count the cells and adjust the cell concentration to 5×10 3 The experimental groups were control group, AMA single treatment group, TAM supernatant treatment group, TAM supernatant and AMA combined treatment group. Add 100 μl of cell suspension to each well of a 96-well plate according to the experimental grouping, and the culture medium of the control group is the stem cell starvation medium. A total of 1 96-well plate was inoculated, 10 duplicate wells were set up for each group, and cultured at 37°C;

[0054] 4) Supplement 50 μl of culture solut...

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Abstract

The invention belongs to the technical field of medicine, and in particular relates to a compound for inhibiting glioma growth, a drug, a method for inhibiting glioma growth and a method for researching and inhibiting glioma growth. The complex is an AMA-CRLR complex, and the AMA-CRLR complex is obtained by combining the AMA with the CRLR. The AMA combines with the CRLR to form the AMA-CRLR complex to inhibit the glioma stem cell (GSC) sphere-forming ability of tumor-associated macrophages (TAM), and inhibit TAM-mediated AKT signaling pathway activation and then Inhibits the growth of glioma. Therefore, the research on the role of AMA-CRLR complex in glioma through ADM-CRLR signaling has important clinical application value and commercial value.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a compound for inhibiting glioma growth, a drug, a method for inhibiting glioma growth and a method for researching and inhibiting glioma growth. Background technique [0002] Glioblastoma (GBM) is the most common primary malignant tumor of the central nervous system. It grows rapidly, recurs easily, and resists conventional treatment. The prognosis of patients is poor. Recent studies have shown that glioma stem cells (GSCs) play a key role in the rapid proliferation, treatment resistance and recurrence of GBM. GSC is a small subset of stem cell-like cells in the heterogeneous GBM cell population, which has the ability of self-renewal, multi-lineage differentiation and tumor reconstruction. [0003] GSCs are in a specific microenvironment and interact with microenvironment components to maintain their own "stemness", promote tumor angiogenesis, immune evasion and th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/23A61P35/00
CPCA61K38/23A61P35/00
Inventor 时雨王文英吴晶晶平轶芳卞修武
Owner THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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