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The application of aryl hydrocarbon receptor (ahr) in predicting the effect of immunotherapy

A technology of immunohistochemistry and immune detection point, which is applied in the field of medicine and can solve the problem of low efficiency of PD-1/PD-L1 antibody

Active Publication Date: 2021-11-09
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem of relatively low efficiency of PD-1 / PD-L1 antibodies in the clinical application process, it is urgent to find new methods to predict the efficacy of PD-1 / PD-L1 antibodies to accurately screen effective cases

Method used

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  • The application of aryl hydrocarbon receptor (ahr) in predicting the effect of immunotherapy
  • The application of aryl hydrocarbon receptor (ahr) in predicting the effect of immunotherapy
  • The application of aryl hydrocarbon receptor (ahr) in predicting the effect of immunotherapy

Examples

Experimental program
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Effect test

Embodiment 1

[0028] The effect of tobacco exposure on PD-L1 expression has not been reported so far. In the present invention, tobacco extract (CES) was prepared by using a smoke generator, and the smoke was dissolved in 50 mL of serum-free 1640 medium. Normal lung epithelial 16HBE cells and non-small cell lung cancer H460 cells were treated with tobacco extract (CES) and tobacco carcinogen benzopyrene (BaP) (sigma). See results Figure 1A ~ Figure 1D . in, Figure 1A , H460 cells and 16HBE cells were treated with tobacco extract, real-time PCR was used to detect the expression of PDL1. Figure 1B , H460 cells and 16HBE cells were treated with tobacco extract, and the expression of PDL1 was detected by flow cytometry. Figure 1C , H460 cells and 16HBE cells were treated with different concentrations of BaP, and the expression of PDL1 was detected by real-time PCR. Figure 1D , H460 cells and 16HBE cells were treated with 5 μM BaP, the cells were collected at different time periods, and...

Embodiment 2

[0034] In order to further study the role of AhR inhibitors in the occurrence of other cancers, in this example, the mouse colon cancer cell line MC38 with high expression of PD-L1 was inoculated into C57 mice, and the AhR antagonist ANF was given 4 days after the inoculation, and the experiment was completed The tumor tissues of the mice were then removed and photographed. See results Figure 6A , the results showed that AhR inhibitors can significantly inhibit the growth of MC38 in mice, and have a good combined effect with anti-PD-L1 antibodies.

[0035] The mouse fibrosarcoma cell line Ag104ld with high expression of PD-L1 but resistant to PD-L1 antibody was selected to inoculate B6C3F1 mice. Four days after the inoculation, the AhR inhibitor combined with PD-L1 antibody was given. After the experiment, the tumor tissues of the mice were taken out. And take pictures. See results Figure 6B , found that the combination of AhR inhibitor and PD-L1 antibody can reverse PD-L...

Embodiment 3

[0037] This example studies the role of the level of AhR expression in predicting the therapeutic effect of immunotherapy. The lung cancer samples of patients treated with PD-1 antibody were selected, and the relationship between the expression level of AhR and the efficacy of PD-1 antibody was detected by immunohistochemical method. The specific detection method is as follows:

[0038] 1) Dewaxing: Preheat in an oven at 60°C for 30 minutes, then put them in fresh xylene I&II&III for 10 minutes each;

[0039] 2) Hydration: 5 minutes each in absolute ethanol I&II&II, 5 minutes each in 95%, 85%, and 75% ethanol;

[0040] 3) Wash once with PBS;

[0041] 4) Put the slices in the citric acid buffer repairing solution, cook in the microwave for 5 minutes on high heat, and cook on medium and low heat for 15 minutes;

[0042] 5) Cool down to room temperature for about 1 hour;

[0043] 6) Blocking: Add blocking solution dropwise, incubate at 37°C for 30 minutes, and remove excess l...

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Abstract

The present invention provides a new application of aryl hydrocarbon receptor (AhR) in predicting the effect of immunotherapy, specifically, the present invention provides the application of AhR as a target in the development, screening and / or preparation of drugs for treating tumors , also provides the application of the reagents for inhibiting, silencing and / or knocking out AhR in the preparation of drugs for treating tumors, and also provides the use of reagents for detecting the level of AhR in the preparation for predicting the effect of immune checkpoint inhibitors in treating tumors application in agents.

Description

technical field [0001] The present invention relates to a new application of transcription factor aryl hydrocarbon receptor (AhR), in particular, relates to the detection method of aryl hydrocarbon receptor in predicting the treatment effect of immunotherapy drugs and the application as the target of anticancer drugs, which belongs to medicine field. Background technique [0002] Aryl hydrocarbon receptor (AhR) is a transcription factor that plays an important role in regulating the metabolism of polycyclic aromatic hydrocarbons and other compounds in the environment, body immunity, rhythm, reproduction, oxidative stress, etc. by regulating the transcription of downstream genes effect. AhR exists in various tissues and cells such as the lung, liver, kidney, placenta, tonsil, skin, and B lymphocytes of the human body, and the metabolites of tryptophan, heme, and arachidonic acid are its endogenous sex ligand. [0003] AhR is involved in the maintenance and stability of the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
Inventor 周光飚王桂珍张力
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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