Method for screening acute radiation gastrointestinal syndrome treatment targets

A therapeutic target and radioactive technology, applied in the field of biomedicine, can solve the problems of cells susceptible to radiation damage, no treatment for intestinal radiation damage, unpredictable radiation treatment effectiveness, etc., and achieve the effect of promoting stem cell proliferation.

Pending Publication Date: 2020-08-07
SUZHOU UNIV
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Problems solved by technology

However, when URI is knocked out, the β-catenin-c-MYC signaling pathway that was originally inhibited by URI will be activated, and the cells will proliferate rapidly and be more susceptible to radiation damage, resulting in the death of mice from radiation gastrointestinal syndrome
Although this study studied intestinal crypt quiescent stem cells, post-irradiation genetic intervention was not performed, so the effectiveness in radiation rescue cannot be predicted.
[0010] However, in ordinary knockout mice or gene overexpression mice, the target gene is already in a stable knockout or overexpression state, and it is impossible to regulate gene expression after irradiation
At present, the relatively advanced and mature transgenic animal model is the CreERT-loxP conditional overexpression mouse model, but there is no relevant research on the use of the CreERT-loxP conditional overexpression mouse model for the treatment of intestinal radiation injury

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  • Method for screening acute radiation gastrointestinal syndrome treatment targets
  • Method for screening acute radiation gastrointestinal syndrome treatment targets
  • Method for screening acute radiation gastrointestinal syndrome treatment targets

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Construction of CreERT-loxP conditionally overexpressed transgenic mice

[0039] In order to effectively promote the proliferation of crypt quiescent stem cells, this technical scheme uses Bmi1-CreERT; loxP conditionally overexpressed transgenic mice, and genetically intervened the mouse crypt quiescent stem cells, using TIGAR as the target gene, through tamoxifen Induce the expression of TIGAR protein in crypt quiescent stem cells, such as figure 2 shown.

[0040] In mice with the above genotypes, TIGAR can be overexpressed only in crypt quiescent stem cells through Bmi1, a crypt quiescent stem cell-specific promoter.

[0041] The writing method of the above mouse is: Bmi1-creERT; H11-Tigar. Specifically, Bmi1 is a specific promoter of crypt quiescent stem cells. Cre is a gene encoding a recombinase, which can be translated into a recombinase to cut a specific gene sequence. ERT encodes the estrogen receptor. When creERT is translated as a whole, the re...

Embodiment 2

[0044] Example 2: Induction of target gene overexpression after irradiation with ionizing radiation

[0045] Since it takes a certain amount of time from drug injection to the overexpression of TIGAR in crypt quiescent stem cells (for the CreERT-loxP animal model, it is usually 18-24hr), we irradiated the whole abdomen of mice with 15 Gy X-rays ( image 3 ) immediately after intraperitoneal injection of the drug (tamoxifen, single injection, 4.5mg / 20g mouse body weight).

[0046] On the 1st, 3rd, and 5th day after the mice were irradiated, the mice were sacrificed, and the intestinal tissues were made into frozen sections to observe the expression of TIGAR protein in the quiescent stem cells in the crypts, as shown in Figure 4 shown. Since TIGAR and green fluorescent protein (EGFP) are expressed simultaneously during the design and construction of transgenic mice, the expression level of green fluorescent protein can be used to indicate the expression level of TIGAR. One d...

Embodiment 3

[0047] Embodiment 3: Evaluation of Radiation Treatment Effect

[0048] In order to evaluate the radiation rescue effect of TIGAR overexpression, the survival rate of mice and HE staining of intestinal tissue sections were used to evaluate. In the survival rate experiment, mice in the control group (insert the Tigar gene downstream of the loxP-STOP-loxP sequence to obtain the loxP-STOP-loxP-Tigar sequence, insert the above sequence into the H11 site of the mouse genome to obtain the H11-Tigar small rats) and intestinal crypt quiescent stem cell TIGAR overexpressed mice received 15Gy X-ray whole-abdominal irradiation ( image 3 ), intraperitoneal injection of tamoxifen (single injection, 4.5mg / 20g mouse body weight) was carried out immediately after irradiation. After the injection, continue to feed the mice, and observe the survival of the mice, such as Figure 5 shown.

[0049] It can be seen that the mice in the control group (H11-Tigar mice, WT) all died of radiation gast...

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Abstract

The invention discloses a method for screening acute radiation gastrointestinal syndrome treatment targets, and belongs to the technical field of biological medicines. According to the invention, a CreERT-loxP condition is utilized to overexpress a transgenic mouse model, and proliferation of intestinal crypt stationary-phase stem cells is effectively promoted after large-dose ionizing radiation irradiation, so that a treatment target which still has a treatment effect after irradiation for 18-24 hours and aims at the radioactive gastrointestinal syndrome is screened. Only after tamoxifen is injected into a CreERT-loxP condition overexpression transgenic mouse, can gene shearing occur in specific cells, so that gene expression is regulated and controlled. Because of the characteristic, thepractical situation of irradiation first and then treatment after a nuclear accident occurs can be well simulated, so that the practical significance is achieved; and the screened treatment target isdeveloped into a medicine for nuclear accident treatment, and precious time can be gained for nuclear accident treatment.

Description

technical field [0001] The invention relates to a method for screening therapeutic targets for acute radiation gastrointestinal syndrome, belonging to the technical field of biomedicine. Background technique [0002] The degree of damage to different tissues of the human body after exposure to radioactive rays varies. It is generally believed that the sensitivity of cells to radiation is positively correlated with the proliferation rate of cells and negatively correlated with the degree of differentiation of cells. Under physiological conditions, the intestinal epithelium of the human body is rapidly replaced by the proliferation and driving of intestinal stem cells. Since small intestinal crypt stem cells are in a state of high-speed proliferation under physiological conditions, they are extremely vulnerable to radiation damage and lose their original ability to proliferate and divide. The stagnation of stem cell division will lead to the loss of the source of cell renewa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N13/00C12N15/85A01K67/027
CPCC12N13/00C12N15/85A01K67/0278A01K2207/35A01K2217/07A01K2227/105A01K2267/03
Inventor 张昊文邵春林王忠敏刘芬菊林龙昕陈飞张钰烁
Owner SUZHOU UNIV
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