CN107137711B - Application of Kindlin-2 protein as target in preparation of medicine for treating osteoarthritis - Google Patents
Application of Kindlin-2 protein as target in preparation of medicine for treating osteoarthritis Download PDFInfo
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Abstract
The invention relates to an application of Kindlin-2 protein as a target spot in preparation of a medicament for treating osteoarthritis. According to research results, after the Kindlin-2 gene is specifically knocked out, articular cartilage and subchondral bone of a mouse are seriously damaged, the area of the articular cartilage is remarkably reduced, joint swelling is serious, and the pain threshold of the mouse is remarkably reduced. The experimental results of this study suggest that the Kindlin-2 protein can be used as a new target for drugs for treating osteoarthritis.
Description
Technical Field
The invention relates to the field of medicines, in particular to application of Kindlin-2 protein serving as a target spot in preparation of medicines for treating osteoarthritis.
Background
Osteoarthritis is a degenerative arthritis disease, commonly occurs in middle-aged and elderly people, and is often manifested as subchondral sclerosis, chronic synovitis, periarticular osteophyte formation and the like. With the progress of China into the aging society, people suffering from osteoarthritis rise at an incredible speed, and the osteoarthritis is easy to cause pain, walking disorder and the like, so that the life quality of patients is seriously affected. At present, the treatment of osteoarthritis mainly comprises schemes of sports, joint health care, drug pain relief and the like. However, these schemes are usually temporary and permanent, and cannot radically treat osteoarthritis, and the used drugs are often lack of specificity and specificity, so that the development of novel specific drugs for treating osteoarthritis is very urgent.
Disclosure of Invention
Based on the above, there is a need for an application of the Kindlin-2 protein as a target in preparing a medicament for treating osteoarthritis.
Use of a Kindlin-2 protein in the preparation of a medicament for treating osteoarthritis, said medicament targeting the Kindlin-2 protein, said medicament being capable of promoting gene transcription or expression of the Kindlin-2 protein.
In one embodiment, the drug targets the Kindlin-2 protein in chondrocytes.
In one embodiment, the drug is capable of increasing the expression level of Kindlin-2 protein in the chondrocytes in an amount that increases the expression level of Kindlin-2 protein.
A medicament for treating osteoarthritis, wherein the medicament targets the Kindlin-2 protein, and the medicament comprises a component for promoting the gene transcription or expression of the Kindlin-2 protein.
In one embodiment, the drug targets the Kindlin-2 protein in chondrocytes.
In one embodiment, the agent is capable of increasing the amount of Kindlin-2 protein expressed in chondrocytes.
A pharmaceutical composition for treating osteoarthritis, comprising a Kindlin-2 protein promoter and a targeting agent, wherein the Kindlin-2 protein promoter is loaded on the targeting agent, and the targeting agent targets the Kindlin-2 protein.
In one embodiment, the targeting agent is selected from at least one of an AAV virus, miRNA, and polymer micelle.
In one embodiment, the Kindlin-2 protein promoter is a gene expressing the protein of Kindlin-2 and the targeting agent is an AAV virus, and the gene expressing the protein of Kindlin-2 is carried on the AAV virus.
Use of the Kindlin-2 protein in the preparation of a detection reagent for osteoarthritis, wherein the Kindlin-2 protein is used as a biomarker for osteoarthritis.
Experimental results show that after the Kindlin-2 gene is specifically knocked out, articular cartilage and subchondral bone of a mouse are seriously damaged, the area of the articular cartilage is remarkably reduced, joint swelling is serious, and the pain threshold of the mouse is remarkably reduced. The experimental results of this study suggest that the Kindlin-2 protein can be used as a new target for drugs for treating osteoarthritis.
Drawings
FIG. 1 is a graph showing the results of ABG staining of articular cartilage of mice in the experimental group and the control group in example 1;
FIG. 2 is a graph showing the statistical results of measurement of the areas of articular cartilage in tibia and femur using Image Pro plus7 software in the articular cartilage of the experimental group and the control group of mice in example 1;
FIG. 3 is a 3D reconstructed model of the bone joints of the mice in the experimental group and the control group in example 2 after micro CT scan analysis;
FIG. 4 is an external view of knee joints of mice in the experimental group and the control group in example 3;
FIG. 5 is a statistical result of pain threshold values of mice of the experimental group and the control group in example 4.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with examples are described in detail below. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention can be embodied in many different forms than those herein described and one skilled in the art can make similar modifications without departing from the spirit of the invention and it is therefore not limited to the specific embodiments disclosed below.
The use of a Kindlin-2 protein according to an embodiment for the preparation of a medicament for the treatment of osteoarthritis, wherein the medicament targets the Kindlin-2 protein and the medicament is capable of promoting gene transcription or expression of the Kindlin-2 protein.
The Kindlin-2 protein is a focal adhesion protein and is involved in the activation of integrin, and has been shown to play an important role in the elongation of muscle cells. However, the Kindlin-2 protein has less research on target therapy, and no research report on osteoarthritis exists at present. The inventor of the present invention has unexpectedly found that the expression of Kindlin-2 protein has a close correlation with osteoarthritis after experimental studies. After the Kindlin-2 gene is specifically knocked out, articular cartilage and subchondral bone of a mouse are seriously damaged, the area of the articular cartilage is remarkably reduced, joint swelling is serious, and the pain threshold of the mouse is remarkably reduced. The experimental result of the research provides a new target for the medicine for treating osteoarthritis. The drug promotes the gene transcription or expression of the Kindlin-2 protein by targeting the Kindlin-2 protein, thereby assisting in the treatment of osteoarthritis.
Specifically, the "drug for treating osteoarthritis" referred to herein may be a drug prepared for treating a patient who has been diagnosed with osteoarthritis, or a drug prepared for preventing osteoarthritis.
In one embodiment, a Kindlin-2 protein is used in the preparation of a medicament for treating osteoarthritis, the medicament targeting the Kindlin-2 protein in chondrocytes. The chondrocytes are closely related to osteoarthritis, and the Kindlin-2 protein in the chondrocytes is targeted by the medicine, so that the specificity of the medicine can be further enhanced, and side effects can be reduced.
In one embodiment, the drug is specifically targeted to cells positive for Aggrecan (aggrecanase).
In one embodiment, the use of the Kindlin-2 protein in the preparation of a medicament for treating osteoarthritis, the medicament is capable of increasing the expression level of the Kindlin-2 protein in chondrocytes. Research results show that the area of articular cartilage of mice after Kindlin-2 gene specific knockout is obviously reduced, and the research results suggest that articular cartilage is damaged when the secretion of Kindlin-2 protein is reduced, and further symptoms such as osteoarthritis and the like are caused. Therefore, the Kindlin-2 protein can be used as a new target of a medicament for treating osteoarthritis, the expression level of the Kindlin-2 protein can be improved through medicaments, articular chondrocytes are specifically protected, and articular cartilage damage is prevented.
More specifically, in one embodiment, the use of the Kindlin-2 protein in the preparation of a medicament for the treatment of osteoarthritis, the medicament is capable of increasing the expression level of the Kindlin-2 protein in articular chondrocytes.
In conclusion, the results of the studies show that the expression level of Kindlin-2 protein is closely related to osteoarthritis. The Kindlin-2 protein can be used as a new target spot for preparing a medicament for treating osteoarthritis, and has good application prospect in preparing medicaments for preventing or treating osteoarthritis.
Further, the present application provides a drug for treating osteoarthritis according to an embodiment, wherein the drug targets the Kindlin-2 protein, and the drug contains a component that promotes the transcription or expression of the Kindlin-2 protein gene.
The component for promoting the gene transcription or expression of the Kindlin-2 protein in the medicine can improve the expression level of the Kindlin-2 protein, specifically protect articular chondrocytes and prevent articular cartilage from being damaged.
In one embodiment, the agent for treating osteoarthritis targets the Kindlin-2 protein in chondrocytes. Furthermore, the drug can increase the expression level of Kindlin-2 protein in chondrocytes.
More specifically, the chondrocyte may be an articular chondrocyte, and the drug for treating osteoarthritis targets the Kindlin-2 protein in the articular chondrocyte and increases the expression level of the Kindlin-2 protein in the articular chondrocyte.
The medicine takes the Kindlin-2 protein as a therapeutic target, and treats osteoarthritis by promoting the gene transcription or expression of the Kindlin-2 protein. Research results show that when the Kindlin-2 gene is specifically knocked out, the Kindlin-2 protein expression level is reduced, articular cartilage and subchondral bone of a mouse are seriously damaged, the articular cartilage area is remarkably reduced, joint swelling is serious, and the pain threshold of the mouse is remarkably reduced. Therefore, the Kindlin-2 protein gene can be promoted to be transcribed or expressed by medicaments, so that the osteoarthritis can be treated.
Further, the present application also provides a pharmaceutical composition for treating osteoarthritis according to an embodiment, the composition comprising a Kindlin-2 protein promoter and a targeting agent, the Kindlin-2 protein promoter being supported on the targeting agent, and the targeting agent targeting the Kindlin-2 protein.
Wherein, the Kindlin-2 protein promoter is a drug or a reagent capable of improving the expression level of the Kindlin-2 protein, and the targeting agent is a drug or a reagent capable of targeting the Kindlin-2 protein. The Kindlin-2 protein promoter can be loaded on the targeting agent through chemical bond connection or physical modes such as coating and the like.
In one embodiment, the targeting agent may be selected from at least one of AAV virus (Adeno-associated virus), mirna (microrna), and polymer micelles.
Specifically, AAV, miRNA or polymer micelle is used as a carrier to carry the Kindlin-2 protein promoter to a specific part in a body, and the Kindlin-2 protein promoter is released to promote the expression of the Kindlin-2 protein in the body, so that the expression level of the Kindlin-2 protein is increased, and articular cartilage is repaired or protected, thereby treating or preventing osteoarthritis.
In one embodiment, the Kindlin-2 protein promoter is a gene expressing Kindlin-2 protein, the targeting agent is AAV, the gene expressing Kindlin-2 protein is carried on AAV, and the AAV enters the body and expresses Kindlin-2 protein, thereby increasing the expression level of Kindlin-2 protein.
The pharmaceutical composition for treating osteoarthritis comprises a Kindlin-2 protein promoter and a targeting agent, which have synergistic effects, so that the expression level of the Kindlin-2 protein is increased, and further osteoarthritis is prevented or treated.
Further, the present application provides use of the Kindlin-2 protein according to an embodiment in preparing a detection reagent for osteoarthritis, wherein the detection reagent uses the Kindlin-2 protein as a biomarker for osteoarthritis.
Research shows that the expression level of the Kindlin-2 protein is closely related to osteoarthritis, so that the Kindlin-2 protein can be used as a biomarker of osteoarthritis. Specifically, the detection reagent can determine the condition of osteoarthritis from the expression level of Kindlin-2 protein.
In conclusion, the close correlation between the Kindlin-2 protein and osteoarthritis is unexpectedly found after experimental research. After gene specificity is knocked out, the expression level of the Kindlin-2 protein is reduced, so that articular cartilage and subchondral bone of a mouse are seriously damaged, the area of the articular cartilage is remarkably reduced, joint swelling is serious, and the pain threshold of the mouse is remarkably reduced. The experimental result indicates that the Kindlin-2 protein can be used as a new target point for treating osteoarthritis. The Kindlin-2 protein has good application prospect in preparing medicines for treating osteoarthritis or preparing detection reagents for treating osteoarthritis.
The following are specific examples.
The examples, which are not specifically illustrated, employ drugs and equipment, all of which are conventional in the art. The experimental procedures, in which specific conditions are not indicated in the examples, are usually carried out according to conventional conditions, such as those described in the literature, in books, or as recommended by the manufacturer of the kits.
Not specifically stated, WT represents a wild-type control mouse in the following examples, and cKO represents a mouse after Kindlin-2 inducing specific knockdown of Aggrecan-positive chondrocytes using tamoxifen. The experimental mice were provided by the university of lash, usa, and were all male mice.
Example 1
Effect of Kindlin-2 gene knockout on mouse articular cartilage
Two month old experimental mice (cKO) were injected with tamoxifen to induce Kindlin-2 knock-out at a concentration of 10mg/ml, at an injection dose of 0.1ml/10g body weight, and five injections were administered sequentially, one injection per day, and one additional injection was administered one month later. Corn oil injection was performed under the same conditions as the experimental control group (WT). After two months, the mice were sacrificed, the knee joints were removed, fixed with formalin, decalcified with 10% EDTA for 21 days, embedded with paraffin, sectioned, and stained with ABG (Alcian blue and safranin-O/fast green staining, Alcian blue and safranin/fast green staining), with the results shown in fig. 1, in which articular cartilage and subchondral bone were severely damaged after specific knockout of kinglin-2 in Aggrecan positive cells, compared to the control group. The area of articular cartilage in the tibia and femur was measured in figure 1 using Image Pro plus7 software and the statistical results are shown in figure 2.
The results show that the specific knockout of Kindlin-2 in Aggrecan positive cells leads to the remarkable reduction of the articular cartilage area, and the close connection between Kindlin-2 and the development of osteoarthritis is suggested.
Example 2
The Kindlin-2 gene was knocked out in Aggrecan positive cells in experimental mice (cKO) as described in example 1. Corn oil injection was performed under the same conditions as the experimental control group (WT). After two months, the mice were sacrificed, the bone joints were removed, fixed with formaldehyde, subjected to micro CT (μ CT) scanning analysis, and then 3D reconstructed, and the results are shown in fig. 3, in which (a) shows a front view of the bone joints of WT type mice, (b) shows a rear view of the bone joints of WT type mice, (c) shows a front view of the bone joints of cKO type mice, and (D) shows a rear view of the bone joints of cKO type mice.
micro-CT (μ CT) scanning showed that after Kindlin-2 was specifically knocked out in Aggrecan positive cells, the structure and morphology of the bone joint was severely damaged. Suggesting that osteoarthritis can be prevented or treated by a drug which promotes the transcription or expression of the gene of Kindlin-2 protein.
Example 3
The Kindlin-2 gene was knocked out in Aggrecan positive cells in experimental mice (cKO) as described in example 1. Corn oil injection was performed under the same conditions as the experimental control group (WT). After two months, the mice were sacrificed and the knee joints were removed. As shown in FIG. 4, mice had severe knee swelling after specific Kindlin-2 knock-out in Aggrecan positive cells.
Example 4
Behavioral testing for pain index
Mechanical force-induced pain was measured according to the von Frey sensitivity assay, and the test mice were allowed to become familiar with the environment prior to the von Frey test. In making the measurements, the hind paw of the mouse was poked from below using a calibrated von Frey filament, and then a force with drawwalthreshold (50%) was calculated according to the iterative approach. The statistical results of pain threshold values of the experimental group mouse (cKO) and the control group mouse are shown in FIG. 5. It was shown that the pain threshold of mice was significantly decreased following specific knockout of Kindlin-2 in Aggrecan positive cells.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (1)
- Use of the Kindlin-2 protein in the preparation of a detection reagent for osteoarthritis, wherein the detection reagent uses the Kindlin-2 protein as a biomarker for osteoarthritis.
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| CN112843235B (en) * | 2021-01-27 | 2023-03-31 | 南方科技大学 | Application of reagent for inhibiting Kindlin-2 protein expression level in liver cells in preparation of fatty liver treatment product |
| CN115105597A (en) * | 2022-07-25 | 2022-09-27 | 南方科技大学 | Application of Kindlin-2 as target in treating osteoporosis and preventing cancer bone metastasis |
Citations (2)
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|---|---|---|---|---|
| WO2012028703A1 (en) * | 2010-09-02 | 2012-03-08 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Method for the prognosis of the progression of cancer |
| CN104407151A (en) * | 2014-11-19 | 2015-03-11 | 汕头大学医学院 | Kit integrating three proteins such as Kindlin-2, Myosin-9 and Annexin II for prognosis evaluation of patient suffering from esophageal squamous cell carcinoma |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012028703A1 (en) * | 2010-09-02 | 2012-03-08 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Method for the prognosis of the progression of cancer |
| CN104407151A (en) * | 2014-11-19 | 2015-03-11 | 汕头大学医学院 | Kit integrating three proteins such as Kindlin-2, Myosin-9 and Annexin II for prognosis evaluation of patient suffering from esophageal squamous cell carcinoma |
Non-Patent Citations (5)
| Title |
|---|
| Kindlin-2 controls TGF-beta signalling and Sox9 expression to regulate chondrogenesis;Chuanyue Wu et al.;《Nature Communication》;20150707;第1页摘要 * |
| Kindlin-2 inhibits serous epithelial ovarian cancer peritoneal dissemination and predicts patient outcomes;Caixia Ren et al.;《Biochemical and Biophysical Research Communications》;20140227;第446卷;第187-194页 * |
| Kindlin家族成员研究进展;熊盈等;《生物化学与生物物理进展》;20101231;第32卷(第12期);第1265-1270页 * |
| Mediation of Rac1 Activation by Kindlin-2: An Essential Function in Osteoblast Adhesion, Spreading, and Proliferation;Gil-Yong Jung et al.;《Journal of Cellular Biochemistry》;20110516;第112卷;第2541-2548页 * |
| 整合素相互作用蛋白Kindlin家族的生物学功能研究进展;杨玫等;《解剖学报》;20141231;第45卷(第6期);第865-869页 * |
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