Application of OLA1 as target spot in preparation of medicine for inhibiting myocardial fibrosis
By inhibiting OLA1 gene expression, OLA1-targeted drugs were developed, which solved the problem of insufficient treatment for myocardial fibrosis in existing technologies and achieved effective inhibition of myocardial fibrosis and protection of cardiac function.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- NANTONG UNIV
- Filing Date
- 2026-02-11
- Publication Date
- 2026-05-12
AI Technical Summary
Existing technologies offer limited and unsatisfactory treatment options for myocardial fibrosis, necessitating the development of new therapeutic targets to effectively inhibit myocardial fibrosis.
By using OLA1 as a target, drugs that inhibit myocardial fibrosis can be developed by inhibiting the expression of the OLA1 gene through siRNA, CRISPR, or small molecule compounds.
It significantly inhibits myocardial fibrosis, reduces the expression of fibrosis markers in cardiac tissue, improves cardiac function, reduces cardiac dysfunction, and provides a new and effective means of treating myocardial fibrosis.
Smart Images
Figure CN122005803A_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of biomedical technology, and in particular to the application of OLA1 as a target in the preparation of drugs that inhibit myocardial fibrosis. Background Technology
[0002] Myocardial fibrosis is a common pathophysiological process in cardiomyopathy, characterized by excessive deposition and abnormal remodeling of cardiac extracellular matrix (ECM) components. Under normal circumstances, the ECM is mainly composed of collagen, elastin, proteoglycans, and glycosaminoglycans, which work together to maintain the structure and function of the heart. However, in the progression of various heart diseases (such as hypertensive heart disease, ischemic heart disease, myocarditis, and myocardial hypertrophy), myocardial fibrosis gradually occurs and worsens, leading to increased cardiac stiffness and decreased compliance, ultimately potentially causing serious consequences such as heart failure, severely impacting patients' quality of life and survival rate.
[0003] The pathogenesis of myocardial fibrosis is complex, involving the interaction of multiple cell types and signaling pathways. Among these, cardiomyocyte damage and death are key factors in initiating myocardial fibrosis. When cardiomyocytes are stimulated by factors such as ischemia, hypoxia, and inflammation, they release a series of cytokines and chemokines, such as transforming growth factor-β (TGF-β), angiotensin II (Ang II), and interleukin-1 (IL-1). These factors can activate cardiac fibroblasts, transforming them from a quiescent state into an activated state with synthetic and secretory functions, leading to the synthesis and secretion of large amounts of extracellular matrix components, especially collagen. Furthermore, oxidative stress, endoplasmic reticulum stress, and mechanical stress also play important roles in the process of myocardial fibrosis, promoting the excessive accumulation and remodeling of the extracellular matrix through multiple pathways.
[0004] Currently, clinical treatment options for myocardial fibrosis are relatively limited and the effects are not ideal. Traditional treatment strategies mainly include treating the underlying disease, such as controlling blood pressure, improving myocardial blood supply, and anti-inflammation. For example, for hypertensive patients, antihypertensive drugs (such as ACE inhibitors and angiotensin receptor blockers) can slow the progression of myocardial fibrosis to some extent; for patients with ischemic heart disease, coronary intervention or coronary artery bypass surgery can improve myocardial blood supply and reduce myocardial ischemia damage, thereby inhibiting myocardial fibrosis to some extent. In addition, some drugs such as aldosterone antagonists and beta-blockers are also used to treat myocardial fibrosis, but the mechanisms of action of these drugs are mainly through improving cardiac hemodynamics or reducing excessive activation of the neuroendocrine system. Current treatments have a weak direct inhibitory effect on myocardial fibrosis itself and still have many limitations. Therefore, it is urgent to discover new therapeutic targets for the treatment of myocardial fibrosis. Summary of the Invention
[0005] The purpose of this application is to provide a new therapeutic target that can effectively inhibit myocardial fibrosis through the discovery of a new therapeutic target.
[0006] To achieve the above objectives, this application provides the following technical solution:
[0007] The application of OLA1 as a target in the preparation of drugs to inhibit myocardial fibrosis, wherein the sequence of OLA1 is shown in SEQ ID NO: 01.
[0008] Preferably, the drug is used to inhibit the expression of the OLA1 gene.
[0009] Preferably, the drug contains at least one of siRNA, CRISPR, and small molecule compounds for inhibiting the expression of the OLA1 gene.
[0010] Preferably, the small molecule compound is selected from RNA interference agents, antisense oligonucleotides, or OLA1 protein inhibitors.
[0011] Preferably, the drug also includes other medically acceptable adjuvants.
[0012] This application also provides a drug for inhibiting myocardial fibrosis, said drug inhibiting myocardial fibrosis by inhibiting the expression of OLA1.
[0013] Preferably, the drug contains at least one of siRNA, CRISPR, and small molecule compounds for inhibiting the expression of the OLA1 gene.
[0014] Preferably, the drug also includes other medically acceptable adjuvants.
[0015] Compared with the prior art, this application has at least the following beneficial effects:
[0016] This application reveals for the first time the direct link between the OLA1 gene and the pathological process of myocardial fibrosis. Through cardiomyocyte-specific OLA1 gene knockout experiments, it directly demonstrates that inhibiting OLA1 expression can significantly improve the prognosis of myocardial fibrosis. In the myocardial fibrosis model, the degree of myocardial fibrosis in gene knockout mice was significantly milder than that in the control group. This core result not only clarifies the key role of the OLA1 gene in the fibrotic process but also directly supports its effectiveness as a therapeutic target, providing a new direction for the development of targeted drugs for myocardial fibrosis.
[0017] This application demonstrates through specific verification experiments that inhibiting the OLA1 gene can effectively block the abnormalities in core pathological markers caused by myocardial fibrosis. Specifically, it significantly inhibits the elevated gene expression levels of the fibrosis markers COL1A1 and COL3A1 in cardiac tissue. The overexpression of these two markers is a key molecular characteristic of abnormal extracellular matrix deposition and an important basis for assessing the severity of fibrosis. This effect indicates that the OLA1 inhibition strategy can directly target the core aspects of the fibrosis pathological process, producing a targeted improvement effect on the pathological changes in cardiac tissue caused by the disease.
[0018] Furthermore, inhibiting the OLA1 gene can significantly reduce cardiac dysfunction caused by myocardial fibrosis, an effect with significant clinical implications. Cardiac dysfunction is one of the most serious complications of myocardial fibrosis, directly impacting patients' quality of life and survival. Experiments using small animal M-mode ultrasound confirmed that gene knockout mice showed better results than the control group in key cardiac function indicators such as ejection fraction (EF%) and left ventricular shortening fraction (FS%). This means that a treatment strategy targeting OLA1 can not only inhibit fibrosis itself but also directly protect the heart's pumping function, providing an effective means for the prevention and treatment of myocardial fibrosis-related cardiac damage and greatly enhancing the clinical application value of this treatment approach. Attached Figure Description
[0019] Figure 1 This is a schematic diagram of constructing a cardiomyocyte-specific OLA1 gene knockout mouse using the Cre-Loxp system according to one embodiment of the present invention; gene identification and Western blot verification of knockout efficiency.
[0020] Figure 2 This is a schematic diagram of masson staining in one embodiment of the present invention.
[0021] Figure 3 In one embodiment of the present invention, RT-qPCR was used to detect the mRNA levels of myocardial fibrosis markers COL1A1 and COL3A1 in mouse heart tissue.
[0022] Figure 4 In one embodiment of the present invention, small animal M-mode ultrasound is used to detect cardiac function indicators such as ejection fraction (EF%) and left ventricular shortening fraction (FS%) in mice.
[0023] (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001) Detailed Implementation
[0024] This application provides the use of OLA1 as a target in the preparation of drugs to inhibit myocardial fibrosis, wherein the drugs are used to inhibit the expression of the OLA1 gene, and the sequence of OLA1 is shown in SEQ ID NO: 01.
[0025] In one embodiment, the drug comprises at least one of siRNA, CRISPR, and a small molecule compound for inhibiting the expression of the OLA1 gene.
[0026] The small molecule compound is selected from RNA interference agents, antisense oligonucleotides, or OLA1 protein inhibitors.
[0027] The drug also includes other medically acceptable adjuvants.
[0028] In addition, this application also provides a drug for inhibiting myocardial fibrosis, which inhibits myocardial fibrosis by inhibiting the expression of OLA1.
[0029] The above content will be explained in conjunction with specific verification experiments below.
[0030] I. Experimental Materials and Sources
[0031]
[0032] II. Experimental Procedure
[0033] 1. Establishment of a mouse model of myocardial fibrosis induced by transverse aortic arch constriction (TAC)
[0034] (1) Grouping of animal models:
[0035] Select 8-week-old male C57 BL / 6 Ckmm-Cre; OLA1 flox / flox and C57 BL / 6 OLA1 flox / flox Ten mice were randomly assigned to either aortic arch coarctation (TAC) or sham surgery and were divided into four groups: OLA1 flox / flox +Sham group, OLA1 flox / flox +TAC group, Ckmm-Cre; OLA1 flox / flox +Sham group, Ckmm-Cre; OLA1 flox / flox +TAC group.
[0036] (2) Constructing a model of myocardial hypertrophy:
[0037] A. All instruments were autoclaved and sterilized the day before the experiment.
[0038] B. Fast all experimental animals for 12 hours before the experiment. During the experiment, mice were anesthetized with 5% isoflurane. After the mice were anesthetized, they were transferred to a heating pad and kept under anesthesia with 1.5% isoflurane. The limbs were fixed with medical tape and all the hair on the mice's chest was shaved off.
[0039] C. Using scissors, make an incision in the middle of the mouse's neck (0.5 cm above the upper edge of the sternum to 0.5 cm below the upper edge of the sternum), bluntly dissect the thyroid gland and neck muscles, expose the mouse's trachea, and then cut the sternum about 0.5-1 cm along the midline of the neck.
[0040] D. Use a homemade fixation device to hook the mouse's sternum, bluntly dissect the thymus, expose the neck blood vessels, use a needle to pick out the aortic arch on the right side of the left common carotid artery, use forceps to hold the needle and place a No. 6 suture under the aortic arch and pull it out (avoid puncturing blood vessels and avoid pneumothorax).
[0041] E. Use a 1 mL syringe needle (27 G) to make a shim and place it parallel to the aortic arch. After ligation, remove the shim.
[0042] F. If it is a sham surgery group, then only the sternum needs to be cut open and no ligation is required.
[0043] G. Turn off the gas anesthesia device, place the mouse on the heating pad, and wait for it to wake up naturally.
[0044] 2. Six weeks after TAC, the effect of OLA1 gene deficiency on TAC-induced myocardial fibrosis in mice was determined by masson staining and RT-qPCR.
[0045] III. Analysis of Experimental Results:
[0046] Please see Figure 1 Gene identification and Western blot techniques were used to identify the OLA1 knockout efficiency in cardiomyocytes.
[0047] like Figure 2 As shown, OLA1 in a TAC-induced myocardial fibrosis mouse model flox / flox The group with the most severe TAC-induced myocardial fibrosis was Ckm-Cre lacking OLA1; OLA1 flox / flox The group inhibited TAC-induced myocardial fibrosis.
[0048] Please see Figure 3 As can be seen from the figure, OLA1 flox / flox The +TAC group showed the highest expression levels of the myocardial fibrosis markers COL1A1 and COL3A1 in cardiac tissue, while the Ckmm-Cre group lacked OLA1; OLA1 flox / flox The expression levels of the myocardial fibrosis markers COL1A1 and COL3A1 were significantly reduced in the cardiac tissue of the +TAC group.
[0049] like Figure 4 As shown, with Ckmm-Cre; OLA1 flox / flox +TAC group mice compared to OLA1 flox / flox The +TAC group mice showed a more pronounced decline in cardiac function due to myocardial fibrosis.
[0050] Based on the above conclusions, this application verifies through in vivo experiments that cardiomyocyte-specific OLA1 gene knockout mice exhibit milder myocardial fibrosis after TAC.
[0051] In summary, this application, through specific validation experiments, demonstrated that in a myocardial fibrosis model, cardiomyocyte-specific OLA1 gene knockout mice exhibited milder myocardial fibrosis and decreased cardiac function compared to control mice. This result not only demonstrates the crucial role of the OLA1 gene in the pathological progression of myocardial fibrosis but also clearly supports its effectiveness as a therapeutic target. Further experimental results showed that inhibiting the OLA1 gene effectively suppressed the elevated expression levels of fibrosis markers COL1A1 and COL3A1 genes in cardiac tissue caused by myocardial fibrosis. These changes in physiological indicators are important evidence for assessing disease severity and treatment efficacy, indicating that OLA1 gene inhibition can have a positive impact on the cardiac pathological changes induced by myocardial fibrosis. More importantly, inhibiting the OLA1 gene also significantly reduced cardiac function decline caused by myocardial fibrosis. Decreased cardiac function is one of the serious complications caused by myocardial fibrosis, often leading to poor patient prognosis. This finding suggests that the strategy of inhibiting the OLA1 gene may provide a new means for the prevention and treatment of cardiac damage associated with myocardial fibrosis. Based on the above results, this application reveals the crucial role of the OLA1 gene in the pathological process of myocardial fibrosis and confirms its potential clinical value as a therapeutic target. This discovery provides a foundation and new direction for developing novel targeted drugs and improving the treatment efficacy of myocardial fibrosis.
Claims
1. The application of OLA1 as a target in the preparation of drugs to inhibit myocardial fibrosis, characterized in that: The sequence of OLA1 is shown in SEQ ID NO:
01.
2. The application of OLA1 as a target in the preparation of drugs for inhibiting myocardial fibrosis according to claim 1, characterized in that: The drug is used to inhibit the expression of the OLA1 gene.
3. The application of OLA1 as a target in the preparation of drugs for inhibiting myocardial fibrosis according to claim 2, characterized in that: The drug contains at least one of siRNA, CRISPR, and small molecule compounds for inhibiting the expression of the OLA1 gene.
4. The application of OLA1 as a target in the preparation of drugs for inhibiting myocardial fibrosis according to claim 3, characterized in that: The small molecule compound is selected from RNA interference agents, antisense oligonucleotides, or OLA1 protein inhibitors.
5. The application of OLA1 as a target in the preparation of drugs for inhibiting myocardial fibrosis according to claim 4, characterized in that: The drug also includes other medically acceptable adjuvants.
6. A drug for inhibiting myocardial fibrosis, characterized in that: The drug inhibits myocardial fibrosis by suppressing the expression of OLA1.
7. A drug for inhibiting myocardial fibrosis according to claim 6, characterized in that: The drug contains at least one of siRNA, CRISPR, and small molecule compounds for inhibiting the expression of the OLA1 gene.
8. A drug for inhibiting myocardial fibrosis according to claim 7, characterized in that: The drug also includes other medically acceptable adjuvants.