Application of PRDX1 in preparation of sensitizer or reversion drug for ovarian cancer treatment drug

Abstract

The invention relates to application of PRDX1 in preparation of a sensitizer or a drug for reversing resistance of ovarian cancer drugs. The invention shows that the sensitivity of the PARP inhibitor for the ovarian cancer can be remarkably improved by reducing the PRDX1, a new solution is provided for the improvement of the maintenance and treatment effect of the PARP inhibitor drug-resistant ovarian cancer by related research results, and a product taking the PRDX1 as a target is provided for improving the drug resistance of the PARP inhibitor.

Description

Technical Field

[0001] The present invention belongs to the technical field of sensitizers, and particularly relates to the application of PRDX1 in preparing a sensitizer for treating ovarian cancer drugs or a drug for reversing drug resistance. Background Art

[0002] Malignant ovarian tumors account for 23-27% of female genital tract malignancies, ranking third in terms of incidence and highest in terms of mortality. Since the ovaries are retroperitoneal organs, ovarian tumors have a high degree of concealment, and about 70% of ovarian cancer patients are already in the advanced stage when the disease is discovered. The initial treatment mode for ovarian cancer is cytoreductive surgery and platinum-based combination chemotherapy. Although most patients can achieve clinical remission after initial treatment, 70% of the patients still relapse within 2-3 years, and the 5-year survival rate has been hovering around 40%. In recent years, with the iterative update of high-throughput whole-genome sequencing technology and the development and clinical transformation of composite biomarkers, maintenance treatment based on targeted drugs has become an important strategy for delaying disease recurrence.

[0003] Multiple high-level evidence-based medical evidences show that for patients with newly diagnosed advanced high-grade serous and endometrioid ovarian cancer who achieve complete remission or partial remission after initial chemotherapy, the application of polyADP-ribose polymerase (PARP) inhibitor for first-line maintenance treatment can significantly prolong the progression-free survival (PFS) time, especially for patients with BRCAm or / and homologous recombination repair deficiency (HRD). However, for HRD-negative patients, who account for about 50% of ovarian cancer patients, the efficacy of PARP inhibitors is relatively limited. In addition, the acquired resistance in the clinical application of PARP inhibitors has become increasingly prominent, which has become a huge obstacle to further expanding its clinical potential. Therefore, the phenomenon and mechanism of PARP inhibitor resistance have attracted more and more attention from domestic and foreign researchers.

[0004] Redox reaction is a basic intracellular biological reaction process that plays a key role in cell metabolism, cell differentiation, immune response, and various chemical transformations. Researchers have observed in vitro that various tumor cell lines, including melanoma, colon cancer, neuroblastoma, and ovarian cancer, can produce a large amount of hydrogen peroxide (H2O2) at a rate of up to 0.5 nmol / (10^4 cells·h). In eukaryotic cells, a variety of antioxidant factors constitute the cell defense system, including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidases (GPxs) with substrate affinity for hydroperoxides, and the peroxiredoxin 1 (PRDX1) / thioredoxin (TXN) system, as well as non-enzymatic preparations such as vitamins C and E. During the cancer process, antioxidant enzymes play an important dual role. On the one hand, antioxidant enzymes can effectively prevent DNA oxidation caused by the accumulation of harmful oxidants ADDIN EN.CITEADDIN EN.CITE.DATA ; on the other hand, researchers have demonstrated that a variety of tumor cells rely on upregulated expression of antioxidant enzymes for tumor progression and metastasis. Therefore, therapies involving antioxidant, pro-oxidant, or targeting antioxidant enzyme expression may have certain prospects in tumor treatment.

[0005] The peroxiredoxin family (PRDXs) is a thiol-dependent peroxidase superfamily. Among the six members of the PRDXs family, peroxiredoxin 1 (PRDX1) has the highest abundance in various mammalian tissues, making it an indispensable role in redox homeostasis. Previous studies have observed elevated expression levels of PRDX1 in various cancer types, suggesting that PRDX1 may be involved in tumorigenesis. Further research has shown that in addition to its initial role as an antioxidant to protect cells from oxidative stress damage, PRDX1 is also involved in regulating other cellular behaviors such as apoptosis, cell cycle progression, DNA damage response, and anti-tumor immunity. In addition, targeting PRDX1 has been proven effective in preclinical studies of various cancers, indicating that anti-PRDX1 therapy has great therapeutic potential. Recently, researchers have confirmed that celastrol is a covalent inhibitor of PRDX1 and constructed a celastrol derivative with reduced nephrotoxicity, which has shown good performance in the treatment of intestinal cancer.

[0006] Mitophagy is a selective autophagy process that maintains cellular energy homeostasis and metabolic balance by clearing damaged or dysfunctional mitochondria. Mitophagy may play different roles in different stages and different types of tumor cells: Mitophagy can inhibit tumor development by removing dysfunctional mitochondria, while eliminating dysfunctional mitochondria also helps protect the tumor mitochondrial functional network to maintain the unlimited growth of tumor cells.

[0007] Currently, in female reproductive system malignancies, there are few studies on mitophagy. Especially in the field of ovarian cancer, only a few studies have deeply discussed the role of mitophagy in tumorigenesis and development. Therefore, further studying the mechanism of PARP inhibitor resistance and exploring the connection between PARP inhibitor resistance and mitophagy, so as to develop new targeted therapies or combination therapy strategies, has very important clinical significance. Summary of the Invention

[0008] The technical problem to be solved by the present invention is to provide the application of PRDX1 in preparing a sensitizer or a resistance reversing agent for drugs for treating ovarian cancer, indicating that downregulating PRDX1 can significantly increase the sensitivity of ovarian cancer to PARP inhibitors. The relevant research results will provide a new solution for improving the maintenance treatment efficacy of PARP inhibitor-resistant ovarian cancer, and propose products targeting PRDX1 for improving the problem of PARP inhibitor resistance.

[0009] The present invention provides the application of PRDX1 in preparing a sensitizer or a resistance reversing agent for drugs for treating ovarian cancer.

[0010] Further, the drug for treating ovarian cancer is a PARP inhibitor.

[0011] Further, the drug also contains a pharmaceutically acceptable carrier and / or excipient.

[0012] Further, the pharmaceutically acceptable carrier and / or excipient includes at least one of a diluent, a binder, a surfactant, a humectant, an adsorption carrier, a lubricant, a filler, and a disintegrant.

[0013] Further, the dosage form of the drug includes at least one of tablets, pills, powders, solutions, suspensions, emulsions, and granules.

[0014] Beneficial effects The present invention indicates that downregulating PRDX1 can significantly increase the sensitivity of ovarian cancer to PARP inhibitors. The relevant research results will provide a new solution for improving the maintenance treatment efficacy of PARP inhibitor-resistant ovarian cancer, and propose products targeting PRDX1 for improving the problem of PARP inhibitor resistance. Brief Description of the Drawings

[0015] Figure 1 For clinical database - related analysis. A: Data in the CPTAC dataset indicate that PRDX1 is highly expressed in OC compared with normal ovarian tissue; B: In ovarian cancer samples from the UALCAN database, the high expression of PRDX1 in tumor tissues is stage - dependent; C: In ovarian cancer samples from the UALCAN database, the high expression of PRDX1 in tumor tissues is grade - dependent; D: In the TCGA - GTEx combined database samples, PRDX1 is highly expressed in various tumor tissues compared with normal tissues; E, F: In patients with high - grade serous ovarian cancer in the Kaplan–Meier plotter database, high expression of PRDX1 is associated with poor OS. E is for all patients, and F only includes patients who have undergone debulking surgery; G: The relative mRNA expression levels of PRDX1 in various ovarian cancer cell lines compared with the normal ovarian epithelial cell line IOSE - 80; H: The protein expression of PRDX1 in various ovarian cancer cell lines and the normal ovarian epithelial cell line IOSE - 80; * P <0.05, ** P <0.01, *** P <0.001.

[0016] Figure 2 For cell phenotype analysis after PRDX1 knockdown or overexpression. A: Changes in the cell growth curve after down - regulating or overexpressing PRDX1 in the SKOV3, SKOV3 - NR, and A2780 cell lines; B: Changes in nilaparib IC 50 after down - regulating or overexpressing PRDX1 in the SKOV3, SKOV3 - NR, and A2780 cell lines; C: After down - regulating PRDX1 in the SKOV3 - NR and A2780 cell lines and stimulating with nilaparib, the nuclear γ - H2AX foci increased significantly, and the formation of nuclear RAD51 foci was significantly blocked; D: After down - regulating PRDX1 in the SKOV3, SKOV3 - NR, and A2780 cell lines and stimulating with nilaparib, the percentage of trailing DNA in the comet assay increased and the trailing length increased; E: Detection of the positive correlation between the nilaparib IC 50 value and PRDX1 expression in organoids constructed from tumor tissues of patients with high - grade serous ovarian cancer; F: Detection of the dual - drug combination score (ZIP synergy score) by co - treating the SKOV3 - NR cell line with nilaparib and the PRDX1 inhibitor Celastrol. * P <0.05, ** P <0.01, *** P <0.001.

[0017] Figure 3After overexpressing PRDX1 in the HEK-293t cell line, mitochondria and cytosolic fractions were isolated and bound to the PRDX1 antibody respectively for IP-MS analysis. A: Venn diagram of IP-bound proteins in each group; B: Protein-protein interaction network analysis of differentially expressed proteins between the HEK-293t-oePRDX1-mito and HEK-293t-oeNC-mito groups; C: KEGG analysis of the HEK-293t-oePRDX1-mito and HEK-293t-oeNC-mito groups; D: In the public database GSE249514, GO enrichment analysis of differentially expressed genes in transcriptomes of niraparib-sensitive and -resistant prostate cancer cells. Genes upregulated in niraparib-resistant prostate cancer cells can be enriched in multiple pathways related to mitochondria.

[0018] Figure 4 Transmission electron microscopy was used to observe the morphology of mitochondria. Specific embodiments

[0019] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. In addition, it should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

[0020] Example 1 In vitro experiments demonstrated the role of PRDX1 in PARP inhibitor resistance in ovarian cancer: Using the sequencing analysis results of the stably constructed CRISPR-Cas9 knockout cell line SKOV3-GeCKO after treatment with niraparib and amplification, combined with public databases and literature retrieval, the PRDX1 gene ( Figure 1 A-H) was screened out.

[0021] Verification that knocking out PRDX1 enhances the sensitivity to niraparib: Using small interfering RNA transfection technology, the cell growth was detected by the CCK-8 method in SKOV3, SKOV3 niraparib-resistant strain (SKOV3-NR), and A2780 cells respectively; the niraparib IC50 values of each group were calculated ( Figure 2 A, B); immunofluorescence was used to detect the expression of γ-H2AX foci and RAD51 in the nucleus to observe the DNA damage repair situation ( Figure 2 C); comet electrophoresis was used to detect the DNA damage of cells ( Figure 2 D); in SKOV3, SKOV3-NR, and A2780 cells, the dual-drug synergy scores (ZIP synergy score) of the PRDX1 inhibitor and niraparib were measured respectively ( Figure 2F). Forty ovarian cancer organoid models were constructed, and the drug sensitivity experiment of niraparib and transcriptome sequencing were performed to determine the expression level of PRDX1, and the correlation between the expression level of PRDX1 and the sensitivity to niraparib was analyzed ( Figure 2 E). Organoids with high and low expression of PRDX1 were selected. Before and after using niraparib, the cell growth was detected by ATP luminescence method, and the IC50 values of each group were calculated. It was Figure 2 found that after knocking down PRDX1, the growth rates of all three cell lines slowed down, the IC50 for niraparib decreased, and the degree of DNA damage increased after treatment with niraparib.

[0022] Prove the molecular mechanism of PRDX1 regulating PARP inhibitor resistance: Explore and verify the interacting proteins of PRDX1: Overexpress PRDX1 in the tool cell HEK-293T, isolate mitochondria and cytosolic fractions, detect PRDX1-binding proteins by IP-MS, and perform KEGG and GO analyses ( Figure 3 A-D), and it was found that PRDX1 directly binds to TUFM in mitochondria. Recent research results have shown that TUFM is involved in the Parkin-PINK1-mediated mitophagy pathway, suggesting that PRDX1 may affect mitophagy by regulating TUFM. Observation under transmission electron microscope of SKOV3-NR cells and SKOV3-NR-shPRDX1 cells after treatment with niraparib showed ( Figure 4 ) that after downregulating PRDX1, the mitochondrial morphology shrank and the cristae structure disappeared.

[0023] This invention shows that downregulating PRDX1 can significantly increase the sensitivity of ovarian cancer to PARP inhibitors. The relevant research results will provide a new solution for improving the maintenance treatment efficacy of PARP inhibitor-resistant ovarian cancer, and propose products targeting PRDX1 for improving the problem of PARP inhibitor resistance.

Claims

1. Use of PRDX1 in preparing a sensitizer or a reversal agent for a drug for treating ovarian cancer.

2. The application according to claim 1, wherein: The drug for treating ovarian cancer is a PARP inhibitor.

3. The application according to claim 1, characterized in that: The drug further comprises a pharmaceutically acceptable carrier and / or excipient.

4. The application according to claim 3, characterized in that: The pharmaceutically acceptable carrier and / or excipient includes at least one of a diluent, a binder, a surfactant, a humectant, an adsorption carrier, a lubricant, a filler, and a disintegrant.

5. The application according to claim 1, characterized in that: The dosage form of the drug includes at least one of tablets, pills, powders, solutions, suspensions, emulsions, and granules.

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