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KR-20260067994-A - Composition targeting glycosylation for preventing or treating osteoarthritis

KR20260067994AKR 20260067994 AKR20260067994 AKR 20260067994AKR-20260067994-A

Abstract

The present specification relates to a composition for the prevention or treatment of osteoarthritis targeting glycosylation, and more specifically, said composition can inhibit the abnormal accumulation of UDP-GlcNAc in the cytoplasm (e.g., cytoplasm of chondrocytes) and restore the normal distribution of UDP-GlcNAc to the endoplasmic reticulum and/or Golgi apparatus, thereby inhibiting excessive O-GlcNAc formation caused by cytoplasmic UDP-GlcNAc accumulation in chondrocytes, and ultimately preventing or treating osteoarthritis.

Inventors

  • 김진홍
  • 강찬희
  • 강동현
  • 이지연

Assignees

  • 서울대학교산학협력단

Dates

Publication Date
20260513
Application Date
20251030
Priority Date
20241105

Claims (15)

  1. A composition for the prevention or treatment of osteoarthritis, comprising: a preparation that increases the expression or activity of a gene belonging to solute carrier family 35 (SLC35); a protein encoded by said gene; or an inhibitor that inhibits the accumulation of uridine diphosphate N-acetylglucosamine (UDP-GlcNAc) in the cytoplasm of chondrocytes.
  2. A composition for the prevention or treatment of osteoarthritis according to claim 1, wherein the protein encoded by the gene moves uridine diphosphate N-acetylglucosamine within the cytoplasm of chondrocytes to the Golgi apparatus or endoplasmic reticulum.
  3. A composition according to claim 1, wherein the gene is at least one selected from the group consisting of the SLC35A2 gene, SLC35A3 gene, SLC35A5 gene, SLC35B4 gene, SLC35D1 gene, and SLC35D2 gene.
  4. A composition for the prevention or treatment of osteoarthritis according to claim 1, wherein the agent that increases the expression or activity of the gene is at least one selected from the group consisting of an antagonist small molecule compound, a vector, a plasmid, a viral vector, an adenovirus vector, an adeno-associated virus vector, a lentivirus vector, mRNA, mRNA wrapped in lipid nanoparticles (LNP), liposomes, and nanoparticles.
  5. The composition of claim 1, wherein the composition is administered simultaneously, separately, or sequentially in combination with at least one of a GATA4 inhibitor; or an O-linked-N-acetylglucosaminylation transferase (O-GlcNAc transferase, OGT) inhibitor.
  6. A composition according to claim 5, wherein the GATA4 inhibitor is 2-(1,3-benzodioxol-5-ylmethyl)butanedioic acid, a salt thereof, a hydrate thereof, or a solvate thereof; Gata4 siRNA consisting of the nucleotide sequences of SEQ ID NO. 1 (5′-GAGCAAACCAGAGCCUAGAdTdT-3′) and SEQ ID NO. 2 (5′-UCUAGGCUCUGGUUUGCUCdTdT-3'); or Gata4 siRNA consisting of the nucleotide sequences of SEQ ID NO. 3 (5′-CAGAUGUUACUGAAUGCUUdTdT-3′) and SEQ ID NO. 4 (5′-AAGCAUUCAGUAACAUCUGdTdT-3′).
  7. A composition according to claim 5, wherein the OGT inhibitor is 3-[2-(1-adamantyl)ethyl]-2-(4-chlorophenyl)imino-4-oxo-1,3-thiazinane-6-carboxylic acid, a salt thereof, a hydrate thereof, or a solvate thereof.
  8. A composition according to claim 1, wherein the osteoarthritis is post-traumatic osteoarthritis, inflammatory osteoarthritis, or degenerative osteoarthritis.
  9. A composition for diagnosing or predicting the prognosis of osteoarthritis, comprising: a preparation for measuring the expression level of a gene belonging to solute carrier family 35 (SLC35); a preparation for measuring the level of a protein encoded by said gene; or a preparation for measuring the level of uridine diphosphate N-acetylglucosamine.
  10. A composition according to claim 9, wherein the gene is at least one selected from the group consisting of the SLC35A2 gene, SLC35A3 gene, SLC35A5 gene, SLC35B4 gene, SLC35D1 gene, and SLC35D2 gene.
  11. A composition according to claim 9, wherein the composition is treated to chondrocytes isolated from a subject and measures i) the expression level of a gene belonging to SLC35 in said chondrocytes or the level of a protein encoded by said gene; or ii) the level of uridine diphosphate N-acetylglucosamine in the cytoplasm of said chondrocytes.
  12. In claim 9, the composition wherein the agent for measuring the expression level of the gene belonging to the SLC35 is a primer pair, a probe, an antisense oligonucleotide, a CRISPR guide RNA, or an aptamer that specifically binds to the mRNA of the gene.
  13. In claim 9, the preparation for measuring the level of the protein encoded by the said gene is a composition that is an antibody, aptamer, protein-binding peptide, protein-binding small molecule, nanobody, antibody fragment, or a functional derivative thereof that specifically binds to said protein.
  14. In claim 9, the composition wherein the preparation for measuring the level of uridine diphosphate N-acetylglucosamine (UDP-GlcNAc) is an analytical reagent for mass spectrometry, an enzyme, a chemical derivatization reagent based on fluorescence or chromogenic reaction, an antibody that specifically binds to a nucleotide-sugar, or an aptamer that specifically binds to a nucleotide-sugar.
  15. A composition for the prevention or treatment of osteoarthritis comprising at least one GATA4 inhibitor selected from the group consisting of 2-(1,3-benzodioxol-5-ylmethyl)butanedioic acid), its salt, its hydrate, or its solvate; or Gata4 siRNA consisting of the nucleotide sequence of SEQ ID NO. 1 (5′-GAGCAAACCAGAGCCUAGAdTdT-3′) and SEQ ID NO. 2 (5′-UCUAGGCUCUGGUUUGCUCdTdT-3').

Description

Composition targeting glycosylation for preventing or treating osteoarthritis This specification relates to a composition for the prevention or treatment of osteoarthritis targeting glycosylation, and a composition for the diagnosis or prognosis prediction of osteoarthritis. Osteoarthritis (OA) is a major cause of disability, with the number of patients increasing by 48% worldwide between 1990 and 2019. Various etiological risk factors, such as aging, obesity, and mechanical overload, contribute to the development and progression of osteoarthritis. The progression of osteoarthritis is characterized by pathological changes such as cartilage degeneration, subchondral bone sclerosis, osteophyte formation, and synovial inflammation. Osteoarthritis is characterized by the loss of proteoglycans (PGs) in articular cartilage and chronic inflammation. The disease is exacerbated by senescent chondrocytes accumulating in the cartilage, including in the superficial zone, which secrete SASPs containing inflammatory cytokines, chemokines, and cartilage matrix degrading enzymes. Uridine diphosphate N-acetylglucosamine (UDP-GlcNAc) is a sugar precursor constituting the glucosaminoglycan (GAG) chain and serves as a substrate for O-linked N-acetylglucosamine (O-GlcNAcylation). O-GlcNAcylation is a reversible post-translational modification catalyzed by O-GlcNAc transferase (OGT) and removed by O-GlcNAcase (OGA), and is known as an important mechanism integrating nutritional and metabolic states with the regulation of cellular responses. Dysregulated O-GlcNAcylation is associated with various human diseases, including cancer, type 2 diabetes, and cardiovascular disease. Meanwhile, glucosamine is used as a material to efficiently generate UDP-GlcNAc and is widely supplemented with the expectation that it will maintain proteoglycan levels in patients with osteoarthritis and the elderly, but its clinical effects are controversial. Despite the importance of cellular senescence and SASP in the pathogenesis of osteoarthritis, there is currently a lack of effective treatments capable of selectively regulating SASP in senescent chondrocytes. Figure 1a shows a heatmap of the fold change in expression levels of UDP-GlcNAc and UDP-GalNAc transporters in human osteoarthritis cartilage. The publicly available transcriptome datasets GSE16464, GSE43923, GSE64394, GSE113825, GSE117999, GSE178557, and GSE186220 were analyzed. Figure 1b shows a heatmap of the fold change in expression levels of UDP-GlcNAc and UDP-GalNAc transporters in chondrocytes treated with IL-1β. The publicly available transcriptome datasets GSE6119, GSE104793, and GSE163080 were analyzed. Figure 1c shows a heatmap of the fold change in expression levels of UDP-GlcNAc and UDP-GalNAc transporters in the cartilage of osteoarthritis mouse and rat models. Mouse datasets GSE26475, GSE53857, GSE101573, GSE110268, and GSE143447 and rat dataset GSE28958 were analyzed. Figure 1d shows SA-β-Gal staining and quantification of positive rate of C28/I2 chondrocytes treated with 100 nM doxorubicin (left), representative immunofluorescence images and BrdU integrated quantification (middle), and relative mRNA expression levels of CDK inhibitor or Lmnb1 (right). Figure 1e shows SA-β-Gal staining and positive rate quantification of mouse chondrocytes treated with 50 μg/mL bleomycin (left), representative immunofluorescence images and BrdU integrated quantification (middle), and relative mRNA expression levels of CDK inhibitor or Lmnb1 (right). Figure 1f shows the relative mRNA expression levels of UDP-GlcNAc and UDP-GalNAc carriers in C28/I2 chondrocytes treated with doxorubicin. Figure 1g shows the relative mRNA expression levels of UDP-GlcNAc and UDP-GalNAc carriers in mouse chondrocytes treated with bleomycin. Figure 1h shows the results of quantifying UDP-GlcNAc levels in the whole cell (left), endoplasmic reticulum, Golgi apparatus, and cytoplasmic compartment (right) of C28/I2 chondrocytes treated with vehicle, DON, or doxorubicin. Figure 1i shows the results of quantifying UDP-GlcNAc levels in the whole cells (left) and cytoplasm (right) of mouse chondrocytes treated with vehicle, DON, or bleomycin. Figure 1j shows the results of measuring the secretion of sulfated glycoprotein (sGAG) of C28/I2 chondrocytes treated with vehicle or doxorubicin using the sGAG assay. Figure 1k shows the secretion of sulfated glycoprotein (sGAG) from mouse chondrocytes treated with bleomycin. Scale bar: Fig. 1d (left), Fig. 1e (left), 50 μm, Fig. 1d (center), Fig. 1e (center), 25 μm. Fig. 1d-k, data represent mean ± standard error. P values were obtained from two-sided t-tests (Fig. 1d-Fig. 1g, Fig. 1j, Fig. 1k) or one-way ANOVA with Dunnett post-hoc test (Fig. 1h, Fig. 1i). Figure 2a shows representative co-immunofluorescence images of O-GlcNAc and p16 INK4a obtained by staining intact and damaged sections of human osteoarthritis cartilage with Alcian blue and immunostaining them for O-GlcNAc and p16 IN