Search

EP-4737560-A1 - MESENCHYMAL STEM CELL-DERIVED EXTRACELLULAR VESICLES AND USE THEREOF

EP4737560A1EP 4737560 A1EP4737560 A1EP 4737560A1EP-4737560-A1

Abstract

The present invention relates to human mesenchymal stem cell-derived extracellular vesicles and use thereof. The mesenchymal stem cell-derived extracellular vesicles according to the present invention inhibit the expression of fibrosis-related proteins, wound closure, and cell infiltration, which increase because of TGF-β1 treatment in lung epithelial cells. In addition, microRNAs, contained in large amounts in the extracellular vesicles, inhibit gene expression of fibrosis-related proteins in lung epithelial cells and lung fibroblasts. These extracellular vesicles significantly reduce, in lung fibrosis model mice, the expression of fibrosis-related proteins, collagen production and fibrosis regions. Therefore, the extracellular vesicles according to the present invention can be used to prevent and treat lung fibrosis.

Inventors

  • KIM, KYUNG SUK
  • LEE, TAE YONG

Assignees

  • CorestemChemon Inc.

Dates

Publication Date
20260506
Application Date
20240627

Claims (17)

  1. An extracellular vesicle comprising any one overexpressed microRNA selected from the group consisting of hsa-miR-148a-3p, hsa-miR-100-5p, hsa-miR-143-3p, hsa-let-7a-5p, hsa-let-7f-5p, and a combination thereof.
  2. The extracellular vesicle according to claim 1, wherein the extracellular vesicle comprises hsa-miR-148a-3p at 1 to 20% of the total microRNA content of the extracellular vesicle.
  3. The extracellular vesicle according to claim 1, wherein the extracellular vesicle comprises hsa-miR-100-5p at 1 to 20% of the total microRNA content of the extracellular vesicle.
  4. The extracellular vesicle according to claim 1, wherein the extracellular vesicle comprises hsa-miR-143-3p at 0.1 to 10% of the total microRNA content of the extracellular vesicle.
  5. The extracellular vesicle according to claim 1, wherein the extracellular vesicle comprises hsa-let-7a-5p at 0.1 to 10% of the total microRNA content of the extracellular vesicle.
  6. The extracellular vesicle according to claim 1, wherein the extracellular vesicle comprises hsa-let-7f-5p at 0.1 to 10% of the total microRNA content of the extracellular vesicle.
  7. The extracellular vesicle according to claim 1, wherein the microRNA exhibits the fibrosis inhibition activity.
  8. The extracellular vesicle according to claim 1, wherein the extracellular vesicle further comprise any one expressed microRNA selected from the group consisting of hsa-miR-151a-3p, hsa-let-7b-5p, hsa-miR-21-5p, hsa-miR-10a-5p, hsa-miR-92a-3p, hsa-miR-486-5p, hsa-miR-99b-5p, hsa-let-7i-5p, hsa-miR-320a, hsa-miR-409-3p, hsa-miR-10b-5p, hsa-miR-127-3p, hsa-miR-26a-5p, hsa-miR-221-3p, hsa-let-7e-5p, hsa-miR-493-5p, hsa-miR-25-3p, hsa-miR-574-3p, hsa-miR-423-5p, hsa-miR-122-5p, hsa-miR-382-5p, hsa-miR-155-5p, hsa-miR-451a, hsa-miR-30a-3p, hsa-miR-28-3p, hsa-let-7g-5p, hsa-miR-379-5p, hsa-miR-23a-3p, hsa-miR-485-5p, hsa-miR-30a-5p, hsa-miR-30d-5p, hsa-miR-423-3p, hsa-miR-99a-5p, hsa-let-7c-5p, hsa-miR-92b-3p, hsa-miR-323a-3p, hsa-miR-574-5p, hsa-miR-197-3p, hsa-miR-432-5p, hsa-miR-140-3p, hsa-miR-370-3p, hsa-miR-196a-5p, hsa-miR-654-5p, hsa-miR-184, hsa-miR-181a-2-3p, and a combination thereof.
  9. The extracellular vesicle according to claim 1, wherein the extracellular vesicle is isolated from a human-derived stem cell.
  10. The extracellular vesicle according to claim 9, wherein the stem cell is a mesenchymal stem cell.
  11. The extracellular vesicle according to claim 10, wherein the mesenchymal stem cell is derived from umbilical cord, umbilical cord blood, bone marrow, fat, muscle, nerve, skin, amniotic membrane, tooth, hair root cell, or placenta, or is differentiated from an induced pluripotent stem cell.
  12. The extracellular vesicle according to claim 1, wherein the particle size of the extracellular vesicle is 10 nm to 1,000 nm.
  13. A pharmaceutical composition for preventing or treating fibrosis, comprising the extracellular vesicle according to claim 1 as an active ingredient.
  14. The pharmaceutical composition for preventing or treating fibrosis according to claim 13, wherein the fibrosis is pulmonary fibrosis.
  15. The pharmaceutical composition for preventing or treating fibrosis according to claim 13, wherein the pulmonary fibrosis is any one or more selected from the group consisting of idiopathic pulmonary fibrosis, radiation-induced lung injury, nonspecific interstitial pneumonia, acute interstitial pneumonia, cryptogenic organizing pneumonia, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphoid interstitial pneumonia, interstitial pulmonary fibrosis and diffuse pulmonary fibrosis, pulmonary edema, cystic fibrosis, and pulmonary fibrosis caused by metabolic diseases.
  16. A use of the extracellular vesicle according to claim 1 for the prevention or treatment of fibrosis.
  17. A method for preventing or treating fibrosis, comprising administering the extracellular vesicle according to claim 1 to a subject.

Description

Technical Field The present invention relates to a human mesenchymal stem cell-derived extracellular vesicle and a use thereof. Background Art Idiopathic pulmonary fibrosis (IPF) is the most common disease of idiopathic interstitial pneumonia (IIP) within interstitial lung disease (ILD), but its exact cause remains unknown. Previously, chronic inflammation-induced fibrosis was considered the primary cause of idiopathic pulmonary fibrosis. Pirfenidone and nintedanib, approved by the FDA in 2014, are used as therapeutic agents for idiopathic pulmonary fibrosis. However, these therapeutic agents do not address the underlying cause and have been reported to cause tolerability and side effects. In addition, drugs currently under development through clinical trials are single-acting therapeutic agents, raising concerns about efficacy and side effects. Therefore, the development of new therapeutic agents with multiple therapeutic mechanisms is needed. Mesenchymal stem cells have been reported to be effective in treating idiopathic pulmonary fibrosis through their immunomodulatory, anti-inflammatory, and epithelial cell regeneration-promoting effects (Yang et al., Front Cell Dev Biol. (2021) 9:639657, Xie et al., Front Pharmacol. (2021) 11:590972). However, the biggest challenge with stem cells is that their engraftment and survival rates vary across patients, making it difficult to accurately demonstrate their efficacy. Furthermore, there is the potential for stem cells to transform into cancer cells. Therefore, research into treatments for pulmonary fibrosis utilizing mesenchymal stem cells is needed. Detailed Description of Invention Technical Problem Accordingly, the present inventors conducted research to develop a therapeutic agent for pulmonary fibrosis. As a result, the present inventors found that extracellular vesicles isolated from a culture solution of mesenchymal stem cells exhibit an excellent effect in treating pulmonary fibrosis in a pulmonary fibrosis model mouse, thereby completing the present invention. Solution to Problem In order to achieve the above object, in one aspect of the present invention, there is provided an extracellular vesicle comprising any one overexpressed microRNA selected from the group consisting of hsa-miR-148a-3p, hsa-miR-100-5p, hsa-miR-143-3p, hsa-let-7a-5p, hsa-let-7f-5p, and a combination thereof. In another aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating fibrosis, comprising the extracellular vesicle as an active ingredient. In another aspect of the present invention, there is provided a use of the extracellular vesicle for the prevention or treatment of fibrosis. In another aspect of the present invention, there is provided a method for preventing or treating fibrosis, comprising administering the extracellular vesicle to a subject. Effects of Invention The mesenchymal stem cell-derived extracellular vesicles according to the present invention inhibited the expression of fibrosis-related proteins, wound closure, and cell infiltration increased by TGF-β1 treatment in lung epithelial cells. In addition, microRNAs, contained in large amounts in the extracellular vesicles, inhibited gene expression of fibrosis-related proteins in lung epithelial cells and lung fibroblasts. These extracellular vesicles significantly reduced, in pulmonary fibrosis model mice, the expression of fibrosis-related proteins, collagen production and fibrosis regions. Therefore, the extracellular vesicles according to the present invention can be used to prevent and treat pulmonary fibrosis. Brief Description of Drawings Figure 1 is a schematic diagram showing extracellular vesicles secreted from human mesenchymal stem cells (hMSCs) derived from various human tissues and their use in the treatment of pulmonary fibrosis.Figure 2 is a schematic diagram showing a method for isolating human mesenchymal stem cell-derived extracellular vesicles according to one embodiment of the present invention.Figure 3 is a graph and table showing the particle size distribution and concentration of extracellular vesicles (hMSCs-EVs) isolated from a culture solution of mesenchymal stem cells derived from human umbilical cord, using a nanoparticle tracking analyzer, according to one embodiment of the present invention.Figure 4 is a diagram showing the results obtained by observing the double lipid membrane structure of hMSCs-EVs using a cryogenic transmission electron microscope, according to one embodiment of the present invention.Figure 5 is a diagram showing the results obtained by identifying markers in hMSCs-EVs using Western blotting, according to one embodiment of the present invention.Figure 6 is a graph showing the results obtained by identifying the phenotype of hMSCs-EVs using flow cytometry, according to one embodiment of the present invention.Figures 7a and 7b are graphs showing the results obtained by analyzing the intrinsic factors of hMSCs-EVs u