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US-20260124247-A1 - METHODS FOR MAKING AUDITORY PROGENITOR CELLS AND USES THEREOF

US20260124247A1US 20260124247 A1US20260124247 A1US 20260124247A1US-20260124247-A1

Abstract

Methods are described for preparing auditory progenitor cells from gingival mesenchymal cells, for uses such as restoring hearing in hearing impaired individuals.

Inventors

  • Alireza MOSHAVERINIA
  • Sevda POURAGHAEI SEVARI

Assignees

  • THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

Dates

Publication Date
20260507
Application Date
20251106

Claims (20)

  1. 1 . A method of treating hearing loss associated with loss of sensory neurons in a human subject, the method comprising: a. obtaining a population of gingival mesenchymal stem cells (GMSCs); b. optionally expanding the population of GMSCs in vitro; c. encapsulating the population of GMSCs in an elastic three-dimensional scaffold; d. exposing the encapsulated population of GMSCs to a composition comprising one or more growth factors; e. allowing a sufficient period for the population of GMSCs to differentiate towards auditory progenitor cells; f. optionally retrieving the auditory progenitor cells from the scaffold; and g. introducing the auditory progenitor cells into the inner ear of the subject.
  2. 2 . The method of claim 1 wherein the population of GMSCs are obtained from gingival tissue of a donor.
  3. 3 . The method of claim 2 wherein the donor is the human subject.
  4. 4 . The method of claim 1 wherein the elasticity of the scaffold is a Young's modulus between about 1 kPa and about 30 kPa.
  5. 5 . The method of claim 4 wherein the elasticity of the scaffold is a Young's modulus is between about 1 kPa and about 12 kPa.
  6. 6 . The method of claim 4 wherein the elasticity of the scaffold is a Young's modulus is between about 1 kPa and about 6 kPa.
  7. 7 . The method of claim 1 wherein the elasticity of the scaffold is equal to or below an elastic modulus calculated from the slope of the initial portion of the loading curve of RGD-ALG/10-MG depicted in FIG. 3 C .
  8. 8 . The method of claim 1 wherein the elasticity of the scaffold is equal to or below an elastic modulus calculated from the slope of the initial portion of the loading curve of RGD-ALG/20-MG depicted in FIG. 3 C .
  9. 9 . The method of claim 1 wherein the one or more growth factors are selected from basic fibroblast growth factor (bFGF), insulin-like growth factor 1 (IGF-1), epidermal growth factor (EGF), BMP4, FGF-2, SB-431542, CHIR99021, LDN-193189 or any combination thereof.
  10. 10 . The method of claim 1 wherein the one or more growth factors are a combination of basic fibroblast growth factor (bFGF), insulin-like growth factor 1 (IGF-1), and epidermal growth factor (EGF).
  11. 11 . The method of claim 1 wherein the scaffold comprises a hydrogel.
  12. 12 . The method of claim 11 wherein the hydrogel is a nanocomposite hydrogel.
  13. 13 . The method of claim 11 wherein the hydrogel comprises RGD peptide.
  14. 14 . The method of claim 11 wherein the hydrogel comprises alginate crosslinked with a divalent cation.
  15. 15 . The method of claim 11 wherein the hydrogel comprises RGD-alginate.
  16. 16 . The method of claim 11 wherein the hydrogel comprises MATRIGEL.
  17. 17 . The method of claim 16 wherein the hydrogel comprises about at least 10% MATRIGEL.
  18. 18 . The method of claim 17 wherein the hydrogel comprises about 10% to about 20% MATRIGEL.
  19. 19 . The method of claim 11 wherein the hydrogel comprises RDG-alginate and at least 10% MATRIGEL.
  20. 20 . The method of claim 19 wherein the hydrogel comprises RDG-alginate and about 10% to about 20% MATRIGEL.

Description

CROSS REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. patent application Ser. No. 17/640,267, filed Mar. 3, 2022, which is a National Phase Application of PCT International Application No. PCT/US20/50468, International Filing Date Sep. 11, 2020, which claims benefit of U.S. Provisional Patent Application No. 62/899,570, filed Sep. 12, 2019, which are incorporated by reference herein in their entirety. GOVERNMENT SUPPORT This invention was made with government support under DE023825, awarded by the National Institutes of Health. The government has certain rights in the invention. INCORPORATION-BY-REFERENCE OF SEQUENCE LISTING The Sequence Listing XML associated with this application is provided electronically in XML file format and is hereby incorporated by reference into the specification. The name of the XML file containing the Sequence Listing XML is “P-589146-US1_SQL_ST.26_06NOV25.xml”. The XML file is 9,804 bytes, was created on Nov. 6, 2025, and is being submitted electronically via USPTO Patent Center. BACKGROUND Hearing loss is the most common sensory disability in humans, impairing the normal communication of more than 5% of population in industrialized societies. The core component of the cochlea, i.e., the organ of Corti, contains a highly ordered cellular mosaic of sensory hair cells (HCs) and non-sensory supporting cells (SCs). Once the sensory HCs are lost, the individual is shut off irreversibly from the hearing world. According to the World Health Organization (WHO), 466 million individuals are suffering from disabling hearing loss worldwide, and this number is estimated to exceed 900 million individuals by 2050. Hearing loss has a number of causes including genetic diseases, birth abnormalities, infections, certain medications, aging, and exposure to loud noises. Drug therapies (e.g., corticosteroids) and implantable auditory devices are among the current treatment modalities used to combat sensorineural hearing loss, with limited success in treating acute cases; however, there is not any effective medical intervention available for treating chronic hearing loss. A growing body of biotechnological approaches including stem cell and gene therapy have emerged recently and have the potential to regenerate the damaged tissue. However, numerous limitations and obstacles have thus far prevented these developments from being translated into clinical application. These limitations include a lack of ready access to the necessary number of stem cells and concerns over the long-term viability of gene therapy. Therefore, an effective method for the treatment of hearing loss is yet to be discovered. It is toward developing a treatment for hearing loss and the formation and utilization of auditory progenitor cells that the present invention is directed. SUMMARY OF THE INVENTION In one aspect, a method of treating hearing loss associated with loss of sensory neurons in a human subject is provided, the method comprising the steps of: a. obtaining a population of gingival mesenchymal stem cells (GMSCs);b. optionally expanding the population of GMSCs in vitro;c. encapsulating the population of GMSCs in an elastic three-dimensional scaffold;d. exposing the encapsulated population of GMSCs to a composition comprising one or more growth factors;e. allowing a sufficient period for the population of GMSCs to differentiate towards auditory progenitor cells;f. optionally retrieving the auditory progenitor cells from the scaffold; andg. introducing the auditory progenitor cells into the inner ear of the subject. In one embodiment, the population of GMSCs are obtained from gingival tissue of a donor. In one embodiment, the donor is the human subject. In one embodiment, the elasticity of the scaffold is a Young's modulus between about 1 kPa and about 30 kPa. In one embodiment, the elasticity of the scaffold is a Young's modulus is between about 1 kPa and about 12 kPa. In one embodiment, the elasticity of the scaffold is a Young's modulus is between about 1 kPa and about 6 kPa. In one embodiment, the elasticity of the scaffold is equal to or below an elastic modulus calculated from the slope of the initial portion of the loading curve of RGD-ALG/10-MG depicted in FIG. 3C. In one embodiment, the elasticity of the scaffold is equal to or below an elastic modulus calculated from the slope of the initial portion of the loading curve of RGD-ALG/20-MG depicted in FIG. 3C. In one embodiment, the one or more growth factors are selected from basic fibroblast growth factor (bFGF), insulin-like growth factor 1 (IGF-1), epidermal growth factor (EGF), BMP4, FGF-2, SB-431542, CHIR99021, LDN-193189 or any combination thereof. In one embodiment, the one or more growth factors are a combination of basic fibroblast growth factor (bFGF), insulin-like growth factor 1 (IGF-1), and epidermal growth factor (EGF). In one embodiment, the scaffold comprises a hydrogel. In one embodiment, the hydrogel is a nanocomposi