US-12616721-B2 - Stratified squamous epithelial cell normal differentiation and maturation promoting agent, epithelial disease therapeutic agent, and stratified squamous epithelial cell normal differentiation and maturation promoting method

Abstract

The present invention addresses the problem of providing a novel therapeutic agent that is effective against diseases involving stratified squamous epithelial cells such as dry eye syndrome. The present invention is an agent for promoting normal differentiation/maturation of stratified squamous epithelial cells, which comprises a secretion of mesenchymal stem cells. It is preferable that the secretions not include animal serum, that the mesenchymal stem cells be adipose-derived mesenchymal stem cells, umbilical cord-derived mesenchymal stem cells, or bone marrow-derived mesenchymal stem cells, and that the stratified squamous epithelial cells be at least one selected from the group consisting of corneal epithelial cells, conjunctival epithelial cells, epidermal keratinocytes, oral epithelial cells, epiglottic epithelial cells, esophageal epithelial cells, vaginal epithelial cells, vocal fold epithelial cells, nasal epithelial cells, and nasal vestibular epithelial cells.

Inventors

• Kohji Nishida
• Ryuhei Hayashi
• Shun Shibata
• Toru Okubo
• Yoichi Honma

Assignees

• OSAKA UNIVERSITY
• ROHTO PHARMACEUTICAL CO., LTD.

Dates

Publication Date

20260505

Application Date

20190111

Priority Date

20180112

Claims (5)

1. 1 . A method for promoting normal differentiation or maturation of stratified squamous epithelial cells, comprising administering an effective amount of a culture supernatant of adipose-derived mesenchymal stem cells to a subject in need thereof, wherein the culture supernatant does not include animal serum and wherein administration of the culture supernatant results in increased expression of KRT12 in the epithelial cells.
2. 2 . The method for promoting normal differentiation or maturation of stratified squamous epithelial cells according to claim 1 , wherein the stratified squamous epithelial cells comprise at least one type of cells selected from the group consisting of corneal epithelial cells, conjunctival epithelial cells, epidermal keratinocytes, oral epithelial cells, epiglottis epithelial cells, esophageal epithelial cells, vaginal epithelial cells, vocal cord fold epithelial cells, nasal epithelial cells, and nasal vestibular epithelial cells.
3. 3 . A method for treating an epithelial disease involving a tissue having stratified squamous epithelial cells, comprising administering an effective amount of a culture supernatant of mesenchymal stem cells to a subject in need thereof, wherein, the culture supernatant does not include animal serum; the mesenchymal stem cells are adipose-derived mesenchymal stem cells; administration of the culture supernatant results in an increased expression of KRT12 in the epithelial cells; and the stratified squamous epithelial cells comprise at least one type of cells selected from the group consisting of corneal epithelial cells, conjunctival epithelial cells, epidermal keratinocytes, oral epithelial cells, epiglottis epithelial cells, esophageal epithelial cells, vaginal epithelial cells, vocal cord fold epithelial cells, nasal epithelial cells, and nasal vestibular epithelial cells.
4. 4 . The method for treating an epithelial disease involving a tissue having the stratified squamous epithelial cells according to claim 3 , wherein the epithelial disease is a corneal disease, a conjunctival disease, an oral disease, or an epidermal disease.
5. 5 . The method for treating an epithelial disease involving a tissue having the stratified squamous epithelial cells according to claim 4 , wherein the epithelial disease is dry eye, pterygium, scar, EB viral keratitis, corneal epithelial stem cell deficiency, Sjogren's syndrome, or scleroderma.

Description

TECHNICAL FIELD The present invention relates to an agent for promoting normal differentiation/maturation of stratified squamous epithelial cells, an epithelial disease therapeutic agent and a method for promoting normal differentiation/maturation of stratified squamous epithelial cells. BACKGROUND ART The ocular surface plays an important role in protecting the eye from outside infection sources. A major disease of the ocular surface, dry eye, is a chronic disease of tear fluid, corneal epithelium and conjunctival epithelium, caused by qualitative or quantitative abnormity of tear fluid and causes a sense of discomfort in the eye and visual abnormality. Of the dry-eye diseases, serious dry eye such as Sjogren's syndrome significantly decreases quality of life (QOL) and sometimes causes blindness. Note that, the number of Sjogren's syndrome patients in Japan is reported to be 70,000, and probably, 200,000 or more if the number of potential patients are included. Treatments for dry eye conventionally known include a method of placing an eye-drop using artificial tear; a method of using a moisture aid which consists of panels to be provided to both sides of a pair of glasses to prevent evaporation of tear; a punctual occlusion method, which is a method of closing a tear discharge port, punctum, to keep tear fluid in the conjunctival sac; a method of applying hyaluronic acid to keep moisture, and a method of promoting secretion of mucin and a moisture content by administering a drug. However, any one of the methods is unsatisfactory. In the meantime, development of medicines using cells and cell-culture supernatant have been broadly made. For instance, mesenchymal stem cells are precursor cells having pluripotency, which were, for the first time, isolated by Friedenstein from the bone marrow (see, Non Patent Document 1). It has been found that the mesenchymal stem cells are present in a wide variety of tissues such as bone marrow, umbilical cord and adipose tissues, and involved in repair of various tissues and have immunosuppressive effect. Transplantation of mesenchymal stem cells is expected as a novel therapy for various types of intractable diseases (see, Patent Documents 1 and 2). In addition, e.g., a method of using a pharmaceutical composition containing a culture supernatant of mesenchymal stem cells for repairing damaged sites of a target tissue (see, Patent Documents 3 and 4) and a method of using a pharmaceutical composition containing a culture supernatant of mesenchymal stem cells for treating a secondary fibromyalgia caused by diseases or damages (see, Patent Document 5) are also known. Non Patent Document 2 reports that stability of tear fluid can be improved by instilling mesenchymal stem cells themselves in a rat model having dry eye, which is induced by placing a drop of benzalkonium chloride, and discloses that, in this model, mesenchymal stem cells instilled infiltrate into the meibomian glands and conjunctival epithelium, and contribute to improvement of tear fluid stability. PRIOR ART DOCUMENTS Patent Document Patent Document 1: JP 2012-157263 APatent Document 2: JP 2012-508733 APatent Document 3: International Publication No. WO2011/118795Patent Document 4: JP 2010-540662 APatent Document 5: JP 2007-528703 A Non Patent Document Non Patent Document 1: Pittenger F. M. et. al., Science, 1999, 284, pp. 143-147Non Patent Document 2: Beyazyildiz E. et. al. Stem Cells Int. 2014; vol. 2014: 250230 SUMMARY OF INVENTION Technical Problem In the circumstance, it has been desired to develop a therapeutic agent having a higher effect than conventional ones on diseases such as dry eye, in which the stratified squamous epithelial cells are involved, particularly, a therapeutic agent based on a new mechanism of action. Then, an object of the present invention is to provide a novel therapeutic agent effective for a disease such as dry eye, in which stratified squamous epithelial cells are involved. Solution to Problem The present inventors conducted various studies with a view to attaining the object. As a result, they found that normal differentiation/maturation and formation of tight junction between cells can be promoted by adding a secretion from mesenchymal stem cells to corneal epithelial cells, thereby improving barrier function. Based on the finding, the present invention was accomplished. The present invention is summarized as follows. [1] An agent for promoting normal differentiation/maturation of stratified squamous epithelial cells, which comprises a secretion from mesenchymal stem cells.[2] The normal differentiation/maturation-promoting agent according to [1], wherein the secretion does not include animal serum.[3] The normal differentiation/maturation-promoting agent according to [1] or [2], wherein the mesenchymal stem cells are adipose-derived mesenchymal stem cells, umbilical cord-derived mesenchymal stem cells or bone marrow-derived mesenchymal stem cells.[4] The normal differentiatio