Reactivities of a Prostanoid EP2 Agonist, Omidenepag, Are Useful for Distinguishing between 3D Spheroids of Human Orbital Fibroblasts without or with Graves' Orbitopathy.

Yosuke Ida,Megumi Watanabe,Hiroshi Ohguro,Araya Umetsu,Masato Furuhashi,Hanae Ichioka,Fumihito Hikage

doi:10.3390/cells10113196

Yosuke Ida, Megumi Watanabe + Show 5 more

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https://doi.org/10.3390/cells10113196

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Journal: Cells	Publication Date: Nov 16, 2021
Citations: 2	License type: CC BY 4.0

Affiliation: Sapporo Medical University

Abstract

Background. To obtain new insights into the activation of the thyroid-stimulating hormone (TSH) and insulin-like growth factor 1 (IGF-1) receptors in human orbital fibroblasts (n-HOFs), the effects of the prostanoid EP2 agonist, omidenepag (OMD), and a rho-associated coiled-coil-containing protein kinase (ROCK) inhibitor, ripasudil (Rip) were evaluated using three-dimension (3D) n-HOFs spheroids in the absence and presence of the recombinant human TSH receptor antibodies, M22 and IGF-1. Methods. The effects of 100 nM OMD or 10 μM Rip on the physical properties, size, stiffness, and mRNA expression of several extracellular matrix (ECM) molecules, their regulator, inflammatory cytokines, and endoplasmic reticulum (ER) stress-related factors were examined and compared among 3D spheroids of n-HOFs, M22-/IGF-1-activated n-HOFs and GO-related human orbital fibroblasts (GHOFs). Results. The physical properties and mRNA expressions of several genes of the 3D n-HOFs spheroids were significantly and diversely modulated by the presence of OMD or Rip. The OMD-induced effects on M22-/IGF-1-activated n-HOFs were similar to the effects caused by GHOHs, but quite different from those of n-HOFs. Conclusions. The findings presented herein indicate that the changes induced by OMD may be useful in distinguishing between n-HOFs and GHOFs.

Highlights

Graves’ orbitopathy (GO), an autoimmune disease that affects orbital and periorbital tissues, demonstrates several characteristic clinical manifestations, including upper eyelid retraction, edema, and erythema of the periorbital tissues and conjunctivae, as well as exophthalmos [1,2]
Effects of an EP2 Agonist, OMD and the rho-associated coiled-coil-containing protein kinase (ROCK) Inhibitor, Rip on Physical Properties, Size and Stiffness of the 3D Spheroid Obtained from Non-GO-Related Human Orbital Fibroblast (n-Human Orbital Fibroblasts (HOFs)) and GO-Related Human Orbital Fibroblast (GHOF)
We reported that new types of anti-glaucoma drugs, including an EP2 agonist, OMD, and the ROCK inhibitor, Rip, induced different effects from those of the prostaglandin analogues (PGs) in 3D spheroids obtained from GO-related fibroblasts (GHOF) [11]

Summary

Introduction

Graves’ orbitopathy (GO), an autoimmune disease that affects orbital and periorbital tissues, demonstrates several characteristic clinical manifestations, including upper eyelid retraction, edema, and erythema of the periorbital tissues and conjunctivae, as well as exophthalmos [1,2]. To obtain new insights into the activation of the thyroid-stimulating hormone (TSH) and insulin-like growth factor 1 (IGF-1) receptors in human orbital fibroblasts (n-HOFs), the effects of the prostanoid EP2 agonist, omidenepag (OMD), and a rho-associated coiled-coil-containing protein kinase (ROCK) inhibitor, ripasudil (Rip) were evaluated using three-dimension (3D) n-HOFs spheroids in the absence and presence of the recombinant human TSH receptor antibodies, M22 and IGF-1. The effects of 100 nM OMD or 10 μM Rip on the physical properties, size, stiffness, and mRNA expression of several extracellular matrix (ECM) molecules, their regulator, inflammatory cytokines, and endoplasmic reticulum (ER) stress-related factors were examined and compared among 3D spheroids of n-HOFs, M22-/IGF-1-activated n-HOFs and GO-related human orbital fibroblasts (GHOFs).

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