ROCK inhibitor converts corneal endothelial cells into a phenotype capable of regenerating in vivo endothelial tissue.
Corneal endothelial dysfunction accompanied by visual disturbance is a primary indication for corneal transplantation. We previously reported that the adhesion of corneal endothelial cells (CECs) to a substrate was enhanced by the selective ROCK inhibitor Y-27632. It is hypothesized that the inhibition of ROCK signaling may manipulate cell adhesion properties, thus enabling the transplantation of cultivated CECs as a form of regenerative medicine. In the present study, using a rabbit corneal endothelial dysfunction model, the transplantation of CECs in combination with Y-27632 successfully achieved the recovery of corneal transparency. Complications related to cell injection therapy, such as the abnormal deposition of the injected cells as well as the elevation of intraocular pressure, were not observed. Reconstructed corneal endothelium with Y-27632 exhibited a monolayer hexagonal cell shape with a normal expression of function-related markers, such as ZO-1, and Na(+)/K(+)-ATPase, whereas reconstruction without Y-27632 exhibited a stratified fibroblastic phenotype without the expression of markers. Moreover, transplantation of CECs in primates in the presence of the ROCK inhibitor also achieved the recovery of long-term corneal transparency with a monolayer hexagonal cell phenotype at a high cell density. Taken together, these results suggest that the selective ROCK inhibitor Y-27632 enables cultivated CEC-based therapy and that the modulation of Rho-ROCK signaling activity serves to enhance cell engraftment for cell-based regenerative medicine.
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Naoki
N
Okumura
Noriko
N
Koizumi
Morio
M
Ueno
Yuji
Y
Sakamoto
Hiroaki
H
Takahashi
Hideaki
H
Tsuchiya
Junji
J
Hamuro
Shigeru
S
Kinoshita
D000577:Amides / Q000494:pharmacology* / Q000627:therapeutic use; D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D003131:Combined Modality Therapy; D065306:Corneal Injuries; D016039:Corneal Transplantation / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D042783:Endothelial Cells / Q000187:drug effects / Q000502:physiology / Q000637:transplantation; D004728:Endothelium, Corneal / Q000187:drug effects* / Q000293:injuries / Q000502:physiology / Q000637:transplantation; D004791:Enzyme Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D008252:Macaca fascicularis; D010641:Phenotype; D011725:Pyridines / Q000494:pharmacology* / Q000627:therapeutic use; D011817:Rabbits; D012038:Regeneration / Q000187:drug effects* / Q000502:physiology; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
D000577:Amides; D004791:Enzyme Inhibitors; D011725:Pyridines; C108830:Y 27632; D054460:rho-Associated Kinases