ABSTRACT
Background/Aims@#Tacrolimus has been used as an immunosuppressive agent in organ transplantation. Despite the therapeutic benefits, tacrolimus’s use is limited due to its nephrotoxicity. To reduce tacrolimus nephrotoxicity, effective humanized experimental models may be helpful. Here, we modeled tacrolimus nephrotoxicity using kidney organoids derived from human inducible pluripotent stem cells (iPSCs) in vitro. @*Methods@#Kidney organoids were differentiated from the CMC11 iPSC cell line, re-seeded in 96-well plates, and treated with tacrolimus at doses of 0, 30, or 60 μM for 24 hours. This in vitro model was compared to a mouse model of tacrolimus nephrotoxicity and the associated mechanisms were investigated. @*Results@#The size of the kidney organoids and cell viability decreased in dose-dependent manners after treatment with tacrolimus. The number of tubular cells decreased with a loss of polarity, similar to the effects seen in mouse tacrolimus nephrotoxicity. Ultrastructural analysis showed numerous vacuoles in the proximal tubular cells of the kidney organoids treated with tacrolimus. Tacrolimus treatment induced oxidative stress and mitochondrial dysfunction, and autophagic activity was enhanced in the kidney organoids. Rapamycin, an autophagy inducer, accelerated cell death in the kidney organoid model of tacrolimus nephrotoxicity, which was attenuated by treatment with 3-methyladenine, an autophagy inhibitor. These findings indicate that the augmentation of autophagy by rapamycin treatment accelerated tacrolimus nephrotoxicity. @*Conclusions@#Our data suggest that human kidney organoids are an effective in vitro model of tacrolimus nephrotoxicity and that autophagy plays a critical role in tacrolimus nephrotoxicity.
ABSTRACT
The height, width, and cross-sectional area of the vertebral canal and spinal cord along with the area ratio of spinal cord to vertebral canal in the cervical vertebra were evaluated in images obtained using computed tomography (CT). Measurements were taken at the cranial, middle, and caudal point of each cervical vertebra in eight clinically normal small breed dogs (two shih tzu, two miniature schnauzers, and four mixed breed), 10 beagles, and four German shepherds. CT myelography facilitated the delineation of the epidural space, subarachnoid space, and spinal cord except at the caudal portion of the 7th cervical vertebra. The spinal cord had a tendency to have a clear ventral border in the middle portion of the vertebral canal and lateral borders near both end plates. The height, width, and area of the vertebral canal and spinal cord in the cervical vertebra were increased as the size of dog increased. However, the ratio of the spinal cord area to vertebral canal area in the small dogs was higher than that of the larger dogs. Results of the present study could provide basic and quantitative information for CT evaluation of pathologic lesions in the cervical vertebra and spinal cord.