Fig. 1
KPT-9274 causes gender-dependent kidney toxicity and anemia. A 4× and 20× representative images of vehicle control-treated and KPT-9274-treated NSG mouse eyes showing no disruption of retinal epithelial and neuronal cell layers. B Histopathology of stomachs and kidneys of KPT-9274-treated and vehicle-treated NSG mice (representative images at 2× , 40× and 60× magnifications). There is evidence of nephrotoxicity in kidneys from male and female mice treated with KPT-9274, characterized by acute tubular necrosis, tubular atrophy and regenerative hyperplasia. C Precent area with renal injury is estimated for treated NSG mice. D Representative images of TUNEL staining kidney sections and quantification of TUNEL+ cells in treated NSG mice. *p-value < 0.05. E Serum levels of BUN and creatinine and F erythropoietin (EPO) are measured in treated NSG and NRGS mice. *p-value < 0.05; **p-value < 0.01; ****p-value < 0.0001. G The relationship between EPO levels and the severity of renal injury (% renal cortex affected) in NSG mice. H RBC concentrations are measured in treated NSG and NRGS mice. *p-value < 0.05. I KPT-9274 exposure inhibits in vivo bone marrow erythropoiesis. The frequencies of erythroblast subsets (ProE, EryA, EryB and EryC) in treated bone marrow of NRGS mice are measured by CD71 and Ter119 staining. Data are expressed as mean ± SEM. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001. J The levels of SIRT3, acetyl-SOD, full length PARP1, cleaved PARP1, cleaved Caspase-3 and cleaved Caspase-7 after treatment of KPT-9274 at various concentrations in IMCD3 cells as detected by Western blotting. Results are representative of 2 replicates. K ROS levels in IMCD3 cells after treatment of KPT-9274 at various concentrations as measured by CM-H2DCFDA flow cytometric analysis; *p-value < 0.05; **p-value < 0.01. L IMCD3 cell apoptosis being treated with increasing concentrations of KPT-9274 and rescued by SIRT3 activator Honokiol as measured by Annexin V/PI staining. *p-value < 0.05; **p-value < 0.01. ns not significant