Side Effects on the Heart and Skeleton of Growing Mice Attributed to Chronic Imatinib Exposure.

Objectives: Chronic myeloid leukemia (CML) is effectively treated by Imatinib (IM) via inhibition of the BCR-ABL tyrosine kinase. However, also related tyrosine kinases like abl, c-Kit, PDGF-R, and c-FMS are blocked by IM. As shown in adult humans and mice, abl-controlled protein folding as part of the endoplasmatic stress response in heart myoblasts as well as bone “remodeling” depending on PDGF-R and c-FMS is impaired under imatinib exposure (Dewar AL et al 2005, Kerkelä R et al 2006, Fitter S et al 2008). The influence of IM on the growing heart and skeleton of immature animals has not been studied so far. With respect to treatment of pediatric CML we report alterations in these organs of juvenile mice chronically exposed to IM during the growth period.

Methods: From the age of 4–14 weeks (w) [development milestones of mice: weaning 3 w; puberty 7 w; epiphysial lines closure 18 w] C3H/Neu male and female wild-type mice were chronically exposed to IM via the drinking water at concentrations of 500 mg/l (group A), 750 mg/l (group B), and 1000mg/l (group C). Femur length and overall skeletal development was analysed by whole body X-ray analysis using a mammography device. Bone metabolic activity was assessed by total body Na¹⁸F PET and CT after 5w and 10w of exposure using dedicated small animal tomographs. Bone mineral density and microstructure of tibiae were analysed by pQCT and microCT (resolution 12.5μ m) while the number of osteoclasts and resorption lacunae in femora and vertebrae was assessed by histomorphometry. Plasma concentration of IM, osteocalcin, and activity of the tartrate resistant acid phosphatase (TRAP5b) was also determined. The heart was examined histologically and ultrastructurally by electron microscopy.

Results: IM was tolerated well and mean uptake of 80 mg/kg/d

• 110 mg/kg/d

• and 150 mg/kg/d

• resulted in serum levels of 60–674 ng/ml, 36–242 ng/ml and 51–534 ng/ml, respectively.

Body weight gain was delayed in groups B and C until the age of 8 w while no change in overall growth, development and behaviour was observed at 14 w. At higher doses of IM and at younger age there was a non-significant trend to a reduction in femur length. Heart morphological examination exhibited an increased number (p<0.05) of hypertrophic cardiomyocytes (toxic damage) paralleled by ultrastructural alterations in mitochondria, myofibrils, and nucleus. In the skeleton, no significant differences compared to controls concerning ¹⁸F-kinetics and uptake in vertebrae and femura could be demonstrated. However, IM dose-dependently reduced the number of osteoclasts and resorption lacunae (p<0.05); these effects were less pronounced in female mice. Tibia cortical thickness was increased significantly in males by 6.1% (B) and 11.2% (C), respectively, and 7.5% in females (C). By microCT cancellous bone exhibited a significant increase in trabecular bone mass density and volume and number resulting in an increase in trabecular connectivity in males by 63% (B) and 64% (C), respectively, and in females by 22% (B) and 38% (C), respectively. Bone biomarkers indicated a significant reduction of TRAP5b activity while osteocalcin levels remained unchanged.

Conclusion: In juvenile mice, a chronic exposure of IM resulted in toxic damage of the cardiomyocytes at higher dose rates. However, these alterations do not necessarily imply also a functional impairment which can only be studied in vivo. In the skeleton, IM reduced the number of osteoclasts and resorption lacunae in long bones but not in vertebrae. IM showed an antiresorptive effect in cancellous bone and increased cortical thickness and trabecular number by inhibiting the expansion of the marrow cavity. The effects were more pronounced in male mice and at younger age.