Commissioning of a 4D MRI phantom for use in MR-guided radiotherapy.

PURPOSE: Systems for integrated magnetic resonance guided radiation therapy (MRgRT) provide real-time and online MRI guidance for unequaled targeting performance of moving tumors and organs at risk. The clinical introduction of such systems requires dedicated methods for commissioning and routine machine quality assurance (QA). The aim of the study was to develop a commissioning protocol and method for automatic quantification of target motion and geometric accuracy using a 4D MRI motion phantom. MATERIALS AND METHODS: The commissioning was performed on a clinically used 3 T MR scanner. The phantom was positioned on a flat tabletop overlay using an in-house constructed base plate for a quick and reproducible setup. The torso-shaped phantom body, which was filled with mineral oil as signal generating medium, included a 3D grid structure for image distortion analysis and a cylindrical thru-hole in which a software-controlled moving rod with a hypo-intense background gel and a decentralized hyper-intense target simulated 3D organ motion patterns. To allow for sequence optimization, MR relaxometry was performed to determine the longitudinal T1 and transverse T2 relaxation times of both target and background gel in the movable cylinder. The geometric image distortion was determined as the mean and maximum 3D Euclidean distance (Δmean , Δmax ) of grid points determined by nonrigid registration of a 3D spoiled gradient echo MRI scan and a CT scan. Sinusoidal 1D/2D/3D motion trajectories, varying in amplitude and frequency, as well as an exemplary 1D MR navigator diaphragm motion pattern extracted from a healthy volunteer scan, were scanned by means of 2D cine MRI and 4D MRI. Target positions were automatically extracted from 2D cine MRI using an in-house developed software tool. RESULTS: The base plate enabled a reproducible setup with a deviation of <1 mm in all directions. Relaxometry yielded T1 /T2 values for target and background gel of 208.1 ± 2.8/30.5 ± 4.7 ms and 871 ± 36/13.4  ±  1.3 ms, respectively. The 3D geometric image distortion increased with distance from the magnetic isocenter, with Δmean  = 0.58 ± 0.30 mm and Δmax  = 1.31 mm. The frequencies of the reconstructed motion patterns agreed with the preset values within 0.5%, whereas the reconstructed amplitudes showed a maximum deviation to the preset amplitudes of <0.5 mm in AP/LR direction and <0.3 mm in IS direction. CONCLUSION: A method and protocol for commissioning of a 4D MRI motion phantom on a 3 T MR scanner for MRgRT was developed. High-contrast and geometrically reliable 2D cine MR images of the phantom's moving target could be obtained. The preset motion parameters could be extracted with sufficient spatio-temporal accuracy from 2D cine MRI in all motion directions. The overall 3D geometric image distortion of <1.31 mm within the phantom grid confirms geometric accuracy of the clinically utilized 3D spoiled gradient echo sequence. The method developed can be used for routine QA tests of spatio-temporally resolved MRI data in MRgRT.

Effects of combined valproic acid and the epidermal growth factor/vascular endothelial growth factor receptor tyrosine kinase inhibitor AEE788 on renal cell carcinoma cell lines in vitro.

OBJECTIVE: To evaluate adhesion and growth inhibiting effects of the multiple receptor tyrosine kinase inhibitor AEE788 and the histone deacetylase (HDAC) inhibitor valproic acid (VPA) on renal cell carcinoma (RCC) cells. MATERIALS AND METHODS: Caki-1 cells were treated with AEE788 and VPA, either alone or in combination, to investigate RCC cell adhesion to vascular endothelial cells or to immobilized extracellular matrix proteins. Tumour cell proliferation was examined by MTT dye reduction assay. Effects of drug treatment on cell signalling pathways were determined by Western blotting. The expression levels of integrin alpha and beta subtypes were evaluated by flow cytometry (surface expression) and Western blotting (intracellular protein expression). RESULTS: RCC cell treatment with AEE788 and VPA in combination resulted in a stronger inhibition of tumour cell proliferation than that caused by either drug alone. There were also additive effects of the combined treatment on tumour cell adhesion to endothelial cells and to immobilized laminin (but not to immobilized fibronectin and collagen). AEE788 alone or combined with VPA reduced Akt expression and histone H3 acetylation. Both compounds altered integrin alpha and beta subtype expression, in particular alpha1, alpha3 and beta4, and blocked integrin-dependent integrin-linked kinase and focal-adhesion kinase (total and phosphorylated) signalling. CONCLUSIONS: Both AEE788 and VPA profoundly block the interaction of RCC cells with endothelium and extracellular matrix and reduce tumour growth in vitro. Therefore, this combined regimen warrants further preclinical and possible clinical study for treating advanced RCC.

Combining the receptor tyrosine kinase inhibitor AEE788 and the mammalian target of rapamycin (mTOR) inhibitor RAD001 strongly inhibits adhesion and growth of renal cell carcinoma cells.

BACKGROUND: Treatment options for metastatic renal cell carcinoma (RCC) are limited due to resistance to chemo- and radiotherapy. The development of small-molecule multikinase inhibitors has now opened novel treatment options. We evaluated the influence of the receptor tyrosine kinase inhibitor AEE788, applied alone or combined with the mammalian target of rapamycin (mTOR) inhibitor RAD001, on RCC cell adhesion and proliferation in vitro. METHODS: RCC cell lines Caki-1, KTC-26 or A498 were treated with various concentrations of RAD001 or AEE788 and tumor cell proliferation, tumor cell adhesion to vascular endothelial cells or to immobilized extracellular matrix proteins (laminin, collagen, fibronectin) evaluated. The anti-tumoral potential of RAD001 combined with AEE788 was also investigated. Both, asynchronous and synchronized cell cultures were used to subsequently analyze drug induced cell cycle manipulation. Analysis of cell cycle regulating proteins was done by western blotting. RESULTS: RAD001 or AEE788 reduced adhesion of RCC cell lines to vascular endothelium and diminished RCC cell binding to immobilized laminin or collagen. Both drugs blocked RCC cell growth, impaired cell cycle progression and altered the expression level of the cell cycle regulating proteins cdk2, cdk4, cyclin D1, cyclin E and p27. The combination of AEE788 and RAD001 resulted in more pronounced RCC growth inhibition, greater rates of G0/G1 cells and lower rates of S-phase cells than either agent alone. Cell cycle proteins were much more strongly altered when both drugs were used in combination than with single drug application. The synergistic effects were observed in an asynchronous cell culture model, but were more pronounced in synchronous RCC cell cultures. CONCLUSION: Potent anti-tumoral activitites of the multikinase inhibitors AEE788 or RAD001 have been demonstrated. Most importantly, the simultaneous use of both AEE788 and RAD001 offered a distinct combinatorial benefit and thus may provide a therapeutic advantage over either agent employed as a monotherapy for RCC treatment.