Felix Schwab

Selective targeting of genetically engineered mesenchymal stem cells to tumor stroma microenvironments using tissue-specific suicide gene expression suppresses growth of hepatocellular carcinoma

Background:The use of engineered mesenchymal stem cells (MSCs) as therapeutic vehicles for the treatment of experimental pancreatic and breast cancer has been previously demonstrated. The potential application of MSCs for the treatment of hepatocellular carcinoma (HCC) has been controversial. The general approach uses engineered MSCs to target different aspects of tumor biology, including angiogenesis or the fibroblast-like stromal compartment, through the use of tissue-specific expression of therapeutic transgenes. The aim of the present study was (1) to evaluate the effect of exogenously added MSCs on the growth of HCC and (2) the establishment of an MSC-based suicide gene therapy for experimental HCC. Methods:Mesenchymal stem cells were isolated from bone marrow of C57/Bl6 p53−/− mice. The cells were injected into mice with HCC xenografts and the effect on tumor proliferation and angiogenesis was evaluated. The cells were then stably transfected with red fluorescent protein (RFP) or Herpes simplex virus thymidine kinase (HSV-Tk) gene under control of the Tie2 promoter/enhancer or the CCL5 promoter. Mesenchymal stem cells were injected intravenously into mice with orthotopically growing xenografts of HCC and treated with ganciclovir (GCV). Results:Ex vivo examination of hepatic tumors revealed tumor-specific recruitment, enhanced tumor growth, and increased microvessel density after nontherapeutic MSC injections. After their homing to the hepatic xenografts, engineered MSCs demonstrated activation of the Tie2 or CCL5 promoter as shown by RFP expression. Application of CCL5/HSV-TK transfected MSCs in combination with GCV significantly reduced tumor growth by 56.4% as compared with the control group and by 71.6% as compared with nontherapeutic MSC injections. CCL5/HSV-TK+ transfected MSCs proved more potent in tumor inhibition as compared with Tie2/HSV-TK+ MSCs. Conclusion:Exogenously added MSCs are recruited to growing HCC xenografts with concomitant activation of the CCL5 or Tie2 promoters within the MSCs. Stem cell–mediated introduction of suicide genes into the tumor followed by prodrug administration was effective for treatment of experimental HCC and thus may help fill the existing gap in bridging therapies for patients suffering from advanced HCCs.

Diagnosing and mapping pulmonary emphysema on X-ray projection images: incremental value of grating-based X-ray dark-field imaging.

Purpose To assess whether grating-based X-ray dark-field imaging can increase the sensitivity of X-ray projection images in the diagnosis of pulmonary emphysema and allow for a more accurate assessment of emphysema distribution. Materials and Methods Lungs from three mice with pulmonary emphysema and three healthy mice were imaged ex vivo using a laser-driven compact synchrotron X-ray source. Median signal intensities of transmission (T), dark-field (V) and a combined parameter (normalized scatter) were compared between emphysema and control group. To determine the diagnostic value of each parameter in differentiating between healthy and emphysematous lung tissue, a receiver-operating-characteristic (ROC) curve analysis was performed both on a per-pixel and a per-individual basis. Parametric maps of emphysema distribution were generated using transmission, dark-field and normalized scatter signal and correlated with histopathology. Results Transmission values relative to water were higher for emphysematous lungs than for control lungs (1.11 vs. 1.06, p<0.001). There was no difference in median dark-field signal intensities between both groups (0.66 vs. 0.66). Median normalized scatter was significantly lower in the emphysematous lungs compared to controls (4.9 vs. 10.8, p<0.001), and was the best parameter for differentiation of healthy vs. emphysematous lung tissue. In a per-pixel analysis, the area under the ROC curve (AUC) for the normalized scatter value was significantly higher than for transmission (0.86 vs. 0.78, p<0.001) and dark-field value (0.86 vs. 0.52, p<0.001) alone. Normalized scatter showed very high sensitivity for a wide range of specificity values (94% sensitivity at 75% specificity). Using the normalized scatter signal to display the regional distribution of emphysema provides color-coded parametric maps, which show the best correlation with histopathology. Conclusion In a murine model, the complementary information provided by X-ray transmission and dark-field images adds incremental diagnostic value in detecting pulmonary emphysema and visualizing its regional distribution as compared to conventional X-ray projections.

Assessment of pulmonary perfusion with breath-hold and free-breathing dynamic contrast-enhanced magnetic resonance imaging: quantification and reproducibility

ObjectivesThe purpose of this study was to investigate whether quantification of pulmonary perfusion from dynamic contrast-enhanced magnetic resonance imaging (DCE MRI) yields more reproducible results with data acquired during free breathing than with data from conventional breath-hold measurements. Material and MethodsTen healthy male volunteers underwent 2 imaging sessions at a clinical 1.5-T MRI system, separated by a week ± 1 day. Each of these sessions comprised 2 DCE MRI acquisitions: one performed during breath-hold and one during free, shallow breathing; both acquisitions were separated by at least 20 minutes. For all DCE MRI measurements, a standard dose of gadobutrol was used. Breath-hold measurements lasted 53 seconds; free-breathing acquisitions were performed in a total acquisition time of 146 seconds.Lung tissue was segmented automatically to minimize user influence, and pulmonary plasma flow (PPF) and volume (PPV) were quantified on a per-pixel basis with a 1-compartment model. Free-breathing measurements were analyzed twice, (a) including data from the entire acquisition duration and (b) after truncation to the duration of the breath-hold measurements. For further statistical analysis, median values of the resulting parameter maps were determined. To assess intraindividual reproducibility, intraclass correlation coefficients and coefficients of variation between the first and second measurements were calculated for breath-hold, truncated, and full free-breathing measurements, respectively. Differences in the coefficients of variation were assessed with a nonparametric 2-sided paired Wilcoxon signed rank test. ResultsAll 40 measurements were completed successfully. Maps of PPF and PPV could be calculated from both measurement techniques; PPF and PPV in the breath-hold measurements were significantly lower (P < 0.001) than in truncated and full free-breathing measurements. Both evaluations of the free-breathing measurements yielded higher intraclass correlation coefficients and lower coefficients of variation between the first and second measurements than the breath-hold measurements. ConclusionsBesides offering substantially higher patient comfort, free-breathing DCE MRI acquisitions allow for pixelwise quantification of pulmonary perfusion and hence generation of parameter maps. Moreover, quantitative perfusion estimates derived from free-breathing DCE MRI measurements have better reproducibility than estimates from the conventionally used breath-hold measurements.

Combined Diffusion-Weighted, Blood Oxygen Level–Dependent, and Dynamic Contrast-Enhanced MRI for Characterization and Differentiation of Renal Cell Carcinoma

PURPOSE To investigate a multiparametric magnetic resonance imaging (MRI) approach comprising diffusion-weighted imaging (DWI), blood oxygen-dependent (BOLD), and dynamic contrast-enhanced (DCE) MRI for characterization and differentiation of primary renal cell carcinoma (RCC). MATERIAL AND METHODS Fourteen patients with clear-cell carcinoma and four patients with papillary RCC were examined with DWI, BOLD MRI, and DCE MRI at 1.5T. The apparent diffusion coefficient (ADC) was calculated with a monoexponential decay. The spin-dephasing rate R2* was derived from parametric R2* maps. DCE-MRI was analyzed using a two-compartment exchange model allowing separation of perfusion (plasma flow [FP] and plasma volume [VP]), permeability (permeability surface area product [PS]), and extravascular extracellular volume (VE). Statistical analysis was performed with Wilcoxon signed-rank test, Pearsons correlation coefficient, and receiver operating characteristic curve analysis. RESULTS Clear-cell RCC showed higher ADC and lower R2* compared to papillary subtypes, but differences were not significant. FP of clear-cell subtypes was significantly higher than in papillary RCC. Perfusion parameters showed moderate but significant inverse correlation with R2*. VE showed moderate inverse correlation with ADC. Fp and Vp showed best sensitivity for histological differentiation. CONCLUSION Multiparametric MRI comprising DWI, BOLD, and DCE MRI is feasible for assessment of primary RCC. BOLD moderately correlates to DCE MRI-derived perfusion. ADC shows moderate correlation to the extracellular volume, but does not correlate to tumor oxygenation or perfusion. In this preliminary study DCE-MRI appeared superior to BOLD and DWI for histological differentiation.

Lung tumors on multimodal radiographs derived from grating-based X-ray imaging – A feasibility study

PURPOSE The purpose of this study was to assess whether grating-based X-ray imaging may have a role in imaging of pulmonary nodules on radiographs. MATERIALS AND METHODS A mouse lung containing multiple lung tumors was imaged using a small-animal scanner with a conventional X-ray source and a grating interferometer for phase-contrast imaging. We qualitatively compared the signal characteristics of lung nodules on transmission, dark-field and phase-contrast images. Furthermore, we quantitatively compared signal characteristics of lung tumors and the adjacent lung tissue and calculated the corresponding contrast-to-noise ratios. RESULTS Of the 5 tumors visualized on the transmission image, 3/5 tumors were clearly visualized and 1 tumor was faintly visualized in the dark-field image as areas of decreased small angle scattering. In the phase-contrast images, 3/5 tumors were clearly visualized, while the remaining 2 tumors were faintly visualized by the phase-shift occurring at their edges. No additional tumors were visualized in either the dark-field or phase-contrast images. Compared to the adjacent lung tissue, lung tumors were characterized by a significant decrease in transmission signal (median 0.86 vs. 0.91, p = 0.04) and increase in dark-field signal (median 0.71 vs. 0.65, p = 0.04). Median contrast-to-noise ratios for the visualization of lung nodules were 4.4 for transmission images and 1.7 for dark-field images (p = 0.04). CONCLUSION Lung nodules can be visualized on all three radiograph modalities derived from grating-based X-ray imaging. However, our initial data suggest that grating-based multimodal X-ray imaging does not increase the sensitivity of chest radiographs for the detection of lung nodules.

High-resolution cine MRI with TGRAPPA for fast assessment of left ventricular function at 3 Tesla

PURPOSE To implement and evaluate the accuracy of multislice dual-breath hold cine MR for analysis of global systolic and diastolic left ventricular function at 3T. MATERIALS AND METHODS 25 patients referred to cardiac MR underwent cine imaging at 3T (MAGNETOM Verio) using prospective triggered SSFP (TR 3.1 ms; TE 1.4 ms; FA 60°). Analysis of LV function was performed using a standard non-accelerated single-slice approach (STD) with multiple breath-holds and an accelerated multi-slice technique (TGRAPPA; R=4) encompassing the ventricles with 5 slices/breath-hold. Parameters of spatial and temporal resolution were kept identical (pixel: 1.9 × 2.5 mm(2); temporal resolution: 47 ms). Data of both acquisition techniques were analyzed by two readers using semiautomatic algorithms (syngoARGUS) with respect to EDV, ESV, EF, myocardial mass (MM), peak filling rate (PFR) and peak ejection rate (PER) including assessment of interobserver agreement. RESULTS Volumetric results of the TGRAPPA approach did not show significant differences to the STD approach for left ventricular ejection fraction (62.3 ± 10.6 vs. 61.0 ± 8.4, P=0.2), end-diastolic volume (135.8 ± 47.5 vs. 130.8 ± 46.4, P=0.07), endsystolic volume (53.0 ± 29.7 vs. 53.1 ± 32.7, P=0.99) and myocardial mass (114.2 ± 32.5 vs. 114.6±30.6, P=0.9). Moreover, a comparison of peak ejection rate (601.3 ± 190.2 vs. 590.8 ± 218.2, P=0.8) and peak filling rate (535.1±191.2 vs. 535.4 ± 210.7, P=0.99) did not reveal significant differences between the two groups. Limits in interobserver agreement were low for all systolic and diastolic parameters in both groups (P ≥ 0.05). Total acquisition time for STD was 273 ± 124 s and 34 ± 5 s for TGRAPPA (P ≤ 0.001). Evaluation time for standard and multislice approach was equal (10.8 ± 1.4 vs. 9.8 ± 2.1 min; P=0.08).

Tracer kinetic modeling in myocardial perfusion quantification using MRI

To investigate and compare several quantification methods of myocardial perfusion measurements, paying special attention to the relation between the techniques and the required measurement duration.

Free breathing real-time cardiac cine imaging with improved spatial resolution at 3 T.

ObjectivesThe aim of this study was to evaluate free-breathing single-shot real-time cine imaging for functional cardiac imaging at 3 T with increased spatial resolution. Special emphasis of this study was placed on the influence of parallel imaging techniques. Materials and MethodsGradient echo phantom images were acquired with GRAPPA and modified SENSE reconstruction using both integrated and separate reference scans as well as TGRAPPA and TSENSE. In vivo measurements were performed for GRAPPA reconstruction with an integrated and a separate reference scan, as well as TGRAPPA using balanced steady-state free precession protocols. Three clinical protocols, rtLRInt (Tres = 51.3 milliseconds; voxel, 2.5 × 5.0 × 10 mm3), rtMRSep (Tres = 48.8 milliseconds; voxel, 1.9 × 3.1 × 10 mm3), and rtHRSep (Tres = 48.3 milliseconds; voxel, 1.6 × 2.6 × 10 mm3), were investigated on 20 volunteers using GRAPPA reconstruction with internal as well as separate reference scans. End-diastolic volume, end-systolic volume, ejection fraction, peak ejection rate, peak filling rate, and myocardial mass were evaluated for the left ventricle and compared with an electrocardiogram-triggered segmented readout cine protocol used as standard of reference. All studies were performed at 3 T. ResultsPhantom and in vivo data demonstrate that the combination of GRAPPA reconstruction with a separate reference scan provides an optimal compromise of image quality as well as spatial and temporal resolution. Functional values (P values) for the standard of reference, rtLRInt, rtMRSep, and rtHRSep end-diastolic volume were 141 ± 24 mL, 138 ± 21 mL, 138 ± 19 mL, and 128 ± 33 mL, respectively (P = 0.7, 0.7, 0.4); end-systolic volume, 55 ± 15 mL, 61 ± 14 mL, 58 ± 12 mL, and 55 ± 20 mL, respectively (P = 0.23, 0.43, 0.62); ejection fraction, 61% ± 5%, 57% ± 5%, 58% ± 4%, and 56% ± 8%, respectively (P = 0.01, 0.11, 0.06); peak ejection rate, 481 ± 73 mL/s, 425 ± 62 mL/s, 434 ± 67 mL/s, and 381 ± 86 mL/s, respectively (P = 0.03, 0.04, 0.01); peak filling rate, 555 ± 80 mL/s, 480 ± 70 mL/s, 500 ± 70 mL/s, and 438 ± 108 mL/s, respectively (P= 0.007, 0.05, 0.004); and myocardial mass, 137 ± 26 g, 141 ± 25 g, 141 ± 23 g, and 130 ± 31 g, respectively (P = 0.62, 0.54, 0.99). ConclusionsUsing a separate reference scan and high acceleration factors up to R = 6, single-shot real-time cardiac imaging offers adequate temporal and spatial resolution for accurate assessment of global left ventricular function in free breathing with short examination times.

Feasibility of free-breathing, GRAPPA-based, real-time cardiac cine assessment of left-ventricular function in cardiovascular patients at 3T.

OBJECTIVES To determine the feasibility of free-breathing, GRAPPA-based, real-time (RT) cine 3T cardiac magnetic resonance imaging (MRI) with high acceleration factors for the assessment of left-ventricular function in a cohort of patients as compared to conventional segmented cine imaging. MATERIALS AND METHODS In this prospective cohort study, subjects with various cardiac conditions underwent MRI involving two RT cine sequences (high resolution and low resolution) and standard segmented cine imaging. Standard qualitative and quantitative parameters of left-ventricular function were quantified. RESULTS Among 25 subjects, 24 were included in the analysis (mean age: 50.5±21 years, 67% male, 25% with cardiomyopathy). RT cine derived quantitative parameters of volumes and left ventricular mass were strongly correlated with segmented cine imaging (intraclass correlation coefficient [ICC]: >0.72 for both RT cines) but correlation for peak ejection and filling rates were moderate to poor for both RT cines (ICC<0.40). Similarly, RT cines significantly underestimated peak ejection and filling rates (>103.2±178 ml/s). Among patient-related factors, heart rate was strongly predictive for deviation of measurements (p<0.05). CONCLUSIONS RT cine MRI at 3T is feasible for qualitative and quantitative assessment of left ventricular function for low and high-resolution sequences but results in significant underestimation of systolic function, peak ejection and filling rates.

Diagnosing and mapping pulmonary emphysema on X-ray projection images

To assess whether grating-based X-ray dark-field imaging can increase the sensitivity of X-ray projection images in the diagnosis of pulmonary emphysema and allow for a more accurate assessment of emphysema distribution. Lungs from three mice with pulmonary emphysema and three healthy mice were imaged ex vivo using a laser-driven compact synchrotron X-ray source. Median signal intensities of transmission (T), dark-field (V) and a combined parameter (normalized scatter) were compared between emphysema and control group. To determine the diagnostic value of each parameter in differentiating between healthy and emphysematous lung tissue, a receiver-operating-characteristic (ROC) curve analysis was performed both on a per-pixel and a per-individual basis. Parametric maps of emphysema distribution were generated using transmission, dark-field and normalized scatter signal and correlated with histopathology. Transmission values relative to water were higher for emphysematous lungs than for control lungs (1.11 vs. 1.06, p<0.001). There was no difference in median dark-field signal intensities between both groups (0.66 vs. 0.66). Median normalized scatter was significantly lower in the emphysematous lungs compared to controls (4.9 vs. 10.8, p<0.001), and was the best parameter for differentiation of healthy vs. emphysematous lung tissue. In a per-pixel analysis, the area under the ROC curve (AUC) for the normalized scatter value was significantly higher than for transmission (0.86 vs. 0.78, p<0.001) and dark-field value (0.86 vs. 0.52, p<0.001) alone. Normalized scatter showed very high sensitivity for a wide range of specificity values (94% sensitivity at 75% specificity). Using the normalized scatter signal to display the regional distribution of emphysema provides color-coded parametric maps, which show the best correlation with histopathology. In a murine model, the complementary information provided by X-ray transmission and dark-field images adds incremental diagnostic value in detecting pulmonary emphysema and visualizing its regional distribution as compared to conventional X-ray projections.