G. W. Kauffmann

Molecular profiling of angiogenesis with targeted ultrasound imaging: early assessment of antiangiogenic therapy effects

Molecular ultrasound is capable of elucidating the expression of angiogenic markers in vivo. However, the capability of the method for volumetric “multitarget quantification” and for the assessment of antiangiogenic therapy response has rather been investigated. Therefore, we generated cyanoacrylate microbubbles linked to vascular endothelial growth factor receptor 2 (VEGFR2) and αvβ3 integrin binding ligands and quantified their accumulation in squamous cell carcinoma xenografts (HaCaT-ras-A-5RT3) in mice with the quantitative volumetric ultrasound scanning technique, sensitive particle acoustic quantification. Specificity of VEGFR2 and αvβ3 integrin binding microbubbles was shown, and changes in marker expression during matrix metalloproteinase inhibitor treatment were investigated. In tumors, accumulation of targeted microbubbles was significantly higher compared with nonspecific ones and could be inhibited competitively by addition of the free ligand in excess. Also, multimarker imaging could successfully be done during the same imaging session. Molecular ultrasound further indicated a significant increase of VEGFR2 and αvβ3 integrin expression during tumor growth and a considerable decrease in both marker densities after matrix metalloproteinase inhibitor treatment. Histologic data suggested that the increasing VEGFR2 and αvβ3 integrin concentrations in tumors during growth are related to an up-regulation of its expression by the endothelial cells, whereas its decrease under therapy is more related to the decreasing relative vessel density. In conclusion, targeted ultrasound appears feasible for the longitudinal molecular profiling of tumor angiogenesis and for the sensitive assessment of therapy effects in vivo. [Mol Cancer Ther 2008;7(1):101–9]

Preoperative Staging of Renal Cell Carcinoma With Inferior Vena Cava Thrombus Using Multidetector Ct and Mri: Prospective Study With Histopathological Correlation

Objective: To evaluate the accuracy of multidetector computed tomography (CT) and magnetic resonance imaging (MRI) in staging and estimating renal carcinomas with caval thrombus. Methods: Initially, 23 patients with suspected caval thrombi were admitted into this prospective study. Triphasic CT imaging was performed using a multidetector CT with a reconstructed slice thickness of 2 mm. 3D CT reconstructions were used to improve surgical planning. MRI protocol included: a transversal T1-weighted GE sequence with and without Gd-DTPA, a transversal T2-weighted respiratory-gated TSE, and a coronal T1-weighted GE sequence with Gd-DTPA and fat saturation. In addition, a multiphase 3D angiography was performed after Gd-DTPA injection. Patients were divided into 3 groups: caval thrombus below the insertion of the hepatic veins, within the intrahepatic vena cava, and intra-atrial extension. The results the tumor thrombus extension and staging results of 2 independent readers were correlated with surgical and histopathological staging. Results: Of the 23 patients admitted, CT and MR scans of 14/13 patients respectively were correlated with histopathological workup. CT thrombus detection sensitivity and specificity for both readers was 0.93 and 0.8 respectively. MRI sensitivity and specificity for both readers was 1.0/0.85 and 0.75. Readers I and II evaluated the uppermost extension of the cranial tumor thrombus by both CT and MRI. CT and MR accuracy was 78% and 72%, 88% and 76% respectively. Conclusion: In cases of a suspected tumor thrombus, MRI and multidetector CT imaging showed similar staging results. Consequently, these staging modalities can be used to assess the extension of the tumor thrombus.

Vessel Fractions in Tumor Xenografts Depicted by Flow-or Contrast-Sensitive Three-Dimensional High-Frequency Doppler Ultrasound Respond Differently to Antiangiogenic Treatment

High-frequency volumetric Power Doppler ultrasound (HF-VPDU) captures flow-dependent signals in blood vessels and can be used to assess antiangiogenic therapy effects in rodent tumors. However, the sensitivity is limited to vessels larger than capillaries. Contrast-enhanced HF-VPDU reveals all perfused vessels by assessing stimulated acoustic emissions from disintegrating microbubbles. Thus, we investigated whether flow-sensitive and contrast-enhanced HF-VPDU can depict different vessel fractions and assess their early response to antiangiogenic therapy. Mice with A431 tumors were scanned before and after administration of polybutylcyanoacrylate microbubbles by HF-VPDU. Animals received either antiangiogenic treatment (SU11248) or a control substance and were imaged repeatedly over 9 days. At each time point, tumors were removed for immunohistochemical analysis. During growth of untreated tumors, vascularization decreased correspondingly on flow-sensitive and contrast-enhanced scans. Treated tumors showed a significantly (P < 0.05) stronger decline in vascularization than controls, which was more pronounced in contrast-enhanced scans. Surprisingly, whereas vascularization remained low in contrast-enhanced scans, flow-sensitive ultrasound indicated a reincrease after day 6 with a higher vascularization than the controls at day 9. Histologic evaluation indicated that immature vessels degraded markedly on therapy, whereas large mature vessels on the tumor periphery were more therapy resistant and drew closer due to tumor shrinkage. In conclusion, contrast-enhanced HF-VPDU and flow-sensitive HF-VPDU are both capable of assessing the effects of antiangiogenic therapy. Because contrast-sensitive ultrasound is more sensitive for small immature vessels and flow-sensitive ultrasound mostly captures large vessels at the tumor periphery, the combination of both methods can provide evidence of vascular maturity in tumors.

Pharmacodynamics of streptavidin-coated cyanoacrylate microbubbles designed for molecular ultrasound imaging

Objectives:To assess the pharmacodynamic behavior of cyanoacrylate, streptavidin-coated microbubbles (MBs) and to investigate their suitability for molecular ultrasound imaging. Materials and Methods:Biodistribution of MBs was analyzed in tumor-bearing mice using &ggr;-counting, immunohistochemistry, flow cytometry, and ultrasound. Further, vascular endothelial growth factor receptor 2-antibody coupled MBs were used to image tumor neovasculature. Results:After 1 minute >90% of MBs were cleared from the blood and pooled in the lungs, liver, and spleen. Subsequently, within 1 hour a decent reincrease of MB-concentration was observed in the blood. The remaining MBs were removed by liver and spleen macrophages. About 30% of the phagocytosed MBs were intact after 48 hours. Shell fragments were found in the kidneys only. No relevant MB-accumulation was observed in tumors. In contrast, vascular endothelial growth factor receptor 2-specific MBs accumulated significantly within the tumor vasculature (P < 0.05). Conclusions:The pharmacokinetic behavior of streptavidin-coated cyanoacrylate MBs has been studied. In this context, the low amount of MBs in tumors after >5 minutes is beneficial for specific targeting of angiogenesis.

Prolonged survival following palliative renal tumor embolization by capillary occlusion

Nine patients with renal cell carcinoma and severe hematuria were palliatively treated with a new type of angioocclusion: The concept of capillary embolization. The so-called occlusion gel Ethibloc was used as embolizing agent. Each patient was followed up until death or for at least 4 years. All patients had a stage T3 or T4 tumor, 3 patients had metastases to multiple organs, 3 had lung metastases, and 3 were free of metastatic disease. In all cases, very high volumes (14–40 ml) of the embolizing agent were necessary to achieve total occlusion of the entire arterial compartment. Patients without metastatic disease had a mean survival time of 6 years and 4 months, all of them without signs of malignant disease. Patients with metastases had a mean survival time of 3 years. Compared with the natural history of renal cell carcinoma treated otherwise, this represents a substantial prolongation of survival time. Contrary to other angioocclusive treatment modalities, the concept of capillary occlusion with Ethibloc seem to achieve total tumor destruction.

New coil concept for endoluminal MR imaging

Our aim was to conduct a prospective study to evaluate staging accuracy of a new coil concept for endoluminal magnetic resonance imaging (MRI) on ex vivo gastric carcinomas. Twenty-eight consecutive patients referred to surgery with a clinically proven primary gastric malignancy were included. Surgical specimens were examined with a foldable and self-expanding loop coil (8-cm diameter) at 1.5 Tesla immediately after total gastrectomy. T1- and T2-weighted and opposed-phase sequences (axial, frontal sections; 3- to 4-mm slice thickness) were acquired. Investigators blinded to any patient information analyzed signal intensity of normal gastric wall, gastric tumor, and lymph nodes. Findings were compared with histopathological staging. On surgical specimens, 2–5 gastric wall layers could be visualized. All gastric tumors (26 carcinomas, two lymphomas) were identified on endoluminal MR data (100%). Overall accuracy for T staging was 75% (18/24); sensitivity to detect serosal involvement was 80% and specificity 89%. N staging correlated in 58% (14/24) with histopathology (N+ versus N−). The endoluminal coil concept is feasible and applicable for an ex vivo setting. Endoluminal MR data provided sufficient detail for gastric wall layer differentiation, and therefore, identification of T stages in gastric carcinoma is possible. Further investigations in in vivo settings should explore the potential of our coil concept for endoluminal MR imaging.

Präoperatives Staging von Nierenzellkarzinomen mit Kavazapfen: welches diagnostische Verfahren?

ZusammenfassungZiel dieser Untersuchung war die Evaluation der optimalen und effektiven Diagnostik beim präoperativen Staging von Nierenzellkarzinomen mit Kavazapfen. Ist der Einsatz der MRT gerechtfertigt? Es wurden 7 Nierenzellkarzinome der Tumorstadien T3b und T3c präoperativ in der CT und MRT untersucht und das Staging mit dem histopathologischen Ergebnis korreliert. In der MRT wurden 4 von 7 Kavazapfen in ihrer Ausdehnung korrekt und sicher beurteilt, in der CT keiner korrekt und sicher. Die MRT mit Gadolinium ist der CT im Staging von Nierenzellkarzinomen der hohen Tumorstadien überlegen und kann hier die Kavographie ersetzen. Die MRT ist in den Fällen, in denen sonographisch ein hohes Tumorstadium mit Kavazapfen vermutet wird, als präoperative Diagnostik gerechtfertigt.SummaryTo evaluate whether MRI is usefull in staging renal cell carcinomas with caval thrombus the accuracy of computed tomography (CT) and magnetic resonance imaging (MRI) in staging renal tumors with caval thrombus were preoperatively examined. Tumor staging by CT and MR imaging were correlated with histopathological tumor stadium. In MRI 4 out of 7 thrombi were correctly diagnosed with high accuracy, in CT none. In advanced renal carcinoma MRI with Gadolinium was superior to CT imaging, especially in diagnosing tumor thrombus. Consequently the extent of tumor thrombus may be assessed by MRI which therefore may replace conventional cavography.

Assessment of hepatic perfusion in transplanted livers by pharmacokinetic analysis of dynamic magnetic resonance measurements

Objective:The purpose of this study was to validate the assessment of hepatic perfusion by pharmacokinetic analysis of dynamic contrast-enhanced magnetic resonance image series. Materials and Methods:Dynamic measurements were performed with a saturation recovery turbo fast low angle shot (ie, FLASH) sequence over the course of approximately 4 minutes in 17 patients with transplanted livers. By pharmacokinetic analysis using an open 2-compartment model, we estimated and correlated an amplitude of signal enhancement, A, and the perfusion rate, kp, with invasive perfusion measurements from implanted thermo-diffusion probes (FTDP). Results:Data analysis for segment IV of the transplanted livers yielded a mean blood flow of 81 ± 19 mL/min/100g and a mean perfusion rate of 13 ± 6 minutes−1. There was a significant correlation between FTDP and kp (rS = 0.64, P = 0.01) but not with A. Conclusions:Although our open 2-compartment model oversimplifies the complexity of hepatic perfusion, it allows a numerically robust estimation of regional blood flow per unit of blood volume. Thus, dynamic magnetic resonance imaging represents a noninvasive method to assess hepatic perfusion rate which can be visualized in color coded images.

Experimental study of the effectiveness of capillary embolization using contrast-enhanced Ethibloc.

In experimental and clinical use, Ethibloc in combination with 40% glucose preinjection has proven to be of major advantage in tumor embolization. However, its low radiographic contrast is a limiting factor in monitoring its vascular distribution and venous propagation. Various contrast media were tested in order to enhance this contrast in laboratory and animal experiments. Normal rat kidneys (N = 96) and renal tumors, induced by Dimethylnitrosamine (N = 66) were tested as in previous studies. Ethibloc-N was produced by substituting Lipiodol for poppy seed oil which is an ingredient of the original Ethibloc. This proved to be the only embolization medium that combined the excellent properties of the original Ethibloc with increased contrast. All other embolization media tested resulted in new complications such as under-or overembolization and pulmonary embolism.

Efficacy and Safety of Percutaneous Transhepatic Portal Embolization before Right Liver Resection Using an Ethibloc/Lipiodol Mixture: A Single-Center Experience

Aim: The purpose of this study was to evaluate the safety and efficacy of percutaneous transhepatic portal vein embolization of the right portal vein with an Ethibloc/Lipiodol mixture to induce hypertrophy of the left liver lobe in patients with primarily unresectable liver tumor. Methods: 15 patients (8 primary liver tumors, 7 liver metastases) underwent portal vein embolization. Liver volumetry, duration of hospitalization, complication rates, relevant laboratory values were documented. Results: In 13/15 patients (84.6%) embolization could be performed with a median of 8.8 ml (range 1.5–28 ml) Ethibloc/Lipiodol. One minor procedure-related complication (subcapsular hematoma) occurred, which did not affect the two-step liver resection. No patient developed acute liver failure after embolization or liver resection. The volume of the left liver lobe increased significantly (p = 0.0015) by 25% from a median of 750 ml (587–1,114 ml) to 967 ml (597–1,249 ml). 11/13 (81.8%) of the embolized patients underwent liver resection at a median of 49 days after embolization. Median hospitalization time was 4 days after embolization and 7 days after liver resection. Median overall survival of the 11 operated patients was 376 days. Conclusion: Percutaneous transhepatic portal vein embolization using an Ethibloc/Lipiodol mixture is a safe, feasible, and efficient interventional procedure.