Spatiotemporal heterogeneity of tumor vasculature during tumor growth and antiangiogenic treatment: MRI assessment using permeability and blood volume parameters.

Tumor heterogeneity is an important concept when assessing intratumoral variety in vascular phenotypes and responses to antiangiogenic treatment. This study explored spatiotemporal heterogeneity of vascular alterations in C6 glioma mice during tumor growth and antiangiogenic treatment on serial MR examinations (days 0, 4, and 7 from initiation of vehicle or multireceptor tyrosine kinase inhibitor administration). Transvascular permeability (TP) was quantified on dynamic-contrast-enhanced MRI (DCE-MRI) using extravascular extracellular agent (Gd-DOTA); blood volume (BV) was estimated using intravascular T2 agent (SPION). With regard to region-dependent variability in vascular phenotypes, the control group demonstrated higher TP in the tumor center than in the periphery, and greater BV in the tumor periphery than in the center. This distribution pattern became more apparent with tumor growth. Antiangiogenic treatment effect was regionally heterogeneous: in the tumor center, treatment significantly suppressed the increase in TP and decrease in BV (ie, typical temporal change in the control group); in the tumor periphery, treatment-induced vascular alterations were insignificant and BV remained high. On histopathological examination, the control group showed greater CD31, VEGFR2, Ki67, and NG2 expression in the tumor periphery than in the center. After treatment, CD31 and Ki67 expression was significantly suppressed only in the tumor center, whereas VEGFR2 and α-caspase 3 expression was decreased and NG2 expression was increased in the entire tumor. These results demonstrate that MRI can reliably depict spatial heterogeneity in tumor vascular phenotypes and antiangiogenic treatment effects. Preserved angiogenic activity (high BV on MRI and high CD31) and proliferation (high Ki67) in the tumor periphery after treatment may provide insights into the mechanism of tumor resistance to antiangiogenic treatment.

The Effects of Breathing Motion on DCE-MRI Images: Phantom Studies Simulating Respiratory Motion to Compare CAIPIRINHA-VIBE, Radial-VIBE, and Conventional VIBE.

OBJECTIVE: To compare the breathing effects on dynamic contrast-enhanced (DCE)-MRI between controlled aliasing in parallel imaging results in higher acceleration (CAIPIRINHA)-volumetric interpolated breath-hold examination (VIBE), radial VIBE with k-space-weighted image contrast view-sharing (radial-VIBE), and conventional VIBE (c-VIBE) sequences using a dedicated phantom experiment. MATERIALS AND METHODS: We developed a moving platform to simulate breathing motion. We conducted dynamic scanning on a 3T machine (MAGNETOM Skyra, Siemens Healthcare) using CAIPIRINHA-VIBE, radial-VIBE, and c-VIBE for six minutes per sequence. We acquired MRI images of the phantom in both static and moving modes, and we also obtained motion-corrected images for the motion mode. We compared the signal stability and signal-to-noise ratio (SNR) of each sequence according to motion state and used the coefficients of variation (CoV) to determine the degree of signal stability. RESULTS: With motion, CAIPIRINHA-VIBE showed the best image quality, and the motion correction aligned the images very well. The CoV (%) of CAIPIRINHA-VIBE in the moving mode (18.65) decreased significantly after the motion correction (2.56) (p < 0.001). In contrast, c-VIBE showed severe breathing motion artifacts that did not improve after motion correction. For radial-VIBE, the position of the phantom in the images did not change during motion, but streak artifacts significantly degraded image quality, also after motion correction. In addition, SNR increased in both CAIPIRINHA-VIBE (from 3.37 to 9.41, p < 0.001) and radial-VIBE (from 4.3 to 4.96, p < 0.001) after motion correction. CONCLUSION: CAIPIRINHA-VIBE performed best for free-breathing DCE-MRI after motion correction, with excellent image quality.

Intravoxel incoherent motion MRI for monitoring the therapeutic response of hepatocellular carcinoma to sorafenib treatment in mouse xenograft tumor models.

Background With the introduction of targeted therapies, there has been a growing need for non-invasive imaging methods which accurately evaluate therapeutic effects and overcome the limitations of tumor size-based therapeutic response assessments. Purpose To assess diagnostic values of intra-voxel incoherent motion (IVIM) imaging in evaluating therapeutic effects of sorafenib on hepatocellular carcinoma (HCC) using mouse xenograft model. Material and Methods Twenty-four mice bearing Huh-7 were divided into a control group and two treatment groups received sorafenib doses of 5 mg/kg (5 mg-Tx) or 30 mg/kg (30 mg-Tx). IVIM imaging was performed using 10 b-values (0-900 s/mm2). The apparent diffusion coefficient (ADC), diffusion coefficient ( D), and perfusion fraction ( f) were measured for whole tumors and tumor periphery. Changes between baseline and post-treatment parameters ( Δ ADC, Δ D, and Δ f) were calculated, and these parameters were compared with microvessel density (MVD) and area of tumor cell death. Results The post-treatment f and Δ f for tumor periphery were significantly higher in control group, followed by 5 mg-Tx and 30 mg-Tx ( P < 0.001). MVD showed significant positive correlation with post-treatment f ( r = 0.584, P = 0.003) and negative correlation with D ( r = -0.495, P = 0.014) for tumor periphery, while no parameter showed significant correlation with area of tumor cell death. Conclusion The f is significantly correlated with MVD of HCC, and could potentially be used to evaluate the anti-angiogenic effects of sorafenib.

Perfusion Assessment Using Intravoxel Incoherent Motion-Based Analysis of Diffusion-Weighted Magnetic Resonance Imaging: Validation Through Phantom Experiments.

OBJECTIVES: The aims of this study were to demonstrate the theoretical meaning of intravoxel incoherent motion (IVIM) parameters and to compare the robustness of 2 biexponential fitting methods through magnetic resonance experiments using IVIM phantoms. MATERIALS AND METHODS: Intravoxel incoherent motion imaging was performed on a 3 T magnetic resonance imaging scanner using 15 b values (0-800 s/mm) for 4 phantoms with different area fractions of the flowing water compartment (FWC%), at the infusion flow rates of 0, 1, 2, and 3 mL/min. Images were quantitatively analyzed using monoexponential free biexponential, and segmented biexponential fitting models. RESULTS: There were some inconsistent variations in Dslow with changing flow rates. The perfusion fraction, f, showed a significant positive correlation with the flow rate for both the free and segmented fitting methods (ρ = 0.838 to 0.969; P < 0.001). The fast diffusion coefficient, Dfast, had a significant positive correlation with the flow rate for segmented fitting (ρ = 0.745 to 0.969; P < 0.001), although it showed an inverse correlation with the flow rate for free fitting (ρ = -0.527 to -0.791; P ≤ 0.017). Significant positive correlations with the FWC% of the phantoms were noted for f (P = 0.510 for free fitting and P = 0.545 for segmented fitting, P < 0.001). CONCLUSIONS: The IVIM model allows for an approximate segmentation of molecular diffusion and perfusion, with a minor contribution of the perfusion effect on Dslow. The f and Dfast can provide a rough estimation of the flow fraction and flow velocity. Segmented fitting may be a more robust method than free fitting for calculating the IVIM parameters, especially for Dfast.

Comparison of CAIPIRINHA-VIBE, Radial-VIBE, and conventional VIBE sequences for dynamic contrast-enhanced (DCE) MRI: A validation study using a DCE-MRI phantom.

OBJECTIVE: To validate radial acquisition of volumetric interpolated breath hold examination (Radial-VIBE) and the controlled aliasing in parallel imaging results in higher acceleration (CAIPIRINHA-VIBE) sequences for dynamic contrast-enhanced MRI (DCE-MRI) by comparing them to conventional VIBE sequence using a phantom. METHODS: On a DCE-MRI phantom containing various concentrations of NiCl2 solutions, six minutes of dynamic series and T1 mapping with variable flip angle methods were acquired using conventional VIBE, Radial-VIBE, and CAIPIRINHA-VIBE sequences on 3.0-T scanners. Signal stability and signal linearity were tested for dynamic series and the precision of R1 values were tested for T1 mapping series. The scans were repeatedly performed at two weeks and three months to test repeatability/reproducibility, assessed by within-subject coefficient of variation (WSCV). RESULTS: Signal stability over six minutes was excellent in all three sequences. Regarding the signal linearity, CAIPIRINHA-VIBE demonstrated the highest linear correlation (r=0.963), followed by conventional VIBE (r=0.959) and Radial-VIBE (r=0.953). Regarding the R1 precision, CAIPIRINHA-VIBE (r=0.985) was the most accurate, followed by conventional VIBE (r=0.861) and Radial-VIBE (r=0.442). CAIPIRINHA-VIBE showed excellent repeatability/reproducibility (WSCV, 1.79-6.71%) compared with Radial-VIBE (WSCV, 2.04-67.2%) and conventional VIBE (WSCV, 3.4-31.9%). CONCLUSION: In terms of signal stability, signal linearity, R1 precision, and repeatability/reproducibility, CAIPIRINHA-VIBE demonstrated outstanding performance for DCE-MRI compared with Radial-VIBE and conventional VIBE.

Feasibility of test-bolus DCE-MRI using CAIPIRINHA-VIBE for the evaluation of pancreatic malignancies.

OBJECTIVES: To evaluate the feasibility of test-bolus dynamic contrast-enhanced (DCE) MRI with CAIPIRINHA-VIBE for pancreatic malignancies. METHODS: Thirty-two patients underwent DCE-MRI with CAIPIRINHA-VIBE after injection of 2 mL gadolinium. From the resulting time-intensity curve (TIC), we estimated the arterial (AP) and portal venous phase (PVP) scan timing for subsequent multiphasic MRI. DCE-MRI perfusion maps were generated, and perfusion parameters were calculated. The image quality was rated on a 5-point scale (1: poor, 5: excellent). Goodness-of-fit of the TIC was evaluated by Pearson's χ2 test. RESULTS: Test-bolus DCE-MRIs with high temporal (3 s) and spatial resolution (1 × 1 × 4 mm3) were acquired with good-quality perfusion maps of Ktrans and iAUC (mean score 4.313 ± 0.535 and 4.125 ± 0.554, respectively). The mean χ2 values for fitted TICs were 0.115 ± 0.082 for the pancreatic parenchyma and 0.784 ± 0.074 for pancreatic malignancies, indicating an acceptable goodness-of-fit. Test-bolus DCE-MRI was highly accurate in estimating the proper timing of AP (90.6 %) and PVP (100 %) of subsequent multiphasic MRI. Between pancreatic adenocarcinomas and neuroendocrine tumours, there were significant differences in the Ktrans (0.073 ± 0.058 vs. 0.308 ± 0.062, respectively; p = 0.007) and iAUC (1.501 ± 0.828 vs. 3.378 ± 0.378, respectively; p = 0.045). CONCLUSIONS: Test-bolus DCE-MRI using CAIPIRINHA-VIBE is feasible for incorporating perfusion analysis of pancreatic tumours into routine multiphasic MRI. KEY POINTS: • Test-bolus DCE-MRI using CAIPIRINHA-VIBE is feasible for perfusion analysis of pancreatic tumours. • CAIPIRINHA-VIBE enables DCE-MRI with high temporal and spatial resolution. • Test-bolus DCE-MRI is highly accurate in estimating the proper timing of multiphasic MRI.

Simultaneous evaluation of vascular morphology, blood volume and transvascular permeability using SPION-based, dual-contrast MRI: imaging optimization and feasibility test.

Exploiting ultrashort-T(E) (UTE) MRI, T1-weighted positive contrast can be obtained from superparamagnetic iron oxide nanoparticles (SPIONs), which are widely used as a robust T2-weighted, negative contrast agent on conventional MR images. Our study was designed (a) to optimize the dual-contrast MRI method using SPIONs and (b) to validate the feasibility of simultaneously evaluating the vascular morphology, blood volume and transvascular permeability using the dual-contrast effect of SPIONs. All studies were conducted using 3 T MRI. According to numerical simulation, 0.15 mM was the optimal blood SPION concentration for visualizing the positive contrast effect using UTE MRI (T(E) = 0.09 ms), and a flip angle of 40° could provide sufficient SPION-induced enhancement and acceptable measurement noise for UTE MR angiography. A pharmacokinetic study showed that this concentration can be steadily maintained from 30 to 360 min after the injection of 29 mg/kg of SPIONs. An in vivo study using these settings displayed image quality and CNR of SPION-enhanced UTE MR angiography (image quality score 3.5; CNR 146) comparable to those of the conventional, Gd-enhanced method (image quality score 3.8; CNR 148) (p > 0.05). Using dual-contrast MR images obtained from SPION-enhanced UTE and conventional spin- and gradient-echo methods, the transvascular permeability (water exchange index 1.76-1.77), cerebral blood volume (2.58-2.60%) and vessel caliber index (3.06-3.10) could be consistently quantified (coefficient of variation less than 9.6%; Bland-Altman 95% limits of agreement 0.886-1.111) and were similar to the literature values. Therefore, using the optimized setting of combined SPION-based MRI techniques, the vascular morphology, blood volume and transvascular permeability can be comprehensively evaluated during a single session of MR examination.

LHT7, a chemically modified heparin, inhibits multiple stages of angiogenesis by blocking VEGF, FGF2 and PDGF-B signaling pathways.

Despite the therapeutic benefits of the angiogenesis inhibitors shown in the clinics, they have encountered an unexpected limitation by the occurrence of acquired resistance. Although the mechanism of the resistance is not clear so far, the upregulation of alternative angiogenic pathways and stabilization of endothelium by mural cells were reported to be responsible. Therefore, blocking multiple angiogenic pathways that are crucial in tumor angiogenesis has been highlighted to overcome such limitations. To develop an angiogenesis inhibitor that could block multiple angiogenic factors, heparin is an excellent lead compound since wide array of angiogenic factors are heparin-binding proteins. In previous study, we reported a heparin-derived angiogenesis inhibitor, LHT7, as a potent angiogenesis inhibitor and showed that it blocked VEGF signaling pathway. Here we show that LHT7 could block the fibroblast growth factor 2 (FGF2) and platelet-derived growth factor B (PDGF-B) in addition to VEGF. Simultaneous blockade of these angiogenic factors resulted in inhibition of multiple stages of the angiogenic process, including initial angiogenic response to maturation of the endothelium by pericyte coverage in vitro. In addition, the treatment of LHT7 in vivo did not show any sign of vascular normalization and directly led to decreased blood perfusion throughout the tumor. Our findings show that LHT7 could effectively inhibit tumor angiogenesis by blocking multiple stages of the angiogenesis, and could potentially be used to overcome the resistance.

Sunitinib--CLIO conjugate: a VEGFR/PDGFR-targeting active MR probe.

PURPOSE: This study was conducted to evaluate feasibility of sunitinib-CLIO conjugate as a vascular endothelial growth factor receptor/platelet-derived growth factor receptor (VEGFR/PDGFR)-specific magnetic resonance (MR) probe. PROCEDURE: VEGFR/PDGFR-targeting MR probe was synthesized by conjugating cross-linked iron-oxide (CLIO) with tyrosine-kinase inhibitor (sunitinib). In VEGFR/PDGFR-positive (U118MG) and VEGFR/PDGFR-negative (HT29) cells and tumor models, conjugate-driven ΔR 2 was estimated, while CLIO was used as control. Prussian-blue staining was performed for quantifying the amount of tumor-binding conjugates. RESULTS: ΔR 2 between sunitinib-CLIO-treated and non-treated cells was greater in U118MG (mean, 2.1/s) than in HT29 cells (1.0/s). In in vivo study, conjugate induced a greater ΔR 2 in U118MG (11.2/s) than HT29 tumors (5.9/s). Conjugate-induced R 2 changes were not correlated with degree of Gd-DTPA enhancement, demonstrating that tumor binding of sunitinib-CLIO was independent of enhanced permeability and retention effect. % area of Prussian-blue staining was greater in U118MG (8.5 %) than in HT29 (1.4 %). CONCLUSIONS: Sunitinib-CLIO conjugate can be used as an active MR probe for quantifying VEGFR/PDGFR.

Dynamic contrast-enhanced MRI for monitoring antiangiogenic treatment: determination of accurate and reliable perfusion parameters in a longitudinal study of a mouse xenograft model.

OBJECTIVE: To determine the reliable perfusion parameters in dynamic contrast-enhanced MRI (DCE-MRI) for the monitoring antiangiogenic treatment in mice. MATERIALS AND METHODS: Mice, with U-118 MG tumor, were treated with either saline (n = 3) or antiangiogenic agent (sunitinib, n = 8). Before (day 0) and after (days 2, 8, 15, 25) treatment, DCE examinations using correlations of perfusion parameters (Kep, Kel, and A(H) from two compartment model; time to peak, initial slope and % enhancement from time-intensity curve analysis) were evaluated. RESULTS: Tumor growth rate was found to be 129% ± 28 in control group, -33% ± 11 in four mice with sunitinib-treatment (tumor regression) and 47% ± 15 in four with sunitinib-treatment (growth retardation). Kep (r = 0.80) and initial slope (r = 0.84) showed strong positive correlation to the initial tumor volume (p < 0.05). In control mice, tumor regression group and growth retardation group animals, Kep (r : 0.75, 0.78, 0.81, 0.69) and initial slope (r : 0.79, 0.65, 0.67, 0.84) showed significant correlation with tumor volume (p < 0.01). In four mice with tumor re-growth, Kep and initial slope increased 20% or greater at earlier (n = 2) than or same periods (n = 2) to when the tumor started to re-grow with 20% or greater growth rate. CONCLUSION: Kep and initial slope may a reliable parameters for monitoring the response of antiangiogenic treatment.

Is apparent diffusion coefficient reliable and accurate for monitoring effects of antiangiogenic treatment in a longitudinal study?

PURPOSE: To evaluate the reliability and accuracy of the apparent diffusion coefficient (ADC) for monitoring antiangiogenic treatment in a longitudinal study. MATERIALS AND METHODS: Tumor volume and ADC were monitored by T2-weighted magnetic resonance imaging (MRI) and diffusion-weighted MRI, respectively, in 18 mice with angiogenesis-dependent tumors (U118MG) before (day 0) and after 2, 7, 14, and 21 days of administration of the antiangiogenic agent sunitinib maleate (n = 12) or vehicle (n = 6). Percent changes in tumor volume and ADC were calculated and correlations between tumor volume and ADC were evaluated. RESULTS: Tumor volume and ADC showed a negative correlation at 69 of the 72 (96%) follow-up measurements. In the 13 mice with tumor regrowth, ADC started to decrease before (27%) or at the same time (73%) as tumor regrowth. Pretreatment ADC and percent change in ADC change on days 0-2 were similar in mice with positive and negative responses to treatment (0.851 vs. 0.999, 24% vs. 16%). Percent change of ADC showed significant negative correlation with percent change in tumor volume in both the control (r = -0.69) and treated (r = -0.65) groups. CONCLUSION: Percent change in ADC is a reliable and accurate marker for monitoring the effects of antiangiogenic treatment, whereas pretreatment ADC and early changes in ADC (ie, days 0-2) are limited in predicting treatment outcome.

Prostate cancer detection on dynamic contrast-enhanced MRI: computer-aided diagnosis versus single perfusion parameter maps.

OBJECTIVE: The purpose of this article is to assess the value of computer-aided diagnosis (CAD) for prostate cancer detection on dynamic contrast-enhanced MRI (DCE-MRI). MATERIALS AND METHODS: DCE-MRI examinations of 42 patients with prostate cancer were used to generate perfusion parameters, including baseline and peak signal intensities, initial slope, maximum slope within the initial 50 seconds after the contrast injection (slope(50)), wash-in rate, washout rate, time to peak, percentage of relative enhancement, percentage enhancement ratio, time of arrival, efflux rate constant from the extravascular extracellular space to the blood plasma (k(ep)), first-order rate constant for eliminating gadopentetate dimeglumine from the blood plasma (k(el)), and constant depending on the properties of the tissue and represented by the size of the extravascular extracellular space (A(H)). CAD for cancer detection was established by comprehensive evaluation of parameters using a support vector machine. The diagnostic accuracy of single perfusion parameters was estimated using receiver operating characteristic analysis, which determined threshold and parametric maps for cancer detection. The diagnostic performance of CAD for cancer detection was compared with those of T2-weighted imaging (T2WI) and single perfusion parameter maps, using histologic results as the reference standard. RESULTS: The accuracy, sensitivity, and specificity of CAD were 83%, 77%, and 77%, respectively, in the entire prostate; 77%, 91%, and 64%, respectively, in the transitional zone; and 89%, 89%, and 89%, respectively, in the peripheral zone. Values for k(ep), k(el), initial slope, slope(50), wash-in rate, washout rate, and time to peak showed greater area under the curve values (0.803-0.888) than did the other parameters (0.545-0.665) (p < 0.01) and were compared with values for CAD. In the entire prostate, accuracy was greater for CAD than for all perfusion parameters or T2WI (63-77%); sensitivity was greater for CAD than for T2WI, initial slope, wash-in rate, slope(50), and washout rate (38-77%); and specificity was greater for CAD than for T2WI, k(ep), k(el), and time to peak (59-68%) (p < 0.05). CONCLUSION: CAD can improve the diagnostic performance of DCE-MRI in prostate cancer detection, which may vary according to zonal anatomy.

Diagnosis of lymph node metastasis in uterine cervical cancer: usefulness of computer-aided diagnosis with comprehensive evaluation of MR images and clinical findings.

BACKGROUND: Lymph node (LN) status is an important parameter for determining the treatment strategy and for predicting the prognosis for patients with uterine cervical cancer. Computer-aided diagnosis (CAD) can be feasible for differentiating metastatic from non-metastatic lymph nodes in patients with uterine cervical cancer. PURPOSE: To determine the usefulness of CAD that comprehensively evaluates MR images and clinical findings for detecting LN metastasis in uterine cervical cancer. MATERIAL AND METHODS: In 680 LNs from 143 patients who underwent radical hysterectomy for uterine cervical cancer, the CAD system using the Bayesian classifier estimated the probability of metastasis based on MR findings and clinical findings. We compared the diagnostic accuracy for detecting metastatic LNs in the CAD and MR findings. RESULTS: Metastasis was diagnosed in 70 (12%) LNs from 34 (24%) patients. The area under ROC curves of CAD (0.924) was greater than those of the mean ADC (0.854), minimum ADC (0.849), maximum ADC (0.827), short-axis diameter (0.856) and long-axis diameter (0.753) (P < 0.05). The specificity and accuracy of the CAD (86%, 86%) were greater than those of the mean ADC (77%, 77%), maximum ADC (77%, 77%), minimum ADC (68%, 70%), and short-axis diameter (65%, 67%) (P < 0.05). CONCLUSION: CAD system can improve the diagnostic performance of MR for detecting metastatic LNs in uterine cervical cancer.

Feasibility of FAIR imaging for evaluating tumor perfusion.

PURPOSE: To evaluate the feasibility of flow-sensitive alternating inversion recovery (FAIR) for measuring blood flow in tumor models. MATERIALS AND METHODS: In eight mice tumor models, FAIR and dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) was performed. The reliability for measuring blood flow on FAIR was evaluated using the coefficient of variation of blood flow on psoas muscle. Three regions of interest (ROIs) were drawn in the peripheral, intermediate, and central portions within each tumor. The location of ROI was the same on FAIR and DCE-MR images. The correlation between the blood flow on FAIR and perfusion-related parameters on DCE-MRI was evaluated using the Pearson correlation coefficient. RESULTS: The coefficient of variation for measuring blood flow was 9.8%. Blood flow on FAIR showed a strong correlation with Kep (r = 0.77), percent relative enhancement (r = 0.73), and percent enhancement ratio (r = 0.81). The mean values of blood flow (mL/100 g/min) (358 vs. 207), Kep (sec(-) (1)) (7.46 vs. 1.31), percent relative enhancement (179% vs. 134%), and percent enhancement ratio (42% vs. 26%) were greater in the peripheral portion than in the central portion (P < 0.01). CONCLUSION: As blood flow measurement on FAIR is reliable and closely related with that on DCE-MR, FAIR is feasible for measuring tumor blood flow.