Jens-Peter Kühn

Visualization of Hepatic Uptake Transporter Function in Healthy Subjects by Using Gadoxetic Acid–enhanced MR Imaging

PURPOSE To determine if genetic polymorphisms of liver-specific human organic anion transporting polypeptide (OATP) 1B1 and OATP1B3 influence cellular uptake of gadoxetic acid in vitro and if functionally relevant polymorphisms are confounders for liver enhancement by gadoxetic acid in healthy subjects. MATERIALS AND METHODS This study received ethics approval, and all subjects provided written informed consent. Cellular uptake of gadoxetic acid by OATP1B1 and OATP1B3 and their frequent genetic variants was measured by using stable transfected embryonic kidney HEK293 cells. Liver signal intensity at gadoxetic acid-enhanced MR imaging and pharmacokinetics of gadoxetic acid were evaluated in 36 healthy carriers of SLCO1B1/1B3 wild-type alleles (n = 10), SLCO1B1*1b/*1b (n = 8), SLCO1B1*15/*15 (n = 7), SLCO1B1*5/*15 (n = 1), SLCO1B1*1a/*5 (n = 6), and SLCO1B3*4/*4 (n = 4) by using T1-weighted MR imaging and liquid chromatography tandem mass spectrometry. RESULTS Transport activity for gadoxetic acid was increased in cells transfected with SLCO1B1c.388A>G (12.8 pmol/[mg·min]6 3.53, P = .001) but decreased in cells with SLCO1B1c.388A>G/521T>C (3.11 pmol/[mg·min] ± 0.918, P = .004) compared with cells with nonvariant transporter (6.32 pmol/[mg·min] ± 2.73). Compared with activity of cells transfected with the nonvariant SLCO1B3 (7.43 pmol/[mg·min] ± 2.43), SLCO1B3c.699G>A was a gain-of-function variant (15.1 pmol/[mg·min] ± 5.52, P = .002), whereas SLCO1B3c.334T>G (0.364 pmol/[mg·min] ± 0.125, P = .0001) and SLCO1B3c.1564G>T (0.295 pmol/[mg·min] ± 0.247, P = .0001) were variants with lower function. Liver enhancement with gadoxetic acid was reduced in subjects with OATP1B1*1a/*5 compared with wild-type subjects and those with OATP1B1*1b/*1b (area under enhancement curve, 3-480 minutes in arbitrary units [au]; 20.7 au ± 6.85 vs 36.5 au ± 8.08 [P = .006] vs 34.6 au ± 8.92 [P = .026]). The OATP1B3*4 polymorphism was not of functional relevance. No pharmacokinetic characteristics of gadoxetic acid were influenced by genetic polymorphisms of OATP1B1 and OATP1B3. CONCLUSION Liver-specific OATP1B1 and OATP1B3 are uptake carriers for gadoxetic acid in subjects. Genetic polymorphisms of OATP1B1 are signal confounders in gadoxetic acid-enhanced liver MR imaging.

Quantitative chemical shift-encoded MRI is an accurate method to quantify hepatic steatosis.

To compare the accuracy of liver fat quantification using a three‐echo chemical shift‐encoded magnetic resonance imaging (MRI) technique without and with correction for confounders with spectroscopy (MRS) as the reference standard.

Noninvasive quantification of hepatic fat content using three-echo dixon magnetic resonance imaging with correction for T2* relaxation effects.

Objective:To investigate three-echo T2*-corrected Dixon magnetic resonance imaging (MRI) for noninvasively estimating hepatic fat content (HFC) compared with biopsy. Materials and Methods:One hundred patients (50 men, 50 women; mean age, 57.7 ± 14.2 years) underwent clinically indicated liver core biopsy (102 valid tissue samples) and liver MRI 24 to 72 hours later. MRI was performed at 1.5T (Magnetom Avanto, Siemens Healthcare, Erlangen, Germany) using Dixon imaging with T2* correction (work in progress, WIP-432.rev.1, Siemens Healthcare). An ultrafast breath-hold three-echo 3D-gradient echo sequence with TR/TE1/TE2/TE3 of 11/2.4/4.8/9.6 milliseconds, and online calculation of T2*-corrected water images (signal intensities of water [SIW]), fat images (SIF), and fat content map (SIFAT = 10 × SIF/(SIW + SIF)) was used. SIs of the calculated fat content map (SIFAT) were verified using the histologically quantified HFC (HFC(path)). Spearman correlation for HFC(path) and SIFAT was calculated. Stage of fibrosis, hepatic iron content, and patterns of liver fat (macrovesicular, microvesicular, mixed) and their influence on predicting HFC by MRI were determined. Results:Correlation between SIFAT and HFC(path) was rspearman = 0.89. Agreement between HFC predicted by MRI and HFC(path) calculated by nonlinear saturation-growth regression was rspearman = 0.89. Kruskal-Wallis analysis revealed no significant difference for SIFAT across fibrosis grades (P = 0.90) and liver iron content (P = 0.76). Regarding the cellular architecture of liver fat, the microvesicular pattern showed lower mean ranks in SI than macrovesicular and mixed patterns (P = 0.01). Conclusion:T2*-corrected Dixon MRI is a noninvasive tool for estimating HFC, showing excellent correlation with liver biopsy without being limited by liver iron content and fibrosis/cirrhosis.

Quantification of Hepatic Steatosis With Dual-Energy Computed Tomography: Comparison With Tissue Reference Standards and Quantitative Magnetic Resonance Imaging in the ob/ob Mouse

ObjectiveThe aim of this study was to compare dual-energy computed tomography (DECT) and magnetic resonance imaging (MRI) for fat quantification using tissue triglyceride concentration and histology as references in an animal model of hepatic steatosis. Materials and MethodsThis animal study was approved by our institution’s Research Animal Resource Center. After validation of DECT and MRI using a phantom consisting of different triglyceride concentrations, a leptin-deficient obese mouse model (ob/ob) was used for this study. Twenty mice were divided into 3 groups based on expected levels of hepatic steatosis: low (n = 6), medium (n = 7), and high (n = 7) fat. After MRI at 3 T, a DECT scan was immediately performed. The caudate lobe of the liver was harvested and analyzed for triglyceride concentration using a colorimetric assay. The left lateral lobe was also extracted for histology. Magnetic resonance imaging fat-fraction (FF) and DECT measurements (attenuation, fat density, and effective atomic number) were compared with triglycerides and histology. ResultsPhantom results demonstrated excellent correlation between triglyceride content and each of the MRI and DECT measurements (r2 ≥ 0.96, P ⩽ 0.003). In vivo, however, excellent triglyceride correlation was observed only with attenuation (r2 = 0.89, P < 0.001) and MRI-FF (r2 = 0.92, P < 0.001). Strong correlation existed between attenuation and MRI-FF (r2 = 0.86, P < 0.001). Nonlinear correlation with histology was also excellent for attenuation and MRI-FF. ConclusionsDual-energy computed tomography (CT) data generated by the current Gemstone Spectral Imaging analysis tool do not improve the accuracy of fat quantification in the liver beyond what CT attenuation can already provide. Furthermore, MRI may provide an excellent reference standard for liver fat quantification when validating new CT or DECT methods in human subjects.

Normal Dynamic MRI Enhancement Patterns of the Upper Abdominal Organs: Gadoxetic Acid Compared With Gadobutrol

OBJECTIVE The purpose of this study was to investigate whether, at dynamic MRI of the upper abdominal organs, contrast enhancement with gadoxetic acid, a hepatobiliary contrast agent, is comparable with that achieved with an extracellular contrast agent. SUBJECTS AND METHODS Dynamic gadoxetic acid-enhanced MRI of the pancreas, spleen, kidney, liver, and abdominal aorta was performed on 50 patients; dynamic gadobutrol-enhanced MRI was performed on a control group of 50 patients; and the images were compared. Dynamic imaging with a T1-weighted volumetric interpolated breath-hold examination gradient-echo sequence (TR/TE, 3.35/1.35; flip angle, 12 degrees ) was performed before and 20 (arterial phase), 55 (portal venous phase), and 90 (hepatic venous phase) seconds after bolus injection of gadoxetic acid (0.25 mmol/mL) or gadobutrol (1.0 mmol/mL). Signal-to-noise ratios and enhancement indexes were calculated for each organ and time. RESULTS All MR images in both groups were of diagnostic quality. During the early dynamic phases, significantly lower mean enhancement indexes were found in the gadoxetic acid group than in the gadobutrol group in the pancreas (portal venous phase, 0.66, 1.39, p <or= 0.001; hepatic venous phase, 0.51, 1.36, p <or= 0.001), spleen (portal venous phase, 1.54, 2.41, p <or= 0.001; hepatic venous phase, 1.19, 2.23, p <or= 0.001), renal cortex (portal venous phase, 1.76, 2.63, p <or= 0.001; hepatic venous phase, 1.60, 2.63, p <or= 0.001), and liver (portal venous phase, 0.76, 0.94, p = 0.016; hepatic venous phase, 0.76, 1.04, p <or= 0.001). In the abdominal aorta, the mean enhancement index was greater after bolus injection of gadoxetic acid (arterial phase, 3.33, 2.24, p <or= 0.005). CONCLUSION Early dynamic MRI of the upper abdominal organs, especially the spleen, pancreas, and kidney, benefits from the higher gadolinium concentration of gadobutrol than in the organ-specific contrast agent gadoxetic acid. Higher protein binding resulting in increased relaxivity of gadoxetic acid compensates for the low gadolinium concentration in the abdominal aorta.

Intragastric Volume Changes after Intake of a High-Caloric, High-Fat Standard Breakfast in Healthy Human Subjects Investigated by MRI

The aim of this magnetic resonance imaging (MRI) study was to investigate gastric emptying after intake of a high-caloric and high-fat standard meal as recommended by FDA and EMA for food-effect bioavailability and fed bioequivalence studies. Twelve healthy human subjects (7 male, 5 female) received the standard meal after an overnight fast. MRI was performed before as well as 15, 25, 35, 45, 55, 65, 105, 195, 275, and 375 min after meal intake using strong T2-weighted sequences and chemical shift imaging. In addition, 30 min after the beginning of meal intake subjects ingested 240 mL of water representing the recommended coadministration of water during drug intake. Gastric content volume was assessed using T2-weighted images, and fat fraction was estimated using a calculation of fat fraction in chemical shift imaging. In addition, the existence of a mechanism allowing fast gastric emptying of water in the fed state was investigated. After a lag phase of 50-90 min, gastric content volume decreased constantly with a rate of 1.7 mL/min. The water ingested 30 min after the start of the meal intake directly reached the antrum and subsequently was emptied quickly from the human stomach. Complete gastric emptying within 6 h was observed in only one out of 12 subjects. The fat fraction of the intragastric chyme decreased from 9.5% directly after meal intake to 6.3% at the end of the experiments. Moreover, the fat fraction in fundus was significantly higher compared to the antrum. This study contributes fundamental data for the assessment of food effects of solid oral dosage forms.

Comparative In Vitro Study on Magnetic Iron Oxide Nanoparticles for MRI Tracking of Adipose Tissue-Derived Progenitor Cells

Magnetic resonance imaging (MRI) using measurement of the transverse relaxation time (R2*) is to be considered as a promising approach for cell tracking experiments to evaluate the fate of transplanted progenitor cells and develop successful cell therapies for tissue engineering. While the relationship between core composition of nanoparticles and their MRI properties is well studied, little is known about possible effects on progenitor cells. This in vitro study aims at comparing two magnetic iron oxide nanoparticle types, single vs. multi-core nanoparticles, regarding their physico-chemical characteristics, effects on cellular behavior of adipose tissue-derived stem cells (ASC) like differentiation and proliferation as well as their detection and quantification by means of MRI. Quantification of both nanoparticle types revealed a linear correlation between labeling concentration and R2* values. However, according to core composition, different levels of labeling concentrations were needed to achieve comparable R2* values. Cell viability was not altered for all labeling concentrations, whereas the proliferation rate increased with increasing labeling concentrations. Likewise, deposition of lipid droplets as well as matrix calcification revealed to be highly dose-dependent particularly regarding multi-core nanoparticle-labeled cells. Synthesis of cartilage matrix proteins and mRNA expression of collagen type II was also highly dependent on nanoparticle labeling. In general, the differentiation potential was decreased with increasing labeling concentrations. This in vitro study provides the proof of principle for further in vivo tracking experiments of progenitor cells using nanoparticles with different core compositions but also provides striking evidence that combined testing of biological and MRI properties is advisable as improved MRI properties of multi-core nanoparticles may result in altered cell functions.

R2* estimation using “in-phase” echoes in the presence of fat: The effects of complex spectrum of fat

To investigate R2* mapping robustness in the presence of fat using in‐phase echoes, without and with spectral modeling of fat (single‐peak and multipeak models, respectively), using varying numbers of echoes.

Navigating the human gastrointestinal tract for oral drug delivery: Uncharted waters and new frontiers☆

Many concepts of oral drug delivery are based on our comprehension of human gastrointestinal physiology. Unfortunately, we tend to oversimplify the complex interplay between the various physiological factors in the human gut and, in particular, the dynamics of these transit conditions to which oral dosage forms are exposed. Recent advances in spatial and temporal resolution of medical instrumentation as well as improved access to these technologies have facilitated clinical trials to characterize the dynamic processes within the human gastrointestinal tract. These studies have shown that highly relevant parameters such as fluid volumes, dosage form movement, and pH values in the lumen of the upper GI tract are very dynamic. As a result of these new insights into the human gastrointestinal environment, some common concepts and ideas of oral drug delivery are no longer valid and have to be reviewed in order to ensure efficacy and safety of oral drug therapy.

Secretin-stimulated MRCP in volunteers: assessment of safety, duct visualization, and pancreatic exocrine function.

OBJECTIVE The objective of our study was to investigate secretin-stimulated MRCP in terms of the safety of secretin, improvement of duct visualization, and assessment of pancreatic exocrine function. MATERIALS AND METHODS Eight hundred sixteen volunteers (370 women and 446 men; mean age, 49.7 ± 13.1 [SD] years) underwent 3D MRCP before and after secretin stimulation (1 U/kg of body weight) at 1.5 T. For the first 2 hours after secretin injection, subjects were evaluated for adverse reactions. Improvement of duct visualization after secretin stimulation was subjectively evaluated by two readers and was quantified by duct diameter measurements. Pancreatic exocrine function was evaluated subjectively by two readers according to the duodenal filling and was quantified using calibrated volumetric measurements of total excreted volume and pancreatic flow output. RESULTS Two subjects (0.2%) showed flushing (minor adverse reaction). Duct visualization after secretin injection was improved for reader 1 in 468 (57.4%) and for reader 2 in 478 (58.6%) subjects, was unchanged for reader 1 in 324 (39.7%) and for reader 2 in 315 (38.6%) subjects, and was worse for reader 1 in 24 (2.9%) and reader 2 in 23 (2.8%) subjects (interrater agreement, κ = 0.925). Main pancreatic duct diameters increased significantly after secretin stimulation: pancreatic head, 10.5% (mean); body, 12.5%; and tail, 7.7%. Pancreatic exocrine function evaluated according to assessment of duodenal filling was as follows: grade 0 (restricted function) in 0.7% of subjects by both readers, grade 1 (reduced function) in 4.8% of subjects by reader 1 and 4.5% of subjects by reader 2, grade 2 (low-grade reduced function) in 31.1% of subjects by reader 1 and 26.5% of subjects by reader 2, and grade 3 (physiologic function) in 63.4% of subjects by reader 1 and 68.3% of subjects by reader 2 (interrater agreement, κ = 0.838). The mean total excreted volume was 111.8 ± 49.8 (SD) mL, and the mean pancreatic flow output was 9.6 ± 4.2 mL/min. CONCLUSION Secretin-stimulated MRCP moderately improves main pancreatic duct visualization and allows noninvasive quantification of pancreatic exocrine function with a negligible risk of side effects.