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Background: Prostate cancer tissues are inherently heterogeneous, which presents a challenge for metabolic profiling using traditional bulk analysis methods that produce an averaged profile. The aim of this study was therefore to spatially detect metabolites and lipids on prostate tissue sections by using mass spectrometry imaging (MSI), a method that facilitates molecular imaging of heterogeneous tissue sections, which can subsequently be related to the histology of the same section., Methods: Here, we simultaneously obtained metabolic and lipidomic profiles in different prostate tissue types using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MSI. Both positive and negative ion mode were applied to analyze consecutive sections from 45 fresh-frozen human prostate tissue samples (N = 15 patients). Mass identification was performed with tandem MS., Results: Pairwise comparisons of cancer, non-cancer epithelium, and stroma revealed several metabolic differences between the tissue types. We detected increased levels of metabolites crucial for lipid metabolism in cancer, including metabolites involved in the carnitine shuttle, which facilitates fatty acid oxidation, and building blocks needed for lipid synthesis. Metabolites associated with healthy prostate functions, including citrate, aspartate, zinc, and spermine had lower levels in cancer compared to non-cancer epithelium. Profiling of stroma revealed higher levels of important energy metabolites, such as ADP, ATP, and glucose, and higher levels of the antioxidant taurine compared to cancer and non-cancer epithelium., Conclusions: This study shows that specific tissue compartments within prostate cancer samples have distinct metabolic profiles and pinpoint the advantage of methodology providing spatial information compared to bulk analysis. We identified several differential metabolites and lipids that have potential to be developed further as diagnostic and prognostic biomarkers for prostate cancer. Spatial and rapid detection of cancer-related analytes showcases MALDI-TOF MSI as a promising and innovative diagnostic tool for the clinic.

T 2 -weighted (T 2 W) MRI provides high spatial resolution and tissue-specific contrast, but it is predominantly used for qualitative evaluation of prostate anatomy and anomalies. This retrospective multicenter study evaluated the potential of T 2 W image-derived textural features for quantitative assessment of peripheral zone prostate cancer (PCa) aggressiveness. A standardized preoperative multiparametric MRI was performed on 87 PCa patients across 6 institutions. T 2 W intensity and apparent diffusion coefficient (ADC) histogram, and T 2 W textural features were computed from tumor volumes annotated based on whole-mount histology. Spearman correlations were used to evaluate association between textural features and PCa grade groups (i.e. 1-5). Feature utility in differentiating and classifying low-(grade group 1) vs. intermediate/high-(grade group ≥ 2) aggressive cancers was evaluated using Mann-Whitney U-tests, and a support vector machine classifier employing "hold-one-institution-out" cross-validation scheme, respectively. Textural features indicating image homogeneity and disorder/complexity correlated significantly (p < 0.05) with PCa grade groups. In the intermediate/high-aggressive cancers, textural homogeneity and disorder/complexity were significantly lower and higher, respectively, compared to the low-aggressive cancers. The mean classification accuracy across the centers was highest for the combined ADC and T 2 W intensity-textural features (84%) compared to ADC histogram (75%), T 2 W histogram (72%), T 2 W textural (72%) features alone or T 2 W histogram and texture (77%), T 2 W and ADC histogram (79%) combined. Texture analysis of T 2 W images provides quantitative information or features that are associated with peripheral zone PCa aggressiveness and can augment their classification.

Objective: Simultaneous PET/MRI combines soft-tissue contrast of MRI with high molecular sensitivity of PET in one session. The aim of this prospective study was to evaluate detection rates of recurrent prostate cancer by 18 F-fluciclovine PET/MRI., Methods: Patients with biochemical recurrence (BCR) or persistently detectable prostate specific antigen (PSA), were examined with simultaneous 18 F-fluciclovine PET/MRI. Multiparametric MRI (mpMRI) and PET/MRI were scored on a 3-point scale (1-negative, 2-equivocal, 3-recurrence/metastasis) and detection rates (number of patients with suspicious findings divided by total number of patients) were reported. Detection rates were further stratified based on PSA level, PSA doubling time (PSAdt), primary treatment and inclusion criteria (PSA persistence, European Association of Urology (EAU) Low-Risk BCR and EAU High-Risk BCR). A detailed investigation of lesions with discrepancy between mpMRI and PET/MRI scores was performed to evaluate the incremental value of PET/MRI to mpMRI. The impact of the added PET acquisition on further follow-up and treatment was evaluated retrospectively., Results: Among patients eligible for analysis (n=84), 54 lesions were detected in 38 patients by either mpMRI or PET/MRI. Detection rates were 41.7% for mpMRI and 39.3% for PET/MRI (score 2 and 3 considered positive). There were no significant differences in detection rates for mpMRI versus PET/MRI. Disease detection rates were higher in patients with PSA≥1ng/mL than in patients with lower PSA levels but did not differ between patients with PSAdt above versus below 6 months. Detection rates in patients with primary radiation therapy were higher than in patients with primary surgery. Patients categorized as EAU Low-Risk BCR had a detection rate of 0% both for mpMRI and PET/MRI. For 15 lesions (27.8% of all lesions) there was a discrepancy between mpMRI score and PET/MRI score. Of these, 10 lesions scored as 2-equivocal by mpMRI were changed to a more definite score (n=4 score 1 and n=6 score 3) based on the added PET acquisition. Furthermore, for 4 of 10 patients with discrepancy between mpMRI and PET/MRI scores, the added PET acquisition had affected the treatment choice., Conclusion: Combined 18 F-fluciclovine PET/MRI can detect lesions suspicious for recurrent prostate cancer in patients with a range of PSA levels. Combined PET/MRI may be useful to select patients for appropriate treatment, but is of limited use at low PSA values or in patients classified as EAU Low-Risk BCR, and the clinical value of 18 F-fluciclovine PET/MRI in this study was too low to justify routine clinical use., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Selnæs, Krüger-Stokke, Elschot, Johansen, Steen, Langørgen, Aksnessæther, Indrebø, Sjøbakk, Tessem, Moestue, Knobel, Tandstad, Bertilsson, Solberg and Bathen.)

Computer-aided detection and diagnosis (CAD) systems have the potential to improve robustness and efficiency compared to traditional radiological reading of magnetic resonance imaging (MRI). Fully automated segmentation of the prostate is a crucial step of CAD for prostate cancer, but visual inspection is still required to detect poorly segmented cases. The aim of this work was therefore to establish a fully automated quality control (QC) system for prostate segmentation based on T2-weighted MRI. Four different deep learning-based segmentation methods were used to segment the prostate for 585 patients. First order, shape and textural radiomics features were extracted from the segmented prostate masks. A reference quality score (QS) was calculated for each automated segmentation in comparison to a manual segmentation. A least absolute shrinkage and selection operator (LASSO) was trained and optimized on a randomly assigned training dataset (N = 1756, 439 cases from each segmentation method) to build a generalizable linear regression model based on the radiomics features that best estimated the reference QS. Subsequently, the model was used to estimate the QSs for an independent testing dataset (N = 584, 146 cases from each segmentation method). The mean ± standard deviation absolute error between the estimated and reference QSs was 5.47 ± 6.33 on a scale from 0 to 100. In addition, we found a strong correlation between the estimated and reference QSs (rho = 0.70). In conclusion, we developed an automated QC system that may be helpful for evaluating the quality of automated prostate segmentations.

Purpose: To evaluate different non-Gaussian representations for the diffusion-weighted imaging (DWI) signal in the b-value range 200 to 3000 s/mm 2 in benign and malignant breast lesions., Methods: Forty-three patients diagnosed with benign (n = 18) or malignant (n = 25) tumors of the breast underwent DWI (b-values 200, 600, 1200, 1800, 2400, and 3000 s/mm 2 ). Six different representations were fit to the average signal from regions of interest (ROIs) at different b-value ranges. Quality of fit was assessed by the corrected Akaike information criterion (AICc), and the Friedman test was used for assessing representation ranks. The area under the curve (AUC) of receiver operating characteristic curves were used to evaluate the power of derived parameters to differentiate between malignant and benign lesions. The lesion ROI was divided in central and peripheral parts to assess potential effect of heterogeneity. Sensitivity to noise-floor correction was also evaluated., Results: The Padé exponent was ranked as the best based on AICc, whereas 3 models (kurtosis, fractional, and biexponential) achieved the highest AUC = 0.99 for lesion differentiation. The monoexponential model at b max = 600 s/mm 2 already provides AUC = 0.96, with considerably shorter acquisition time and simpler analysis. Significant differences between central and peripheral parts of lesions were found in malignant lesions. The mono- and biexponential models were most stable against varying degrees of noise-floor correction., Conclusion: Non-Gaussian representations are required for fitting of the DWI curve at high b-values in breast lesions. However, the added clinical value from the high b-value data for differentiation of benign and malignant lesions is not clear., (© 2020 The Authors. Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine.)

Background: The prognosis for women with locally advanced breast cancer (LABC) is poor and there is a need for better treatment stratification. Gray-level co-occurrence matrix (GLCM) texture analysis of magnetic resonance (MR) images has been shown to predict pathological response and could become useful in stratifying patients to more targeted treatments., Purpose: To evaluate the ability of GLCM textural features obtained before neoadjuvant chemotherapy to predict overall survival (OS) seven years after diagnosis of patients with LABC., Material and Methods: This retrospective study includes data from 55 patients with LABC. GLCM textural features were extracted from segmented tumors in pre-treatment dynamic contrast-enhanced 3-T MR images. Prediction of OS by GLCM textural features was assessed and compared to predictions using traditional clinical variables., Results: Linear mixed-effect models showed significant differences in five GLCM features (f 1 , f 2 , f 5 , f 10 , f 11 ) between survivors and non-survivors. Using discriminant analysis for prediction of survival, GLCM features from 2 min post-contrast images achieved a classification accuracy of 73% ( P  < 0.001), whereas traditional prognostic factors resulted in a classification accuracy of 67% ( P  = 0.005). Using a combination of both yielded the highest classification accuracy (78%, P  < 0.001). Median values for features f 1 , f 2 , f 10 , and f 11 provided significantly different survival curves in Kaplan-Meier analysis., Conclusion: This study shows a clear association between textural features from post-contrast images obtained before neoadjuvant chemotherapy and OS seven years after diagnosis. Further studies in larger cohorts should be undertaken to investigate how this prognostic information can be used to benefit treatment stratification.

Background: Increased deposition and reorientation of stromal collagen fibers are associated with breast cancer progression and invasiveness. Diffusion-weighted imaging (DWI) may be sensitive to the collagen fiber organization in the stroma and could provide important biomarkers for breast cancer characterization., Purpose: To understand how collagen fibers influence water diffusion in vivo and evaluate the relationship between collagen content and the apparent diffusion coefficient (ADC) and the signal fractions of the biexponential model using a high b-value scheme., Study Type: Prospective., Subjects/specimens: Forty-five patients with benign (n = 8), malignant (n = 36), and ductal carcinoma in situ (n = 1) breast tumors. Lesions and normal fibroglandular tissue (n = 9) were analyzed using sections of formalin-fixed, paraffin-embedded tissue stained with hematoxylin, erythrosine, and saffron., Field Strength/sequence: MRI (3T) protocols: Protocol I: Twice-refocused spin-echo echo-planar imaging with: echo time (TE) 85 msec; repetition time (TR) 9300/11600 msec; matrix 90 × 90 × 60; voxel size 2 × 2 × 2.5 mm 3 ; b-values: 0 and 700 s/mm 2 . Protocol II: Stejskal-Tanner spin-echo echo-planar imaging with: TE: 88 msec; TR: 10600/11800 msec, matrix 90 × 90 × 60; voxel size 2 × 2 × 2.5 mm 3 ; b-values [0, 200, 600, 1200, 1800, 2400, 3000] s/mm 2 ., Assessment: Area fractions of cellular and collagen content in histologic sections were quantified using whole-slide image analysis and compared with the corresponding DWI parameters., Statistical Tests: Correlations were assessed using Pearson's r. Univariate analysis of group median values was done using the Mann-Whitney U-test., Results: Collagen content correlated with the fast signal fraction (r = 0.67, P < 0.001) and ADC (r = 0.58, P < 0.001) and was lower (P < 0.05) in malignant lesions than benign and normal tissues. Cellular content correlated inversely with the fast signal fraction (r = -0.67, P < 0.001) and ADC (r = -0.61, P < 0.001) and was different (P < 0.05) between malignant, benign, and normal tissues., Data Conclusion: Our findings suggest stromal collagen content increases diffusivity observed by MRI and is associated with higher ADC and fast signal fraction of the biexponential model., Level of Evidence: 3 Technical Efficacy Stage: 3 J. Magn. Reson. Imaging 2020;51:1868-1878., (© 2019 The Authors. Journal of Magnetic Resonance Imaging published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine.)

Background: Relative enhanced diffusivity (RED) is a potential biomarker for indirectly measuring perfusion in tissue using diffusion-weighted magnetic resonance imaging (MRI) with 3 b values., Purpose: To optimize the RED MRI protocol for the prostate, and to investigate its potential for prostate cancer (PCa) diagnosis., Study Type: Prospective., Population: Ten asymptomatic healthy volunteers and 35 patients with clinical suspicion of PCa., Sequence: 3T T 2 - and diffusion-weighted MRI with b values: b = 0, 50, [100], 150, [200], 250, [300], 400, 800 s/mm 2 (values in brackets were only used for patients)., Assessment: Monte Carlo simulations were performed to assess noise sensitivity of RED as a function of intermediate b value. Volunteers were scanned 3 times to assess repeatability of RED. Patient data were used to investigate RED's potential for discriminating between biopsy-confirmed cancer and healthy tissue, and between true and false positive radiological findings., Statistical Tests: Within-subject coefficient of variation (WCV) to assess repeatability and receiver-operating characteristic curve analysis and logistic regression to assess diagnostic performance of RED., Results: The repeatability was acceptable (WCV = 0.2-0.3) for all intermediate b values tested, apart from b = 50 s/mm 2 (WCV = 0.3-0.4). The simulated RED values agreed well with the experimental data, showing that an intermediate b value between 150-250 s/mm 2 minimizes noise sensitivity in both peripheral zone (PZ) and transition zone (TZ). RED calculated with the b values 0, 150 and 800 s/mm 2 was significantly higher in tumors than in healthy tissue in both PZ (P < 0.001, area under the curve [AUC] = 0.85) and PZ + TZ (P < 0.001, AUC = 0.84). RED was shown to aid apparent diffusion coefficient (ADC) in differentiating between false-positive findings and true-positive PCa in the PZ (AUC; RED = 0.71, ADC = 0.74, RED+ADC = 0.77)., Data Conclusion: RED is a repeatable biomarker that may have value for prostate cancer diagnosis. An intermediate b value in the range of 150-250 s/mm 2 minimizes the influence of noise and maximizes repeatability., Level of Evidence: 2 Technical Efficacy Stage: 1 J. Magn. Reson. Imaging 2020;51:1900-1910., (© 2019 The Authors. Journal of Magnetic Resonance Imaging published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine.)

Objectives. Heart failure (HF) impairs resting myocardial energetics, myocardial mitochondrial performance, and maximal oxygen uptake (VO 2max ). Exercise training is included in most rehabilitation programs and benefits HF patients. However, the effect of exercise intensity on cardiac mitochondrial respiration and concentrations of the key bioenergetic metabolites phosphocreatine (PCr), adenosine triphosphate (ATP), and inorganic phosphate (Pi) is unclear. This study aimed to investigate the effects of exercise training at different intensities in rats with HF. Methods. Rats underwent myocardial infarction or sham operations and were divided into three subgroups: sedentary, moderate intensity, or high intensity. The impact of HF and 6 weeks of exercise training on energy metabolism was evaluated by 31 P magnetic resonance spectroscopy and mitochondrial respirometry. The concentrations of PCr, ATP, and Pi were quantified by magnetic resonance spectroscopy. VO 2max was measured by treadmill respirometry. Results. Exercise training increased VO 2max in sham and HF. PCr/ATP ratio was reduced in HF ( p  < .01) and remained unchanged by exercise training. PCr concentration was significantly lower in HF compared to sham ( p  < .01). Moderate and high-intensity exercise training increased ATP in HF and sham. HF impaired complex I (CI) and complex II ( p  = .034) respiration. High-intensity exercise training recovered CI respiration in HF rats compared to HF sedentary ( p  = .014). Conclusions. Exercise training improved cardiac performance, as indicated by increased VO 2max and higher exercise capacity, without changing the myocardial PCr/ATP ratio. These observations suggest that the PCr/ATP biomarker is not suited to evaluate the beneficial effects of exercise training in the heart. The exact mechanisms require further investigations, as exercise training did increase ATP levels and CI respiration.

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Objectives: To investigate the reliability of simultaneous positron emission tomography and magnetic resonance imaging (PET/MRI)-derived biomarkers using semi-automated Gaussian mixture model (GMM) segmentation on PET images, against conventional manual tumor segmentation on dynamic contrast-enhanced (DCE) images., Materials and Methods: Twenty-four breast cancer patients underwent PET/MRI (following 18F-fluorodeoxyglucose (18F-FDG) injection) at baseline and during neoadjuvant treatment, yielding 53 data sets (24 untreated, 29 treated). Two-dimensional tumor segmentation was performed manually on DCE-MRI images (manual DCE) and using GMM with corresponding PET images (GMM-PET). Tumor area and mean apparent diffusion coefficient (ADC) derived from both segmentation methods were compared, and spatial overlap between the segmentations was assessed with Dice similarity coefficient and center-of-gravity displacement., Results: No significant differences were observed between mean ADC and tumor area derived from manual DCE segmentation and GMM-PET. There were strong positive correlations for tumor area and ADC derived from manual DCE and GMM-PET for untreated and treated lesions. The mean Dice score for GMM-PET was 0.770 and 0.649 for untreated and treated lesions, respectively., Discussion: Using PET/MRI, tumor area and mean ADC value estimated with a GMM-PET can replicate manual DCE tumor definition from MRI for monitoring neoadjuvant treatment response in breast cancer.

Levels of zinc, along with its mechanistically related metabolites citrate and aspartate, are widely reported as reduced in prostate cancer compared to healthy tissue and are therefore pointed out as potential cancer biomarkers. Previously, it has only been possible to analyze zinc and metabolites by separate detection methods. Through matrix-assisted laser desorption/ionization mass spectrometry imaging (MSI), we were for the first time able to demonstrate, in two different sample sets ( n = 45 and n = 4), the simultaneous spatial detection of zinc, in the form of ZnCl 3 - , together with citrate, aspartate, and N -acetylaspartate on human prostate cancer tissues. The reliability of the ZnCl 3 - detection was validated by total zinc determination using laser ablation inductively coupled plasma MSI on adjacent serial tissue sections. Zinc, citrate, and aspartate were correlated with each other (range r = 0.46 to 0.74) and showed a significant reduction in cancer compared to non-cancer epithelium ( p < 0.05, log 2 fold change range: -0.423 to -0.987), while no significant difference between cancer and stroma tissue was found. Simultaneous spatial detection of zinc and its metabolites is not only a valuable tool for analyzing the role of zinc in prostate metabolism but might also provide a fast and simple method to detect zinc, citrate, and aspartate levels as a biomarker signature for prostate cancer diagnostics and prognostics.

Angiogenesis is necessary for tumor growth and has been targeted in breast cancer; however, it is unclear which patients will respond and benefit from antiangiogenic therapy. We report noninvasive monitoring of patient response to neoadjuvant chemotherapy given alone or in combination with anti-vascular endothelial growth factor (bevacizumab) in a randomized clinical trial. At four time points during neoadjuvant chemotherapy ± bevacizumab of receptor tyrosine-protein kinase erbB-2-negative breast cancers, we measured metabolites and inflammation-related markers in patient's serum. We report significant changes in the levels of several molecules induced by bevacizumab, the most prominent being an increase in pentraxin 3 (PTX3) and von Willebrand factor (VWF). Serum levels of AXL, VWF and pulmonary and activation-regulated cytokine (PARC/CCL18) reflected response to chemotherapy alone or in combination with bevacizumab. We further analyzed serum cytokines in relation to tumor characteristics such as gene expression, tumor metabolites and tumor infiltrating leukocytes. We found that VWF and growth-differentiation factor 15 tumor mRNA levels correlated with their respective serum protein levels suggesting that these cytokines may be produced by tumors and outflow to the bloodstream while influencing the tumor microenvironment locally. Finally, we used binomial logistic regression which allowed to predict patient's response using only 10 noninvasive biomarkers. Our study highlights the potential of monitoring circulating levels of cytokines and metabolites during breast cancer therapy., (© 2019 The Authors. International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.)

Background: The metabolic consequences of preoperative carbohydrate load in breast cancer patients are not known. The present explorative study investigated the systemic and tumor metabolic changes after preoperative per-oral carbohydrate load and their influence on tumor characteristics and survival., Methods: The study setting was on university hospital level with primary and secondary care functions in south-west Norway. Serum and tumor tissue were sampled from a population-based cohort of 60 patients with operable breast cancer who were randomized to either per-oral carbohydrate load (preOp™; n = 25) or standard pre-operative fasting (n = 35) before surgery. Magnetic resonance (MR) metabolomics was performed on serum samples from all patients and high-resolution magic angle spinning (HR-MAS) MR analysis on 13 tumor samples available from the fasting group and 16 tumor samples from the carbohydrate group., Results: Fourteen of 28 metabolites were differently expressed between fasting and carbohydrate groups. Partial least squares discriminant analysis showed a significant difference in the metabolic profile between the fasting and carbohydrate groups, compatible with the endocrine effects of insulin (i.e., increased serum-lactate and pyruvate and decreased ketone bodies and amino acids in the carbohydrate group). Among ER-positive tumors (n = 18), glutathione was significantly elevated in the carbohydrate group compared to the fasting group (p = 0.002), with a positive correlation between preoperative S-insulin levels and the glutathione content in tumors (r = 0.680; p = 0.002). In all tumors (n = 29), glutamate was increased in tumors with high proliferation (t-test; p = 0.009), independent of intervention group. Moreover, there was a positive correlation between tumor size and proliferation markers in the carbohydrate group only. Patients with ER-positive / T2 tumors and high tumor glutathione (≥1.09), high S-lactate (≥56.9), and high S-pyruvate (≥12.5) had inferior clinical outcomes regarding relapse-free survival, breast cancer-specific survival, and overall survival. Moreover, Integrated Pathway Analysis (IPA) in serum revealed activation of five major anabolic metabolic networks contributing to proliferation and growth., Conclusions: Preoperative carbohydrate load increases systemic levels of lactate and pyruvate and tumor levels of glutathione and glutamate in ER-positive patients. These biological changes may contribute to the inferior clinical outcomes observed in luminal T2 breast cancer patients., Trial of Registration: ClinicalTrials.gov; NCT03886389. Retrospectively registered March 22, 2019.

Background: Metabolomic characterization of tumours can potentially improve prediction of cancer prognosis and treatment response. Here, we describe efforts to validate previous metabolomic findings using a historical cohort of breast cancer patients and discuss challenges with using older biobanks collected with non-standardized sampling procedures. Methods: In total, 100 primary breast cancer samples were analysed by high-resolution magic angle spinning magnetic resonance spectroscopy (HR MAS MRS) and subsequently examined by histology. Metabolomic profiles were related to the presence of cancer tissue, hormone receptor status, T-stage, N-stage, and survival. RNA integrity number (RIN) and metabolomic profiles were compared with an ongoing breast cancer biobank. Results: The 100 samples had a median RIN of 4.3, while the ongoing biobank had a significantly higher median RIN of 6.3 ( p = 5.86 × 10 -7 ). A low RIN was associated with changes in choline-containing metabolites and creatine, and the samples in the older biobank showed metabolic differences previously associated with tissue degradation. The association between metabolomic profile and oestrogen receptor status was in accordance with previous findings, however, with a lower classification accuracy. Conclusions: Our findings highlight the importance of standardized biobanking procedures in breast cancer metabolomics studies.

Metabolomics analysis of biofluids is a feasible tool for disease characterization and monitoring due to its minimally invasive nature. To reduce unwanted variation in biobanks and clinical studies, it is important to determine the effect of external factors on metabolic profiles of biofluids. In this study we examined the effect of sample collection and sample processing procedures on NMR measured serum lipoproteins and small-molecule metabolites in serum and urine, using a cohort of men diagnosed with either prostate cancer or benign prostatic hyperplasia. We determined day-to-day reliability of metabolites by systematic sample collection at two different days, in both fasting and non-fasting conditions. Study participants received prostate massage the first day to assess the differences between urine with and without prostate secretions. Further, metabolic differences between first-void and mid-stream urine samples, and the effect of centrifugation of urine samples before storage were assessed. Our results show that day-to-day reliability is highly variable between metabolites in both serum and urine, while lipoprotein subfractions possess high reliability. Further, fasting status clearly influenced the metabolite concentrations, demonstrating the importance of keeping this condition constant within a study cohort. Day-to-day reliabilities were however comparable in fasting and non-fasting samples. Urine sampling procedures such as sampling of first-void or mid-stream urine, and centrifugation or not before sample storage, were shown to only have minimal effect on the overall metabolic profile, and is thus unlikely to constitute a confounder in clinical studies utilizing NMR derived metabolomics., (Copyright © 2019. Published by Elsevier B.V.)

Background: Diffusion-weighted MRI (DWI) has potential to noninvasively characterize breast cancer lesions; models such as intravoxel incoherent motion (IVIM) provide pseudodiffusion parameters that reflect tissue perfusion, but are dependent on the details of acquisition and analysis strategy., Purpose: To examine the effect of fitting algorithms, including conventional least-squares (LSQ) and segmented (SEG) methods as well as Bayesian methods with global shrinkage (BSP) and local spatial (FBM) priors, on the power of IVIM parameters to differentiate benign and malignant breast lesions., Study Type: Prospective patient study., Subjects: 61 patients with confirmed breast lesions., Field Strength/sequence: DWI (bipolar SE-EPI, 13 b values) was included in a clinical MR protocol including T 2 -weighted and dynamic contrast-enhanced MRI on a 3T scanner., Assessment: The IVIM model was fitted voxelwise in lesion regions of interest (ROIs), and derived parameters were compared across methods within benign and malignant subgroups (correlation, coefficients of variation). Area under receiver operator characteristic curves (ROC AUCs) were calculated to determine discriminatory power of parameter combinations from all fitting methods., Statistical Tests: Kruskal-Wallis, Mann-Whitney, Pearson correlation., Results: All methods provided useful IVIM parameters; D was well-correlated across all methods (r > 0.8), with a wider range for f and D* (0.3-0.7). Fitting methods gave detectable differences in parameters, but all showed increased f and decreased D in malign lesions. D was the most discriminatory single parameter, with LSQ performing least well (AUC 0.83). In general, ROC AUCs were maximized by the inclusion of pseudodiffusion parameters, and by the use of Bayesian methods incorporating prior information (maximum AUC of 0.92 for BSP)., Data Conclusion: DWI performs well at classifying breast lesions, but careful consideration of analysis procedure can improve performance. D is the most discriminatory single parameter, but including pseudodiffusion parameters (f and D*) increases ROC AUC. Bayesian methods outperformed conventional least-squares and segmented fitting methods for breast lesion classification., Level of Evidence: 3 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2019;50:1478-1488., (© 2019 International Society for Magnetic Resonance in Medicine.)

Clinical end-stage parameters define the pregnancy disorders preeclampsia and fetal growth restriction while classification of the underlying placental dysfunction is missing and urgently needed. Flt-1 (FMS-like tyrosine kinase receptor 1) is the most promising placenta-derived predictive biomarker for preeclampsia. We aimed to classify placental dysfunction in preeclampsia and fetal growth restriction at delivery by metabolic profiling and authenticate the biomarker Flt-1 for placental dysfunction. We studied 143 pregnancies with or without preeclampsia and/or fetal growth restriction delivered by cesarean section. Metabolic placenta profiles were created by high-resolution magic angle spinning nuclear magnetic resonance spectroscopy and the resulting placental phenotypes obtained by hierarchical clustering. Placental Flt-1 expression (membrane-bound and soluble isoforms combined) and maternal serum Flt-1 expression (soluble isoforms) were analyzed by immunohistochemistry and ELISA, respectively. We identified 3 distinct placenta groups by 21 metabolites and diagnostic outcome parameters; normal placentas, moderate placental dysfunction, and severe placental dysfunction. Increased placental Flt-1 was associated with severe placental dysfunction, and increased serum Flt-1 was associated with moderate and severe placental dysfunction. The preeclamptic pregnancies with and without placental dysfunction could be distinguished by 5 metabolites and placental Flt-1. Placental Flt-1 alone could separate normal pregnancies with and without placental dysfunction. In conclusion, metabolomics could classify placental dysfunction and provide information not identified by traditional diagnostics and metabolites with biomarker potential were identified. Flt-1 was confirmed as precision biomarker for placental dysfunction, substantiating its usefulness for identification of high-risk pregnancies for preeclampsia and fetal growth restriction with placental involvement.

Patients with locally advanced breast cancer have a worse prognosis compared to patients with localized tumors and require neoadjuvant treatment before surgery. The aim of this study was to characterize the systemic metabolic effect of neoadjuvant chemotherapy in patients with large primary breast cancers and to relate these changes to treatment response and long-term survival. This study included 132 patients with large primary breast tumors randomized to receive neoadjuvant chemotherapy with or without the addition of the antiangiogenic drug Bevacizumab. Tumor biopsies and serum were collected before and during treatment and, serum additionally 6 weeks after surgery. Samples were analyzed by nuclear magnetic resonance spectroscopy (NMR). Correlation analysis showed low correlations between metabolites measured in cancer tissue and serum. Multilevel partial least squares discriminant analysis (PLS-DA) showed clear changes in serum metabolite levels during treatment ( p -values ≤ 0.001), including unfavorable changes in lipid levels. PLS-DA revealed metabolic differences between tissue samples from survivors and nonsurvivors collected 12 weeks into treatment with an accuracy of 72% ( p -value = 0.005); however, this was not evident in serum samples. Our results demonstrate a potential clinical application for serum-metabolomics for patient monitoring during and after treatment, and indicate potential for tissue NMR spectroscopy for predicting patient survival.

Metabolic profiling of biofluids by nuclear magnetic resonance (NMR) spectroscopy serves as an important tool in disease characterization, and its accuracy largely depends on the quality of samples. We aimed to explore possible effects of repeated freeze-thaw cycles (FTCs) on concentrations of lipoprotein parameters in serum and metabolite concentrations in serum and urine samples. After one to five FTCs, serum and urine samples ( n = 20) were analyzed by NMR spectroscopy, and 112 lipoprotein parameters, 20 serum metabolites, and 35 urine metabolites were quantified by a commercial analytical platform. Principal component analysis showed no systematic changes related to FTCs, and samples from the same donor were closely clustered, showing a higher between-subject variation than within-subject variation. The coefficients of variation were small (medians of 4.3%, 11.0%, and 4.9%  for lipoprotein parameters and serum and urine metabolites, respectively). Minor, but significant accumulated freeze-thaw effects were observed for 32 lipoprotein parameters and one serum metabolite (acetic acid) when comparing FTC1 to further FTCs. Remaining lipoprotein and metabolite concentrations showed no significant change. In conclusion, five FTCs did not significantly alter the concentrations of urine metabolites and introduced only minor changes to serum lipoprotein parameters and metabolites evaluated by the NMR-based platform.

This article presents an overview of the current literature on PET imaging with prostate-specific membrane antigen ligands, especially focusing on the potential role of simultaneous PET/magnetic resonance imaging for the planning of salvage radiotherapy in patients with prostate cancer with biochemical recurrence after radical prostatectomy., (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background: Oxygen is a liberally dosed medicine; however, too much oxygen can be harmful. In certain situations, treatment with high oxygen concentration is necessary, e.g. after cardiopulmonary resuscitation. The amount of oxygen and duration of hyperoxia causing pulmonary damage is not fully elucidated. The aim of this study was to investigate pathophysiological and metabolite changes in lung tissue during hyperoxia while the lungs were kept open under constant low pressure., Methods: Seven pigs were exposed to 100% oxygen for five hours, using an apneic oxygenation technique with one long uninterrupted inspiration, while carbon dioxide was removed with an interventional lung assist. Arterial blood samples were collected every 30 minutes. Lung biopsies were obtained before and after hyperoxia. Microscopy and high-resolution magic angle spinning nuclear magnetic resonance spectroscopy were used to detect possible pathological and metabolite changes, respectively. Unsupervised multivariate analysis of variance and paired sample tests were performed. A two-tailed p-value ≤ 0.05 was considered significant., Results: No significant changes in arterial pH, and partial pressure of carbon dioxide, and no clear histopathological changes were observed after hyperoxia. While blood glucose and lactate levels changed to a minor degree, their levels dropped significantly in the lung after hyperoxia (p ≤ 0.04). Reduced levels of antioxidants (p ≤ 0.05), tricarboxylic acid cycle and energy (p ≤ 0.04) metabolites and increased levels of several amino acids (p ≤ 0.05) were also detected., Conclusion: Despite no histological changes, tissue metabolites were altered, indicating that exposure to hyperoxia affects lung tissue matrix on a molecular basis., (© 2019 The Authors.)

Background and Aims: Physical activity is favourably associated with certain markers of lipid metabolism. The relationship of physical activity with lipoprotein particle profiles in children is not known. Here we examine cross-sectional associations between objectively measured physical activity and sedentary time with serum markers of lipoprotein metabolism., Methods: Our cohort included 880 children (49.0% girls, mean age 10.2 years). Physical activity intensity and time spent sedentary were measured objectively using accelerometers. 30 measures of lipoprotein metabolism were quantified using nuclear magnetic resonance spectroscopy. Multiple linear regression models adjusted for age, sex, sexual maturity and socioeconomic status were used to determine associations of physical activity and sedentary time with lipoprotein measures. Additional models were adjusted for adiposity. Isotemporal substitution models quantified theoretical associations of replacing 30 min of sedentary time with 30 min of moderate- to vigorous-intensity physical activity (MVPA)., Results: Time spent in MVPA was associated with a favourable lipoprotein profile independent of sedentary time. There were inverse associations with a number of lipoprotein measures, including most apolipoprotein B-containing lipoprotein subclasses and triglyceride measures, the ratio of total to high-density lipoprotein (HDL) cholesterol, and non-HDL cholesterol concentration. There were positive associations with larger HDL subclasses, HDL cholesterol concentration and particle size. Reallocating 30 min of sedentary time to MVPA had broadly similar associations. Sedentary time was only partly and weakly associated with an unfavourable lipoprotein profile., Conclusions: Physical activity of at least moderate-intensity is associated with a favourable lipoprotein profile in schoolchildren, independent of time spent sedentary, adiposity and other confounders., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Effective transverse relaxivity of gadolinium-based contrast agents is often neglected in dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI). Here, we assess time and tissue dependence of R2* enhancement and its impact on pharmacokinetic parameter quantification and treatment monitoring. Multiecho DCE-MRI was performed at 7 T on mice bearing subcutaneous TOV-21G human ovarian cancer xenografts (n = 8) and on the transgenic adenocarcinoma of the mouse prostate (TRAMP) model (n = 7). Subsequently, the TOV-21G tumor-bearing mice were treated with bevacizumab and rescanned 2 days later. Pharmacokinetic analysis (extended Tofts model) was performed using either the first echo signal only (standard single-echo DCE-MRI) or the estimated signal at TE = 0 derived from exponential fitting of R2* relaxation (R2*-corrected). Neglecting R2* enhancement causes underestimation of Gd-DOTA concentration (peak enhancement underestimated by 9.4%-16% in TOV-21G tumors and 13%-20% in TRAMP prostates). Median K trans and v e were underestimated in every mouse (TOV-21G K trans : 11%-19%, TOV-21G v e : 5.3%-8.9%; TRAMP K trans : 8.6%-19%, TRAMP v e : 12%-21%). Bevacizumab treatment reduced K trans in all TOV-21G tumors after 48 hours. Treatment effect was significantly greater in all tumors after R2* correction (median change of -0.050 min -1 in R2*-corrected K trans vs. -0.037 min -1 in uncorrected K trans ). R2* enhancement in DCE-MRI is both time- and tissue-dependent and may not be negligible at 7 T in tissue with high K trans . This has consequences for the use of K trans and other DCE-MRI parameters as biomarkers, because treatment effect size can be underestimated when R2* enhancement is neglected., Competing Interests: Conflict of Interest: None reported., (© 2019 The Authors. Published by Grapho Publications, LLC.)

In the preceding decades, molecular characterization has revolutionized breast cancer (BC) research and therapeutic approaches. Presented herein, an unbiased analysis of breast tumor proteomes, inclusive of 9995 proteins quantified across all tumors, for the first time recapitulates BC subtypes. Additionally, poor-prognosis basal-like and luminal B tumors are further subdivided by immune component infiltration, suggesting the current classification is incomplete. Proteome-based networks distinguish functional protein modules for breast tumor groups, with co-expression of EGFR and MET marking ductal carcinoma in situ regions of normal-like tumors and lending to a more accurate classification of this poorly defined subtype. Genes included within prognostic mRNA panels have significantly higher than average mRNA-protein correlations, and gene copy number alterations are dampened at the protein-level; underscoring the value of proteome quantification for prognostication and phenotypic classification. Furthermore, protein products mapping to non-coding genomic regions are identified; highlighting a potential new class of tumor-specific immunotherapeutic targets.

The metabolism and performance of myocardial and skeletal muscle are impaired in heart failure (HF) patients. Exercise training improves the performance and benefits the quality of life in HF patients. The purpose of the present study was to determine the metabolic profiles in myocardial and skeletal muscle in HF and exercise training using MRS, and thus to identify targets for clinical MRS in vivo. After surgically establishing HF in rats, we randomized the rats to exercise training programs of different intensities. After the final training session, rats were sacrificed and tissues from the myocardial and skeletal muscle were extracted. Magnetic resonance spectra were acquired from these extracts, and principal component and metabolic enrichment analysis were used to assess the differences in metabolic profiles. The results indicated that HF affected myocardial metabolism by changing multiple metabolites, whereas it had a limited effect on skeletal muscle metabolism. Moreover, exercise training mainly altered the metabolite distribution in skeletal muscle, indicating regulation of metabolic pathways of taurine and hypotaurine metabolism and carnitine synthesis.

NMR-based metabolomics has shown promise in the diagnosis of diseases as it enables identification and quantification of metabolic biomarkers. Using high-resolution magic-angle-spinning (HR-MAS) NMR spectroscopy, metabolic profiles from intact tissue specimens can be obtained with high spectral resolution. In addition, HR-MAS NMR requires minimal sample preparation and the sample is kept intact for subsequent analyses. In this chapter, we describe a typical protocol for NMR-based metabolomics of tissue samples. We cover all major steps ranging from tissue sample collection to determination of biomarkers, including experimental precautions taken to ensure reproducible and reliable reporting of data in the area of clinical application.

Tumor hypoxia contributes to therapy resistance and metastatic progression of locally advanced rectal cancer (LARC). We postulated that the tumor mitochondrial metabolism, manifested by reactive oxygen species (ROS) and mitochondrial DNA (mtDNA) damage, reflects how hypoxic conditions connect to cancer-induced systemic inflammation and poor outcome. Levels of ROS and mtDNA damage were analyzed in three colorectal cancer (CRC) cell lines cultured for 24 hours under normoxia (21% O 2 ) or hypoxia (0.2% O 2 ) and serum sampled at the time of diagnosis from 35 LARC patients participating in a prospective therapy study. Compared with normoxia, ROS were significantly repressed and mtDNA damage was significantly enhanced in the hypoxic CRC cell lines; hence, a low ratio of ROS to mtDNA damage was an indicator of hypoxic conditions. In the LARC patients, low serum ROS were associated with elevated levels of circulating carcinoembryonic antigen and tumor choline concentration, both indicative of unfavorable biology, as well as adverse progression-free and overall survival. A low ratio of ROS to mtDNA damage in serum was associated with poor local tumor response to the neoadjuvant treatment and, of note, elevated systemic inflammation factors (C-reactive protein, the interleukin-1 receptor antagonist, and factors involved in tumor necrosis factor signaling), indicating that deficient treatment response locally and detrimental inflammation systemically link to a hypoxic mitochondrial metabolism. In conclusion, serum ROS and damaged mtDNA may be markers of the mitochondrial metabolism driven by the state of oxygenation of the primary tumor and possibly implicated in systemic inflammation and adverse outcome of LARC., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)

Purpose: Exercise training increases aerobic capacity and is beneficial for health, whereas low aerobic exercise capacity is a strong independent predictor of cardiovascular disease and premature death. The purpose of the present study was to determine the metabolic profiles in a rat model of inborn low versus high capacity runners (LCR/HCR) and to determine the effect of inborn aerobic capacity, aging, and exercise training on skeletal muscle metabolic profile., Methods: LCR/HCR rats were randomized to high intensity low volume interval treadmill training twice a week or sedentary control for 3 or 11 months before they were sacrificed, at 9 and 18 months of age, respectively. Magnetic resonance spectra were acquired from soleus muscle extracts, and partial least square discriminative analysis was used to determine the differences in metabolic profile., Results: Sedentary HCR rats had 54% and 30% higher VO2max compared to sedentary LCR rats at 9 months and 18 months, respectively. In HCR, exercise increased running speed significantly, and VO2max was higher at age of 9 months, compared to sedentary counterparts. In LCR, changes were small and did not reach the level of significance. The metabolic profile was significantly different in the LCR sedentary group compared to the HCR sedentary group at the age of 9 and 18 months, with higher glutamine and glutamate levels (9 months) and lower lactate level (18 months) in HCR. Irrespective of fitness level, aging was associated with increased soleus muscle concentrations of glycerophosphocholine and glucose. Interval training did not influence metabolic profiles in LCR or HCR rats at any age., Conclusion: Differences in inborn aerobic capacity gave the most marked contrasts in metabolic profile, there were also some changes with ageing. Low volume high intensity interval training twice a week had no detectable effect on metabolic profile., Competing Interests: The authors have declared that no competing interests exist.

The objective of this study was to evaluate the effect of including bone in Dixon-based attenuation correction for 18 F-fluciclovine PET/MRI of primary and recurrent prostate cancer. Methods: 18 F-fluciclovine PET data from 2 PET/MRI studies-one for staging of high-risk prostate cancer (28 patients) and one for diagnosis of recurrent prostate cancer (81 patients)-were reconstructed with a 4-compartment (reference) and 5-compartment attenuation map. In the latter, continuous linear attenuation coefficients for bone were included by coregistration with an atlas. The SUV max and mean 50% isocontour SUV (SUV iso ) of primary, locally recurrent, and metastatic lesions were compared between the 2 reconstruction methods using linear mixed-effects models. In addition, mean SUVs were obtained from bone marrow in the third lumbar vertebra (L3) to investigate the effect of including bone attenuation on lesion-to-bone marrow SUV ratios (SUVR max and SUVR iso ; recurrence study only). The 5-compartment attenuation maps were visually compared with the in-phase Dixon MR images for evaluation of bone registration errors near the lesions. P values of less than 0.05 were considered significant. Results: Sixty-two lesions from 39 patients were evaluated. Bone registration errors were found near 19 (31%) of these lesions. In the remaining 8 primary prostate tumors, 7 locally recurrent lesions, and 28 lymph node metastases without bone registration errors, use of the 5-compartment attenuation map was associated with small but significant increases in SUV max (2.5%; 95% confidence interval [CI], 2.0%-3.0%; P < 0.001) and SUV iso (2.5%; 95% CI, 1.9%-3.0%; P < 0.001), but not SUVR max (0.2%; 95% CI, -0.5%-0.9%; P = 0.604) and SUVR iso (0.2%; 95% CI -0.6%-1.0%; P = 0.581), in comparison to the 4-compartment attenuation map. Conclusion: The investigated method for atlas-based inclusion of bone in 18 F-fluciclovine PET/MRI attenuation correction has only a small effect on the SUVs of soft-tissue prostate cancer lesions, and no effect on their lesion-to-bone marrow SUVRs when using signal from L3 as a reference. The attenuation maps should always be checked for registration artifacts for lesions in or close to the bones., (© 2018 by the Society of Nuclear Medicine and Molecular Imaging.)

Purpose: To investigate the associations of metabolite levels derived from magnetic resonance spectroscopic imaging (MRSI) and 18 F-fluciclovine positron emission tomography (PET) with prostate tissue characteristics. Methods: In a cohort of 19 high-risk prostate cancer patients that underwent simultaneous PET/MRI, we evaluated the diagnostic performance of MRSI and PET for discrimination of aggressive cancer lesions from healthy tissue and benign lesions. Data analysis comprised calculations of correlations of mean standardized uptake values (SUV mean ), maximum SUV (SUV max ), and the MRSI-derived ratio of (total choline + spermine + creatine) to citrate (CSC/C). Whole-mount histopathology was used as gold standard. Results: The results showed a moderate significant correlation between both SUVmean and SUVmax with CSC/C ratio. Conclusions: We demonstrated that the simultaneous acquisition of 18 F-fluciclovine PET and MRSI with an integrated PET/MRI system is feasible and a combination of these imaging modalities has potential to improve the diagnostic sensitivity and specificity of prostate cancer lesions.

Background: Breast cancer treatment has metabolic side effects, potentially affecting risk of cardiovascular disease (CVD) and recurrence. We aimed to compare alterations in serum metabolites and lipoproteins during treatment between recipients and non-recipients of chemotherapy, and describe metabolite profiles associated with treatment-related weight gain., Methods: This pilot study includes 60 stage I/II breast cancer patients who underwent surgery and were treated according to national guidelines. Serum sampled pre-surgery and after 6 and 12 months was analysed by MR spectroscopy and mass spectrometry. In all, 170 metabolites and 105 lipoprotein subfractions were quantified., Results: The metabolite and lipoprotein profiles of chemotherapy recipients and non-recipients changed significantly 6 months after surgery (p < 0.001). Kynurenine, the lipid signal at 1.55-1.60 ppm, ADMA, 2 phosphatidylcholines (PC aa C38:3, PC ae C42:1), alpha-aminoadipic acid, hexoses and sphingolipids were increased in chemotherapy recipients after 6 months. VLDL and small dense LDL increased after 6 months, while HDL decreased, with triglyceride enrichment in HDL and LDL. At baseline, weight gainers had less acylcarnitines, phosphatidylcholines, lyso-phosphatidylcholines and sphingolipids, and showed an inflammatory lipid profile., Conclusion: Chemotherapy recipients exhibit metabolic changes associated with inflammation, altered immune response and increased risk of CVD. Altered lipid metabolism may predispose for treatment-related weight gain.

Reactive stroma is a tissue feature commonly observed in the tumor microenvironment of prostate cancer and has previously been associated with more aggressive tumors. The aim of this study was to detect differentially expressed genes and metabolites according to reactive stroma content measured on the exact same prostate cancer tissue sample. Reactive stroma was evaluated using histopathology from 108 fresh frozen prostate cancer samples gathered from 43 patients after prostatectomy (Biobank1). A subset of the samples was analyzed both for metabolic (n = 85) and transcriptomic alterations (n = 78) using high resolution magic angle spinning magnetic resonance spectroscopy (HR-MAS MRS) and RNA microarray, respectively. Recurrence-free survival was assessed in patients with clinical follow-up of minimum five years (n = 38) using biochemical recurrence (BCR) as endpoint. Multivariate metabolomics and gene expression analysis compared low (≤15%) against high reactive stroma content (≥16%). High reactive stroma content was associated with BCR in prostate cancer patients even when accounting for the influence of Grade Group (Cox hazard proportional analysis, p = 0.013). In samples with high reactive stroma content, metabolites and genes linked to immune functions and extracellular matrix (ECM) remodeling were significantly upregulated. Future validation of these findings is important to reveal novel biomarkers and drug targets connected to immune mechanisms and ECM in prostate cancer. The fact that high reactive stroma grading is connected to BCR adds further support for the clinical integration of this histopathological evaluation.

Objective: To investigate the diagnostic potential of simultaneous 18 F-fluciclovine PET/MRI for pelvic lymph node (LN) staging in patients with high-risk prostate cancer., Methods: High-risk prostate cancer patients (n=28) underwent simultaneous 18 F-fluciclovine PET/MRI prior to surgery. LNs were removed according to a predefined template of eight regions. PET and MR images were evaluated for presence of LN metastases according to these regions. Sensitivity/specificity for detection of LN metastases were calculated on patient and region basis. Sizes of LN metastases in regions with positive and negative imaging findings were compared with linear mixed models. Clinical parameters of PET-positive and -negative stage N1 patients were compared with the Mann-Whitney U test., Results: Patient- and region-based sensitivity/specificity for detection of pelvic LN metastases was 40 %/87.5 % and 35 %/95.7 %, respectively, for MRI and 40 %/100 % and 30 %/100 %, respectively, for PET. LN metastases in true-positive regions were significantly larger than metastases in false-negative regions. PET-positive stage N1 patients had higher metastatic burden than PET-negative N1 patients., Conclusion: Simultaneous 18 F-fluciclovine PET/MRI provides high specificity but low sensitivity for detection of LN metastases in high-risk prostate cancer patients. 18 F-Fluciclovine PET/MRI scan positive for LN metastases indicates higher metastatic burden than negative scan., Key Points: • 18 F-Fluciclovine PET/MRI has high specificity for detection of lymph node metastasis. • 18 F-Fluciclovine PET/MRI lacks sensitivity to replace ePLND. • 18 F-Fluciclovine PET/MRI may be used to aid surgery and select adjuvant therapy. • 18 F-Fluciclovine PET-positive patients have more extensive disease than PET-negative patients. • Size of metastatic lymph nodes is an important factor for detection.

Background: Steady state susceptibility contrast (SSC)-MRI provides information on vascular morphology but is a rarely used method., Purpose: To investigate the utility of the ultrasmall superparamagnetic iron oxide particles (USPIOs) GEH121333 for measuring tumor response to bevacizumab and compare this with gadolinium-based DCE-MRI., Study Type: Prospective preclinical animal model study., Animal Model: Mice bearing subcutaneous TOV-21G human ovarian cancer xenografts treated with bevacizumab (n = 9) or saline (n = 9)., Field Strength/sequence: Imaging was performed on a 7T Bruker Biospec. For SSC-MRI with GEH121333 we acquired R 1 -maps (RARE-sequence with variable TR), R 2 -maps (multi-spin echo), and R2*-maps (multi-gradient echo). Additionally, R 1 and R 2 maps were measured on the days after USPIO injection. For DCE-MRI with gadodiamide we acquired 200 T 1 -weighted images (RARE-sequence)., Assessment: ΔR 1 , ΔR 2 , and ΔR2* maps were computed from SSC-MRI. DCE-MRI was analysed using the extended Tofts model., Statistical Tests: Results from pre- and 3 days posttreatment SSC-MRI were compared using paired-sample t-tests. Treatment and control groups were compared using independent sample t-tests. Performance of SSC- and DCE-MRI was compared using multivariate partial least squares discriminant analysis., Results: Already one day after treatment and USPIO injection, R 1 and R 2 values were lower in treated (R 1  = 0.49 ± 0.03s -1 , R 2  = 23.07 ± 1.49s -1 ) compared with control tumors (R 1  = 0.52 ± 0.02s -1 , R 2  = 24.98 ± 1.01s -1 ), indicating lower USPIO accumulation. Posttreatment SSC-MRI displayed significantly decreased tumor blood volume (change in ΔR 2  = -0.43 ± 0.26s -1 , P = 0.001) and vessel density (change in Q = -0.032 ± 0.020s -1/3 , P = 0.002). DCE-MRI showed among others lower K trans in treated tumors (control = 0.064 ± 0.011min -1 , tx = 0.046 ± 0.008min -1 , P = 0.002). Multivariate analysis suggests that SSC-MRI was slightly inferior to DCE-MRI in distinguishing treated from control tumors (accuracy = 75%, P = 0.058 versus 80%, P = 0.028), but a combination of both was best (accuracy = 85%; P = 0.003)., Data Conclusion: SSC-MRI with GEH121333 is sensitive to early (<24 h) and late changes in tumor vasculature. SSC-MRI and DCE-MRI provide complementary information and can be used to assess different aspects of vascular responses to anti-angiogenic therapies., Level of Evidence: 1 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2018;47:1589-1600., (© 2017 International Society for Magnetic Resonance in Medicine.)

Objective: To explore the relationship between relative enhanced diffusivity (RED) and intravoxel incoherent motion (IVIM), as well as the impact of noise and the choice of intermediate diffusion weighting (b value) on the RED parameter., Materials and Methods: A mathematical derivation was performed to cast RED in terms of the IVIM parameters. Noise analysis and b value optimization was conducted by using Monte Carlo calculations to generate diffusion-weighted imaging data appropriate to breast and liver tissue at three different signal-to-noise ratios., Results: RED was shown to be approximately linearly proportional to the IVIM parameter f, inversely proportional to D and to follow an inverse exponential decay with respect to D*. The choice of intermediate b value was shown to be important in minimizing the impact of noise on RED and in maximizing its discriminatory power. RED was shown to be essentially a reparameterization of the IVIM estimates for f and D obtained with three b values., Conclusion: RED imaging in the breast and liver should be performed with intermediate b values of 100 and 50 s/mm 2 , respectively. Future clinical studies involving RED should also estimate the IVIM parameters f and D using three b values for comparison.

Background: Diffusion-weighted MRI (DWI) is currently one of the fastest developing MRI-based techniques in oncology. Histogram properties from model fitting of DWI are useful features for differentiation of lesions, and classification can potentially be improved by machine learning., Purpose: To evaluate classification of malignant and benign tumors and breast cancer subtypes using support vector machine (SVM)., Study Type: Prospective., Subjects: Fifty-one patients with benign (n = 23) and malignant (n = 28) breast tumors (26 ER+, whereof six were HER2+)., Field Strength/sequence: Patients were imaged with DW-MRI (3T) using twice refocused spin-echo echo-planar imaging with echo time / repetition time (TR/TE) = 9000/86 msec, 90 × 90 matrix size, 2 × 2 mm in-plane resolution, 2.5 mm slice thickness, and 13 b-values., Assessment: Apparent diffusion coefficient (ADC), relative enhanced diffusivity (RED), and the intravoxel incoherent motion (IVIM) parameters diffusivity (D), pseudo-diffusivity (D*), and perfusion fraction (f) were calculated. The histogram properties (median, mean, standard deviation, skewness, kurtosis) were used as features in SVM (10-fold cross-validation) for differentiation of lesions and subtyping., Statistical Tests: Accuracies of the SVM classifications were calculated to find the combination of features with highest prediction accuracy. Mann-Whitney tests were performed for univariate comparisons., Results: For benign versus malignant tumors, univariate analysis found 11 histogram properties to be significant differentiators. Using SVM, the highest accuracy (0.96) was achieved from a single feature (mean of RED), or from three feature combinations of IVIM or ADC. Combining features from all models gave perfect classification. No single feature predicted HER2 status of ER + tumors (univariate or SVM), although high accuracy (0.90) was achieved with SVM combining several features. Importantly, these features had to include higher-order statistics (kurtosis and skewness), indicating the importance to account for heterogeneity., Data Conclusion: Our findings suggest that SVM, using features from a combination of diffusion models, improves prediction accuracy for differentiation of benign versus malignant breast tumors, and may further assist in subtyping of breast cancer., Level of Evidence: 3 Technical Efficacy: Stage 3 J. Magn. Reson. Imaging 2018;47:1205-1216., (© 2017 International Society for Magnetic Resonance in Medicine.)

The objective of this study was to investigate whether quantitative imaging features derived from combined 18 F-fluciclovine PET/multiparametric MRI show potential for detection and characterization of primary prostate cancer. Methods: Twenty-eight patients diagnosed with high-risk prostate cancer underwent simultaneous 18 F-fluciclovine PET/MRI before radical prostatectomy. Volumes of interest (VOIs) for prostate tumors, benign prostatic hyperplasia (BPH) nodules, prostatitis, and healthy tissue were delineated on T2-weighted images, using histology as a reference. Tumor VOIs were marked as high-grade (≥Gleason grade group 3) or not. MRI and PET features were extracted on the voxel and VOI levels. Partial least-squared discriminant analysis (PLS-DA) with double leave-one-patient-out cross-validation was performed to distinguish tumors from benign tissue (BPH, prostatitis, or healthy tissue) and high-grade tumors from other tissue (low-grade tumors or benign tissue). The performance levels of PET, MRI, and combined PET/MRI features were compared using the area under the receiver-operating-characteristic curve (AUC). Results: Voxel and VOI features were extracted from 40 tumor VOIs (26 high-grade), 36 BPH VOIs, 6 prostatitis VOIs, and 37 healthy-tissue VOIs. PET/MRI performed better than MRI and PET alone for distinguishing tumors from benign tissue (AUCs of 87%, 81%, and 83%, respectively, at the voxel level and 96%, 93%, and 93%, respectively, at the VOI level) and high-grade tumors from other tissue (AUCs of 85%, 79%, and 81%, respectively, at the voxel level and 93%, 93%, and 91%, respectively, at the VOI level). T2-weighted MRI, diffusion-weighted MRI, and PET features were the most important for classification. Conclusion: Combined 18 F-fluciclovine PET/multiparametric MRI shows potential for improving detection and characterization of high-risk prostate cancer, in comparison to MRI and PET alone., (© 2018 by the Society of Nuclear Medicine and Molecular Imaging.)

Purpose: To evaluate the effect of correction for B 0 inhomogeneity-induced geometric distortion in echo-planar diffusion-weighted imaging on quantitative apparent diffusion coefficient (ADC) analysis in multiparametric prostate MRI., Methods: Geometric distortion correction was performed in echo-planar diffusion-weighted images (b = 0, 50, 400, 800 s/mm 2 ) of 28 patients, using two b 0 scans with opposing phase-encoding polarities. Histology-matched tumor and healthy tissue volumes of interest delineated on T 2 -weighted images were mapped to the nondistortion-corrected and distortion-corrected data sets by resampling with and without spatial coregistration. The ADC values were calculated on the volume and voxel level. The effect of distortion correction on ADC quantification and tissue classification was evaluated using linear-mixed models and logistic regression, respectively., Results: Without coregistration, the absolute differences in tumor ADC (range: 0.0002-0.189 mm 2 /s×10 -3 (volume level); 0.014-0.493 mm 2 /s×10 -3 (voxel level)) between the nondistortion-corrected and distortion-corrected were significantly associated (P < 0.05) with distortion distance (mean: 1.4 ± 1.3 mm; range: 0.3-5.3 mm). No significant associations were found upon coregistration; however, in patients with high rectal gas residue, distortion correction resulted in improved spatial representation and significantly better classification of healthy versus tumor voxels (P < 0.05)., Conclusions: Geometric distortion correction in DWI could improve quantitative ADC analysis in multiparametric prostate MRI. Magn Reson Med 79:2524-2532, 2018. © 2017 International Society for Magnetic Resonance in Medicine., (© 2017 International Society for Magnetic Resonance in Medicine.)

Background: The relationship between cholesterol and prostate cancer has been extensively studied for decades, where high levels of cellular cholesterol are generally associated with cancer progression and less favorable outcomes. However, the role of in vivo cellular cholesterol synthesis in this process is unclear, and data on the transcriptional activity of cholesterol synthesis pathway genes in tissue from prostate cancer patients are inconsistent., Methods: A common problem with cancer tissue data from patient cohorts is the presence of heterogeneous tissue which confounds molecular analysis of the samples. In this study we present a general method to minimize systematic confounding from stroma tissue in any prostate cancer cohort comparing prostate cancer and normal samples. In particular we use samples assessed by histopathology to identify genes enriched and depleted in prostate stroma. These genes are then used to assess stroma content in tissue samples from other prostate cancer cohorts where no histopathology is available. Differential expression analysis is performed by comparing cancer and normal samples where the average stroma content has been balanced between the sample groups. In total we analyzed seven patient cohorts with prostate cancer consisting of 1713 prostate cancer and 230 normal tissue samples., Results: When stroma confounding was minimized, differential gene expression analysis over all cohorts showed robust and consistent downregulation of nearly all genes in the cholesterol synthesis pathway. Additional Gene Ontology analysis also identified cholesterol synthesis as the most significantly altered metabolic pathway in prostate cancer at the transcriptional level., Conclusion: The surprising observation that cholesterol synthesis genes are downregulated in prostate cancer is important for our understanding of how prostate cancer cells regulate cholesterol levels in vivo. Moreover, we show that tissue heterogeneity explains the lack of consistency in previous expression analysis of cholesterol synthesis genes in prostate cancer.

This review describes the current status of NMR-based metabolomics of biofluids with respect to cancer risk assessment, detection, disease characterization, prognosis, and treatment monitoring. While the metabolism of cancer cells is altered compared with that of non-proliferating cells, the metabolome of blood and urine reflects the entire organism. We conclude that many studies show impressive associations between biofluid metabolomics and cancer progression, but translation to clinical practice is currently hindered by lack of validation, difficulties in biological interpretation, and non-standardized analytical procedures., (Copyright © 2018 John Wiley & Sons, Ltd.)

Background In vivo magnetic resonance spectroscopy (MRS) enables non-invasive measurements of tumor metabolites. Choline-containing metabolites play a key role in tumor metabolism. Purpose To explore whether preoperative MRS-derived tumor choline levels are associated with clinical and histological features in endometrial carcinomas. Material and Methods Preoperative pelvic magnetic resonance imaging (MRI) (1.5T), including structural and diffusion-weighted imaging and localized multivoxel proton MR (1H-MR) spectroscopy, was performed in 77 prospectively included patients with histologically confirmed endometrial carcinomas. Relative levels of total choline-containing metabolites (tCho) in tumor and myometrium were measured using the ratios: tCho/Creatine; tCho/Water; and tCho/Noise. MRS parameters were analyzed in relation to histological subtype and grade, surgicopathological staging parameters, MRI-measured tumor volume, and tumor apparent diffusion coefficient (ADC) value and clinical outcome. Results Tumor tissue had significantly higher ratios for tCho/Creatine, tCho/Water, and tCho/Noise than normal myometrial tissue ( P < 0.001 for all). High tumor tCho/Water ratio was significantly associated with high tumor grade in endometrioid tumors ( P = 0.02). Tumor tCho/Creatine ratio was positively correlated to MRI-measured tumor volume (r s  = 0.25; P = 0.03). Conclusion High choline levels in tumor are associated with high-risk features. In vivo MRS may potentially aid in the preoperative risk stratification in endometrial cancer.

This corrects the article DOI: 10.1038/bjc.2017.346.

Docetaxel is the chemotherapeutic choice for metastatic hormone-refractory prostate cancer, however, it only marginally improves the survival rate. The purpose of the present study was to examine if a peptide targeting the cellular scaffold protein PCNA could improve docetaxel's efficacy. We found that docetaxel given in combination with a cell penetrating peptide containing the AlkB homolog 2 PCNA interacting motif (APIM-peptide), reduced the prostate volume and limited prostate cancer regrowth in vivo in the immunocompetent transgenic adenocarcinoma model of prostate cancer (TRAMP). In accordance with this, we found that the APIM-peptide enhanced the efficacy of docetaxel in vitro . Gene expression analysis on prostate cancer cell lines indicated that the combination of docetaxel and APIM-peptide alters expression of genes involved in cellular signaling, apoptosis, and prostate cancer development. These changes were not detected in single agent treated cells. Our results suggest that targeting PCNA and thereby affecting multiple cellular pathways simultaneously has the potential to improve docetaxel therapy of advanced prostate cancer., Competing Interests: CONFLICTS OF INTEREST APIM Therapeutics is a spin-off company of the Norwegian University of Science and Technology, and has co-funded this study. Professor Marit Otterlei is an inventor, minority shareholder and CSO of this company. Patent application no: PCT/GB2009/000489 “New PCNA interacting motif”, filed on February 20, 2009. There are no further patents, products or development or marked products to declare. The other authors declare no conflicts of interest.

Prostate cancer is the second most common malignancy, and the fifth leading cause of cancer-related death among men, worldwide. A major unsolved clinical challenge in prostate cancer is the ability to accurately distinguish indolent cancer types from the aggressive ones. Reprogramming of metabolism is now a widely accepted hallmark of cancer development, where cancer cells must be able to convert nutrients to biomass while maintaining energy production. Metabolomics is the large-scale study of small molecules, commonly known as metabolites, within cells, biofluids, tissues, or organisms. Nuclear magnetic resonance (NMR) spectroscopy is commonly applied in metabolomics studies of cancer. This chapter provides protocols for NMR-based metabolomics of cell cultures, biofluids (serum and urine), and intact tissue, with concurrent advice for optimal biobanking and sample preparation procedures.

Metabolic profiles reflect biological conditions as a result of biochemical changes within a living system. It is therefore possible to associate metabolic signatures with clinical endpoints of diseases, such as breast cancer. Nuclear magnetic resonance (NMR) spectroscopy is one of the most common techniques used for metabolic profiling, and produces high dimensional datasets from which meaningful biological information can be extracted. Here, we present an overview of data analysis techniques used to achieve this, describing key steps in the procedure. Moreover, examples of clinical endpoints of interest are provided. Although these are specific for breast cancer, the procedures for the analysis of NMR spectra as described here are applicable to any type of cancer and to other diseases.

Diffusion-weighted magnetic resonance imaging (DWI) enables non-invasive, quantitative staging of prostate cancer via measurement of the apparent diffusion coefficient (ADC) of water within tissues. In cancer, more advanced disease is often characterized by higher cellular density (cellularity), which is generally accepted to correspond to a lower measured ADC. A quantitative relationship between tissue structure and in vivo measurements of ADC has yet to be determined for prostate cancer. In this study, we establish a theoretical framework for relating ADC measurements with tissue cellularity and the proportion of space occupied by prostate lumina, both of which are estimated through automatic image processing of whole-slide digital histology samples taken from a cohort of six healthy mice and nine transgenic adenocarcinoma of the mouse prostate (TRAMP) mice. We demonstrate that a significant inverse relationship exists between ADC and tissue cellularity that is well characterized by our model, and that a decrease of the luminal space within the prostate is associated with a decrease in ADC and more aggressive tumor subtype. The parameters estimated from our model in this mouse cohort predict the diffusion coefficient of water within the prostate-tissue to be 2.18 × 10 -3  mm 2 /s (95% CI: 1.90, 2.55). This value is significantly lower than the diffusion coefficient of free water at body temperature suggesting that the presence of organelles and macromolecules within tissues can drastically hinder the random motion of water molecules within prostate tissue. We validate the assumptions made by our model using novel in silico analysis of whole-slide histology to provide the simulated ADC (sADC); this is demonstrated to have a significant positive correlation with in vivo measured ADC (r 2  = 0.55) in our mouse population. The estimation of the structural properties of prostate tissue is vital for predicting and staging cancer aggressiveness, but prostate tissue biopsies are painful, invasive, and are prone to complications such as sepsis. The developments made in this study provide the possibility of estimating the structural properties of prostate tissue via non-invasive virtual biopsies from MRI, minimizing the need for multiple tissue biopsies and allowing sequential measurements to be made for prostate cancer monitoring.

Background: Robust biomarkers that identify prostate cancer patients with high risk of recurrence will improve personalised cancer care. In this study, we investigated whether tissue metabolites detectable by high-resolution magic angle spinning magnetic resonance spectroscopy (HR-MAS MRS) were associated with recurrence following radical prostatectomy., Methods: We performed a retrospective ex vivo study using HR-MAS MRS on tissue samples from 110 radical prostatectomy specimens obtained from three different Norwegian cohorts collected between 2002 and 2010. At the time of analysis, 50 patients had experienced prostate cancer recurrence. Associations between metabolites, clinicopathological variables, and recurrence-free survival were evaluated using Cox proportional hazards regression modelling, Kaplan-Meier survival analyses and concordance index (C-index)., Results: High intratumoural spermine and citrate concentrations were associated with longer recurrence-free survival, whereas high (total-choline+creatine)/spermine (tChoCre/Spm) and higher (total-choline+creatine)/citrate (tChoCre/Cit) ratios were associated with shorter time to recurrence. Spermine concentration and tChoCre/Spm were independently associated with recurrence in multivariate Cox proportional hazards modelling after adjusting for clinically relevant risk factors (C-index: 0.769; HR: 0.72; P=0.016 and C-index: 0.765; HR: 1.43; P=0.014, respectively)., Conclusions: Spermine concentration and tChoCre/Spm ratio in prostatectomy specimens were independent prognostic markers of recurrence. These metabolites can be noninvasively measured in vivo and may thus offer predictive value to establish preoperative risk assessment nomograms.

Increased knowledge of the molecular differences between indolent and aggressive prostate cancer is needed for improved risk stratification and treatment selection. Secreted frizzled-related protein 4 (SFRP4) is a modulator of the cancer-associated Wnt pathway, and previously suggested as a potential marker for prostate cancer aggressiveness. In this study, we investigated and validated the association between SFRP4 gene expression and aggressiveness in nine independent cohorts (n = 2157). By differential expression and combined meta-analysis of all cohorts, we detected significantly higher SFRP4 expression in cancer compared with normal samples, and in high (3-5) compared with low (1-2) Grade Group samples. SFRP4 expression was a significant predictor of biochemical recurrence in six of seven cohorts and in the overall analysis, and was a significant predictor of metastatic event in one cohort. In our study cohort, where metabolic information was available, SFRP4 expression correlated significantly with the concentrations of citrate and spermine, two previously suggested biomarkers for aggressive prostate cancer. SFRP4 immunohistochemistry in an independent cohort (n = 33) was not associated with aggressiveness. To conclude, high SFRP4 gene expression is associated with high Grade Group and recurrent prostate cancer after surgery. Future studies investigating the mechanistic and clinical usefulness of SFRP4 in prostate cancer are warranted.