Hyperpolarized MRI Research Articles

Abstract 6035: Glutaminase inhibitors block metabolic switch in prostate cancer and prevents anti-androgen treatment resistance

Abstract Background: All prostate cancers first manifest as androgen dependent tumors and most patients are initially treated with surgery or radiation. About 30% of these patients return to clinic with recurrent prostate cancer that initially responds to androgen ablation therapy (ADT). Unfortunately, almost all of them fail ADT and develop castrate resistant disease (CRPC) that responds poorly to most cancer therapies and progress to often lethal metastatic CRPC (mCRPC). There are accumulating evidence that at a certain state of progression of hormone sensitive prostate cancer (HSPC) to CRPC, prostate cancer cells adopt to androgen signaling axis targeted therapy (ASI) by switching their metabolism from glycolysis to mitochondrial oxidative phosphorylation (OXPHOS). High OXPHOS activity induces mitochondrial dysfunction, autophagy, cancer invasion, and metastasis. A mitochondrial enzyme glutaminase (GLS) produces L-glutamate from L-glutamine. L-glutamate is utilized in mitochondrial metabolism for energy production through TCA cycle. GLS inhibitors CB-839 and IACS-6274 are being clinically developed for treatment of various human malignancies. We have shown that growth of prostate cancer patient derived xenografts (PDX) is markedly inhibited by a combination treatment of a clinically used ASI enzalutamide (ENZA) followed by a GLS inhibitor. Methods: We followed the growth of one HSPC and two CRPC PDX tumors in mice and collected tumor tissues and circulating human tumor cells (CTC) from mouse blood samples at sacrifice. We used super resolution STED confocal microscopy to image mitochondria in the tissue samples and in the CTCs, desorption ionization mass spectrometry (DESI-MS) imaging of multiple metabolites in the tissue samples as well as hyperpolarized MRI (HP-MRI) of the xenografted tumors to follow the changes in cellular central carbon metabolism in vivo. Results: We observed that growth of PDXs developing ENZA resistance is markedly inhibited when ENZA is followed by adding GLS inhibitors with a concomitant reduction in OXPHOS activity. In addition, the ex vivo and in vivo methods introduced to monitor tumor metabolic activities can be easily translated into clinic to monitor the metabolic status of the tumors for early detection of resistance development and as a pharmacodynamic marker for metabolic inhibitor therapy. Conclusion: GLS inhibitors added to ASI could be effective against CRPC and mCRPC developing resistance to ASI treatment. Citation Format: Hirak Subhra Basu, Meredith Spradlin, Tian Weihua, Jose Enriquez, Pratip Bhattacharya, Livia Eberlin. Glutaminase inhibitors block metabolic switch in prostate cancer and prevents anti-androgen treatment resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6035.

Abstract 3585: Probing the relationship between renal carcinoma perfusion, hypoxia, and metabolism during response to VEGFR inhibition and at resistance

Abstract Clear cell renal cell carcinoma (ccRCC) is the most common primary kidney cancer. VEGF-receptor tyrosine kinase inhibitors remain a second-line or concurrent (with immunotherapy) treatment option for advanced RCC, but resistance inevitably occurs. There remains a need for biomarkers predictive of response to VEGFR targeting. Anti-VEGFR therapy-induced hypoxia can result in changes to both tumor metabolism and vascularization that contribute to the aggressiveness of treatment-resistant tumors. In this study we used in vivo hyperpolarized 13C-tert-butanol, a freely-diffusible novel MRI tracer that provides high-SNR tissue perfusion mapping, and hyperpolarized 13C pyruvate MRI, which enables real-time investigation of glycolytic lactate production, to understand the evolution of RCC perfusion and metabolism in response to VEGFR targeting, while correlating to histologic measures of tumor proliferation (Ki67) and vascularization (CD34). A498, a human ccRCC cell line, was implanted subcutaneously into 12-week-old nude mice. When tumors reached 15mm, the mice were placed into three arms, untreated (UT), sunitinib sensitive (SS), and sunitinib resistant (SR). SS mice were treated with sunitinib (25 mg/kg via gavage) then the mice were imaged with MRI at 48 hours. SR mice were treated with sunitinib daily until 2mm of re-growth was detected, then MRI was performed. UT mice were imaged when tumors reached 20mm. Pimonidazole infusion immediately after MRI, then the tumors were harvested. Histology included Ki67, CD34, and pimonidazole (hypoxia) staining. Ki67 expression was greater in the UT group compared to SS (37.56±5.21 vs 26.23±3.78, p=0.004), but not compared to SR (37.56±5.21 vs 34.32±1.12, p=0.93). The UT group demonstrated greater vascularization with CD-34 vessel counting than SS (UT 6.70±1.81 vs. SS 2.39±1.16, p=0.002), while vascularization was increased in SR compared to SS (SR 3.82±1.81 vs SS 2.39±1.16, p=0.05). Ki67 expression was positively correlated with CD34 expression. MR-measured tumor perfusion of the SS group was greater than UT (0.0086±0.0056 vs 0.0035±0.0026, p=0.0001) and SR (0.0086±0.0056 vs 0.0043±0.0033, p=0.0003). Tumor lactate production in the SS group was lower than UT (0.752±0.268 vs 0.609±0.188, p=0.04) and SR (0.752±0.268 vs 0.593±0.246, p=0.006). However, tumor perfusion was negatively correlated with lactate production ratio only in the SR group. Untreated and sunitinib-resistent RCC tumors are associated with increased perfusion and vascularity, and decreased lactate production. An inverse relationship between lactate production and perfusion in SR may be attributable to vascular normalization. Analysis of the spatial relationship between tumor metabolism and perfusion will further yield additional detail on the interplay between hypoxia and metabolism. Citation Format: Qianhui Dou, Patricia Coutinho de Souza, Xiaoen Wang, Aaron K. Grant, Leo L. Tsai. Probing the relationship between renal carcinoma perfusion, hypoxia, and metabolism during response to VEGFR inhibition and at resistance. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3585.

Inter- and intravisit repeatability of free-breathing MRI in pediatric cystic fibrosis lung disease.

The purpose of this study is to assess the intra- and interscan repeatability of free-breathing phase-resolved functional lung (PREFUL) MRI in stable pediatric cystic fibrosis (CF) lung disease in comparison to static breath-hold hyperpolarized 129-xenon MRI (Xe-MRI) and pulmonary function tests. Free-breathing 1-hydrogen MRI and Xe-MRI were acquired from 15 stable pediatric CF patients and seven healthy age-matched participants on two visits, 1 month apart. Same-visit MRI scans were also performed on a subgroup of the CF patients. Following the PREFUL algorithm, regional ventilation (RVent) and regional flow volume loop cross-correlation maps were determined from the free-breathing data. Ventilation defect percentage (VDP) was determined from RVent maps (VDPRVent ), regional flow volume loop cross-correlation maps (VDPCC ), VDPRVent ∪ VDPCC , and multi-slice Xe-MRI. Repeatability was evaluated using Bland-Altman analysis, coefficient of repeatability (CR), and intraclass correlation. Minimal bias and no significant differences were reported for all PREFUL MRI and Xe-MRI VDP parameters between intra- and intervisits (all P > 0.05). Repeatability of VDPRVent , VDPCC , VDPRVent ∪ VDPCC , and multi-slice Xe-MRI were lower between the two-visit scans (CR=14.81%, 15.36%, 16.19%, and 9.32%, respectively) in comparison to the same-day scans (CR=3.38%, 2.90%, 1.90%, and 3.92%, respectively). pulmonary function tests showed high interscan repeatability relative to PREFUL MRI and Xe-MRI. PREFUL MRI, similar to Xe-MRI, showed high intravisit repeatability but moderate intervisit repeatability in CF, which may be due to inherent disease instability, even in stable patients. Thus, PREFUL MRI may be considered a suitable outcome measure for future treatment response studies.

Proton-Only Sensing of Hyperpolarized [1,2-13C2]Pyruvate.

Hyperpolarized MRI is emerging as a next-generation molecular imaging modality that can detect metabolic transformations in real time deep inside tissue and organs. 13C-hyperpolarized pyruvate is the leading hyperpolarized contrast agent that can probe cellular energetics in real time. Currently, hyperpolarized MRI requires specialized "multinuclear" MRI scanners that have the ability to excite and detect 13C signals. The objective of this work is the development of an approach that works on conventional (i.e., proton-only) MRI systems while taking advantage of long-lived 13C hyperpolarization. The long-lived singlet state of [1,2-13C2]pyruvate is hyperpolarized with parahydrogen in reversible exchange, and subsequently, the polarization is transferred from the 13C2 spin pair to the methyl protons of pyruvate for detection. This polarization transfer is accomplished with spin-lock induced crossing pulses that are only applied to the methyl protons yet access the hyperpolarization stored in the 13C2 singlet state. Theory and first experimental demonstrations are provided for our method, which obviates 13C excitation and detection for proton sensing of 13C-hyperpolarized pyruvate with an overall experimental-polarization transfer efficiency of ∼22% versus a theoretically predicted polarization transfer efficiency of 25%.

Imaging biomarkers of lung ventilation in interstitial lung disease from 129Xe and oxygen enhanced 1H MRI

PurposeTo compare imaging biomarkers from hyperpolarised 129Xe ventilation MRI and dynamic oxygen-enhanced MRI (OE-MRI) with standard pulmonary function tests (PFT) in interstitial lung disease (ILD) patients. To evaluate if biomarkers can separate ILD subtypes and detect early signs of disease resolution or progression. Study typeProspective longitudinal. PopulationForty-one ILD (fourteen idiopathic pulmonary fibrosis (IPF), eleven hypersensitivity pneumonitis (HP), eleven drug-induced ILD (DI-ILD), five connective tissue disease related-ILD (CTD-ILD)) patients and ten healthy volunteers imaged at visit 1. Thirty-four ILD patients completed visit 2 (eleven IPF, eight HP, ten DIILD, five CTD-ILD) after 6 or 26 weeks. Field strength/sequenceMRI was performed at 1.5 T, including inversion recovery T1 mapping, dynamic MRI acquisition with varying oxygen levels, and hyperpolarised 129Xe ventilation MRI. Subjects underwent standard spirometry and gas transfer testing. AssessmentFive 1H MRI and two 129Xe MRI ventilation metrics were compared with spirometry and gas transfer measurements. Statistical testTo evaluate differences at visit 1 among subgroups: ANOVA or Kruskal-Wallis rank tests with correction for multiple comparisons. To assess the relationships between imaging biomarkers, PFT, age and gender, at visit 1 and for the change between visit 1 and 2: Pearson correlations and multilinear regression models. ResultsThe global PFT tests could not distinguish ILD subtypes. Percentage ventilated volumes were lower in ILD patients than in HVs when measured with 129Xe MRI (HV 97.4 ± 2.6, CTD-ILD: 91.0 ± 4.8 p = 0.017, DI-ILD 90.1 ± 7.4 p = 0.003, HP 92.6 ± 4.0 p = 0.013, IPF 88.1 ± 6.5 p < 0.001), but not with OE-MRI. 129Xe reported more heterogeneous ventilation in DI-ILD and IPF than in HV, and OE-MRI reported more heterogeneous ventilation in DI-ILD and IPF than in HP or CTD-ILD. The longitudinal changes reported by the imaging biomarkers did not correlate with the PFT changes between visits. Data conclusionNeither 129Xe ventilation nor OE-MRI biomarkers investigated in this study were able to differentiate between ILD subtypes, suggesting that ventilation-only biomarkers are not indicated for this task. Limited but progressive loss of ventilated volume as measured by 129Xe-MRI may be present as the biomarker of focal disease progresses. OE-MRI biomarkers are feasible in ILD patients and do not correlate strongly with PFT. Both OE-MRI and 129Xe MRI revealed more spatially heterogeneous ventilation in DI-ILD and IPF.

Electroplated waveguides to enhance DNP and EPR spectra of silicon and diamond particles.

Electroplating the waveguide of a 7 T polarizer in a simple innovative way increased microwave power delivered to the sample by 3.1 dB. Silicon particles, while interesting for hyperpolarized MRI applications, are challenging to polarize due to inefficient microwave multipliers at the electron Larmor frequency at high magnetic fields and fast electronic relaxation times. Improving microwave transmission directly translates to more efficient EPR excitation at high-field, low-temperature conditions and promises faster and higher Si polarization buildup through dynamic nuclear polarization (DNP).

Mapping Alveolar Oxygen Partial Pressure in COPD Using Hyperpolarized Helium-3: The Multi-Ethnic Study of Atherosclerosis (MESA) COPD Study.

Chronic obstructive pulmonary disease (COPD) and emphysema are characterized by functional and structural damage which increases the spaces for gaseous diffusion and impairs oxygen exchange. Here we explore the potential for hyperpolarized (HP) 3He MRI to characterize lung structure and function in a large-scale population-based study. Participants (n = 54) from the Multi-Ethnic Study of Atherosclerosis (MESA) COPD Study, a nested case-control study of COPD among participants with 10+ packyears underwent HP 3He MRI measuring pAO2, apparent diffusion coefficient (ADC), and ventilation. HP MRI measures were compared to full-lung CT and pulmonary function testing. High ADC values (>0.4 cm2/s) correlated with emphysema and heterogeneity in pAO2 measurements. Strong correlations were found between the heterogeneity of global pAO2 as summarized by its standard deviation (SD) (p < 0.0002) and non-physiologic pAO2 values (p < 0.0001) with percent emphysema on CT. A regional study revealed a strong association between pAO2 SD and visual emphysema severity (p < 0.003) and an association with the paraseptal emphysema subtype (p < 0.04) after adjustment for demographics and smoking status. HP noble gas pAO2 heterogeneity and the fraction of non-physiological pAO2 results increase in mild to moderate COPD. Measurements of pAO2 are sensitive to regional emphysematous damage detected by CT and may be used to probe pulmonary emphysema subtypes. HP noble gas lung MRI provides non-invasive information about COPD severity and lung function without ionizing radiation.

Signal enhancement of hyperpolarized 15 N sites in solution-increase in solid-state polarization at 3.35 T and prolongation of relaxation in deuterated water mixtures.

Hyperpolarized 15 N sites have been found to be promising for generating long-lived hyperpolarized states in solution, and present a promising approach for utilizing dissolution-dynamic nuclear polarization (dDNP)-driven hyperpolarized MRI for imaging in biology and medicine. Specifically, 15 N sites with directly bound protons were shown to be useful when dissolved in D2 O. The purpose of the current study was to further characterize and increase the visibility of such 15 N sites in solutions that mimic an intravenous injection during the first cardiac pass in terms of their H2 O:D2 O composition. The T1 values of hyperpolarized 15 N in [15 N2 ]urea and [15 N]NH4 Cl demonstrated similar dependences on the H2 O:D2 O composition of the solution, with a T1 of about 140 s in 100% D2 O, about twofold shortening in 90% and 80% D2 O, and about threefold shortening in 50% D2 O. [13 C]urea was found to be a useful solid-state 13 C marker for qualitative monitoring of the 15 N polarization process in a commercial pre-clinical dDNP device. Adding trace amounts of Gd3+ to the polarization formulation led to higher solid-state polarization of [13 C]urea and to higher polarization levels of [15 N2 ]urea in solution.

Imaging Glioblastoma Metabolism by Using Hyperpolarized [1-13C]Pyruvate Demonstrates Heterogeneity in Lactate Labeling: A Proof of Principle Study.

PurposeTo evaluate glioblastoma (GBM) metabolism by using hyperpolarized carbon 13 (13C) MRI to monitor the exchange of the hyperpolarized 13C label between injected [1-13C]pyruvate and tumor lactate and bicarbonate.Materials and MethodsIn this prospective study, seven treatment-naive patients (age [mean ± SD], 60 years ± 11; five men) with GBM were imaged at 3 T by using a dual-tuned 13C–hydrogen 1 head coil. Hyperpolarized [1-13C]pyruvate was injected, and signal was acquired by using a dynamic MRI spiral sequence. Metabolism was assessed within the tumor, in the normal-appearing brain parenchyma (NABP), and in healthy volunteers by using paired or unpaired t tests and a Wilcoxon signed rank test. The Spearman ρ correlation coefficient was used to correlate metabolite labeling with lactate dehydrogenase A (LDH-A) expression and some immunohistochemical markers. The Benjamini-Hochberg procedure was used to correct for multiple comparisons.ResultsThe bicarbonate-to-pyruvate (BP) ratio was lower in the tumor than in the contralateral NABP (P < .01). The tumor lactate-to-pyruvate (LP) ratio was not different from that in the NABP (P = .38). The LP and BP ratios in the NABP were higher than those observed previously in healthy volunteers (P < .05). Tumor lactate and bicarbonate signal intensities were strongly correlated with the pyruvate signal intensity (ρ = 0.92, P < .001, and ρ = 0.66, P < .001, respectively), and the LP ratio was weakly correlated with LDH-A expression in biopsy samples (ρ = 0.43, P = .04).ConclusionHyperpolarized 13C MRI demonstrated variation in lactate labeling in GBM, both within and between tumors. In contrast, bicarbonate labeling was consistently lower in tumors than in the surrounding NABP.Keywords: Hyperpolarized 13C MRI, Glioblastoma, Metabolism, Cancer, MRI, Neuro-oncology Supplemental material is available for this article. Published under a CC BY 4.0 license.

Quantification of lung ventilation defects on hyperpolarized MRI: The Multi-Ethnic Study of Atherosclerosis (MESA) COPD study

PurposeTo develop an end-to-end deep learning (DL) framework to segment ventilation defects on pulmonary hyperpolarized MRI. Materials and methodsThe Multi-Ethnic Study of Atherosclerosis Chronic Obstructive Pulmonary Disease (COPD) study is a nested longitudinal case-control study in older smokers. Between February 2016 and July 2017, 56 participants (age, mean ± SD, 74 ± 8 years; 34 men) underwent same breath-hold proton (1H) and helium (3He) MRI, which were annotated for non-ventilated, hypo-ventilated, and normal-ventilated lungs. In this retrospective DL study, 820 1H and 3He slices from 42/56 (75%) participants were randomly selected for training, with the remaining 14/56 (25%) for test. Full lung masks were segmented using a traditional U-Net on 1H MRI and were imported into a cascaded U-Net, which were used to segment ventilation defects on 3He MRI. Models were trained with conventional data augmentation (DA) and generative adversarial networks (GAN)-DA. ResultsConventional-DA improved 1H and 3He MRI segmentation over the non-DA model (P = 0.007 to 0.03) but GAN-DA did not yield further improvement. The cascaded U-Net improved non-ventilated lung segmentation (P < 0.005). Dice similarity coefficients (DSC) between manually and DL-segmented full lung, non-ventilated, hypo-ventilated, and normal-ventilated regions were 0.965 ± 0.010, 0.840 ± 0.057, 0.715 ± 0.175, and 0.883 ± 0.060, respectively. We observed no statistically significant difference in DCSs between participants with and without COPD (P = 0.41, 0.06, and 0.18 for non-ventilated, hypo-ventilated, and normal-ventilated regions, respectively). ConclusionThe proposed cascaded U-Net framework generated fully-automated segmentation of ventilation defects on 3He MRI among older smokers with and without COPD that is consistent with our reference method.

Large-scale investigation of deep learning approaches for ventilated lung segmentation using multi-nuclear hyperpolarized gas MRI

Respiratory diseases are leading causes of mortality and morbidity worldwide. Pulmonary imaging is an essential component of the diagnosis, treatment planning, monitoring, and treatment assessment of respiratory diseases. Insights into numerous pulmonary pathologies can be gleaned from functional lung MRI techniques. These include hyperpolarized gas ventilation MRI, which enables visualization and quantification of regional lung ventilation with high spatial resolution. Segmentation of the ventilated lung is required to calculate clinically relevant biomarkers. Recent research in deep learning (DL) has shown promising results for numerous segmentation problems. Here, we evaluate several 3D convolutional neural networks to segment ventilated lung regions on hyperpolarized gas MRI scans. The dataset consists of 759 helium-3 (3He) or xenon-129 (129Xe) volumetric scans and corresponding expert segmentations from 341 healthy subjects and patients with a wide range of pathologies. We evaluated segmentation performance for several DL experimental methods via overlap, distance and error metrics and compared them to conventional segmentation methods, namely, spatial fuzzy c-means (SFCM) and K-means clustering. We observed that training on combined 3He and 129Xe MRI scans using a 3D nn-UNet outperformed other DL methods, achieving a mean ± SD Dice coefficient of 0.963 ± 0.018, average boundary Hausdorff distance of 1.505 ± 0.969 mm, Hausdorff 95th percentile of 5.754 ± 6.621 mm and relative error of 0.075 ± 0.039. Moreover, limited differences in performance were observed between 129Xe and 3He scans in the testing set. Combined training on 129Xe and 3He yielded statistically significant improvements over the conventional methods (p < 0.0001). In addition, we observed very strong correlation and agreement between DL and expert segmentations, with Pearson correlation of 0.99 (p < 0.0001) and Bland–Altman bias of − 0.8%. The DL approach evaluated provides accurate, robust and rapid segmentations of ventilated lung regions and successfully excludes non-lung regions such as the airways and artefacts. This approach is expected to eliminate the need for, or significantly reduce, subsequent time-consuming manual editing.

Quantitative assessment of regional lung ventilation in emphysematous mice using hyperpolarized 129Xe MRI with a continuous flow hyperpolarizing system

BackgroundLung ventilation function in small animals can be assessed by using hyperpolarized gas MRI. For these experiments a free breathing protocol is generally preferred to mechanical ventilation as mechanical ventilation can often lead to ventilation lung injury, while the need to maintain a gas reservoir may lead to a partial reduction of the polarization. PurposeTo evaluate regional lung ventilation of mice by a simple but fast method under free breathing and give evidence for effectiveness with an elastase instilled emphysematous mice. Animal modelEmphysematous mice. Materials and methodsA Look-Locker based saturation recovery sequence was developed for continuous flow hyperpolarized (CF-HP) 129Xe gas experiments, and the apparent gas-exchange rate, k’, was measured by the analysis of the saturation recovery curve. ResultsIn mice with elastase-induced mild emphysema, reductions of 15–30% in k’ values were observed as the results of lesion-induced changes in the lung. Data conclusionThe proposed method was applied to an emphysematous model mice and ventilation dysfunctions have been approved as a definite decrease in k’ values, supporting the usefulness for a non-invasive assessment of the lung functions in preclinical study by the CF-HP 129Xe experiments.

LED-Based Photo-CIDNP Hyperpolarization Enables 19F MR Imaging and 19F NMR Spectroscopy of 3-Fluoro-DL-tyrosine at 0.6 T

Although 19F has high potential to serve as a background-free molecular marker in bioimaging, the molar amount of marker substance is often too small to enable 19F MR imaging or 19F NMR spectroscopy with a sufficiently high signal-to-noise ratio (SNR). Hyperpolarization methods such as parahydrogen-based hyperpolarization or dynamic nuclear polarization (DNP) can significantly improve the SNR, but require expensive and complex sample preparation and the removal of toxic catalysts and solvents. Therefore, we used the biologically compatible model of the fluorinated amino acid 3-Fluoro-DL-tyrosine with riboflavin 5ʹ-monophosphate (FMN) as a chromophore dissolved in D2O with 3.4% H2Odest., allowing to transform light energy into hyperpolarization of the 19F nucleus via photo-chemically induced dynamic nuclear polarization (photo-CIDNP). We used a low-cost high-power blue LED to illuminate the sample replacing traditionally used laser excitation, which is both potentially harmful and costly. For the first time, we present results of hyperpolarized 19F MRI and 19F NMR performed with a low-cost 0.6 T benchtop MRI system. The device allowed simultaneous dual-channel 1H/19F NMR. 19F imaging was performed with a (0.94 mm)2 in-plane resolution. This enabled the spatial resolution of different degrees of hyperpolarization within the sample. We estimated the photo-CIDNP-based 19F signal enhancement at 0.6 T to be approximately 465. FMN did not bleach out even after multiple excitations, so that the signal-to-noise ratio could be further improved by averaging hyperpolarized signals. The results show that the easy-to-use experimental setup has a high potential to serve as an efficient preclinical tool for hyperpolarization studies in bioimaging.

Deuterium metabolic imaging and hyperpolarized 13C-MRI of the normal human brain at clinical field strength reveals differential cerebral metabolism

Deuterium metabolic imaging (DMI) and hyperpolarized 13C-pyruvate MRI (13C-HPMRI) are two emerging methods for non-invasive and non-ionizing imaging of tissue metabolism. Imaging cerebral metabolism has potential applications in cancer, neurodegeneration, multiple sclerosis, traumatic brain injury, stroke, and inborn errors of metabolism. Here we directly compare these two non-invasive methods at 3 T for the first time in humans and show how they simultaneously probe both oxidative and non-oxidative metabolism. DMI was undertaken 1–2 h after oral administration of [6,6′-2H2]glucose, and 13C-MRI was performed immediately following intravenous injection of hyperpolarized [1–13C]pyruvate in ten and nine normal volunteers within each arm respectively. DMI was used to generate maps of deuterium-labelled water, glucose, lactate, and glutamate/glutamine (Glx) and the spectral separation demonstrated that DMI is feasible at 3 T. 13C-HPMRI generated maps of hyperpolarized carbon-13 labelled pyruvate, lactate, and bicarbonate. The ratio of 13C-lactate/13C-bicarbonate (mean 3.7 ± 1.2) acquired with 13C-HPMRI was higher than the equivalent 2H-lactate/2H-Glx ratio (mean 0.18 ± 0.09) acquired using DMI. These differences can be explained by the route of administering each probe, the timing of imaging after ingestion or injection, as well as the biological differences in cerebral uptake and cellular physiology between the two molecules. The results demonstrate these two metabolic imaging methods provide different yet complementary readouts of oxidative and reductive metabolism within a clinically feasible timescale. Furthermore, as DMI was undertaken at a clinical field strength within a ten-minute scan time, it demonstrates its potential as a routine clinical tool in the future.

Whole-Abdomen Metabolic Imaging of Healthy Volunteers Using Hyperpolarized [1-13 C]pyruvate MRI.

Hyperpolarized 13 C MRI quantitatively measures enzyme-catalyzed metabolism in cancer and metabolic diseases. Whole-abdomen imaging will permit dynamic metabolic imaging of several abdominal organs simultaneously in healthy and diseased subjects. Image hyperpolarized [1-13 C]pyruvate and products in the abdomens of healthy volunteers, overcoming challenges of motion, magnetic field variations, and spatial coverage. Compare hyperpolarized [1-13 C]pyruvate metabolism across abdominal organs of healthy volunteers. Prospective technical development. A total of 13 healthy volunteers (8 male), 21-64 years (median 36). A 3 T. Proton: T1 -weighted spoiled gradient echo, T2 -weighted single-shot fast spin echo, multiecho fat/water imaging. Carbon-13: echo-planar spectroscopic imaging, metabolite-specific echo-planar imaging. Transmit magnetic field was measured. Variations in main magnetic field (ΔB0 ) determined using multiecho proton acquisitions were compared to carbon-13 acquisitions. Changes in ΔB0 were measured after localized shimming. Improvements in metabolite signal-to-noise ratio were calculated. Whole-organ regions of interests were drawn over the liver, spleen, pancreas, and kidneys by a single investigator. Metabolite signals, time-to-peak, decay times, and mean first-order rate constants for pyruvate-to-lactate (kPL ) and alanine (kPA ) conversion were measured in each organ. Linear regression, one-sample Kolmogorov-Smirnov tests, paired t-tests, one-way ANOVA, Tukey's multiple comparisons tests. P ≤ 0.05 considered statistically significant. Proton ΔB0 maps correlated with carbon-13 ΔB0 maps (slope=0.93, y-intercept=-2.88, R2 =0.73). Localized shimming resulted in mean frequency offset within ±25 Hz for all organs. Metabolite SNR significantly increased after denoising. Mean kPL and kPA were highest in liver, followed by pancreas, spleen, and kidneys (all comparisons with liver were significant). Whole-abdomen coverage with hyperpolarized carbon-13 MRI was feasible despite technical challenges. Multiecho gradient echo 1 H acquisitions accurately predicted chemical shifts observed using carbon-13 spectroscopy. Carbon-13 acquisitions benefited from local shimming. Metabolite energetics in the abdomen compiled for healthy volunteers can be used to design larger clinical trials in patients with metabolic diseases. 2 TECHNICAL EFFICACY: Stage 1.

Computational investigation of fitting for calculation of signal dynamics from hyperpolarized xenon-129 Gas MRI

Aim: Computational fitting methods were investigated to determine the most accurate fitting approach for the calculation of dynamic hyperpolarized MRI parameters.&#x0D; Materials and Methods: The signal decay of a time-series Hyperpolarized xenon gas MRI phantom was fitted to Bloch equations using three methods varying the fitting parameters for calculation of flip angle, α, and longitudinal relaxation time, T1. The first fitting method used an initial calculation of α before the fitting process. The second and third techniques used direct fitting of signal decay equations with and without upper-lower boundaries for calculation of α, and T1. Wilcoxon signed-rank test was used to investigate the statistical significance of the calculated parameters.&#x0D; Results: The first approach was the most accurate fitting technique that allowed direct calculation of α=8.65° in agreement to the third approach α=8.73±0.78°, 8.75±0.12°, 8.67±0.05°. Additionally, the standard deviation of the calculated T1 was lower than 1% (T1=103.2±0.04s) which was significantly more accurate than the second method (T1=90±30.2s and 135.7±10.3s) and the third method (T1=101.4±5.1s and 113.5±16.1s).&#x0D; Conclusion: The first technique provides repeatable and reliable calculation of signal decay parameters including α and T1 from the dynamic hyperpolarized gas MR images and more accurate than direct fitting methods.

Application of a 2D frequency encoding sectoral approach to hyperpolarized 129Xe MRI at low field

Inhaled hyperpolarized 129Xe MRI is a non-invasive and radiation risk free lung imaging method, which can directly measure the business unit of the lung where gas exchange occurs: the alveoli and acinar ducts (lung function). Currently, three imaging approaches have been demonstrated to be useful for hyperpolarized 129Xe MR in lungs: Fast Gradient Recalled Echo (FGRE), Radial Projection Reconstruction (PR), and spiral/cones. Typically, non-Cartesian acquisitions such as PR and spiral/cones require specific data post-processing, such as interpolating, regridding, and density-weighting procedures for image reconstruction, which often leads to smoothing effects and resolution degradation. On the other hand, Cartesian methods such as FGRE are not short-echo time (TE) methods; they suffer from imaging gradient-induced diffusion-weighting of the k-space center, and employ a significant number of radio-frequency (RF) pulses.Due to the non-renewable magnetization of the hyperpolarized media, the use of a large number of RF pulses (FGRE/PR) required for full k-space coverage is a significant limitation, especially for low field (<0.5 T) hyperpolarized gas MRI.We demonstrate an ultra-fast, purely frequency-encoded, Cartesian pulse sequence called Frequency-Encoding Sectoral (FES), which takes advantage of the long T2* of hyperpolarized 129Xe gas at low field strength (0.074 T). In contrast to PR/FGRE, it uses a much smaller number of RF pulses, and consequently maximizes image Signal-to-Noise Ratio (SNR) while shortening acquisition time. Additionally, FES does not suffer from non-uniform T2* decay leading to image blurring; a common issue with interleaved spirals/cones. The Cartesian k-space coverage of the proposed FES method does not require specific k-space data post-processing, unlike PR/FGRE and spiral/cones methods.Proton scans were used to compare the FES sequence to both FGRE and Phase Encoding Sectoral, in terms of their SNR values and imaging efficiency estimates. Using FES, proton and hyperpolarized 129Xe images were acquired from a custom hollow acrylic phantom (0.04L) and two normal rats (129Xe only), utilizing both single-breath and multiple-breath schemes. For the 129Xe phantom images, the apparent diffusion coefficient, T1, and T2* relaxation maps were acquired and generated. Blurring due to the T2* decay and B0 field variation were simulated to estimate dependence of the image resolution on the duration of the data acquisition windows (i.e. sector length), and temperature-induced resonance frequency shift from the low field magnet hardware.

Temperature Cycling Enables Efficient 13C SABRE-SHEATH Hyperpolarization and Imaging of [1-13C]-Pyruvate.

Molecular metabolic imaging in humans is dominated by positron emission tomography (PET). An emerging nonionizing alternative is hyperpolarized MRI of 13C-pyruvate, which is innocuous and has a central role in metabolism. However, similar to PET, hyperpolarized MRI with dissolution dynamic nuclear polarization (d-DNP) is complex costly, and requires significant infrastructure. In contrast, Signal Amplification By Reversible Exchange (SABRE) is a fast, cheap, and scalable hyperpolarization technique. SABRE in SHield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH) can transfer polarization from parahydrogen to 13C in pyruvate; however, polarization levels remained low relative to d-DNP (1.7% with SABRE-SHEATH versus ≈60% with DNP). Here we introduce a temperature cycling method for SABRE-SHEATH that enables >10% polarization on [1-13C]-pyruvate, sufficient for successful in vivo experiments. First, at lower temperatures, ≈20% polarization is accumulated on SABRE catalyst-bound pyruvate, which is released into free pyruvate at elevated temperatures. A kinetic model of differential equations is developed that explains this effect and characterizes critical relaxation and buildup parameters. With the large polarization, we demonstrate the first 13C pyruvate images with a cryogen-free MRI system operated at 1.5 T, illustrating that inexpensive hyperpolarization methods can be combined with low-cost MRI systems to obtain a broadly available, yet highly sensitive metabolic imaging platform.

Mucus Plugs in Asthma at CT Associated with Regional Ventilation Defects at 3He MRI.

Background Airway mucus plugs in asthma are associated with exacerbation frequency, increased eosinophilia, and reduced lung function. The relationship between mucus plugs and spatially overlapping ventilation abnormalities observed at hyperpolarized gas MRI has not been assessed quantitatively. Purpose To assess regional associations between CT mucus plugs scored by individual bronchopulmonary segment and corresponding measurements of segmental ventilation defect percentage (VDP) at hyperpolarized helium 3 (3He) MRI. Materials and Methods In this secondary analysis of a Health Insurance Portability and Accountability Act-compliant prospective observational cohort, participants in the Severe Asthma Research Program (SARP) III (NCT01760915) between December 2012 and August 2015 underwent hyperpolarized 3He MRI to determine segmental VDP. Segmental mucus plugs at CT were scored by two readers, with segments scored as plugged only if both readers agreed independently. A linear mixed-effects model controlling for interpatient variability was then used to assess differences in VDP in plugged versus plug-free segments. Results Forty-four participants with asthma were assessed (mean age ± standard deviation, 47 years ± 15; 29 women): 19 with mild-to-moderate asthma and 25 with severe asthma. Mucus plugs were observed in 49 total bronchopulmonary segments across eight of 44 patients. Segments containing mucus plugs had a median segmental VDP of 25.9% (25th-75th percentile, 7.3%-38.3%) versus 1.4% (25th-75th percentile, 0.1%-5.2%; P < .001) in plug-free segments. Similarly, the model estimated a segmental VDP of 18.9% (95% CI: 15.7, 22.2) for mucus-plugged segments versus 5.1% (95% CI: 3.3, 7.0) for plug-free segments (P < .001). Participants with one or more mucus plugs had a median whole-lung VDP of 11.1% (25th-75th percentile, 7.1%-18.9%) versus 3.1% (25th-75th percentile, 1.1%-4.4%) in those without plugs (P < .001). Conclusion Airway mucus plugging at CT was associated with reduced ventilation in the same bronchopulmonary segment at hyperpolarized helium 3 MRI, suggesting that mucus plugging may be an important cause of ventilation defects in asthma. © RSNA, 2021 Online supplemental material is available for this article.

Accumulation of Intracellular L-Lactate and Irreversible Disruption of Mitochondrial Membrane Potential upon Dual Inhibition of Oxphos and Lactate Transporter MCT-1 Induce Synthetic Lethality in T-ALL Via Mitochondrial Exhaustion

Metabolic reprogramming is recognized as one of the key hallmarks in acquiring aggressive phenotype and chemoresistance in solid tumors and hematologic malignancies.We have previously demonstrated that T-ALL are characterized by significant dependency on oxidative phosphorylation (OxPhos) with ability to utilize glutamine either in oxidative or reductive directions of TCA cycle, when mitochondria are blocked by Complex I Inhibitor (Baran N, et al. ASH 2020). To survive upon Complex I blockade leukemic cells require functional monocarboxylate transporter MCT1, that enables excretion of lactate and permissive pyruvate flux (Fig.1 a).Here we show that metabolic intervention utilizing OxPhos blockade can be potentiated by targeting MCT1 transporter and propose a novel metabolic synthetic lethality that could be exploited to eradicate T-ALL and other OxPhos-dependent malignancies.We first demonstrated that Complex I inhibition leads to increased MCT1 expression; on the contrary, MCT1 transporter blockade forces cells to increase OxPhos. In turn, the combinatorial therapy with Complex I inhibitor (IACS-010759) and MCT1 inhibitor (AZD3965) causes loss of ATP content (Fig. 1b), significant reduction of cell number and massive induction of apoptosis. Mechanistically, the combination treatment further reduced oxygen consumption rate (OCR) (Fig. 1c) and increased extracellular acidification rate, as measured by Seahorse. In concert with those results, dual inhibition led to TCA blockade, accumulation of intracellular lactate and depletion of glutamine, cystathionine and glutathione, indicating severe disruption of redox balance as measured by mass spectrometry and confirmed by significant accumulation of intracellular and mitochondrial reactive oxygen species (ROS) (Fig. 1d), loss of mitochondrial membrane potential (ΔΨ) (Fig. 1e) and subsequent mitochondria swelling. RNAseq data showed simultaneous upregulation of glycolysis and glutathione-related processes as possible mechanisms of metabolic compensation, yet strong upregulation of genes regulating apoptosis related to mitochondria dysfunction (Fig. 1f).Real-time hyperpolarized MRI based metabolic imaging studies with [1-13C]-pyruvate in patient-derived xenografts in vivo revealed significant decrease of lactate-to-pyruvate ratio in mice treated with AZD3965 or IACS-010759 alone, and in mice treated with drug combination. [13C]-Glucose isotope tracing analysis in patient-derived xenografts in vivo revealed an increased intracellular trapping of lactate as a marker of treatment effectiveness in mice subjected to dual blockade.While MCT1 inhibition induced only moderate reduction of leukemia growth in vitro and tumor burden in vivo, combination with IACS-010759 depleted significantly both, circulating and marrow/spleen/liver resident leukemia cells. Mechanistically, inhibition of MCT1 by AZD3965 therapy in leukemia-bearing mice led to lactate accumulation, OCR increase, moderate ROS production and mitochondrial membrane hyperpolarization, while Complex I blockade resulted in upregulation of MCT-1, reduction of OCR, lactate production and increase of ROS ; consequently, combinatorial therapy caused complete mitochondria shut-down and drastic inhibition of tumor growth both in vitro and in vivo in two xenografts models and led to significant extension of overall survival (p<0.0001) (Fig. 1g). In summary, these results demonstrate a novel synthetic vulnerability of concomitant blockade of OxPhos and MCT-1, uncovering metabolic checkpoints that can ultimately translate into successful therapies in T-ALL and OxPhos-dependent malignancies. [Display omitted] DisclosuresSkwarska: Halilovich E, Wang Y, Morris E, Konopleva M, Skwarska A.: Patents & Royalties: Combination of a MCL-1 inhibitor and midostaurin, uses and pharmaceutical composition thereof.. Konopleva: Reata Pharmaceuticals: Current holder of stock options in a privately-held company, Patents & Royalties: intellectual property rights; Rafael Pharmaceuticals: Other: grant support, Research Funding; Stemline Therapeutics: Research Funding; Eli Lilly: Patents & Royalties: intellectual property rights, Research Funding; Ascentage: Other: grant support, Research Funding; Genentech: Consultancy, Honoraria, Other: grant support, Research Funding; Ablynx: Other: grant support, Research Funding; AstraZeneca: Other: grant support, Research Funding; AbbVie: Consultancy, Honoraria, Other: Grant Support, Research Funding; Novartis: Other: research funding pending, Patents & Royalties: intellectual property rights; Cellectis: Other: grant support; Sanofi: Other: grant support, Research Funding; KisoJi: Research Funding; Calithera: Other: grant support, Research Funding; Forty Seven: Other: grant support, Research Funding; Agios: Other: grant support, Research Funding; F. Hoffmann-La Roche: Consultancy, Honoraria, Other: grant support.