Hyperpolarized 13C pyruvate magnetic resonance spectroscopy for in vivo metabolic phenotyping of rat HCC

AbstractThe in vivo assessment of tissue metabolism represents a novel strategy for the evaluation of oncologic disease. Hepatocellular carcinoma (HCC) is a high-prevalence, high-mortality tumor entity often discovered at a late stage. Recent evidence indicates that survival differences depend on metabolic alterations in tumor tissue, with particular focus on glucose metabolism and lactate production. Here, we present an in vivo imaging technique for metabolic tumor phenotyping in rat models of HCC. Endogenous HCC was induced in Wistar rats by oral diethyl-nitrosamine administration. Peak lactate-to-alanine signal ratios (L/A) were assessed with hyperpolarized magnetic resonance spectroscopic imaging (HPMRSI) after [1-13C]pyruvate injection. Cell lines were derived from a subset of primary tumors, re-implanted in nude rats, and assessed in vivo with dynamic hyperpolarized magnetic resonance spectroscopy (HPMRS) after [1-13C]pyruvate injection and kinetic modelling of pyruvate metabolism, taking into account systemic lactate production and recirculation. For ex vivo validation, enzyme activity and metabolite concentrations were spectroscopically quantified in cell and tumor tissue extracts. Mean peak L/A was higher in endogenous HCC compared to non-tumorous tissue. Dynamic HPMRS revealed higher pyruvate-to-lactate conversion rates (kpl) and lactate signal in subcutaneous tumors derived from high L/A tumor cells, consistent with ex vivo measurements of higher lactate dehydrogenase (LDH) levels in these cells. In conclusion, HPMRS and HPMRSI reveal distinct tumor phenotypes corresponding to differences in glycolytic metabolism in HCC tumor tissue.

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BIMG-08. DEUTERIUM MAGNETIC RESONANCE SPECTROSCOPY USING 2H-PYRUVATE ALLOWS NON-INVASIVE IN VIVO IMAGING OF TERT EXPRESSION IN BRAIN TUMORS

Neuro-Oncology Advances ◽

10.1093/noajnl/vdab024.007 ◽

2021 ◽

Vol 3 (Supplement_1) ◽

pp. i2-i2

Author(s):

Georgios Batsios ◽

Celine Taglang ◽

Meryssa Tran ◽

Anne Marie Gillespie ◽

Joseph Costello ◽

...

Keyword(s):

Magnetic Resonance ◽

Magnetic Resonance Spectroscopy ◽

In Vivo Imaging ◽

Lactate Production ◽

Telomere Maintenance ◽

Resonance Spectroscopy ◽

Low Grade ◽

Non Invasive ◽

Tert Expression

Abstract Telomere shortening constitutes a natural barrier to uncontrolled proliferation and all tumors must find a mechanism of maintaining telomere length. Most human tumors, including high-grade primary glioblastomas (GBMs) and low-grade oligodendrogliomas (LGOGs) achieve telomere maintenance via reactivation of the expression of telomerase reverse transcriptase (TERT), which is silenced in normal somatic cells. TERT expression is, therefore, a driver of tumor proliferation and, due to this essential role, TERT is also a therapeutic target. However, non-invasive methods of imaging TERT are lacking. The goal of this study was to identify magnetic resonance spectroscopy (MRS)-detectable metabolic biomarkers of TERT expression that will enable non-invasive visualization of tumor burden in LGOGs and GBMs. First, we silenced TERT expression by RNA interference in patient-derived LGOG (SF10417, BT88) and GBM (GS2) models. Our results linked TERT silencing to significant reductions in steady-state levels of NADH in all models. NADH is essential for the conversion of pyruvate to lactate, suggesting that measuring pyruvate flux to lactate could be useful for imaging TERT status. Recently, deuterium (2H)-MRS has emerged as a novel, clinically translatable method of monitoring metabolic fluxes in vivo. However, to date, studies have solely examined 2H-glucose and the use of [U-2H]pyruvate for non-invasive 2H-MRS has not been tested. Following intravenous injection of a bolus of [U-2H]pyruvate, lactate production was higher in mice bearing orthotopic LGOG (BT88 and SF10417) and GBM (GS2) tumor xenografts relative to tumor-free mice, suggesting that [U-2H]pyruvate has the potential to monitor TERT expression in vivo. In summary, our study, for the first time, shows the feasibility and utility of [U-2H]pyruvate for in vivo imaging. Importantly, since 2H-MRS can be implemented on clinical scanners, our results provide a novel, non-invasive method of integrating information regarding a fundamental cancer hallmark, i.e. TERT, into glioma patient management.

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BIOM-10. PRECLINICAL PLATFORM FOR THE IDENTIFICATION OF DEUTERIUM MAGNETIC RESONANCE SPECTROSCOPY-BASED BIOMARKERS OF BRAIN TUMOR METABOLISM

Neuro-Oncology ◽

10.1093/neuonc/noab196.041 ◽

2021 ◽

Vol 23 (Supplement_6) ◽

pp. vi12-vi12

Author(s):

Georgios Batsios ◽

Meryssa Tran ◽

Céline Taglang ◽

Anne Marie Gillespie ◽

Sabrina Ronen ◽

...

Keyword(s):

Brain Tumor ◽

Magnetic Resonance ◽

Magnetic Resonance Spectroscopy ◽

Brain Tumors ◽

Treatment Response ◽

Lactate Production ◽

Cell Suspensions ◽

Response To Therapy ◽

Resonance Spectroscopy

Abstract Metabolic reprogramming is a fundamental hallmark of cancer, which can be exploited for non-invasive tumor imaging. Deuterium magnetic resonance spectroscopy (2H-MRS) recently emerged as a novel, translational method of interrogating flux from 2H-labeled substrates to metabolic products. However, to date, preclinical studies have been performed in vivo, an endeavor which suffers from low-throughput and potential wastage of animal life, especially when considering studies of treatment response. Developing in vitro assays for monitoring metabolism of 2H-labeled substrates will enhance throughput, lead to the rapid evaluation of new 2H-based probes and enable identification of treatment response biomarkers, thereby allowing the best 2H-based probes to be translated for further in vivo assessment. The goal of this study was to develop a preclinical cell-based platform for quantifying metabolism of 2H-labeled probes in brain tumor models. Since the Warburg effect, which is characterized by elevated glycolytic production of lactate, is a metabolic phenotype of cancer, including brain tumors, we examined metabolism of 2H-glucose or 2H-pyruvate in patient-derived glioblastoma (GBM6) and oligodendroglioma (BT88) cells and compared to normal human astrocytes (NHACONTROL). Following incubation in media containing [6,6’-2H]glucose or [U-2H]pyruvate, 2H-MR spectra obtained from live cell suspensions showed elevated 2H-lactate production in GBM6 and BT88 cells relative to NHACONTROL. Importantly, 2H-lactate production from [6,6’-2H]glucose or from [U-2H]pyruvate was reduced in GBM6 or BT88 cells subjected to irradiation and temozolomide, which is standard of care for glioma patients, pointing to the utility of this method for detecting response to therapy. Collectively, we have, for the first time, demonstrated the ability to quantify metabolism of 2H-MRS probes in live cell suspensions and validated the utility of our assay for differentiating tumor from normal cells and assessing response to therapy. Our studies will expedite the identification of novel 2H-MRS probes for imaging brain tumors and potentially other types of cancer.

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Proton magnetic resonance spectroscopy of the brain of a neonate with nonketotic hyperglycinemia: in vivo–in vitro (ex vivo) correlation

European Radiology ◽

10.1007/s003300101073 ◽

2001 ◽

Vol 12 (4) ◽

pp. 858-861 ◽

Cited By ~ 49

Author(s):

T. Huisman ◽

T. Thiel ◽

B. Steinmann ◽

G. Zeilinger ◽

E. Martin

Keyword(s):

Magnetic Resonance ◽

Magnetic Resonance Spectroscopy ◽

Proton Magnetic Resonance ◽

Proton Magnetic Resonance Spectroscopy ◽

Ex Vivo ◽

Resonance Spectroscopy ◽

Nonketotic Hyperglycinemia ◽

The Brain

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Proton Magnetic Resonance Spectroscopy Characterization of Rathke’s Cleft Cysts (RCCs): Relevance to the Differential Diagnosis of Pituitary Adenomas and RCCs

Cancers ◽

10.3390/cancers12020360 ◽

2020 ◽

Vol 12 (2) ◽

pp. 360

Author(s):

Omkar B. Ijare ◽

Martyn A. Sharpe ◽

David S. Baskin ◽

Kumar Pichumani

Keyword(s):

Differential Diagnosis ◽

Magnetic Resonance ◽

Magnetic Resonance Spectroscopy ◽

Pituitary Adenomas ◽

Ex Vivo ◽

Medical Professionals ◽

Resonance Spectroscopy ◽

Rathke's Cleft Cysts ◽

Rathke's Cleft

Background: Rathke’s Cleft Cysts (RCCs) are rare epithelial cysts arising from remnants of the Rathke pouch in the pituitary gland. A subset of these lesions enlarge and produce a mass effect with consequent hypopituitarism, and may result in visual loss. Moreover, some RCCs with a high intra-cystic protein content may mimic cystic pituitary adenoma, which makes their differential diagnosis ambiguous. Currently, medical professionals have no definitive way to distinguish RCCs from pituitary adenomas. Therefore, preoperative confirmation of RCCs would be of help to medical professionals for the management and proper surgical decision making. The goal of this study is to identify molecular markers in RCCs. Methods: We characterized aqueous and chloroform extracts of surgically resected RCCs and pituitary adenomas using ex vivo 1H NMR spectroscopy. Results: All RCCs exclusively showed the presence of mucopolysaccharides which are glycosaminoglycans (GAGs) made up of disaccharides of aminosugars and uronic sugars. Conclusion: GAGs can be used as metabolite marker for the detection of RCCs and this knowledge will lay the groundwork for the development of a non-invasive, in vivo magnetic resonance spectroscopy methodology for the differential diagnosis of RCCs and pituitary adenomas using clinical MRI scanners.

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1H magnetic resonance spectroscopy of preinvasive and invasive cervical cancer: In vivo-ex vivo profiles and effect of tumor load

Journal of Magnetic Resonance Imaging ◽

10.1002/jmri.20012 ◽

2004 ◽

Vol 19 (3) ◽

pp. 356-364 ◽

Cited By ~ 44

Author(s):

Marrita M. Mahon ◽

I. Jane Cox ◽

Roberto Dina ◽

W. Patrick Soutter FRCOG ◽

G. Angus McIndoe MRCOG ◽

...

Keyword(s):

Cervical Cancer ◽

Magnetic Resonance ◽

Magnetic Resonance Spectroscopy ◽

Ex Vivo ◽

Invasive Cervical Cancer ◽

Resonance Spectroscopy ◽

1H Magnetic Resonance Spectroscopy ◽

Tumor Load

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In Vivo and Ex Vivo Magnetic Resonance Spectroscopy in the Characterization of Hemangioblastoma Cyst Fluid

Journal of Computer Assisted Tomography ◽

10.1097/rct.0b013e3182a9103a ◽

2014 ◽

Vol Publish Ahead of Print ◽

Author(s):

Girolamo Crisi ◽

Silvano Filice ◽

Thelma A. Pertinhez ◽

Elisa Ventura ◽

Franco Servadei

Keyword(s):

Magnetic Resonance ◽

Magnetic Resonance Spectroscopy ◽

Ex Vivo ◽

Cyst Fluid ◽

Resonance Spectroscopy

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Metabolic Profiling of the Rat Liver After Chronic Ingestion of Alpha-Naphthylisothiocyanate Using In Vivo and Ex Vivo Magnetic Resonance Spectroscopy

Toxicological Sciences ◽

10.1093/toxsci/kfs019 ◽

2012 ◽

Vol 126 (2) ◽

pp. 306-316 ◽

Cited By ~ 4

Author(s):

B. S. Solanky ◽

G. J. Sanchez-Canon ◽

J. F. L. Cobbold ◽

S. D. Taylor-Robinson ◽

J. D. Bell ◽

...

Keyword(s):

Magnetic Resonance ◽

Magnetic Resonance Spectroscopy ◽

Rat Liver ◽

Metabolic Profiling ◽

Ex Vivo ◽

Resonance Spectroscopy ◽

Chronic Ingestion

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P103 Metabolic profiling of the rat liver after chronic ingestion of -naphthylisothiocyanate using in vivo and ex vivo magnetic resonance spectroscopy

Gut ◽

10.1136/gutjnl-2011-300857a.103 ◽

2011 ◽

Vol 60 (Suppl 2) ◽

pp. A48-A48

Author(s):

B. S. Solanky ◽

G. J. Sanchez-Canon ◽

J. F. L. Cobbold ◽

S. D. Taylor-Robinson ◽

J. D. Bell ◽

...

Keyword(s):

Magnetic Resonance ◽

Magnetic Resonance Spectroscopy ◽

Rat Liver ◽

Metabolic Profiling ◽

Ex Vivo ◽

Resonance Spectroscopy ◽

Chronic Ingestion

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In Vivo and Ex Vivo Magnetic Resonance Spectroscopy of the Infarct and the Subventricular Zone in Experimental Stroke

Journal of Cerebral Blood Flow & Metabolism ◽

10.1038/jcbfm.2014.257 ◽

2015 ◽

Vol 35 (5) ◽

pp. 828-834 ◽

Cited By ~ 11

Author(s):

Elena Jiménez-Xarrié ◽

Myriam Davila ◽

Sara Gil-Perotín ◽

Andrés Jurado-Rodríguez ◽

Ana Paula Candiota ◽

...

Keyword(s):

Ischemic Stroke ◽

Magnetic Resonance ◽

Magnetic Resonance Spectroscopy ◽

Subventricular Zone ◽

Ex Vivo ◽

Fold Increase ◽

Resonance Spectroscopy ◽

Magic Angle ◽

Metabolic Pattern

Ex vivo high-resolution magic-angle spinning (HRMAS) provides metabolic information with higher sensitivity and spectral resolution than in vivo magnetic resonance spectroscopy (MRS). Therefore, we used both techniques to better characterize the metabolic pattern of the infarct and the neural progenitor cells (NPCs) in the ipsilateral subventricular zone (SVZi). Ischemic stroke rats were divided into three groups: G0 (non-stroke controls, n = 6), G1 (day 1 after stroke, n = 6), and G7 (days 6 to 8 after stroke, n =12). All the rats underwent MRS. Three rats per group were analyzed by HRMAS. The remaining rats were used for immunohistochemical studies. In the infarct, both techniques detected significant metabolic changes. The most relevant change was in mobile lipids (2.80 ppm) in the G7 group (a 5.53- and a 3.95-fold increase by MRS and HRMAS, respectively). In the SVZi, MRS did not detect any significant metabolic change. However, HRMAS detected a 2.70-fold increase in lactate and a 0.68-fold decrease in N-acetylaspartate in the G1 group. None of the metabolites correlated with the 1.37-fold increase in NPCs detected by immunohistochemistry in the G7 group. In conclusion, HRMAS improves the metabolic characterization of the brain in experimental ischemic stroke. However, none of the metabolites qualifies as a surrogate biomarker of NPCs.

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