50 results on '"Witney TH"'
Search Results
2. (SiFA)SeFe: A Hydrophilic Silicon-Based Fluoride Acceptor Enabling Versatile Peptidic Radiohybrid Tracers.
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Deiser S, Fenzl S, König V, Drexler M, Smith LM, George ME, Beck R, Witney TH, Inoue S, and Casini A
- Subjects
- Animals, Humans, Mice, Cell Line, Tumor, Chorioallantoic Membrane metabolism, Fluorides chemistry, Fluorine Radioisotopes chemistry, Lutetium chemistry, Peptides chemistry, Positron-Emission Tomography, Radioisotopes chemistry, Radiopharmaceuticals chemistry, Radiopharmaceuticals chemical synthesis, Radiopharmaceuticals pharmacokinetics, Silicon chemistry, Tissue Distribution, Coordination Complexes chemical synthesis, Coordination Complexes chemistry, Coordination Complexes pharmacology, Iron Compounds chemistry, Hydrophobic and Hydrophilic Interactions, Receptors, Somatostatin metabolism
- Abstract
The radiohybrid (rh) concept to design targeted (and chemically identical) radiotracers for imaging or radionuclide therapy of tumors has gained momentum. For this strategy, a new bifunctional Silicon-based Fluoride Acceptor (SiFA) moiety (SiFA)SeFe was synthesized, endowed with improved hydrophilicity and high versatility of integration into rh-compounds. Preliminary radiolabeling and stability studies under different conditions were conducted using model bioconjugate peptides. Further, three somatostatin receptor 2 (sstR2)-targeted rh-compounds ( (SiFA)SeFe-rhTATE1-3 , TATE = (Tyr
3 )-octreotate) were developed. Compound (SiFA)SeFe-rhTATE3 , enables labeling with18 F for PET imaging or chelation of177 Lu for therapy. The rh-compounds possess comparable receptor binding affinity and in vitro performance as good as the clinically proven gold standards. SstR2-specificity was further shown for (SiFA)SeFe-rhTATE2 using the chicken chorioallantoic membrane (CAM) model. The biodistribution of two compounds in mice showed high accumulation in tumors and excretion via the kidneys, demonstrating the clinical applicability of the (SiFA)SeFe moiety.- Published
- 2024
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3. Clinical [ 18 F]FSPG Positron Emission Tomography Imaging Reveals Heterogeneity in Tumor-Associated System x c - Activity.
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Sharkey AR, Koglin N, Mittra ES, Han S, Cook GJR, and Witney TH
- Abstract
Background: (4 S )-4-(3-[
18 F]fluoropropyl)-L-glutamic acid ([18 F]FSPG) positron emission tomography/computed tomography (PET/CT) provides a readout of system xc - transport activity and has been used for cancer detection in clinical studies of different cancer types. As system xc - provides the rate-limiting precursor for glutathione biosynthesis, an abundant antioxidant, [18 F]FSPG imaging may additionally provide important prognostic information. Here, we performed an analysis of [18 F]FSPG radiotracer distribution between primary tumors, metastases, and normal organs from cancer patients. We further assessed the heterogeneity of [18 F]FSPG retention between cancer types, and between and within individuals., Methods: This retrospective analysis of prospectively collected data compared [18 F]FSPG PET/CT in subjects with head and neck squamous cell cancer (HNSCC, n = 5) and non-small-cell lung cancer (NSCLC, n = 10), scanned at different institutions. Using semi-automated regions of interest drawn around tumors and metastases, the maximum standardized uptake value (SUVmax ), SUVmean , SUV standard deviation and SUVpeak were measured. [18 F]FSPG time-activity curves (TACs) for normal organs, primary tumors and metastases were subsequently compared to18 F-2-fluoro-2-deoxy-D-glucose ([18 F]FDG) PET/CT at 60 min post injection (p.i.)., Results: The mean administered activity of [18 F]FSPG was 309.3 ± 9.1 MBq in subjects with NSCLC and 285.1 ± 11.3 MBq in those with HNSCC. The biodistribution of [18 F]FSPG in both cohorts showed similar TACs in healthy organs from cancer patients. There was no statistically significant overall difference in the average SUVmax of tumor lesions at 60 min p.i. for NSCLC (8.1 ± 7.1) compared to HNSCC (6.0 ± 4.1; p = 0.29) for [18 F]FSPG. However, there was heterogeneous retention between and within cancer types; the SUVmax at 60 min p.i. ranged from 1.4 to 23.7 in NSCLC and 3.1-12.1 in HNSCC., Conclusion: [18 F]FSPG PET/CT imaging from both NSCLC and HNSCC cohorts showed the same normal-tissue biodistribution, but marked tumor heterogeneity across subjects and between lesions. Despite rapid elimination through the urinary tract and low normal-background tissue retention, the diagnostic potential of [18 F]FSPG was limited by variability in tumor retention. As [18 F]FSPG retention is mediated by the tumor's antioxidant capacity and response to oxidative stress, this heterogeneity may provide important insights into an individual tumor's response or resistance to therapy.- Published
- 2024
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4. Theranostics - a sure cure for cancer after 100 years?
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Song Y, Zou J, Castellanos EA, Matsuura N, Ronald JA, Shuhendler A, Weber WA, Gilad AA, Müller C, Witney TH, and Chen X
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- Humans, Drug Delivery Systems methods, Nanomedicine methods, History, 20th Century, Animals, History, 21st Century, Neoplasms therapy, Neoplasms diagnosis, Theranostic Nanomedicine methods, Precision Medicine methods
- Abstract
Cancer has remained a formidable challenge in medicine and has claimed an enormous number of lives worldwide. Theranostics, combining diagnostic methods with personalized therapeutic approaches, shows huge potential to advance the battle against cancer. This review aims to provide an overview of theranostics in oncology: exploring its history, current advances, challenges, and prospects. We present the fundamental evolution of theranostics from radiotherapeutics, cellular therapeutics, and nanotherapeutics, showcasing critical milestones in the last decade. From the early concept of targeted drug delivery to the emergence of personalized medicine, theranostics has benefited from advances in imaging technologies, molecular biology, and nanomedicine. Furthermore, we emphasize pertinent illustrations showcasing that revolutionary strategies in cancer management enhance diagnostic accuracy and provide targeted therapies customized for individual patients, thereby facilitating the implementation of personalized medicine. Finally, we describe future perspectives on current challenges, emerging topics, and advances in the field., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)
- Published
- 2024
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5. Is there a role for [ 18 F]-FMISO PET to guide dose adaptive radiotherapy in head and neck cancer? A review of the literature.
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Sambasivan K, Barrington SF, Connor SE, Witney TH, Blower PJ, and Urbano TG
- Abstract
Purpose: Hypoxia is a major cause of radioresistance in head and neck cancer (HNC), resulting in treatment failure and disease recurrence.
18 F-fluoromisonidazole [18 F]FMISO PET has been proposed as a means of localising intratumoural hypoxia in HNC so that radiotherapy can be specifically escalated in hypoxic regions. This concept may not be deliverable in routine clinical practice, however, given that [18 F]FMISO PET is costly, time consuming and difficult to access. The aim of this review was to summarise clinical studies involving [18 F]FMISO PET to ascertain whether it can be used to guide radiotherapy treatment in HNC., Methods: A comprehensive literature search was conducted on PubMed and Web of Science databases. Studies investigating [18 F]FMISO PET in newly diagnosed HNC patients were considered eligible for review., Results: We found the following important results from our literature review: 1)Studies have focussed on comparing [18 F]FMISO PET to other hypoxia biomarkers, but currently there is no evidence of a strong correlation between [18 F]FMISO and these biomarkers.2)The results of [18 F]FMISO PET imaging are not necessarily repeatable, and the location of uptake may vary during treatment.3)Tumour recurrences do not always occur within the pretreatment hypoxic volume on [18 F]FMISO PET.4)Dose modification studies using [18 F]FMISO PET are in a pilot phase and so far, none have demonstrated the efficacy of radiotherapy dose painting according to [18 F]FMISO uptake on PET., Conclusions: Our results suggest it is unlikely [18 F]FMISO PET will be suitable for radiotherapy dose adaptation in HNC in a routine clinical setting. Part of the problem is that hypoxia is a dynamic phenomenon, and thus difficult to delineate on a single scan. Currently, it is anticipated that [18 F]FMISO PET will remain useful within the research setting only., Competing Interests: Competing Interests: “The authors have no relevant financial or non-financial interests to disclose.”- Published
- 2024
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6. mRNA therapy corrects defective glutathione metabolism and restores ureagenesis in preclinical argininosuccinic aciduria.
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Gurung S, Timmermand OV, Perocheau D, Gil-Martinez AL, Minnion M, Touramanidou L, Fang S, Messina M, Khalil Y, Spiewak J, Barber AR, Edwards RS, Pinto PL, Finn PF, Cavedon A, Siddiqui S, Rice L, Martini PGV, Ridout D, Heywood W, Hargreaves I, Heales S, Mills PB, Waddington SN, Gissen P, Eaton S, Ryten M, Feelisch M, Frassetto A, Witney TH, and Baruteau J
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- Adult, Humans, Animals, Mice, Cysteine, Glutathione, Metabolomics, Argininosuccinic Aciduria genetics, Argininosuccinic Aciduria therapy, Liver Diseases
- Abstract
The urea cycle enzyme argininosuccinate lyase (ASL) enables the clearance of neurotoxic ammonia and the biosynthesis of arginine. Patients with ASL deficiency present with argininosuccinic aciduria, an inherited metabolic disease with hyperammonemia and a systemic phenotype coinciding with neurocognitive impairment and chronic liver disease. Here, we describe the dysregulation of glutathione biosynthesis and upstream cysteine utilization in ASL-deficient patients and mice using targeted metabolomics and in vivo positron emission tomography (PET) imaging using ( S )-4-(3-
18 F-fluoropropyl)-l-glutamate ([18 F]FSPG). Up-regulation of cysteine metabolism contrasted with glutathione depletion and down-regulated antioxidant pathways. To assess hepatic glutathione dysregulation and liver disease, we present [18 F]FSPG PET as a noninvasive diagnostic tool to monitor therapeutic response in argininosuccinic aciduria. Human hASL mRNA encapsulated in lipid nanoparticles improved glutathione metabolism and chronic liver disease. In addition, hASL mRNA therapy corrected and rescued the neonatal and adult Asl-deficient mouse phenotypes, respectively, enhancing ureagenesis. These findings provide mechanistic insights in liver glutathione metabolism and support clinical translation of mRNA therapy for argininosuccinic aciduria.- Published
- 2024
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7. Imaging the Tumor Antioxidant Response with [ 18 F]FSPG PET.
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Timmermand OV and Witney TH
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- Humans, Cystine, Positron-Emission Tomography methods, Glutamic Acid, Antioxidants, Neoplasms diagnostic imaging
- Abstract
(4S)-4-(3-[
18 F]Fluoropropyl)-L-glutamic acid ([18 F]FSPG) is a flourine-18 labeled glutamate analog that enables the noninvasive in vivo imaging of cellular redox status. [18 F]FSPG is transported across the cell membrane by the cystine/glutamate antiporter, system xc - , whose expression is upregulated in multiple cancer types. The requirement of cystine for the biosynthesis of glutathione, a major antioxidant, connects [18 F]FSPG tissue retention to the intracellular redox response via system xc - activity. We herein describe the use of [18 F]FSPG positron emission tomography (PET) to image the tumor antioxidant response and highlight key methodological considerations., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)- Published
- 2024
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8. [ 18 F]FSPG-PET provides an early marker of radiotherapy response in head and neck squamous cell cancer.
- Author
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Sambasivan K, Tyrrell WE, Farooq R, Mynerich J, Edwards RS, Tanc M, Urbano TG, and Witney TH
- Abstract
The ability to image early treatment response to radiotherapy in head and neck squamous cell carcinoma (HNSCC) will enable the identification of radioresistant tumor volumes suitable for treatment intensification. Here, we propose the system x
c - radiotracer (4 S )-4-(3-[18 F]fluoropropyl)-L-glutamate ([18 F]FSPG) as a non-invasive method to monitor radiation response in HNSCC. We assessed temporal changes in cell death, antioxidant status, and [18 F]FSPG retention following a single dose of 10 Gy irradiation in FaDU HNSCC cells. Next, using a fractionated course of radiotherapy, we assessed tumor volume changes and performed [18 F]FSPG-PET imaging in FaDU-bearing mouse xenografts, followed by ex vivo response assessment. In cells, 10 Gy irradiation reduced [18 F]FSPG retention, coinciding with the induction of apoptosis and the production of reactive oxygen species. In vivo, [18 F]FSPG tumor retention was halved seven days after the start of treatment, which preceded radiotherapy-induced tumor shrinkage, thereby confirming [18 F]FSPG-PET as an early and sensitive marker of radiation response., Competing Interests: Competing interestsT.H.W. is the Editor-in-Chief of npj Imaging. He was blinded to this manuscript's editorial and peer review process. All other authors declare no competing financial or non-financial interests., (© The Author(s) 2024.)- Published
- 2024
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9. Imaging the master regulator of the antioxidant response in non-small cell lung cancer with positron emission tomography.
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Greenwood HE, Edwards RS, Tyrrell WE, Barber AR, Baark F, Tanc M, Khalil E, Falzone A, Ward NP, DeBlasi JM, Torrente L, Pearce DR, Firth G, Smith LM, Timmermand OV, Huebner A, George ME, Swanton C, Hynds RE, DeNicola GM, and Witney TH
- Abstract
Mutations in the NRF2-KEAP1 pathway are common in non-small cell lung cancer (NSCLC) and confer broad-spectrum therapeutic resistance, leading to poor outcomes. The cystine/glutamate antiporter, system x
c - , is one of the >200 cytoprotective proteins controlled by NRF2, which can be non-invasively imaged by ( S )-4-(3-18 F-fluoropropyl)-l-glutamate ([18 F]FSPG) positron emission tomography (PET). Through genetic and pharmacologic manipulation, we show that [18 F]FSPG provides a sensitive and specific marker of NRF2 activation in advanced preclinical models of NSCLC. We validate imaging readouts with metabolomic measurements of system xc - activity and their coupling to intracellular glutathione concentration. A redox gene signature was measured in patients from the TRACERx 421 cohort, suggesting an opportunity for patient stratification prior to imaging. Furthermore, we reveal that system xc - is a metabolic vulnerability that can be therapeutically targeted for sustained tumour growth suppression in aggressive NSCLC. Our results establish [18 F]FSPG as predictive marker of therapy resistance in NSCLC and provide the basis for the clinical evaluation of both imaging and therapeutic agents that target this important antioxidant pathway.- Published
- 2023
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10. The chicken chorioallantoic membrane as a low-cost, high-throughput model for cancer imaging.
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Smith LM, Greenwood HE, Tyrrell WE, Edwards RS, de Santis V, Baark F, Firth G, Tanc M, Terry SYA, Herrmann A, Southworth R, and Witney TH
- Abstract
Mouse models are invaluable tools for radiotracer development and validation. They are, however, expensive, low throughput, and are constrained by animal welfare considerations. Here, we assessed the chicken chorioallantoic membrane (CAM) as an alternative to mice for preclinical cancer imaging studies. NCI-H460 FLuc cells grown in Matrigel on the CAM formed vascularized tumors of reproducible size without compromising embryo viability. By designing a simple method for vessel cannulation it was possible to perform dynamic PET imaging in ovo, producing high tumor-to-background signal for both
18 F-2-fluoro-2-deoxy-D-glucose (18 F-FDG) and (4S)-4-(3-18 F-fluoropropyl)-L-glutamate (18 F-FSPG). The pattern of18 F-FDG tumor uptake were similar in ovo and in vivo, although tumor-associated radioactivity was higher in the CAM-grown tumors over the 60 min imaging time course. Additionally,18 F-FSPG provided an early marker of both treatment response to external beam radiotherapy and target inhibition in ovo. Overall, the CAM provided a low-cost alternative to tumor xenograft mouse models which may broaden access to PET and SPECT imaging and have utility across multiple applications., Competing Interests: Competing Interests The authors declare no competing interests.- Published
- 2023
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11. Is System x c - a Suitable Target for Tumour Detection and Response Assessment with Imaging?
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Sharkey AR, Witney TH, and Cook GJR
- Abstract
System x
c - is upregulated in cancer cells and can be imaged using novel radiotracers, most commonly with (4S)-4-(3-[18 F]fluoropropyl)-L-glutamic acid (18 F-FSPG). The aim of this review was to summarise the use of18 F-FSPG in humans, explore the benefits and limitations of18 F-FSPG, and assess the potential for further use of18 F-FSPG in cancer patients. To date, ten papers have described the use of18 F-FSPG in human cancers. These studies involved small numbers of patients (range 1-26) and assessed the use of18 F-FSPG as a general oncological diagnostic agent across different cancer types. These clinical trials were contrasting in their findings, limiting the scope of18 F-FSPG PET/CT as a purely diagnostic agent, primarily due to heterogeneity of18 F-FSPG retention both between cancer types and patients. Despite these limitations, a potential further application for18 F-FSPG is in the assessment of early treatment response and prediction of treatment resistance. Animal models of cancer have shown that changes in18 F-FSPG retention following effective therapy precede glycolytic changes, as indicated by18 F-FDG, and changes in tumour volume, as measured by CT. If these results could be replicated in human clinical trials, imaging with18 F-FSPG PET/CT would offer an exciting route towards addressing the currently unmet clinical needs of treatment resistance prediction and early imaging assessment of therapy response., Competing Interests: The authors declare no conflicts of interest.- Published
- 2023
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12. A prodrug strategy for the in vivo imaging of aldehyde dehydrogenase activity.
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Pereira R, Flaherty RL, Edwards RS, Greenwood HE, Shuhendler AJ, and Witney TH
- Abstract
Therapy resistance is one of the biggest challenges facing clinical oncology. Despite a revolution in new anti-cancer drugs targeting multiple components of the tumour microenvironment, acquired or innate resistance frequently blunts the efficacy of these treatments. Non-invasive identification of drug-resistant tumours will enable modification of the patient treatment pathway through the selection of appropriate second-line treatments. Here, we have designed a prodrug radiotracer for the non-invasive imaging of aldehyde dehydrogenase 1A1 (ALDH1A1) activity. Elevated ALDH1A1 activity is a marker of drug-resistant cancer cells, modelled here with matched cisplatin-sensitive and -resistant human SKOV3 ovarian cancer cells. The aromatic aldehyde of our prodrug radiotracer was intracellularly liberated by esterase cleavage of the geminal diacetate and specifically trapped by ALDH through its conversion to the charged carboxylic acid. Through this mechanism of action, ALDH-specific retention of our prodrug radiotracer in the drug-resistant tumour cells was twice as high as the drug-sensitive cells. Acylal masking of the aldehyde afforded a modest protection from oxidation in the blood, which was substantially improved in carrier-added experiments. In vivo positron emission tomography imaging of tumour-bearing mice produced high tumour-to-background images and radiotracer uptake in high ALDH-expressing organs but was unable to differentiate between drug-sensitive and drug-resistant tumours. Alternative strategies to protect the labile aldehyde are currently under investigation., Competing Interests: THW has received commercial funding from GlaxoSmithKline and Life Molecular Imaging. The other authors have no conflicts of interest to declare., (This journal is © The Royal Society of Chemistry.)
- Published
- 2022
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13. Radiotracer stereochemistry affects substrate affinity and kinetics for improved imaging of system x C - in tumors.
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Greenwood HE, Edwards R, Koglin N, Berndt M, Baark F, Kim J, Firth G, Khalil E, Mueller A, and Witney TH
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- Animals, Cell Line, Tumor, Female, Glutamic Acid, Humans, Kinetics, Mice, Positron Emission Tomography Computed Tomography methods, Positron-Emission Tomography methods, Radiopharmaceuticals pharmacokinetics, Lung Neoplasms diagnostic imaging, Ovarian Neoplasms metabolism
- Abstract
Amino acid utilization is perturbed in cancer cells, which rewire their metabolism to support cell survival and proliferation. This metabolic reprogramming can be exploited for diagnostic purposes through positron emission tomography imaging of fluorine-18 labeled amino acids. Despite its promise, little is known regarding transporter-recognition of non-natural amino acid stereoisomers or their utility for cancer imaging. We report here the synthesis and in vivo characterization of a radiolabeled amino acid ( R )-4-(3-
18 F-fluoropropyl)-ʟ-glutamate ([18 F]FRPG) and compared its tumor imaging properties to the 4 S -isomer, [18 F]FSPG. Methods : [18 F]FRPG and [18 F]FSPG uptake was assessed in H460 lung cancer cells, with efflux measured 30 min after removal of exogenous activity. Specificity of [18 F]FRPG for system xC - was further examined following transporter inhibition and blocking studies with system xC - substrates. [18 F]FRPG and [18 F]FSPG pharmacokinetics was next quantified in mice bearing subcutaneous A549, H460, VCAP and PC3 tumors, with mice bearing A549 tumors imaged by PET/CT. To better-understand differential tumor retention, radiometabolite analysis was performed on tissue and blood samples after imaging. Next, [18 F]FRPG and [18 F]FSPG retention in lipopolysaccharide-treated lungs were compared to an orthotopic H460 lung cancer model. Finally, the sensitivity of [18 F]FRPG to manipulation of the redox environment was examined in cell and in vivo models. Results : [18 F]FRPG was specifically transported across the plasma membrane by the cystine/glutamate antiporter system xC - and retained at high levels in multiple tumor models. Conversely, [18 F]FRPG was rapidly extracted from the blood and cleared from tissues with low system xC - expression. Due to its favorable imaging properties, tumor-to-blood ratios ≥10 were achieved with [18 F]FRPG, which were either equal to or greater than [18 F]FSPG. In addition, [18 F]FRPG retention in orthotopic lung tumors with high system xC - expression was 2.5-fold higher than inflamed tissue, allowing for clear tumor visualization. In vivo , [18 F]FRPG and [18 F]FSPG were metabolized to a single species, with [18 F]FRPG showing a higher percentage of parent radiotracer in tumors compared to [18 F]FSPG. [18 F]FRPG was sensitive to redox manipulations and tumor retention was reduced following treatment with liposomal doxorubicin in mice bearing ovarian tumors. Conclusions : Given the fast clearance and low background retention of [18 F]FRPG throughout the body, this radiotracer holds promise for the imaging of system xC - activity and treatment response monitoring in tumors of the thorax, abdomen, and head and neck. [18 F]FRPG PET imaging provides a sensitive noninvasive measure of system xC - and excellent properties for cancer imaging., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)- Published
- 2022
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14. Robust and Facile Automated Radiosynthesis of [ 18 F]FSPG on the GE FASTlab.
- Author
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Edwards R, Greenwood HE, McRobbie G, Khan I, and Witney TH
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- Animals, Fluorides, Mice, Positron-Emission Tomography methods, Radiochemistry methods, Radiopharmaceuticals, Carcinoma, Non-Small-Cell Lung, Lung Neoplasms
- Abstract
Purpose: (S)-4-(3-
18 F-Fluoropropyl)-ʟ-Glutamic Acid ([18 F]FSPG) is a radiolabeled non-natural amino acid that is used for positron emission tomography (PET) imaging of the glutamate/cystine antiporter, system xC - , whose expression is upregulated in many cancer types. To increase the clinical adoption of this radiotracer, reliable and facile automated procedures for [18 F]FSPG production are required. Here, we report a cassette-based method to produce [18 F]FSPG at high radioactivity concentrations from low amounts of starting activity., Procedures: An automated synthesis and purification of [18 F]FSPG was developed using the GE FASTlab. Optimization of the reaction conditions and automated manipulations were performed by measuring the isolated radiochemical yield of [18 F]FSPG and by assessing radiochemical purity using radio-HPLC. Purification of [18 F]FSPG was conducted by trapping and washing of the radiotracer on Oasis MCX SPE cartridges, followed by a reverse elution of [18 F]FSPG in phosphate-buffered saline. Subsequently, the [18 F]FSPG obtained from the optimized process was used to image an animal model of non-small cell lung cancer., Results: The optimized protocol produced [18 F]FSPG in 38.4 ± 2.6 % radiochemical yield and >96 % radiochemical purity with a molar activity of 11.1 ± 7.7 GBq/μmol. Small alterations, including the implementation of a reverse elution and an altered Hypercarb cartridge, led to significant improvements in radiotracer concentration from <10 MBq/ml to >100 MBq/ml. The improved radiotracer concentration allowed for the imaging of up to 20 mice, starting with just 1.5 GBq of [18 F]Fluoride., Conclusions: We have developed a robust and facile method for [18 F]FSPG radiosynthesis in high radiotracer concentration, radiochemical yield, and radiochemical purity. This cassette-based method enabled the production of [18 F]FSPG at radioactive concentrations sufficient to facilitate large-scale preclinical experiments with a single prep of starting activity. The use of a cassette-based radiosynthesis on an automated synthesis module routinely used for clinical production makes the method amenable to rapid and widespread clinical translation., (© 2021. The Author(s).)- Published
- 2021
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15. Latest Advances in Imaging Oxidative Stress in Cancer.
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Greenwood HE and Witney TH
- Abstract
Oxidative stress is the imbalance of harmful reactive oxygen species (ROS) and the action of neutralizing antioxidant mechanisms. If left unchecked, the deleterious effects of oxidative stress result in damage to DNA, proteins, and membranes, ultimately leading to cell death. Tumors are highly proliferative and consequently generate high levels of mitochondrial ROS. To compensate for this and maintain redox homeostasis, cancer cells upregulate protective antioxidant pathways, which are further amplified in drug-resistant tumors. This review provides an overview of the latest molecular imaging techniques designed to image oxidative stress in cancer. New probes can now assess heterogeneous ROS and antioxidant production within tumors and across lesions. Together, the noninvasive imaging of these dynamic processes holds great promise for monitoring response to treatment and predicting drug resistance and may provide insight into the metastatic potential of tumors., (© 2021 by the Society of Nuclear Medicine and Molecular Imaging.)
- Published
- 2021
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16. The chemical tool-kit for molecular imaging with radionuclides in the age of targeted and immune therapy.
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Witney TH and Blower PJ
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- Humans, Immunotherapy methods, Molecular Imaging methods, Neoplasms diagnostic imaging, Neoplasms therapy, Positron Emission Tomography Computed Tomography methods, Positron-Emission Tomography methods, Radioisotopes metabolism, Radiopharmaceuticals metabolism
- Abstract
Nuclear medicine has evolved over the last half-century from a functional imaging modality using a handful of radiopharmaceuticals, many of unknown structure and mechanism of action, into a modern speciality that can properly be described as molecular imaging, with a very large number of specific radioactive probes of known structure that image specific molecular processes. The advances of cancer treatment in recent decades towards targeted and immune therapies, combined with recognition of heterogeneity of cancer cell phenotype among patients, within patients and even within tumours, has created a growing need for personalised molecular imaging to support treatment decision. This article describes the evolution of the present vast range of radioactive probes - radiopharmaceuticals - leveraging a wide variety of chemical disciplines, over the last half century. These radiochemical innovations have been inspired by the need to support personalised medicine and also by the parallel development in development of new radionuclide imaging technologies - from gamma scintigraphy, through single photon emission tomography (SPECT), through the rise of clinical positron emission tomography (PET) and PET-CT, and perhaps in the future, by the advent of total body PET. Thus, in the interdisciplinary world of nuclear medicine and molecular imaging, as quickly as radiochemistry solutions are developed to meet new needs in cancer imaging, new challenges emerge as developments in one contributing technology drive innovations in the others.
- Published
- 2021
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17. Correction: Preclinical Assessment of Carboplatin Treatment Efficacy in Lung Cancer by 18F-ICMT-11-Positron Emission Tomography.
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Witney TH, Fortt R, and Aboagye EO
- Abstract
[This corrects the article DOI: 10.1371/journal.pone.0091694.].
- Published
- 2020
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18. High-Throughput PET/CT Imaging Using a Multiple-Mouse Imaging System.
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Greenwood HE, Nyitrai Z, Mocsai G, Hobor S, and Witney TH
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- Animals, Female, Fluorodeoxyglucose F18, Mice, Mice, Inbred BALB C, Phantoms, Imaging, Positron Emission Tomography Computed Tomography methods
- Abstract
A considerable limitation of current small-animal PET/CT imaging is the low throughput of acquisitions. Consequently, to sufficiently power a study, high costs accumulate. Together with a commercial scanner manufacturer, we developed a 4-bed mouse "hotel" to simultaneously image up to 4 mice, thereby reducing costs and maximizing the efficiency of radiotracer use when compared with scans performed with a single mouse bed. Methods: For physiologic evaluation of the mouse hotel, temperature and anesthesia were tested for uniformity in conjunction with
18 F-FDG PET/CT imaging of mini image-quality phantoms designed to fit the new imaging system. After reconstruction, National Electrical Manufacturers Association NU-4 tests examined uniformity, recovery coefficients, and spillover ratios. To evaluate the mouse hotel under standard in vivo imaging conditions, 4 mice were simultaneously scanned by dynamic18 F-FDG PET/CT over 60 min, and quantified images were compared with those acquired using a single mouse bed. Results: The mouse hotel maintained a constant temperature of 36.8°C ± 0.4°C, with anesthesia distributed evenly to each nose cone (2.9 ± 0.1 L/min). The National Electrical Manufacturers Association tests revealed values within tolerable limits for uniformity, for recovery coefficients in rods larger than 2 mm, and for spillover ratios in the nonradioactive water- and air-filled chambers. There was low variability in radiotracer uptake in all major organs for the mouse hotel versus the single mouse bed. Conclusion: Analysis of images acquired using the mouse hotel confirmed its utility to increase the throughput of small-animal PET imaging without considerable loss of image quality or quantitative precision. In comparison to a single mouse bed, the cost and time associated with each scan were substantially reduced., (© 2020 by the Society of Nuclear Medicine and Molecular Imaging.)- Published
- 2020
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19. Iron(II)/Persulfate Mediated Newman-Kwart Rearrangement.
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Gendron T, Pereira R, Abdi HY, Witney TH, and Årstad E
- Abstract
Herein, we report that iron(II)/ammonium persulfate in aqueous acetonitrile mediates the Newman-Kwart rearrangement of O -aryl carbamothioates. Electron-rich substrates react rapidly under moderate heating to afford the rearranged products in excellent yields. The mild conditions, rapid reaction rates, and suitability for scale up offers immediate practical benefits to access functionalized thiophenols.
- Published
- 2020
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20. One-Pot Radiosynthesis and Biological Evaluation of a Caspase-3 Selective 5-[ 123,125 I]iodo-1,2,3-triazole derived Isatin SPECT Tracer.
- Author
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Glaser M, Rajkumar V, Diocou S, Gendron T, Yan R, Sin PKB, Sander K, Carroll L, Pedley RB, Aboagye EO, Witney TH, and Årstad E
- Subjects
- Animals, Apoptosis genetics, Caspase 3 chemistry, Caspase 3 genetics, Cell Line, Tumor, Copper chemistry, Fluorine Radioisotopes chemistry, Fluorine Radioisotopes pharmacology, Heterografts, Humans, Iodine Radioisotopes chemistry, Isatin chemical synthesis, Isatin pharmacology, Mice, Neoplasms pathology, Neoplasms therapy, Radiopharmaceuticals chemical synthesis, Radiopharmaceuticals pharmacology, Tissue Distribution, Tomography, Emission-Computed, Single-Photon methods, Triazoles chemical synthesis, Triazoles pharmacology, Apoptosis drug effects, Caspase 3 isolation & purification, Iodine Radioisotopes pharmacology, Neoplasms diagnostic imaging
- Abstract
Induction of apoptosis is often necessary for successful cancer therapy, and the non-invasive monitoring of apoptosis post-therapy could assist in clinical decision making. Isatins are a class of compounds that target activated caspase-3 during apoptosis. Here we report the synthesis of the 5-iodo-1,2,3-triazole (FITI) analog of the PET tracer [
18 F]ICMT11 as a candidate tracer for imaging of apoptosis with SPECT, as well as PET. Labelling with radioiodine (123,125 I) was achieved in 55 ± 12% radiochemical yield through a chelator-accelerated one-pot cycloaddition reaction mediated by copper(I) catalysis. The caspase-3 binding affinity and selectivity of FITI compares favourably to that of [18 F]ICMT11 (Ki = 6.1 ± 0.9 nM and 12.4 ± 4.7 nM, respectively). In biodistribution studies, etoposide-induced cell death in a SW1222 xenograft model resulted in a 2-fold increase in tumour uptake of the tracer. However, the tumour uptake was too low to allow in vivo imaging of apoptosis with SPECT.- Published
- 2019
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21. Imaging of T-cells and their responses during anti-cancer immunotherapy.
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Krekorian M, Fruhwirth GO, Srinivas M, Figdor CG, Heskamp S, Witney TH, and Aarntzen EHJG
- Subjects
- Animals, Antibodies, Monoclonal immunology, Humans, Immunotherapy methods, Neoplasms immunology, Neoplasms therapy, T-Lymphocytes, Cytotoxic immunology
- Abstract
Immunotherapy has proven to be an effective approach in a growing number of cancers. Despite durable clinical responses achieved with antibodies targeting immune checkpoint molecules, many patients do not respond. The common denominator for immunotherapies that have successfully been introduced in the clinic is their potential to induce or enhance infiltration of cytotoxic T-cells into the tumour. However, in clinical research the molecules, cells and processes involved in effective responses during immunotherapy remain largely obscure. Therefore, in vivo imaging technologies that interrogate T-cell responses in patients represent a powerful tool to boost further development of immunotherapy. This review comprises a comprehensive analysis of the in vivo imaging technologies that allow the characterisation of T-cell responses induced by anti-cancer immunotherapy, with emphasis on technologies that are clinically available or have high translational potential. Throughout we discuss their respective strengths and weaknesses, providing arguments for selecting the optimal imaging options for future research and patient management., Competing Interests: Competing Interests The authors have declared that no competing interests exists.
- Published
- 2019
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22. Evaluation of integrin αvβ 6 cystine knot PET tracers to detect cancer and idiopathic pulmonary fibrosis.
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Kimura RH, Wang L, Shen B, Huo L, Tummers W, Filipp FV, Guo HH, Haywood T, Abou-Elkacem L, Baratto L, Habte F, Devulapally R, Witney TH, Cheng Y, Tikole S, Chakraborti S, Nix J, Bonagura CA, Hatami N, Mooney JJ, Desai T, Turner S, Gaster RS, Otte A, Visser BC, Poultsides GA, Norton J, Park W, Stolowitz M, Lau K, Yang E, Natarajan A, Ilovich O, Srinivas S, Srinivasan A, Paulmurugan R, Willmann J, Chin FT, Cheng Z, Iagaru A, Li F, and Gambhir SS
- Subjects
- Crystallography, X-Ray, Healthy Volunteers, Humans, Magnetic Resonance Imaging, Positron-Emission Tomography, Antigens, Neoplasm metabolism, Idiopathic Pulmonary Fibrosis diagnosis, Integrins metabolism, Neoplasms diagnosis
- Abstract
Advances in precision molecular imaging promise to transform our ability to detect, diagnose and treat disease. Here, we describe the engineering and validation of a new cystine knot peptide (knottin) that selectively recognizes human integrin αvβ
6 with single-digit nanomolar affinity. We solve its 3D structure by NMR and x-ray crystallography and validate leads with 3 different radiolabels in pre-clinical models of cancer. We evaluate the lead tracer's safety, biodistribution and pharmacokinetics in healthy human volunteers, and show its ability to detect multiple cancers (pancreatic, cervical and lung) in patients at two study locations. Additionally, we demonstrate that the knottin PET tracers can also detect fibrotic lung disease in idiopathic pulmonary fibrosis patients. Our results indicate that these cystine knot PET tracers may have potential utility in multiple disease states that are associated with upregulation of integrin αv β6 .- Published
- 2019
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23. [ 18 F]-SuPAR: A Radiofluorinated Probe for Noninvasive Imaging of DNA Damage-Dependent Poly(ADP-ribose) Polymerase Activity.
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Shuhendler AJ, Cui L, Chen Z, Shen B, Chen M, James ML, Witney TH, Bazalova-Carter M, Gambhir SS, Chin FT, Graves EE, and Rao J
- Subjects
- Animals, Breast Neoplasms diagnostic imaging, Female, Humans, NAD metabolism, Positron-Emission Tomography, Receptors, Urokinase Plasminogen Activator metabolism, Substrate Specificity, Uterine Cervical Neoplasms diagnostic imaging, Xenograft Model Antitumor Assays, DNA Damage, Fluorine Radioisotopes administration & dosage, Poly(ADP-ribose) Polymerases metabolism
- Abstract
Poly(ADP ribose) polymerase (PARP) enzymes generate poly(ADP ribose) post-translational modifications on target proteins for an array of functions centering on DNA and cell stress. PARP isoforms 1 and 2 are critically charged with the surveillance of DNA integrity and are the first line guardians of the genome against DNA breaks. Here we present a novel probe ([
18 F]-SuPAR) for noninvasive imaging of PARP-1/2 activity using positron emission tomography (PET). [18 F]-SuPAR is a radiofluorinated nicotinamide adenine dinucleotide (NAD) analog that can be recognized by PARP-1/2 and incorporated into the long branched polymers of poly(ADP ribose) (PAR). The measurement of PARP-1/2 activity was supported by a reduction of radiotracer uptake in vivo following PARP-1/2 inhibition with talazoparib treatment, a potent PARP inhibitor recently approved by FDA for treatment of breast cancer, as well as ex vivo colocalization of radiotracer analog and poly(ADP ribose). With [18 F]-SuPAR, we were able to map the dose- and time-dependent activation of PARP-1/2 following radiation therapy in breast and cervical cancer xenograft mouse models. Tumor response to therapy was determined by [18 F]-SuPAR PET within 8 h of administration of a single dose of radiation equivalent to one round of stereotactic ablative radiotherapy.- Published
- 2019
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24. Measurement of Tumor Antioxidant Capacity and Prediction of Chemotherapy Resistance in Preclinical Models of Ovarian Cancer by Positron Emission Tomography.
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Greenwood HE, McCormick PN, Gendron T, Glaser M, Pereira R, Maddocks ODK, Sander K, Zhang T, Koglin N, Lythgoe MF, Årstad E, Hochhauser D, and Witney TH
- Subjects
- Animals, Antineoplastic Agents therapeutic use, Cell Line, Tumor, Cystine metabolism, Disease Models, Animal, Female, Humans, Mice, Models, Biological, Neoplasm Grading, Ovarian Neoplasms drug therapy, Positron Emission Tomography Computed Tomography, Radiopharmaceuticals, Reactive Oxygen Species metabolism, Xenograft Model Antitumor Assays, Antineoplastic Agents pharmacology, Antioxidants metabolism, Biomarkers, Drug Resistance, Neoplasm, Ovarian Neoplasms diagnosis, Ovarian Neoplasms metabolism, Positron-Emission Tomography methods
- Abstract
Purpose: Drug resistance is a major obstacle for the effective treatment of patients with high-grade serous ovarian cancer (HGSOC). Currently, there is no satisfactory way to identify patients with HGSOC that are refractive to the standard of care. Here, we propose the system x
c - radiotracer (4 S )-4-(3-[18 F]fluoropropyl)-l-glutamate ([18 F]FSPG) as a non-invasive method to measure upregulated antioxidant pathways present in drug-resistant HGSOC., Experimental Design: Using matched chemotherapy sensitive and resistant ovarian cancer cell lines, we assessed their antioxidant capacity and its relation to [18 F]FSPG uptake, both in cells and in animal models of human ovarian cancer. We identified the mechanisms driving differential [18 F]FSPG cell accumulation and evaluated [18 F]FSPG tumor uptake as predictive marker of treatment response in drug-resistant tumors., Results: High intracellular glutathione (GSH) and low reactive oxygen species corresponded to decreased [18 F]FSPG cell accumulation in drug-resistant versus drug-sensitive cells. Decreased [18 F]FSPG uptake in drug-resistant cells was a consequence of changes in intracellular cystine, a key precursor in GSH biosynthesis. In vivo , [18 F]FSPG uptake was decreased nearly 80% in chemotherapy-resistant A2780 tumors compared with parental drug-sensitive tumors, with nonresponding tumors displaying high levels of oxidized-to-reduced GSH. Treatment of drug-resistant A2780 tumors with doxorubicin resulted in no detectable change in tumor volume, GSH, or [18 F]FSPG uptake., Conclusions: This study demonstrates the ability of [18 F]FSPG to detect upregulated antioxidant pathways present in drug-resistant cancer. [18 F]FSPG may therefore enable the identification of patients with HGSOC that are refractory to standard of care, allowing the transferal of drug-resistant patients to alternative therapies, thereby improving outcomes in this disease., (©2019 American Association for Cancer Research.)- Published
- 2019
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25. Assessment of Tumor Redox Status through ( S )-4-(3-[ 18 F]fluoropropyl)-L-Glutamic Acid PET Imaging of System x c - Activity.
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McCormick PN, Greenwood HE, Glaser M, Maddocks ODK, Gendron T, Sander K, Gowrishankar G, Hoehne A, Zhang T, Shuhendler AJ, Lewis DY, Berndt M, Koglin N, Lythgoe MF, Gambhir SS, Årstad E, and Witney TH
- Subjects
- Acetylcysteine pharmacology, Animals, Apoptosis, Cell Proliferation, Cystadenocarcinoma, Serous diagnostic imaging, Cystadenocarcinoma, Serous drug therapy, Cystadenocarcinoma, Serous metabolism, Female, Free Radical Scavengers pharmacology, Humans, Metabolomics, Mice, Mice, Inbred BALB C, Mice, Nude, Ovarian Neoplasms diagnostic imaging, Ovarian Neoplasms drug therapy, Ovarian Neoplasms metabolism, Oxidation-Reduction, Tumor Cells, Cultured, Xenograft Model Antitumor Assays, tert-Butylhydroperoxide pharmacology, Amino Acid Transport System y+ metabolism, Cystadenocarcinoma, Serous pathology, Fluorine Radioisotopes metabolism, Glutamates metabolism, Ovarian Neoplasms pathology, Positron-Emission Tomography methods, Radiopharmaceuticals metabolism
- Abstract
The cell's endogenous antioxidant system is vital to maintenance of redox homeostasis. Despite its central role in normal and pathophysiology, no noninvasive tools exist to measure this system in patients. The cystine/glutamate antiporter system x
c - maintains the balance between intracellular reactive oxygen species and antioxidant production through the provision of cystine, a key precursor in glutathione biosynthesis. Here, we show that tumor cell retention of a system xc - -specific PET radiotracer, ( S )-4-(3-[18 F]fluoropropyl)-L-glutamic acid ([18 F]FSPG), decreases in proportion to levels of oxidative stress following treatment with a range of redox-active compounds. The decrease in [18 F]FSPG retention correlated with a depletion of intracellular cystine resulting from increased de novo glutathione biosynthesis, shown through [U-13 C6 , U-15 N2 ]cystine isotopic tracing. In vivo , treatment with the chemotherapeutic doxorubicin decreased [18 F]FSPG tumor uptake in a mouse model of ovarian cancer, coinciding with markers of oxidative stress but preceding tumor shrinkage and decreased glucose utilization. Having already been used in pilot clinical trials, [18 F]FSPG PET could be rapidly translated to the clinic as an early redox indicator of tumor response to treatment. SIGNIFICANCE: [18 F]FSPG PET imaging provides a sensitive noninvasive measure of tumor redox status and provides an early marker of tumor response to therapy. See related commentary by Lee et al., p. 701 ., (©2018 American Association for Cancer Research.)- Published
- 2019
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26. Mapping Aldehyde Dehydrogenase 1A1 Activity using an [ 18 F]Substrate-Based Approach.
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Pereira R, Gendron T, Sanghera C, Greenwood HE, Newcombe J, McCormick PN, Sander K, Topf M, Årstad E, and Witney TH
- Subjects
- Animals, Biocatalysis, Enzyme Activation, Fluorine Radioisotopes urine, Liver metabolism, Lung metabolism, Mice, Oxidation-Reduction, Positron-Emission Tomography, Substrate Specificity, Aldehyde Dehydrogenase metabolism
- Abstract
Aldehyde dehydrogenases (ALDHs) catalyze the oxidation of aldehydes to carboxylic acids. Elevated ALDH expression in human cancers is linked to metastases and poor overall survival. Despite ALDH being a poor prognostic factor, the non-invasive assessment of ALDH activity in vivo has not been possible due to a lack of sensitive and translational imaging agents. Presented in this report are the synthesis and biological evaluation of ALDH1A1-selective chemical probes composed of an aromatic aldehyde derived from N,N-diethylamino benzaldehyde (DEAB) linked to a fluorinated pyridine ring either via an amide or amine linkage. Of the focused library of compounds evaluated, N-ethyl-6-(fluoro)-N-(4-formylbenzyl)nicotinamide 4 b was found to have excellent affinity and isozyme selectivity for ALDH1A1 in vitro. Following
18 F-fluorination, [18 F]4 b was taken up by colorectal tumor cells and trapped through the conversion to its18 F-labeled carboxylate product under the action of ALDH. In vivo positron emission tomography revealed high uptake of [18 F]4 b in the lungs and liver, with radioactivity cleared through the urinary tract. Oxidation of [18 F]4 b, however, was observed in vivo, which may limit the tissue penetration of this first-in-class radiotracer., (© 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.)- Published
- 2019
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27. Imaging Cancer Metabolism with Positron Emission Tomography (PET).
- Author
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Witney TH and Lewis DY
- Subjects
- Animals, Biomarkers, Carbon Radioisotopes, Disease Models, Animal, Fluorodeoxyglucose F18, Humans, Image Processing, Computer-Assisted, Imaging, Three-Dimensional, Metabolic Networks and Pathways, Mice, Neoplasms pathology, Positron Emission Tomography Computed Tomography, Radiopharmaceuticals, Energy Metabolism, Neoplasms diagnostic imaging, Neoplasms metabolism, Positron-Emission Tomography methods
- Abstract
Positron emission tomography (PET) enables the noninvasive spatiotemporal analysis of cancer metabolism in vivo. Both natural and nonnatural PET tracers have been developed to assess metabolic pathways during tumorigenesis, cancer progression, and metastasis. Here we describe the dynamic in vivo PET/CT imaging of the glucose analogue [
18 F]fluoro-2-deoxy-D-glucose (FDG), taking into consideration the methodology for alternative metabolic PET substrates.- Published
- 2019
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28. 3'-Deoxy-3'-[ 18 F]Fluorothymidine Uptake Is Related to Thymidine Phosphorylase Expression in Various Experimental Tumor Models.
- Author
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Schelhaas S, Heinzmann K, Honess DJ, Smith DM, Keen H, Heskamp S, Witney TH, Besret L, Doblas S, Griffiths JR, Aboagye EO, and Jacobs AH
- Subjects
- Animals, Dideoxynucleosides chemistry, Humans, Mice, Thymidine metabolism, Dideoxynucleosides pharmacokinetics, Neoplasms, Experimental enzymology, Thymidine Phosphorylase metabolism
- Abstract
Purpose: We recently reported that high thymidine phosphorylase (TP) expression is accompanied by low tumor thymidine concentration and high 3'-deoxy-3'-[
18 F]fluorothymidine ([18 F]FLT) uptake in four untreated lung cancer xenografts. Here, we investigated whether this relationship also holds true for a broader range of tumor models., Procedures: Lysates from n = 15 different tumor models originating from n = 6 institutions were tested for TP and thymidylate synthase (TS) expression using western blots. Results were correlated to [18 F]FLT accumulation in the tumors as determined by positron emission tomography (PET) measurements in the different institutions and to previously published thymidine concentrations., Results: Expression of TP correlated positively with [18 F]FLT SUVmax (ρ = 0.549, P < 0.05). Furthermore, tumors with high TP levels possessed lower levels of thymidine (ρ = - 0.939, P < 0.001)., Conclusions: In a broad range of tumors, [18 F]FLT uptake as measured by PET is substantially influenced by TP expression and tumor thymidine concentrations. These data strengthen the role of TP as factor confounding [18 F]FLT uptake.- Published
- 2018
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29. [ 18 F]FSPG-PET reveals increased cystine/glutamate antiporter (xc-) activity in a mouse model of multiple sclerosis.
- Author
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Hoehne A, James ML, Alam IS, Ronald JA, Schneider B, D'Souza A, Witney TH, Andrews LE, Cropper HC, Behera D, Gowrishankar G, Ding Z, Wyss-Coray T, Chin FT, Biswal S, and Gambhir SS
- Subjects
- Animals, Cells, Cultured, Fluorodeoxyglucose F18 metabolism, Humans, Leukocytes, Mononuclear metabolism, Mice, Mice, Inbred C57BL, Multiple Sclerosis diagnostic imaging, Multiple Sclerosis metabolism, Amino Acid Transport Systems, Acidic metabolism, Encephalomyelitis, Autoimmune, Experimental diagnostic imaging, Encephalomyelitis, Autoimmune, Experimental metabolism, Fluorine Radioisotopes metabolism, Glutamates metabolism, Positron-Emission Tomography methods
- Abstract
Background: The cystine/glutamate antiporter (xc-) has been implicated in several neurological disorders and, specifically, in multiple sclerosis (MS) as a mediator of glutamate excitotoxicity and proinflammatory immune responses. We aimed to evaluate an xc-specific positron emission tomography (PET) radiotracer, (4S)-4-(3-[
18 F]fluoropropyl)-L-glutamate ([18 F]FSPG), for its ability to allow non-invasive monitoring of xc- activity in a mouse model of MS., Methods: Experimental autoimmune encephalomyelitis (EAE) was induced in C57BL/6 mice by subcutaneous injection of myelin oligodendrocyte glycoprotein (MOG35-55 ) peptide in complete Freund's adjuvant (CFA) followed by pertussis toxin. Control mice received CFA emulsion and pertussis toxin without MOG peptide, while a separate cohort of naïve mice received no treatment. PET studies were performed to investigate the kinetics and distribution of [18 F]FSPG in naïve, control, pre-symptomatic, and symptomatic EAE mice, compared to18 F-fluorodeoxyglucose ([18 F]FDG). After final PET scans, each mouse was perfused and radioactivity in dissected tissues was measured using a gamma counter. Central nervous system (CNS) tissues were further analyzed using ex vivo autoradiography or western blot. [18 F]FSPG uptake in human monocytes, and T cells pre- and post-activation was investigated in vitro., Results: [18 F]FSPG was found to be more sensitive than [18 F]FDG at detecting pathological changes in the spinal cord and brain of EAE mice. Even before clinical signs of disease, a small but significant increase in [18 F]FSPG signal was observed in the spinal cord of EAE mice compared to controls. This increase in PET signal became more pronounced in symptomatic EAE mice and was confirmed by ex vivo biodistribution and autoradiography. Likewise, in the brain of symptomatic EAE mice, [18 F]FSPG uptake was significantly higher than controls, with the largest changes observed in the cerebellum. Western blot analyses of CNS tissues revealed a significant correlation between light chain of xc- (xCT) protein levels, the subunit of xc- credited with its transporter activity, and [18 F]FSPG-PET signal. In vitro [18 F]FSPG uptake studies suggest that both activated monocytes and T cells contribute to the observed in vivo PET signal., Conclusion: These data highlight the promise of [18 F]FSPG-PET as a technique to provide insights into neuroimmune interactions in MS and the in vivo role of xc- in the development and progression of this disease, thus warranting further investigation.- Published
- 2018
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30. Correction to: The relationship between endogenous thymidine concentrations and [ 18 F]FLT uptake in a range of preclinical tumour models.
- Author
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Heinzmann K, Honess DJ, Lewis DY, Smith DM, Cawthorne C, Keen H, Heskamp S, Schelhaas S, Witney TH, Soloviev D, Williams KJ, Jacobs AH, Aboagye EO, Griffiths JR, and Brindle KM
- Abstract
Correction: Unfortunately, the original version of Figs. 4, 5 and 6b in the article [1] contained errors in the n numbers as indicated on the columns. Please note that column heights and error bars in the original figures and data in the ESM tables are correct and statistical tests are valid. These corrections do not affect any results or conclusions in this article.
- Published
- 2017
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31. Rapid Imaging of Tumor Cell Death In Vivo Using the C2A Domain of Synaptotagmin-I.
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Neves AA, Xie B, Fawcett S, Alam IS, Witney TH, de Backer MM, Summers J, Hughes W, McGuire S, Soloviev D, Miller J, Howat WJ, Hu DE, Rodrigues TB, Lewis DY, and Brindle KM
- Subjects
- Animals, Biomarkers metabolism, Cell Line, Tumor, Female, Humans, Mice, Mice, Inbred C57BL, Mice, Nude, Neoplasms, Experimental diagnostic imaging, Protein Domains, Radiopharmaceuticals pharmacokinetics, Synaptotagmin I chemistry, Tissue Distribution, Apoptosis, Molecular Imaging methods, Neoplasms, Experimental metabolism, Neoplasms, Experimental pathology, Positron-Emission Tomography methods, Synaptotagmin I pharmacokinetics
- Abstract
Cell death is an important target for imaging the early response of tumors to treatment. We describe here the validation of a phosphatidylserine-binding agent for detecting tumor cell death in vivo based on the C2A domain of synaptotagmin-I. Methods: The capability of near-infrared fluorophore-labeled and
99m Tc- and111 In-labeled derivatives of C2Am for imaging tumor cell death, using planar near-infrared fluorescence imaging and SPECT, respectively, was evaluated in implanted and genetically engineered mouse models of lymphoma and in a human colorectal xenograft. Results: The fluorophore-labeled C2Am derivative showed predominantly renal clearance and high specificity and sensitivity for detecting low levels of tumor cell death (2%-5%). There was a significant correlation ( R > 0.9, P < 0.05) between fluorescently labeled C2Am binding and histologic markers of cell death, including cleaved caspase-3, whereas there was no such correlation with a site-directed mutant of C2Am (iC2Am) that does not bind phosphatidylserine.99m Tc-C2Am and111 In-C2Am also showed favorable biodistribution profiles, with predominantly renal clearance and low nonspecific retention in the liver and spleen at 24 h after probe administration.99m Tc-C2Am and111 In-C2Am generated tumor-to-muscle ratios in drug-treated tumors of 4.3× and 2.2×, respectively, at 2 h and 7.3× and 4.1×, respectively, at 24 h after administration. Conclusion: Given the favorable biodistribution profile of99m Tc- and111 In-labeled C2Am, and their ability to produce rapid and cell death-specific image contrast, these agents have potential for clinical translation., (© 2017 by the Society of Nuclear Medicine and Molecular Imaging.)- Published
- 2017
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32. Reporter gene imaging of targeted T cell immunotherapy in recurrent glioma.
- Author
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Keu KV, Witney TH, Yaghoubi S, Rosenberg J, Kurien A, Magnusson R, Williams J, Habte F, Wagner JR, Forman S, Brown C, Allen-Auerbach M, Czernin J, Tang W, Jensen MC, Badie B, and Gambhir SS
- Subjects
- Aged, Brain Neoplasms therapy, Female, Gene Expression, Genetic Therapy methods, Glioma therapy, Humans, Interleukin-13 metabolism, Male, Middle Aged, Neoplasm Recurrence, Local, Positron-Emission Tomography, Prospective Studies, Thymidine Kinase metabolism, Brain Neoplasms diagnostic imaging, Genes, Reporter, Glioma diagnostic imaging, Immunotherapy methods, Receptors, Antigen, T-Cell metabolism, T-Lymphocytes, Cytotoxic cytology
- Abstract
High-grade gliomas are aggressive cancers that often become rapidly fatal. Immunotherapy using CD8
+ cytotoxic T lymphocytes (CTLs), engineered to express both herpes simplex virus type 1 thymidine kinase (HSV1-TK) and interleukin-13 (IL-13) zetakine chimeric antigen receptor (CAR), is a treatment strategy with considerable potential. To optimize this and related immunotherapies, it would be helpful to monitor CTL viability and trafficking to glioma cells. We show that noninvasive positron emission tomography (PET) imaging with 9-[4-[18 F]fluoro-3-(hydroxymethyl)butyl]guanine ([18 F]FHBG) can track HSV1-tk reporter gene expression present in CAR-engineered CTLs. [18 F]FHBG imaging was safe and enabled the longitudinal imaging of T cells stably transfected with a PET reporter gene in patients. Further optimization of this imaging approach for monitoring in vivo cell trafficking should greatly benefit various cell-based therapies for cancer., Competing Interests: Drs. C. Brown and M. Jensen are inventors on patent application WO 02/088334 held by City of Hope National Medical Center that covers Chimeric Immunoreceptor Useful in Treating Human Cancers. Drs. Yaghoubi and Gambhir are co-founders and equity holders of CellSight Inc. that develops strategies to image cell therapies in humans. Dr. Michael Jensen is on the scientific advisory board and equity holder of Juno Therapeutics, which develops novel cancer immunotherapies., (Copyright © 2017, American Association for the Advancement of Science.)- Published
- 2017
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33. The relationship between endogenous thymidine concentrations and [(18)F]FLT uptake in a range of preclinical tumour models.
- Author
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Heinzmann K, Honess DJ, Lewis DY, Smith DM, Cawthorne C, Keen H, Heskamp S, Schelhaas S, Witney TH, Soloviev D, Williams KJ, Jacobs AH, Aboagye EO, Griffiths JR, and Brindle KM
- Abstract
Background: Recent studies have shown that 3'-deoxy-3'-[(18)F] fluorothymidine ([(18)F]FLT)) uptake depends on endogenous tumour thymidine concentration. The purpose of this study was to investigate tumour thymidine concentrations and whether they correlated with [(18)F]FLT uptake across a broad spectrum of murine cancer models. A modified liquid chromatography-mass spectrometry (LC-MS/MS) method was used to determine endogenous thymidine concentrations in plasma and tissues of tumour-bearing and non-tumour bearing mice and rats. Thymidine concentrations were determined in 22 tumour models, including xenografts, syngeneic and spontaneous tumours, from six research centres, and a subset was compared for [(18)F]FLT uptake, described by the maximum and mean tumour-to-liver uptake ratio (TTL) and SUV., Results: The LC-MS/MS method used to measure thymidine in plasma and tissue was modified to improve sensitivity and reproducibility. Thymidine concentrations determined in the plasma of 7 murine strains and one rat strain were between 0.61 ± 0.12 μM and 2.04 ± 0.64 μM, while the concentrations in 22 tumour models ranged from 0.54 ± 0.17 μM to 20.65 ± 3.65 μM. TTL at 60 min after [(18)F]FLT injection, determined in 14 of the 22 tumour models, ranged from 1.07 ± 0.16 to 5.22 ± 0.83 for the maximum and 0.67 ± 0.17 to 2.10 ± 0.18 for the mean uptake. TTL did not correlate with tumour thymidine concentrations., Conclusions: Endogenous tumour thymidine concentrations alone are not predictive of [(18)F]FLT uptake in murine cancer models.
- Published
- 2016
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34. AshwaMAX and Withaferin A inhibits gliomas in cellular and murine orthotopic models.
- Author
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Chang E, Pohling C, Natarajan A, Witney TH, Kaur J, Xu L, Gowrishankar G, D'Souza AL, Murty S, Schick S, Chen L, Wu N, Khaw P, Mischel P, Abbasi T, Usmani S, Mallick P, and Gambhir SS
- Subjects
- Animals, Antineoplastic Agents pharmacology, Brain Neoplasms metabolism, Brain Neoplasms pathology, Cell Line, Tumor, Cell Proliferation drug effects, Cell Survival drug effects, Disease Models, Animal, Dose-Response Relationship, Drug, ErbB Receptors metabolism, Female, Glioblastoma metabolism, Glioblastoma pathology, Humans, Luminescent Measurements, Mice, Mice, Nude, Neural Stem Cells drug effects, Plant Extracts chemistry, Withanolides pharmacology, Xenograft Model Antitumor Assays, Antineoplastic Agents administration & dosage, Brain Neoplasms drug therapy, Glioblastoma drug therapy, Plant Extracts administration & dosage, Withania chemistry, Withanolides administration & dosage
- Abstract
Glioblastoma multiforme (GBM) is an aggressive, malignant cancer Johnson and O'Neill (J Neurooncol 107: 359-364, 2012). An extract from the winter cherry plant (Withania somnifera ), AshwaMAX, is concentrated (4.3 %) for Withaferin A; a steroidal lactone that inhibits cancer cells Vanden Berghe et al. (Cancer Epidemiol Biomark Prev 23: 1985-1996, 2014). We hypothesized that AshwaMAX could treat GBM and that bioluminescence imaging (BLI) could track oral therapy in orthotopic murine models of glioblastoma. Human parietal-cortical glioblastoma cells (GBM2, GBM39) were isolated from primary tumors while U87-MG was obtained commercially. GBM2 was transduced with lentiviral vectors that express Green Fluorescent Protein (GFP)/firefly luciferase fusion proteins. Mutational, expression and proliferative status of GBMs were studied. Intracranial xenografts of glioblastomas were grown in the right frontal regions of female, nude mice (n = 3-5 per experiment). Tumor growth was followed through BLI. Neurosphere cultures (U87-MG, GBM2 and GBM39) were inhibited by AshwaMAX at IC50 of 1.4, 0.19 and 0.22 µM equivalent respectively and by Withaferin A with IC50 of 0.31, 0.28 and 0.25 µM respectively. Oral gavage, every other day, of AshwaMAX (40 mg/kg per day) significantly reduced bioluminescence signal (n = 3 mice, p < 0.02, four parameter non-linear regression analysis) in preclinical models. After 30 days of treatment, bioluminescent signal increased suggesting onset of resistance. BLI signal for control, vehicle-treated mice increased and then plateaued. Bioluminescent imaging revealed diffuse growth of GBM2 xenografts. With AshwaMAX, GBM neurospheres collapsed at nanomolar concentrations. Oral treatment studies on murine models confirmed that AshwaMAX is effective against orthotopic GBM. AshwaMAX is thus a promising candidate for future clinical translation in patients with GBM.
- Published
- 2016
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35. PET imaging of tumor glycolysis downstream of hexokinase through noninvasive measurement of pyruvate kinase M2.
- Author
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Witney TH, James ML, Shen B, Chang E, Pohling C, Arksey N, Hoehne A, Shuhendler A, Park JH, Bodapati D, Weber J, Gowrishankar G, Rao J, Chin FT, and Gambhir SS
- Subjects
- Brain Neoplasms diagnostic imaging, Brain Neoplasms enzymology, Carbon Radioisotopes, Glycolysis, Humans, Hexokinase metabolism, Positron-Emission Tomography, Pyruvate Kinase metabolism
- Abstract
Cancer cells reprogram their metabolism to meet increased biosynthetic demands, commensurate with elevated rates of replication. Pyruvate kinase M2 (PKM2) catalyzes the final and rate-limiting step in tumor glycolysis, controlling the balance between energy production and the synthesis of metabolic precursors. We report here the synthesis and evaluation of a positron emission tomography (PET) radiotracer, [(11)C]DASA-23, that provides a direct noninvasive measure of PKM2 expression in preclinical models of glioblastoma multiforme (GBM). In vivo, orthotopic U87 and GBM39 patient-derived tumors were clearly delineated from the surrounding normal brain tissue by PET imaging, corresponding to exclusive tumor-associated PKM2 expression. In addition, systemic treatment of mice with the PKM2 activator TEPP-46 resulted in complete abrogation of the PET signal in intracranial GBM39 tumors. Together, these data provide the basis for the clinical evaluation of imaging agents that target this important gatekeeper of tumor glycolysis., (Copyright © 2015, American Association for the Advancement of Science.)
- Published
- 2015
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36. A Systematic Comparison of 18F-C-SNAT to Established Radiotracer Imaging Agents for the Detection of Tumor Response to Treatment.
- Author
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Witney TH, Hoehne A, Reeves RE, Ilovich O, Namavari M, Shen B, Chin FT, Rao J, and Gambhir SS
- Subjects
- Animals, Antineoplastic Agents administration & dosage, Antineoplastic Agents pharmacology, Biological Availability, Cell Death, Cell Line, Tumor, Disease Models, Animal, Female, Humans, Mice, Neoplasms therapy, Treatment Outcome, Tumor Burden drug effects, Benzothiazoles metabolism, Neoplasms diagnosis, Oligopeptides metabolism, Positron-Emission Tomography methods, Radiopharmaceuticals metabolism, Tomography, X-Ray Computed
- Abstract
Purpose: An early readout of tumor response to therapy through measurement of drug or radiation-induced cell death may provide important prognostic indications and improved patient management. It has been shown that the uptake of (18)F-C-SNAT can be used to detect early response to therapy in tumors by positron emission tomography (PET) via a mechanism of caspase-3-triggered nanoaggregation., Experimental Design: Here, we compared the preclinical utility of (18)F-C-SNAT for the detection of drug-induced cell death to clinically evaluated radiotracers, (18)F-FDG, (99m)Tc-Annexin V, and (18)F-ML-10 in tumor cells in culture, and in tumor-bearing mice in vivo., Results: In drug-treated lymphoma cells, (18)F-FDG, (99m)Tc-Annexin V, and (18)F-C-SNAT cell-associated radioactivity correlated well to levels of cell death (R(2) > 0.8; P < 0.001), with no correlation measured for (18)F-ML-10 (R(2) = 0.05; P > 0.05). A similar pattern of response was observed in two human NSCLC cell lines following carboplatin treatment. EL-4 tumor uptake of (99m)Tc-Annexin V and (18)F-C-SNAT were increased 1.4- and 2.1-fold, respectively, in drug-treated versus naïve control animals (P < 0.05), although (99m)Tc-Annexin V binding did not correlate to ex vivo TUNEL staining of tissue sections. A differential response was not observed with either (18)F-FDG or (18)F-ML-10., Conclusions: We have demonstrated here that (18)F-C-SNAT can sensitively detect drug-induced cell death in murine lymphoma and human NSCLC. Despite favorable image contrast obtained with (18)F-C-SNAT, the development of next-generation derivatives, using the same novel and promising uptake mechanism, but displaying improved biodistribution profiles, are warranted for maximum clinical utility., (©2015 American Association for Cancer Research.)
- Published
- 2015
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37. Preclinical evaluation of 3-18F-fluoro-2,2-dimethylpropionic acid as an imaging agent for tumor detection.
- Author
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Witney TH, Pisaneschi F, Alam IS, Trousil S, Kaliszczak M, Twyman F, Brickute D, Nguyen QD, Schug Z, Gottlieb E, and Aboagye EO
- Subjects
- Animals, Cell Line, Tumor, Humans, Mice, Positron-Emission Tomography, Fluorine Radioisotopes, Neoplasms, Experimental diagnostic imaging, Pentanoic Acids, Radiopharmaceuticals
- Abstract
Unlabelled: Deregulated cellular metabolism is a hallmark of many cancers. In addition to increased glycolytic flux, exploited for cancer imaging with (18)F-FDG, tumor cells display aberrant lipid metabolism. Pivalic acid is a short-chain, branched carboxylic acid used to increase oral bioavailability of prodrugs. After prodrug hydrolysis, pivalic acid undergoes intracellular metabolism via the fatty acid oxidation pathway. We have designed a new probe, 3-(18)F-fluoro-2,2-dimethylpropionic acid, also called (18)F-fluoro-pivalic acid ((18)F-FPIA), for the imaging of aberrant lipid metabolism and cancer detection., Methods: Cell intrinsic uptake of (18)F-FPIA was measured in murine EMT6 breast adenocarcinoma cells. In vivo dynamic imaging, time course biodistribution, and radiotracer stability testing were performed. (18)F-FPIA tumor retention was further compared in vivo to (18)F-FDG uptake in several xenograft models and inflammatory tissue., Results: (18)F-FPIA rapidly accumulated in EMT6 breast cancer cells, with retention of intracellular radioactivity predicted to occur via a putative (18)F-FPIA carnitine-ester. The radiotracer was metabolically stable to degradation in mice. In vivo imaging of implanted EMT6 murine and BT474 human breast adenocarcinoma cells by (18)F-FPIA PET showed rapid and extensive tumor localization, reaching 9.1% ± 0.5% and 7.6% ± 1.2% injected dose/g, respectively, at 60 min after injection. Substantial uptake in the cortex of the kidney was seen, with clearance primarily via urinary excretion. Regarding diagnostic utility, uptake of (18)F-FPIA was comparable to that of (18)F-FDG in EMT6 tumors but superior in the DU145 human prostate cancer model (54% higher uptake; P = 0.002). Furthermore, compared with (18)F-FDG, (18)F-FPIA had lower normal-brain uptake resulting in a superior tumor-to-brain ratio (2.5 vs. 1.3 in subcutaneously implanted U87 human glioma tumors; P = 0.001), predicting higher contrast for brain cancer imaging. Both radiotracers showed increased localization in inflammatory tissue., Conclusion: (18)F-FPIA shows promise as an imaging agent for cancer detection and warrants further investigation., (© 2014 by the Society of Nuclear Medicine and Molecular Imaging, Inc.)
- Published
- 2014
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38. Radiolabeled RGD tracer kinetics annotates differential αvβ 3 integrin expression linked to cell intrinsic and vessel expression.
- Author
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Alam IS, Witney TH, Tomasi G, Carroll L, Twyman FJ, Nguyen QD, and Aboagye EO
- Subjects
- Animals, Blotting, Western, Cell Line, Tumor, Female, Flow Cytometry, Humans, Immunohistochemistry, Kinetics, Mice, Inbred BALB C, Mice, Nude, Neoplasms blood supply, Neoplasms diagnostic imaging, Neoplasms pathology, Platelet Endothelial Cell Adhesion Molecule-1 metabolism, Positron-Emission Tomography, Radioactive Tracers, Blood Vessels metabolism, Blood Vessels pathology, Integrin alphaVbeta3 metabolism, Organometallic Compounds, Peptides, Cyclic
- Abstract
Purpose: The purpose of this paper is to study the association between RGD binding kinetics and αvβ3 integrin receptor density in the complex tumor milieu., Procedures: We assessed αvβ3 in vitro and by (68)Ga-DOTA-[c(RGDfK)]2 positron emission tomography (PET) in tumors with varying αvβ3., Results: Intrinsic αvβ3 expression decreased in the order of M21 >>> MDA-MB-231 > M21L in cells. Tumor volume of distribution by PET, V T, was significantly higher in M21 compared to isogenic M21L tumors (0.40 ± 0.01 versus 0.25 ± 0.02; p < 0.01) despite similar microvessel density (MVD) likely due to higher αvβ3. V T for MDA-MB-231 (0.40 ± 0.04) was comparable to M21 despite lower αvβ3 but in keeping with the higher MVD, suggesting superior tracer distribution., Conclusions: This study demonstrates that radioligand binding kinetics of PET data can be used to discriminate tumors with different αvβ3 integrin expression-a key component of the angiogenesis phenotype-in vivo.
- Published
- 2014
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39. Preclinical assessment of carboplatin treatment efficacy in lung cancer by 18F-ICMT-11-positron emission tomography.
- Author
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Witney TH, Fortt R, and Aboagye EO
- Subjects
- Animals, Antineoplastic Agents administration & dosage, Apoptosis, Carboplatin administration & dosage, Caspase 3 metabolism, Caspase 7 metabolism, Cell Death drug effects, Cell Line, Tumor, Disease Models, Animal, Drug Evaluation, Preclinical, Enzyme Activation, Female, Humans, Lung Neoplasms drug therapy, Xenograft Model Antitumor Assays, Antineoplastic Agents pharmacology, Azides metabolism, Carboplatin pharmacology, Indoles metabolism, Lung Neoplasms diagnosis, Positron-Emission Tomography
- Abstract
Tumour response to therapy is assessed primarily in the clinic by monitoring reductions in tumour size. However, this approach lacks sensitivity since in many cases several weeks may elapse before there is evidence of tumour shrinkage. There is therefore a need to develop non-invasive imaging techniques for monitoring tumour treatment response in the clinic. Here, we assessed the pre-clinical utility of (18)F-ICMT-11 positron emission tomography--a method for detecting caspase 3/7 activation--in non-small cell lung cancer (NSCLC). (18)F-ICMT-11 uptake was compared to molecular biochemical measures of cell death in PC9 and A549 NSCLC cells following treatment with carboplatin in vitro and in vivo. Carboplatin-induced apoptosis in the ERCC1 low/mutant EGFR PC9 cells was characterised by time and dose-related increased caspase-3/7 activation, poly-ADP-ribose polymerase cleavage and Annexin V staining. 18F-ICMT-11 uptake was consequently increased up to 14-fold at 200 µM carboplatin compared to vehicle treated cells (P<0.01). In contrast, necrosis was the predominant death mechanism in ERCC1 high/wt EGFR A549 cells and no change in (18)F-ICMT-11 uptake was detected. In vivo, histological analysis of PC9 tumour xenografts indicated high pre-therapy necrosis. A 4.6-fold increase in cleaved caspase-3/7 was measured in non-necrotic regions of PC9 tumours at 48 h post carboplatin therapy. Average PET-derived tumour (18)F-ICMT-11 uptake was insensitive to changes in apoptosis in the presence of substantial pre-existing necrosis. PET-based voxel intensity sorting however, identified intra-tumoural regions of high (18)F-ICMT-11 uptake, enabling accurate assessment of apoptosis and therefore therapy response. In A549 tumours that lacked high pre-therapy necrosis, carboplatin induced growth inhibition that was only minimally associated with apoptosis and thus not detectable by (18)F-ICMT-11 PET.
- Published
- 2014
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40. A novel radiotracer to image glycogen metabolism in tumors by positron emission tomography.
- Author
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Witney TH, Carroll L, Alam IS, Chandrashekran A, Nguyen QD, Sala R, Harris R, DeBerardinis RJ, Agarwal R, and Aboagye EO
- Subjects
- Cell Cycle Checkpoints physiology, Cell Line, Tumor, Fluorodeoxyglucose F18 metabolism, G1 Phase physiology, Humans, Positron-Emission Tomography methods, Radiopharmaceuticals metabolism, Resting Phase, Cell Cycle physiology, rab GTP-Binding Proteins genetics, Glycogen metabolism, Neoplasms diagnosis, Neoplasms metabolism
- Abstract
The high rate of glucose uptake to fuel the bioenergetic and anabolic demands of proliferating cancer cells is well recognized and is exploited with (18)F-2-fluoro-2-deoxy-d-glucose positron emission tomography ((18)F-FDG-PET) to image tumors clinically. In contrast, enhanced glucose storage as glycogen (glycogenesis) in cancer is less well understood and the availability of a noninvasive method to image glycogen in vivo could provide important biologic insights. Here, we demonstrate that (18)F-N-(methyl-(2-fluoroethyl)-1H-[1,2,3]triazole-4-yl)glucosamine ((18)F-NFTG) annotates glycogenesis in cancer cells and tumors in vivo, measured by PET. Specificity of glycogen labeling was demonstrated by isolating (18)F-NFTG-associated glycogen and with stable knockdown of glycogen synthase 1, which inhibited (18)F-NFTG uptake, whereas oncogene (Rab25) activation-associated glycogen synthesis led to increased uptake. We further show that the rate of glycogenesis is cell-cycle regulated, enhanced during the nonproliferative state of cancer cells. We demonstrate that glycogen levels, (18)F-NFTG, but not (18)F-FDG uptake, increase proportionally with cell density and G1-G0 arrest, with potential application in the assessment of activation of oncogenic pathways related to glycogenesis and the detection of posttreatment tumor quiescence., (©2014 AACR)
- Published
- 2014
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41. Magnetic resonance imaging with hyperpolarized [1,4-(13)C2]fumarate allows detection of early renal acute tubular necrosis.
- Author
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Clatworthy MR, Kettunen MI, Hu DE, Mathews RJ, Witney TH, Kennedy BW, Bohndiek SE, Gallagher FA, Jarvis LB, Smith KG, and Brindle KM
- Subjects
- Animals, Carbon Isotopes, Early Diagnosis, Folic Acid, Humans, Kidney abnormalities, Kidney pathology, Kidney physiopathology, Kidney Tubular Necrosis, Acute chemically induced, Kidney Tubular Necrosis, Acute physiopathology, Kinetics, Lupus Nephritis diagnosis, Lupus Nephritis pathology, Malates, Mice, Mice, Inbred C57BL, Pyruvic Acid, Fumarates, Kidney Tubular Necrosis, Acute diagnosis, Magnetic Resonance Imaging methods
- Abstract
Acute kidney injury (AKI) is a common and important medical problem, affecting 10% of hospitalized patients, and it is associated with significant morbidity and mortality. The most frequent cause of AKI is acute tubular necrosis (ATN). Current imaging techniques and biomarkers do not allow ATN to be reliably differentiated from important differential diagnoses, such as acute glomerulonephritis (GN). We investigated whether (13)C magnetic resonance spectroscopic imaging (MRSI) might allow the noninvasive diagnosis of ATN. (13)C MRSI of hyperpolarized [1,4-(13)C(2)]fumarate and pyruvate was used in murine models of ATN and acute GN (NZM2410 mice with lupus nephritis). A significant increase in [1,4-(13)C(2)]malate signal was identified in the kidneys of mice with ATN early in the disease course before the onset of severe histological changes. No such increase in renal [1,4-(13)C(2)]malate was observed in mice with acute GN. The kidney [1-(13)C]pyruvate/[1-(13)C]lactate ratio showed substantial variability and was not significantly decreased in animals with ATN or increased in animals with GN. In conclusion, MRSI of hyperpolarized [1,4-(13)C(2)]fumarate allows the detection of early tubular necrosis and its distinction from glomerular inflammation in murine models. This technique may have the potential to identify a window of therapeutic opportunity in which emerging therapies might be applied to patients with ATN, reducing the need for acute dialysis with its attendant morbidity and cost.
- Published
- 2012
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42. Evaluation of deuterated 18F- and 11C-labeled choline analogs for cancer detection by positron emission tomography.
- Author
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Witney TH, Alam IS, Turton DR, Smith G, Carroll L, Brickute D, Twyman FJ, Nguyen QD, Tomasi G, Awais RO, and Aboagye EO
- Subjects
- Animals, Cell Line, Tumor, Disease Models, Animal, Humans, Kidney metabolism, Kinetics, Male, Melanoma diagnostic imaging, Melanoma metabolism, Mice, Mice, Inbred BALB C, Mice, Nude, Neoplasms metabolism, Oxidation-Reduction, Prostatic Neoplasms diagnostic imaging, Prostatic Neoplasms metabolism, Radioactive Tracers, Carbon Radioisotopes, Choline analogs & derivatives, Choline metabolism, Deuterium, Fluorodeoxyglucose F18, Neoplasms diagnostic imaging, Positron-Emission Tomography
- Abstract
Purpose: (11)C-Choline-positron emission tomography (PET) has been exploited to detect the aberrant choline metabolism in tumors. Radiolabeled choline uptake within the imaging time is primarily a function of transport, phosphorylation, and oxidation. Rapid choline oxidation, however, complicates interpretation of PET data. In this study, we investigated the biologic basis of the oxidation of deuterated choline analogs and assessed their specificity in human tumor xenografts., Experimental Design: (11)C-Choline, (11)C-methyl-[1,2-(2)H(4)]-choline ((11)C-D4-choline), and (18)F-D4-choline were synthesized to permit comparison. Biodistribution, metabolism, small-animal PET studies, and kinetic analysis of tracer uptake were carried out in human colon HCT116 xenograft-bearing mice., Results: Oxidation of choline analogs to betaine was highest with (11)C-choline, with reduced oxidation observed with (11)C-D4-choline and substantially reduced with (18)F-D4-choline, suggesting that both fluorination and deuteration were important for tracer metabolism. Although all tracers were converted intracellularly to labeled phosphocholine (specific signal), the higher rate constants for intracellular retention (K(i) and k(3)) of (11)C-choline and (11)C-D4-choline, compared with (18)F-D4-choline, were explained by the rapid conversion of the nonfluorinated tracers to betaine within HCT116 tumors. Imaging studies showed that the uptake of (18)F-D4-choline in three tumors with similar radiotracer delivery (K(1)) and choline kinase α expression-HCT116, A375, and PC3-M-were the same, suggesting that (18)F-D4-choline has utility for cancer detection irrespective of histologic type., Conclusion: We have shown here that both deuteration and fluorination combine to provide protection against choline oxidation in vivo. (18)F-D4-choline showed the highest selectivity for phosphorylation and warrants clinical evaluation., (©2012 AACR.)
- Published
- 2012
- Full Text
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43. Kinetic modeling of hyperpolarized 13C label exchange between pyruvate and lactate in tumor cells.
- Author
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Witney TH, Kettunen MI, and Brindle KM
- Subjects
- Carbon Isotopes chemistry, Cell Line, Tumor, Humans, Kinetics, Lactic Acid metabolism, Models, Biological, Neoplasms metabolism, Pyruvic Acid metabolism
- Abstract
Measurements of the kinetics of hyperpolarized (13)C label exchange between [1-(13)C]pyruvate and lactate in suspensions of intact and lysed murine lymphoma cells, and in cells in which lactate dehydrogenase expression had been modulated by inhibition of the PI3K pathway, were used to determine quantitatively the role of enzyme activity and membrane transport in controlling isotope flux. Both steps were shown to share in the control of isotope flux in these cells. The kinetics of label exchange were well described by a kinetic model that employed rate constants for the lactate dehydrogenase reaction that had been determined previously from steady state kinetic studies. The enzyme showed pyruvate inhibition in steady state kinetic measurements, which the kinetic model predicted should also be observed in the isotope exchange measurements. However, no such pyruvate inhibition was observed in either intact cells or cell lysates and this could be explained by the much higher enzyme concentrations present in the isotope exchange experiments. The kinetic analysis presented here shows how lactate dehydrogenase activity can be determined from the isotope exchange measurements. The kinetic model should be useful for modeling the exchange reaction in vivo, particularly as this technique progresses to the clinic.
- Published
- 2011
- Full Text
- View/download PDF
44. Detection of tumor response to a vascular disrupting agent by hyperpolarized 13C magnetic resonance spectroscopy.
- Author
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Bohndiek SE, Kettunen MI, Hu DE, Witney TH, Kennedy BW, Gallagher FA, and Brindle KM
- Subjects
- Angiogenesis Inhibitors chemistry, Animals, Carbon Isotopes, Contrast Media, Diffusion Magnetic Resonance Imaging, Fumarates administration & dosage, Fumarates pharmacology, Injections, Intravenous, Mice, Neoplasms enzymology, Neoplasms pathology, Pyruvic Acid administration & dosage, Pyruvic Acid pharmacology, Stilbenes chemistry, Time Factors, Angiogenesis Inhibitors therapeutic use, Magnetic Resonance Spectroscopy methods, Neoplasms blood supply, Neoplasms drug therapy, Neovascularization, Pathologic drug therapy, Stilbenes therapeutic use
- Abstract
Nuclear spin hyperpolarization can dramatically increase the sensitivity of the (13)C magnetic resonance experiment, allowing dynamic measurements of the metabolism of hyperpolarized (13)C-labeled substrates in vivo. Here, we report a preclinical study of the response of lymphoma tumors to the vascular disrupting agent (VDA), combretastatin-A4-phosphate (CA4P), as detected by measuring changes in tumor metabolism of hyperpolarized [1-(13)C]pyruvate and [1,4-(13)C(2)]fumarate. These measurements were compared with dynamic contrast agent-enhanced magnetic resonance imaging (DCE-MRI) measurements of tumor vascular function and diffusion-weighted MRI (DW-MRI) measurements of the tumor cell necrosis that resulted from subsequent loss of tumor perfusion. The rate constant describing flux of hyperpolarized (13)C label between [1-(13)C]pyruvate and lactate was decreased by 34% within 6 hours of CA4P treatment and remained at this lower level at 24 hours. The rate constant describing production of labeled malate from hyperpolarized [1,4-(13)C(2)]fumarate increased 1.6-fold and 2.5-fold at 6 and 24 hours after treatment, respectively, and correlated with the degree of necrosis detected in histologic sections. Although DCE-MRI measurements showed a substantial reduction in perfusion at 6 hours after treatment, which had recovered by 24 hours, DW-MRI showed no change in the apparent diffusion coefficient of tumor water at 6 hours after treatment, although there was a 32% increase at 24 hours (P < 0.02) when regions of extensive necrosis were observed by histology. Measurements of hyperpolarized [1-(13)C]pyruvate and [1,4-(13)C(2)]fumarate metabolism may provide, therefore, a more sustained and sensitive indicator of response to a VDA than DCE-MRI or DW-MRI, respectively., (©2010 AACR.)
- Published
- 2010
- Full Text
- View/download PDF
45. Detecting treatment response in a model of human breast adenocarcinoma using hyperpolarised [1-13C]pyruvate and [1,4-13C2]fumarate.
- Author
-
Witney TH, Kettunen MI, Hu DE, Gallagher FA, Bohndiek SE, Napolitano R, and Brindle KM
- Subjects
- Animals, Calcium Dobesilate therapeutic use, Cell Death drug effects, Cell Line, Tumor, Female, Humans, Mice, Mice, SCID, Adenocarcinoma drug therapy, Breast Neoplasms drug therapy, Carbon Isotopes, Fumarates metabolism, Pyruvic Acid metabolism
- Abstract
Background: The recent introduction of a dynamic nuclear polarisation technique has permitted noninvasive imaging of tumour cell metabolism in vivo following intravenous administration of (13)C-labelled cell substrates., Methods: Changes in hyperpolarised [1-(13)C]pyruvate and [1,4-(13)C(2)]fumarate metabolism were evaluated in both MDA-MB-231 cells and in implanted MDA-MB-231 tumours following doxorubicin treatment., Results: Treatment of MDA-MB-231 cells resulted in the induction of apoptosis, which was accompanied by a decrease in hyperpolarised (13)C label flux between [1-(13)C]pyruvate and lactate, which was correlated with a decrease in the cellular NAD(H) coenzyme pool. There was also an increase in the rate of fumarate conversion to malate, which accompanied the onset of cellular necrosis. In vivo, the decrease in (13)C label exchange between pyruvate and lactate and the increased flux between fumarate and malate, following drug treatment, were shown to occur in the absence of any detectable change in tumour size., Conclusion: We show here that the early responses of a human breast adenocarcinoma tumour model to drug treatment can be followed by administration of both hyperpolarised [1-(13)C]pyruvate and [1,4-(13)C(2)]fumarate. These techniques could be used, therefore, in the clinic to detect the early responses of breast tumours to treatment.
- Published
- 2010
- Full Text
- View/download PDF
46. Imaging tumour cell metabolism using hyperpolarized 13C magnetic resonance spectroscopy.
- Author
-
Witney TH and Brindle KM
- Subjects
- Animals, Humans, L-Lactate Dehydrogenase metabolism, Lactic Acid metabolism, Pyruvic Acid metabolism, Carbon Isotopes analysis, Magnetic Resonance Spectroscopy, Neoplasms metabolism
- Abstract
Patients with similar tumour types frequently show different responses to the same therapy. The development of new treatments would benefit, therefore, from imaging methods that allow an early assessment of treatment response in individual patients, allowing rapid selection of the most effective treatment. We have been using (13)C MRSI (magnetic resonance spectroscopic imaging) of tumour cell metabolism, using hyperpolarized (13)C-labelled cellular metabolites, to detect treatment response. Nuclear spin hyperpolarization can increase sensitivity in the magnetic resonance experiment >10,000 times, allowing us to image labelled cell substrates in vivo and their subsequent metabolism. We showed that exchange of hyperpolarized (13)C label between lactate and pyruvate, catalysed by lactate dehydrogenase, was decreased in treated tumours undergoing drug-induced cell death, and that tissue pH could be imaged from the ratio of the signal intensities of hyperpolarized H(13)CO(3)(-) and (13)CO(2) following intravenous injection of hyperpolarized H(13)CO(3). Tumour cell glutaminase activity, a potential measure of cell proliferation, can be determined using hyperpolarized [5-(13)C]glutamine, and treatment-induced tumour cell necrosis can be imaged in vivo from measurements of the conversion of hyperpolarized [1,4-(13)C(2)]fumarate into malate. Since these substrates are endogenous and, in some cases, have already been safely infused into patients, these techniques have the potential to translate to the clinic.
- Published
- 2010
- Full Text
- View/download PDF
47. Comparison of the C2A domain of synaptotagmin-I and annexin-V as probes for detecting cell death.
- Author
-
Alam IS, Neves AA, Witney TH, Boren J, and Brindle KM
- Subjects
- Animals, Annexin A5 metabolism, Breast Neoplasms diagnosis, Cell Line, Tumor, Female, Fluorescent Dyes chemistry, Humans, Lymphoma diagnosis, Protein Binding, Protein Structure, Tertiary, Rats, Synaptotagmin I metabolism, Annexin A5 chemistry, Cell Death, Synaptotagmin I chemistry
- Abstract
The induction of apoptosis is frequently accompanied by the exposure of phosphatidylserine (PS) on the cell surface, which has been detected using radionuclide and fluorescently labeled derivatives of the PS-binding protein, Annexin V. The fluorescently labeled protein has been used extensively in vitro as a diagnostic reagent for detecting cell death, and radionuclide-labeled derivatives have undergone clinical trials for detecting tumor cell death in vivo following treatment. We show here that the C2A domain of Synaptotagmin-I, which had been fluorescently labeled at a single cysteine residue introduced by site-directed mutagenesis, detected the same levels of cell death as a similarly labeled Annexin-V derivative, in drug-treated murine lymphoma and human breast cancer cell lines in vitro. However, the C2A derivative showed significantly less binding to viable cells and, as a consequence, up to 4-fold more specific binding to apoptotic and necrotic cells when compared with Annexin-V. C2A offers a potential route for the development of a new generation of more specific imaging probes for the detection of tumor cell death in the clinic.
- Published
- 2010
- Full Text
- View/download PDF
48. Magnetization transfer measurements of exchange between hyperpolarized [1-13C]pyruvate and [1-13C]lactate in a murine lymphoma.
- Author
-
Kettunen MI, Hu DE, Witney TH, McLaughlin R, Gallagher FA, Bohndiek SE, Day SE, and Brindle KM
- Subjects
- Animals, Carbon Isotopes metabolism, Female, Mice, Mice, Inbred C57BL, Lactates metabolism, Lymphoma metabolism, Magnetic Resonance Spectroscopy methods, Pyruvates metabolism
- Abstract
Measurements of the conversion of hyperpolarized [1-(13)C]pyruvate into lactate, in the reaction catalyzed by lactate dehydrogenase, have shown promise as a metabolic marker for the presence of disease and response to treatment. However, it is unclear whether this represents net flux of label from pyruvate to lactate or exchange of isotope between metabolites that are close to chemical equilibrium. Using saturation and inversion transfer experiments, we show that there is significant exchange of label between lactate and pyruvate in a murine lymphoma in vivo. The rate constants estimated from the magnetization transfer experiments, at specific points during the time course of label exchange, were similar to those obtained by fitting the changes in peak intensities during the entire exchange time course to a kinetic model for two-site exchange. These magnetization transfer experiments may therefore provide an alternative and more rapid way of estimating flux between pyruvate and lactate to serial measurements of pyruvate and lactate (13)C peak intensities following injection of hyperpolarized [1-(13)C]pyruvate.
- Published
- 2010
- Full Text
- View/download PDF
49. Production of hyperpolarized [1,4-13C2]malate from [1,4-13C2]fumarate is a marker of cell necrosis and treatment response in tumors.
- Author
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Gallagher FA, Kettunen MI, Hu DE, Jensen PR, Zandt RI, Karlsson M, Gisselsson A, Nelson SK, Witney TH, Bohndiek SE, Hansson G, Peitersen T, Lerche MH, and Brindle KM
- Subjects
- Animals, Biomarkers, Tumor chemistry, Biomarkers, Tumor metabolism, Carbon Isotopes chemistry, Carbon Isotopes metabolism, Etoposide therapeutic use, Female, Fumarate Hydratase metabolism, Lymphoma metabolism, Lymphoma pathology, Mice, Mice, Inbred C57BL, Necrosis pathology, Neoplasm Transplantation, Nuclear Magnetic Resonance, Biomolecular, Tissue Extracts metabolism, Treatment Outcome, Antineoplastic Agents, Phytogenic therapeutic use, Fumarates chemistry, Fumarates metabolism, Malates chemistry, Malates metabolism, Necrosis metabolism, Neoplasms drug therapy, Neoplasms metabolism, Neoplasms pathology
- Abstract
Dynamic nuclear polarization of (13)C-labeled cell substrates has been shown to massively increase their sensitivity to detection in NMR experiments. The sensitivity gain is sufficiently large that if these polarized molecules are injected intravenously, their spatial distribution and subsequent conversion into other cell metabolites can be imaged. We have used this method to image the conversion of fumarate to malate in a murine lymphoma tumor in vivo after i.v. injection of hyperpolarized [1,4-(13)C(2)]fumarate. In isolated lymphoma cells, the rate of labeled malate production was unaffected by coadministration of succinate, which competes with fumarate for transport into the cell. There was, however, a correlation with the percentage of cells that had lost plasma membrane integrity, suggesting that the production of labeled malate from fumarate is a sensitive marker of cellular necrosis. Twenty-four hours after treating implanted lymphoma tumors with etoposide, at which point there were significant levels of tumor cell necrosis, there was a 2.4-fold increase in hyperpolarized [1,4-(13)C(2)]malate production compared with the untreated tumors. Therefore, the formation of hyperpolarized (13)C-labeled malate from [1,4-(13)C(2)]fumarate appears to be a sensitive marker of tumor cell death in vivo and could be used to detect the early response of tumors to treatment. Given that fumarate is an endogenous molecule, this technique has the potential to be used clinically.
- Published
- 2009
- Full Text
- View/download PDF
50. A comparison between radiolabeled fluorodeoxyglucose uptake and hyperpolarized (13)C-labeled pyruvate utilization as methods for detecting tumor response to treatment.
- Author
-
Witney TH, Kettunen MI, Day SE, Hu DE, Neves AA, Gallagher FA, Fulton SM, and Brindle KM
- Subjects
- Animals, Antineoplastic Agents, Phytogenic pharmacology, Apoptosis drug effects, Blotting, Western, Carbon Isotopes, Carbon Radioisotopes, Cell Line, Tumor, Female, Flow Cytometry, Glucose Transporter Type 1 metabolism, Glucose Transporter Type 3 metabolism, Lactates metabolism, Lymphoma metabolism, Lymphoma pathology, Magnetic Resonance Spectroscopy, Mice, Mice, Inbred C57BL, Poly(ADP-ribose) Polymerases metabolism, Positron-Emission Tomography methods, Treatment Outcome, Etoposide pharmacology, Fluorodeoxyglucose F18 pharmacokinetics, Lymphoma drug therapy, Pyruvates metabolism
- Abstract
Detection of early tumor responses to treatment can give an indication of clinical outcome. Positron emission tomography measurements of the uptake of the glucose analog, [(18)F] 2-fluoro-2-deoxy-D-glucose (FDG), have demonstrated their potential for detecting early treatment response in the clinic. We have shown recently that (13)C magnetic resonance spectroscopy and spectroscopic imaging measurements of the uptake and conversion of hyperpolarized [1-(13)C]pyruvate into [1-(13)C]lactate can be used to detect treatment response in a murine lymphoma model. The present study compares these magnetic resonance measurements with changes in FDG uptake after chemotherapy. A decrease in FDG uptake was found to precede the decrease in flux of hyperpolarized (13)C label between pyruvate and lactate, both in tumor cells in vitro and in tumors in vivo. However, the magnitude of the decrease in FDG uptake and the decrease in pyruvate to lactate flux was comparable at 24 hours after drug treatment. In cells, the decrease in FDG uptake was shown to correlate with changes in plasma membrane expression of the facilitative glucose transporters, whereas the decrease in pyruvate to lactate flux could be explained by an increase in poly(ADP-ribose) polymerase activity and subsequent depletion of the NAD(H) pool. These results show that measurement of flux between pyruvate and lactate may be an alternative to FDG-positron emission tomography for imaging tumor treatment response in the clinic.
- Published
- 2009
- Full Text
- View/download PDF
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