Dynamic Nuclear Polarization Technique Research Articles

Glycyrrhetinic acid interaction with solvated and free electrons studied by the CIDNP and dissociative electron attachment techniques.

Electron transfer plays a crucial role in living systems, including the generation of reactive oxygen species (ROS). Oxygen acts as the terminal electron acceptor in the respiratory chains of aerobic organisms as well as in some photoinduced processes followed by the formation of ROS. This is why the participation of exogenous antioxidants in electron transfer processes in living systems is of particular interest. In the present study, using chemically induced dynamic nuclear polarization (CIDNP) and dissociative electron attachment (DEA) techniques, we have elucidated the affinity of solvated and free electrons to glycyrrhetinic acid (GA)-the aglicon of glycyrrhizin (the main active component of Licorice root). CIDNP is a powerful instrument to study the mechanisms of electron transfer reactions in solution, but the DEA technique shows its effectiveness in gas phase processes. For CIDNP experiments, the photoionization of the dianion of 5-sulfosalicylic acid (HSSA2-) was used as a model reaction of solvated electron generation. DEA experiments testify that GA molecules are even better electron acceptors than molecular oxygen, at least under gas-phase conditions. In addition, the effect of the solvent on the energetics of the reactants is discussed.

Substrate interaction of graphene quantum dots in solution studied by hyperpolarized NMR

The reversible adsorption of CO2-derived species to graphene quantum dots in CO2 electroreduction reaction was characterized by nuclear spin hyperpolarized NMR. Hyperpolarization of 13C-labeled sodium bicarbonate by the dissolution dynamic nuclear polarization (DNP) technique provided a 5000-fold signal enhancement and enabled real-time and in-situ 13C NMR experiments. The R1 and R2 relaxation rates were measured using time-resolved DNP-NMR experiments, and the bound species were confirmed to be carbonic acid and its conjugates. A significant difference in the relaxation rates of bound and free species was found, indicating a reversible binding. In addition to relaxation rates, kinetic rate constants were fitted from the small-flip angle experiments and compared to theoretical calculations. The pH-dependent adsorption and desorption rate constants were found in the range of 3.5–9.5 and 0.7–2.0 s−1, with the average binding distance at 279 - 291 pm. The DNP-NMR technique was demonstrated as an effective way to study surface adsorption, with potential in electrochemistry and kinetics applications.

Hyperpolarized 13C NMR for longitudinal in-cell metabolism using a mobile 3D cell culture system

Hyperpolarization with the dissolution dynamic nuclear polarization (dDNP) technique yields > 10,000-fold signal increases for NMR-active nuclei (e.g. 13C). Hyperpolarized 13C-labeled metabolic tracer molecules thus allow real-time observations of biochemical pathways in living cellular systems without interfering background. This methodology lends itself to the direct observation of altered intracellular reaction chemistry imparted for instance by drug treatment, infections, or other diseases. A reoccurring challenge for longitudinal cell studies of mammalian cells with NMR and dDNP-NMR is maintaining cell viability in the NMR spectrometer. 3D cell culture methods are increasing in popularity because they provide a physiologically more relevant environment compared to 2D cell cultures. Based on such strategies a mobile 3D culture system was devised. The clinical drug etoposide was used to treat cancer cells (HeLa) and the resulting altered metabolism was measured using hyperpolarized [1–13C]pyruvate. We show that sustaining the cell cultivation in cell incubators and only transferring the cells to the NMR spectrometer for the few minutes required for the dDNP-NMR measurements is an attractive alternative to cell maintenance in the NMR tube. High cell viability is sustained, and experimental throughput is many doubled.

NMR Magnetometer Based on Dynamic Nuclear-Polarization for Low-Strength Magnetic Field Measurement

Nuclear magnetic resonance (NMR) magnetometers are considered due to their ability to map magnetic fields with high precision and calibrate other magnetic field measurement devices. However, the low signal-to-noise ratio of low-strength magnetic fields limits the precision when measuring magnetic fields below 40 mT. Therefore, we developed a new NMR magnetometer that combines the dynamic nuclear polarization (DNP) technique with pulsed NMR. The dynamic pre-polarization technique enhances the SNR under a low magnetic field. Pulsed NMR was used in conjunction with DNP to improve measurement accuracy and speed. The efficacy of this approach was validated through simulation and analysis of the measurement process. Next, a complete set of equipment was constructed, and we successfully measured magnetic fields of 30 mT and 8 mT with an accuracy of only 0.5 Hz (11 nT) at 30 mT (0.4 ppm) and 1 Hz (22 nT) at 8mT (3 ppm).

Photoinduced Processes in Lysine-Tryptophan-Lysine Tripeptide with L and D Tryptophan.

Optical isomers of short peptide Lysine-Tryptophan-Lysine (Lys-{L/D-Trp}-Lys) and Lys-Trp-Lys with an acetate counter-ion were used to study photoinduced intramolecular and intermolecular processes of interest in photobiology. A comparison of L- and D-amino acid reactivity is also the focus of scientists' attention in various specialties because today, the presence of amyloid proteins with D-amino acids in the human brain is considered one of the leading causes of Alzheimer's disease. Since aggregated amyloids, mainly Aβ42, are highly disordered peptides that cannot be studied with traditional NMR and X-ray techniques, it is trending to explore the reasons for differences between L- and D-amino acids using short peptides, as in our article. Using NMR, chemically induced dynamic nuclear polarization (CIDNP) and fluorescence techniques allowed us to detect the influence of tryptophan (Trp) optical configuration on the peptides fluorescence quantum yields, bimolecular quenching rates of Trp excited state, and the photocleavage products formation. Thus, compared with the D-analog, the L-isomer shows a greater Trp excited state quenching efficiency with the electron transfer (ET) mechanism. There are experimental confirmations of the hypothesis about photoinduced ET between Trp and the CONH peptide bond, as well as between Trp and another amide group.

Structure of fungal cell walls and remodeling by antifungal drugs elucidated using solid-state NMR.

Understanding the cell wall structure, dynamics, and mechanisms of adaptations are important for the development of cell wall-targeted drugs for the treatment of fatal infections caused by fungi. We first combined ssNMR, and functional genomics analysis to identify the functionality of cell wall carbohydrates in Aspergillus fumigatus and four mutants depleted of major structural polysaccharides. We revealed a rigid inner core of the cell wall formed by tightly associated chitin and α-1,3-glucan, which are embedded in a soft matrix of β-glucans and capped by a mobile outer shell rich in galactosaminogalactan and galactomannan. The distribution of α-1,3-glucan in chemically and dynamically distinct domains supports its dual functionality in structure and pathogenicity. Also, ssNMR revealed the modification of polymer dynamics which suggests that fungi produce a denser and hydrophobic cell wall as a response to structural defects. Second, we characterized the structural responses of A. fumigatus cell walls to the treatment of antifungal drugs. The drug-treated cell walls are highly hydrophobic and stiff to resist external stress. The fungi also increased the content of chitin and produced new forms of α-glucans to compensate for the loss of β-glucan due to drug inhibition. Third, we found that the crystalline structure of chitin bears intrinsic heterogeneity, which is resistant to caspofungin treatment. We documented the unique structural fingerprints of chitin across different fungal species. Finally, we used an innovative approach combing the ssNMR with the dynamic nuclear polarization technique to characterize the cell wall organization in different morphotypes varied along with the life cycle, revealing a highly conserved carbohydrate core in both conidia and mycelia. These studies and the biophysical techniques yield essential information that can serve as potential targets for discovering novel antifungal compounds with broad spectrums.

Primary Structure of Glycans by NMR Spectroscopy.

Glycans, carbohydrate molecules in the realm of biology, are present as biomedically important glycoconjugates and a characteristic aspect is that their structures in many instances are branched. In determining the primary structure of a glycan, the sugar components including the absolute configuration and ring form, anomeric configuration, linkage(s), sequence, and substituents should be elucidated. Solution state NMR spectroscopy offers a unique opportunity to resolve all these aspects at atomic resolution. During the last two decades, advancement of both NMR experiments and spectrometer hardware have made it possible to unravel carbohydrate structure more efficiently. These developments applicable to glycans include, inter alia, NMR experiments that reduce spectral overlap, use selective excitations, record tilted projections of multidimensional spectra, acquire spectra by multiple receivers, utilize polarization by fast-pulsing techniques, concatenate pulse-sequence modules to acquire several spectra in a single measurement, acquire pure shift correlated spectra devoid of scalar couplings, employ stable isotope labeling to efficiently obtain homo- and/or heteronuclear correlations, as well as those that rely on dipolar cross-correlated interactions for sequential information. Refined computer programs for NMR spin simulation and chemical shift prediction aid the structural elucidation of glycans, which are notorious for their limited spectral dispersion. Hardware developments include cryogenically cold probes and dynamic nuclear polarization techniques, both resulting in enhanced sensitivity as well as ultrahigh field NMR spectrometers with a 1H NMR resonance frequency higher than 1 GHz, thus improving resolution of resonances. Taken together, the developments have made and will in the future make it possible to elucidate carbohydrate structure in great detail, thereby forming the basis for understanding of how glycans interact with other molecules.

New Aspects of the Antioxidant Activity of Glycyrrhizin Revealed by the CIDNP Technique.

Electron transfer plays a crucial role in ROS generation in living systems. Molecular oxygen acts as the terminal electron acceptor in the respiratory chains of aerobic organisms. Two main mechanisms of antioxidant defense by exogenous antioxidants are usually considered. The first is the inhibition of ROS generation, and the second is the trapping of free radicals. In the present study, we have elucidated both these mechanisms of antioxidant activity of glycyrrhizin (GL), the main active component of licorice root, using the chemically induced dynamic nuclear polarization (CIDNP) technique. First, it was shown that GL is capable of capturing a solvated electron, thereby preventing its capture by molecular oxygen. Second, we studied the effect of glycyrrhizin on the behavior of free radicals generated by UV irradiation of xenobiotic, NSAID—naproxen in solution. The structure of the glycyrrhizin paramagnetic intermediates formed after the capture of a solvated electron was established from a photo-CIDNP study of the model system—the dianion of 5-sulfosalicylic acid and DFT calculations.

Evolution of α-synuclein conformation ensemble toward amyloid fibril via liquid-liquid phase separation (LLPS) as investigated by dynamic nuclear polarization-enhanced solid-state MAS NMR

Protein fibrillation and human neurodegenerative diseases, with a profound underlying connection suggested between them, have been the subject of intense investigations in the medical, biophysical and bio-engineering sciences. For gaining the molecular mechanistic insights into such connection, i.e., the cause and effect, atomic-resolution molecular structure information especially on the initial oligomeric states is of paramount importance, not only that on the mature amyloid fibrils. α-Synuclein (αSyn) and its amyloid fibril has a direct relevance to the Parkinson's disease and other synucleinopathies, but what triggers the fibrillation is still not entirely clear. We here describe the liquid-liquid phase separation (LLPS) of αSyn and investigate its conformational evolution from its monomeric state into oligomer state within the early-stage of the phase-separated droplets, mainly using solution and magic-angle spinning (MAS) solid-state nuclear magnetic resonance (NMR) spectroscopies, aided with optical and fluorescent microscopies and CD spectroscopy. Based on the analysis of the intricately broadened shapes of the MAS NMR peaks observed for isotopically 13C-labeled His-50 of αSyn, we show that the distribution of the αSyn conformation is skewed from the initial completely random state to a loose β-rich ensembles at/around His-50 as early as day-3 (d3) within the droplet. This intra-droplet loose β-rich assembly showed a very slow progression until d8, and eventually maturated into ThT-positive, long and unbranched amyloid fibrils after 8 weeks. The obtained information on the evolution of the distribution of the conformation ensemble is unique, and difficult to obtain with X-ray crystallography and cryo-electron microscopy (cryoEM). In particular, the sensitivity-enhanced MAS NMR based on the low-temperature dynamic nuclear polarization (DNP) technique was proven to be a key tool in characterizing the conformational ensemble with dilute protein samples such as the liquid-phase droplets.

Stereoselectivity of Interaction of Nonsteroidal Anti-Inflammatory Drug S-Ketoprofen with L/D-Tryptophan in Phospholipid Membranes.

The mechanisms of stereoselectivity of the interaction of chiral drugs with active sites of enzymes and cell receptors attract significant attention. The first reason is the difference in therapeutic activity of the enantiomers of the common drugs. Another reason is the interest in the role of chiral inversion of amino acids involved in various peptides in the development of many diseases including Alzheimer’s, Parkinson’s, type II diabetes, and a number of other pathological conditions. In our study we use elementary chemical process—electron transfer (ET) to simulate individual stages of ligand–receptor and enzyme–substrate interactions. In particular, previous studies of photoinduced ET in chiral donor-acceptor dyads consisting of the nonsteroidal anti-inflammatory drug (R/S)-ketoprofen and (L)-tryptophan show the stereo and spin selectivity of ET in diastereomers. The present study is devoted to the interaction of (S)-ketoprofen with L- and D-enantiomers of tryptophan in homogeneous aqueous solution and in phospholipid membranes. The study was done using the NMR technique and molecular modeling. These approaches confirm efficient penetration of ketoprofen into the lipid bilayer and binding with tryptophan molecule. The short-lived paramagnetic intermediates formed during the photoinduced ET from electron donor tryptophan to ketoprofen have been detected using the chemically induced dynamic nuclear polarization (CIDNP) technique. It was found that S-ketoprofen interacts stereoselectively with tryptophan enantiomers in the lipid membrane. The formation of the ketyl radical of ketoprofen under irradiation leads to the oxidation of membrane lipids and may be the cause of ketoprofen phototoxicity. However, in contrast to a homogeneous solution in phosphate buffer saline, where the amino acid tryptophan accelerates the photodecomposition of KP due to intramolecular hydrogen transfer, tryptophan in a lipid membrane significantly reduces the rate of photodegradation due to a reversible electron (or hydrogen) transfer reaction. The stereoselectivity in the rate of KP and lipids decomposition under UV irradiation of S-ketoprofen in the presence of tryptophan enantiomers in lipid bilayer has been detected.

Solid-state NMR analysis of unlabeled fungal cell walls from Aspergillus and Candida species

Fungal infections cause high mortality in immunocompromised individuals, which has emerged as a significant threat to human health. The efforts devoted to the development of antifungal agents targeting the cell wall polysaccharides have been hindered by our incomplete picture of the assembly and remodeling of fungal cell walls. High-resolution solid-state nuclear magnetic resonance (ss NMR) studies have substantially revised our understanding of the polymorphic structure of polysaccharides and the nanoscale organization of cell walls in Aspergillus fumigatus and multiple other fungi. However, this approach requires 13C/15N-enrichment of the sample being studied, severely restricting its application. Here we employ the dynamic nuclear polarization (DNP) technique to compare the unlabeled cell wall materials of A. fumigatus and C. albicans prepared using both liquid and solid media. For each fungus, we have identified a highly conserved carbohydrate core for the cell walls of conidia and mycelia, and from liquid and solid cultures. Using samples prepared in different media, the recently identified function of α-glucan, which packs with chitin to form the mechanical centers, has been confirmed through conventional ss NMR measurements of polymer dynamics. These timely efforts not only validate the structural principles recently discovered for A. fumigatus cell walls in different morphological stages, but also open up the possibility of extending the current investigation to other fungal materials and cellular systems that are challenging to label.

A novel strategy for site selective spin-labeling to investigate bioactive entities by DNP and EPR spectroscopy

A novel specific spin-labeling strategy for bioactive molecules is presented for eptifibatide (integrilin) an antiplatelet aggregation inhibitor, which derives from the venom of certain rattlesnakes. By specifically labeling the disulfide bridge this molecule becomes accessible for analytical techniques such as Electron Paramagnetic Resonance (EPR) and solid state Dynamic Nuclear Polarization (DNP). The necessary spin-label was synthesized and inserted into the disulfide bridge of eptifibatide via reductive followed by insertion by a double Michael addition under physiological conditions. This procedure is universally applicable for disulfide containing biomolecules and is expected to preserve their tertiary structure with minimal change due to the small size of the label and restoring of the previous disulfide connection. HPLC and MS analysis show the successful introduction of the spin label and EPR spectroscopy confirms its activity. DNP-enhanced solid state NMR experiments show signal enhancement factors of up to 19 in 13C CP MAS experiments which corresponds to time saving factors of up to 361. This clearly shows the high potential of our new spin labeling strategy for the introduction of site selective radical spin labels into biomolecules and biosolids without compromising its conformational integrity for structural investigations employing solid-state DNP or advanced EPR techniques.

Synthetic Approaches for 15 N-Labeled Hyperpolarized Heterocyclic Molecular Imaging Agents for 15 N NMR Signal Amplification by Reversible Exchange in Microtesla Magnetic Fields.

NMR hyperpolarization techniques enhance nuclear spin polarization by several orders of magnitude resulting in corresponding sensitivity gains. This enormous sensitivity gain enables new applications ranging from studies of small molecules by using high-resolution NMR spectroscopy to real-time metabolic imaging in vivo. Several hyperpolarization techniques exist for hyperpolarization of a large repertoire of nuclear spins, although the 13 C and 15 N sites of biocompatible agents are the key targets due to their widespread use in biochemical pathways. Moreover, their long T1 allows hyperpolarized states to be retained for up to tens of minutes. Signal amplification by reversible exchange (SABRE) is a low-cost and ultrafast hyperpolarization technique that has been shown to be versatile for the hyperpolarization of 15 N nuclei. Although large sensitivity gains are enabled by hyperpolarization, 15 N natural abundance is only ∼0.4 %, so isotopic labeling of the molecules to be hyperpolarized is required in order to take full advantage of the hyperpolarized state. Herein, we describe selected advances in the preparation of 15 N-labeled compounds with the primary emphasis on using these compounds for SABRE polarization in microtesla magnetic fields through spontaneous polarization transfer from parahydrogen. Also, these principles can certainly be applied for hyperpolarization of these emerging contrast agents using dynamic nuclear polarization and other techniques.

Hyperpolarized MRI with silicon micro and nanoparticles: Principles and applications.

Silicon-based micro and nanoparticles are ideally suited for use as biomedical imaging agents because of their biocompatibility, biodegradability, and simple surface chemistry that facilitates drug loading and targeting. A method to hyperpolarize silicon particles using dynamic nuclear polarization (DNP), which increases magnetic resonance (MR) imaging signals by several orders-of-magnitude through enhanced nuclear spin alignment, was developed to allow silicon particles to function as contrast agents for in vivo magnetic resonance imaging. In this review, we describe the application of the DNP technique to silicon particles and nanoparticles for background-free real-time molecular MR imaging. This review provides a summary of the state-of-the-science in silicon particle hyperpolarization with a detailed protocol for hyperpolarizing silicon particles. This information will foster awareness and spur interest in this emerging area of nanoimaging and provide a path to new developments and discoveries to further advance the field. This article is categorized under: Diagnostic Tools > In Vivo Nanodiagnostics and Imaging Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease Therapeutic Approaches and Drug Discovery > Emerging Technologies.

Dynamic Nuclear Polarization Enhanced Nuclear Spin Optical Rotation

AbstractNuclear spin optical rotation (NSOR) has been investigated as a magneto‐optical effect, which holds the potential for applications, including hybrid optical‐nuclear magnetic resonance (NMR) spectroscopy and gradientless imaging. The intrinsic nature of NSOR renders its detection relatively insensitive, which has prevented it moving from a proof of concept to a method supporting chemical characterizations. In this work, the dissolution dynamic nuclear polarization technique is introduced to provide nuclear spin polarization, increasing the signal‐to‐noise ratio by several thousand times. NSOR signals of 1H and 19F nuclei are observed in a single scan for diluted compounds, which has made this effect suitable for the determination of electronic transitions from a specific nucleus in a large molecule.

Open-source, partially 3D-printed, high-pressure (50-bar) liquid-nitrogen-cooled parahydrogen generator.

The signal of magnetic resonance imaging (MRI) can be enhanced by several orders of magnitude using hyperpolarization. In comparison to a broadly used dynamic nuclear polarization (DNP) technique that is already used in clinical trials, the parahydrogen (H-based hyperpolarization approaches are less cost-intensive, are scalable, and offer high throughput. However, a H generator is necessary. Available commercial H generators are relatively expensive (EUR 10 000-150 000). To facilitate the spread of H-based hyperpolarization studies, here we provide the blueprints and 3D models as open-source for a low-cost (EUR ) 50-bar liquid-nitrogen-cooled H generator.

Dynamic Nuclear Polarization Using 3D Aromatic Boron Cluster Radicals

A set of two dodecaborate [B12(OR)12]1- radical cluster anions containing a dense layer of fluorinated end-groups provides nuclear spin hyperpolarization via the dissolution dynamic nuclear polarization (D-DNP) technique. We show that these clusters can enhance 19F nuclear magnetic resonance (NMR) signals. Importantly, given the inherent radical delocalization in dodecaborate-based clusters, these species are compatible with reactive compounds such as Lewis acids, providing ∼1000-2000 times of signal enhancement for B(C6F5)3 in liquid state NMR spectroscopy experiments at 9.4 Tesla. This observation suggests that 3D aromatic radicals can provide advantages over the conventional radical species that are currently used for DNP such as 2,2,6,6-tetramethyl-1-piperidinyloxy (TEMPO) by showing superior chemical compatibility. The ability to hyperpolarize reactive compounds using [B12(OR)12]1- cluster radicals opens up new applications of reaction monitoring by D-DNP NMR spectroscopy, including the observation of catalytically active species in complex reaction mixtures.

Imaging Acute Metabolic Changes in Patients with Mild Traumatic Brain Injury Using Hyperpolarized [1-13C]Pyruvate.

SummaryTraumatic brain injury (TBI) involves complex secondary injury processes following the primary injury. The secondary injury is often associated with rapid metabolic shifts and impaired brain function immediately after the initial tissue damage. Magnetic resonance spectroscopic imaging (MRSI) coupled with hyperpolarization of 13C-labeled substrates provides a unique opportunity to map the metabolic changes in the brain after traumatic injury in real-time without invasive procedures. In this report, we investigated two patients with acute mild TBI (Glasgow coma scale 15) but no anatomical brain injury or hemorrhage. Patients were imaged with hyperpolarized [1-13C]pyruvate MRSI 1 or 6 days after head trauma. Both patients showed significantly reduced bicarbonate (HCO3–) production, and one showed hyperintense lactate production at the injured sites. This study reports the feasibility of imaging altered metabolism using hyperpolarized pyruvate in patients with TBI, demonstrating the translatability and sensitivity of the technology to cerebral metabolic changes after mild TBI.

Dynamic nuclear polarization enhanced neutron crystallography: Amplifying hydrogen in biological crystals.

Dynamic nuclear polarization (DNP) can provide a powerful means to amplify neutron diffraction from biological crystals by 10-100-fold, while simultaneously enhancing the visibility of hydrogen by an order of magnitude. Polarizing the neutron beam and aligning the proton spins in a polarized sample modulates the coherent and incoherent neutron scattering cross-sections of hydrogen, in ideal cases amplifying the coherent scattering by almost an order of magnitude and suppressing the incoherent background to zero. This chapter describes current efforts to develop and apply DNP techniques for spin polarized neutron protein crystallography, highlighting concepts, experimental design, labeling strategies and recent results, as well as considering new strategies for data collection and analysis that these techniques could enable.

Abstract 1944: Detection of metabolic change in glioblastoma after radiotherapy using hyperpolarized 13C-MRI: Glycolytic metabolism in cancer stem cell-like cell-derived tumor model

Abstract OBJECTIVES: Since the cancer stem-like cells (CSCs) play an important role in the resistance against radiation therapy due to its different metabolic profile, investigation of the specific metabolic characteristics of CSCs is valuable. Dynamic nuclear polarization (DNP) technique using 13C-labeled substrates enables magnetic resonance imaging (MRI) to monitor specific enzymatic reactions in tumors. In this study, DNP-MRI along with hyperpolarized [1-13C] pyruvate was conducted to evaluate the metabolic change in glycolytic profiles after irradiation between a glioblastoma cell line and a CSC-derived glioma using orthotopic xenograft mouse model. A microarray analysis was also performed to compare to the imaging study. METHODS: DNP-MRI A 3T MRI scanner along with a custom-made head coil was used. To obtain hyperpolarized [1-13C] pyruvate, 30 μL of [1-13C] pyruvic acid containing 15 mM of OX063 (trityl radical compound) and 2.5 mM of the gadolinium chelate were polarized at 3.35 T and 1.4 K in the Hypersense DNP polarizer. The hyperpolarized sample dissolved in a buffered medium was injected via the tail vein followed by an acquisition of 13C-two-dimensional MR-spectroscopy to measure the pyruvate-to-lactate ratios (Lac/Pyr) in the tumor region and the contralateral normal brain separately. Cell lines A human glioblastoma cell line, U251 and a glioma CSC line, NSC11 were cultured for stereotactic intracranial injection. Animal model Orthotopic brain tumor model was developed in athymic nude mice by intracranial implantation. When the tumor volume reached 50 +/- 10 mm3 on T2-weighted MRI, a sequence of DNP-MRI scans was performed 24 hours before and 6, 16 and 24 after 6-Gy whole-brain irradiation (N = 4 each with a non-irradiated control group). Independently, a microarray analysis using the tumor tissue samples excised 6 and 24 hours after irradiation was performed on Affymetrix Human Genome U133 Plus 2.0 array. RESULTS: DNP-MRI showed a decrease in Lac/Pyr 6 and 16 hours after irradiation compared to the control group in U251 whereas no significant change was observed in the NSC11 tumor. The microarray analysis showed a significant down-regulation of lactate dehydrogenase A (LDH-A) expression in U251 6 and 24 hours after irradiation, consistent with the finding of DNP-MRI. In the microarray analysis, SIRT2 and KLF4 expression also changed in the U251 tumor, suggested to regulate the LDH-A activity/expression. CONCLUSION: DNP-MRI demonstrated the difference in radiation response in glycolytic metabolism between the glioblastoma cell line and the CSC-derived glioma. This technique might be a feasible diagnostic method evaluating metabolic profiles of the brain tumors in the clinical settings of pretreatment and post-treatment. Citation Format: Tatsuya Kawai, Murali C. Krishna, Tamalee Kramp, Jeffery R. Brender, Philip Tofilon, Kevin A. Camphausen. Detection of metabolic change in glioblastoma after radiotherapy using hyperpolarized 13C-MRI: Glycolytic metabolism in cancer stem cell-like cell-derived tumor model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1944.