USE OF NON-LABELED SUGARS AND DETECTION BY MRI FOR ASSESSING TISSUE PERFUSION AND METABOLISM

BACKGROUND

1. Field of Invention

This patent application relates to methods for magnetic resonance imaging and spectroscopy, and more specifically relates to magnetic resonance (MR) methods for assessing tissue metabolism and perfusion using injection of non-toxic and non-labeled substances

2. Discussion of Related Art

Increased glucose uptake is a well-accepted marker of tumor aggression. In general, tumors tend to have higher glucose utilization and uptake than normal tissue. Following malignancy, glucose uptake in tumors increases dramatically. Glucose uptake, as a biomarker, can be measured by Fluorodeoxyglucose Positron Emission Tomography (¹⁸FDG-PET), which has become the radiological modality of choice for detecting tumor malignancy. For example, FDG-PET has been proven suitable for detecting and staging primary breast carcinomas and for monitoring response to chemotherapy, as well as many other cancers. However, ¹⁸FDG is a radioactive substance with a half-life of 110 minutes, thus requiring a fresh supply. The use of radioactivity limits repeated frequent use in the same person. In addition PET has low spatial resolution compared to CT and MRI and negligible inherent tissue contrast, leading to the need for anatomical co-registration using CT or MRI.

Other important biomarkers of malignancy include increased permeability of the vascular bed and increased microvessel density, which can be assessed using dynamic contrast-enhanced MRI (DCE-MRI). So far, DCE-MRI has been used to determine tumor grade, extent of disease, and treatment response. For example, recent multi-center results from the International Breast MR Consortium trial concluded that the combined use of architectural (shape) and dynamic contrast features increased specificity for breast MRI. DCE-MRI has been used clinically to image tumor perfusion parameters such as vascular volume and permeability from kinetic modeling of the DCE signal intensity curve as a function of time after bolus injection. However, DCE-MRI relies on the injection of paramagnetic agents such as Gadolinium-DiethyleneTriaminePentaacetic Acid (GdDTPA) that affect relaxation contrast. Such agents have recently been criticized for safety issues in persons with kidney disease (Thomsen, H. S. Nephrogenic systemic fibrosis: A serious late adverse reaction to gadodiamide. Eur Radiol 16, 2619-2621, (2006)).

Thus, there is a need in the art for a methodology to measure abnormalities in tissue metabolism, tissue perfusion and tissue pH that does not use potentially toxic or radioactive exogenous agents and yet is capable of generating sufficient contrast to probe such metabolic and vascular and chemical properties of tissue. This would be especially important for studying tumor malignancy and for monitoring the effects of cancer treatment. If developed, this methodology may reduce false-positive detection rates for cancer by functioning as an add-on for current high-volume screening approaches and to improve treatment monitoring. Another application is for assessing cardiovascular disease, where changes in tissue perfusion parameters and pH may occur during ischemia.

SUMMARY

A method for magnetic resonance (MR) imaging or spectroscopy on a MR scanner to detect tissue physiological parameters in one or more tissue areas in a human or non-human subject according to an embodiment of the current invention includes administering to the subject a contrast enhancing physiologically tolerable amount of a sugar that is non-labeled, subjecting the subject to an MR procedure capable of generating MR signals encoding at least one tissue area in the subject in which the sugar either passes or is taken up, detecting a temporal variation in the MR signals in the at least one tissue area after the administering the sugar, determining at least one tissue-related parameter from the temporal variation, and ascertaining whether the at least one tissue-related parameter is abnormal.

A method for magnetic resonance (MR) imaging or spectroscopy on a MR scanner according to an embodiment of the current invention includes generating a first recorded MR signal by observing a tissue area of a subject under observation in a magnet adapted to provide a characteristic main magnetic field with a corresponding characteristic water proton resonant frequency, generating a second recorded MR signal by repeating the observing at a later time when the subject's blood sugar level is elevated, detecting a difference between the second recorded MR signal and the first recorded MR signal, and subsequently ascertaining a physiological parameter associated with the tissue area of the subject under observation based on the detected difference. A computer-readable medium according to an embodiment of the current invention includes software instructions, which software instructions when executed by a computer, causes the computer, in conjunction with said MR scanner, to implement methods according to embodiments of the current invention.

A toolkit for imaging a subject in a MR system according to an embodiment of the current invention includes an MR-compatible external device constructed to inject a biocompatible dose of sugar in the subject under observation in the MR system, and a sugar analyzer configured to measure blood glucose levels in the subject before, during, and after the dose of glucose is injected into the subject.

BRIEF DESCRIPTION OF THE DRAWINGS

Further objectives and advantages will become apparent from a consideration of the description, drawings, and examples.

FIG. 1 is a schematic illustration of the procedure of administering the sugar as a contrast agent to the subject, including the monitoring by MRI of the subject, before, during and after administration.

FIG. 2A of a magnetic resonance imaging (MRI) system on which some embodiments of the current invention can be implemented.

FIG. 2B shows a flow chart of a method as an embodiment of the current invention.

FIGS. 3A and 3B show example measured relative water signal intensity as a function of saturation frequency in a chemical exchange saturation transfer (CEST) MRI experiment and the asymmetry in the magnetization transfer ratio (MTR_asym) calculated from it, respectively, of a phantom containing varying concentrations of glucose solutions according to some embodiments of the current invention.

FIG. 3C shows the effect of changes in pH on the glucose-based water CEST MTR_asymsignal, showing a large increase in exchange transfer effect with reduction of pH.

FIG. 4A shows a Magnetization Transfer Ratio (MTR) asymmetry image at offset 1.2 parts-per million (ppm) for a mouse inoculated with two tumor xenografts before glucose infusion.

FIG. 4B shows a MTR asymmetry image at offset 1.2 ppm for a mouse inoculated with xenograft tumor after glucose infusion.

FIG. 4C shows the difference image between FIGS. 4a and 4b.

FIG. 4D shows the respective Z-spectra (relative water signal intensity as a function of saturation frequency) before and after infusion.

FIG. 4E shows the respective MTR asymmetry ratios before and after infusion.

FIG. 4F shows the respective MTR asymmetry histograms before and after infusion for a first region of interest.

FIG. 4G shows the respective MTR asymmetry histograms before and after infusion for a second region of interest.

FIG. 4H shows the respective MTR asymmetry histograms before and after infusion for a third region of interest.

FIG. 5A shows an reconstructed CT image of the mouse skeleton with the locations of the two tumor xenographts (MDA-MB-231 and MCF7) indicated.

FIGS. 5B and 5C show representative ¹⁸F-FDG-PET and post-Gd-DTPA difference images for this mouse.

FIG. 6A shows the MTR asymmetry images as a function of time during D-glucose infusion for the two implanted xenografts during the infusion.

FIG. 6B shows the MTR_asymsignal changes as a function of time during D-glucose infusion for the MDA-MB-231 xenograft in the rim and tumor center

FIG. 6C shows the MTR_asymsignal changes for the MCF7 xenograft in the rim and tumor center

FIG. 7A compares the parameter definitions for kinetic analysis of the MR-CEST glucose method with those of PET 11C-glucose, PET 18FDG glucose and dynamic contrast enhanced MRI.

FIG. 7B compares the working equations for the quantitative analysis of glucoCEST data during glucose perfusion, uptake and metabolism with those for the DCE-MRI method.

FIG. 8 is a schematic illustration of a toolkit for imaging a subject in a MR system according to an embodiment of the current invention.

DETAILED DESCRIPTION

Some embodiments of the current invention are discussed in detail below. In describing embodiments, specific terminology is employed for the sake of clarity. However, the invention is not intended to be limited to the specific terminology so selected. A person skilled in the relevant art will recognize that other equivalent components can be employed and other methods developed without departing from the broad concepts of the current invention. All references cited herein are incorporated by reference as if each had been individually incorporated.

FIG. 1 shows a general scheme of a method according to some embodiments of the invention in which a sugar such as D-glucose or any other suitable biocompatible sugar is administered to a subject in a dose sufficient for changes in MR signals of a subject to be detected. The sugar can be non-labeled according to some embodiments of the current invention. For example, the sugar can be non-radioactive, non-paramagnetic, and not containing non-abundant magnetically enriched isotopes. Such MR signals can be measured before, during and after administration of either a sugar solution (11) or a solid sugar (12) that is either in pure form or in the form of a sugar-containing food. In one embodiment of the invention, administration comprises intravascular administering (13) to the subject of a sugar solution using an injector or infusion pump or any other device for intravascular administration known to those skilled in the field. In another embodiment, the invention employs oral administration and ingestion (14) of a sugar solution or a solid pure sugar or a sugar-containing food. In yet another embodiment, any other method of administration known to those skilled in the field can be used. Administration can be done inside or outside the MRI scanner, but the subject (16) needs to be in the scanner (17) to generate the MR signals that will be used to determine the physiologic parameters. Detection of the administered sugar can employ MR procedures capable of generating signals from the changes in the water resonance due to the presence of the exchangeable protons in said sugar.

FIG. 2A shows an example of a magnetic resonance imaging (MRI) system on which the MRI methodologies according to some embodiments of the current invention can be implemented.

The MRI system 100 includes a magnetic resonance scanner 101, a data storage unit 108, and a signal processing unit 109. Magnetic resonance scanner 101 has a main magnet 105 mounted on base 104 that provides a substantially uniform main magnetic field Bo for a subject 102 under observation on scanner bed 103, a gradient system 106 that provides a perturbation of the main magnetic field Bo to encode spatial information of the constituent water molecules within a region of interest of subject 102 under observation, and a radio-frequency (RF) coil system 107 to transmit electromagnetic waves and to receive magnetic resonance signals from subject 102. MRI system 100 may be a human scanner or and experimental scanner for animals or phantoms containing chemicals.

Subject 102 may be one of an animal, a human, or combinations thereof. Subject 102 is capable of receiving a dose of biocompatible sugar as described in FIG. 1.

The RF coil system 107 may transmit one or more radio frequency (RF) pulses into a tissue area of the subject 102 and monitor a responsive signal induction in order to detect the presence of exchangeable protons. The tissue area, can be, for example, a brain, an esophagus, a breast, a pancreas, a small intestine, a colon, a rectum, a liver, a kidney, a prostate, a uterus, a testicle, a muscle, a joint, etc. The tissue area may be normal or have a physiologic abnormality.

The MR approach used for the detection of the sugar(s) can include any pulse sequences sensitizing the water signal to the presence of the sugar, including but not limited to sequences sensitive to exchange transfer effects in order to detect the presence of exchanging hydroxyl protons. Such exchange-transfer based MR can be done either using a chemical exchange saturation transfer (CEST) pulse sequence or a frequency labeled exchange transfer (FLEX) sequence or similar approaches known to those skilled in the field, or by using changes in relaxation due to the presence of exchangeable protons in the sugar. The CEST or FLEX approaches may include, but are not limited to, the assessment of differences in water signal intensity due to saturation or selective excitation and frequency labeling at different NMR frequencies or ranges of frequencies. Such differences can be expressed, for example, as magnetization transfer asymmetry ratios as known by those skilled in the field.

Such sensitization to exchange can be combined with any possible MRI detection scheme. When performing dynamic imaging to measure changes in sugar concentration as a function of time this will most likely be a fast imaging detection scheme, including, but not limited to, echo planar imaging or fast gradient echo managing. When measuring static or steady state changes in sugar concentration this will most likely employ higher resolution approaches, including but not limited to, FLAIR, MPRAGE and other MRI pulse sequences.

Controller 108 and Data storage unit 109 is in communication with signal processing unit 110 to store the recorded signals. Signal processing unit 110 is in communication with the MR scanner 101. Signal processing unit 110 may be partially or totally incorporated within a structure housing magnetic resonance scanner 101. Signal processing unit 110 may be at least partially incorporated in a workstation that is structurally separate from and in communication with magnetic resonance scanner 101. Signal processing unit 110 may be incorporated in a workstation that is structurally separate from and in communication with magnetic resonance scanner 101.

Signal processing unit 110 may process the recorded signals before and after a dose of sugar is administered to the subject 102 under observation to introduce a change in concentration of exchangeable protons to the tissue area. The processing may comprise analyzing the difference in the recorded signals before and after administration to ascertain a physiologic parameter associated with the tissue area of the subject 102. The physiologic parameter may include, for example, tissue uptake or metabolism or a perfusion parameter (blood flow, blood volume, blood transit time, tissue permeability) or tissue pH.

The processed results may be presented to a human observer by an output device in communication with signal processing unit 110. For example, the output device may be a display device, such as, for example, viewing station 111 or a console station 112 or a printer. FIG. 2B shows a flow chart of a method as an embodiment of the current invention. In block 201, a tissue area of a subject 102 is being monitored. The subject 102 may be under observation in the magnet of MR scanner 101 constructed for MR imaging or spectroscopy. The MR scanner can be used to monitor either static (at a certain time point after administration) or dynamic sugar-based water signal changes, by collecting one or more images or spectra while the substance is passing through the body and taken up by it. Such signals can be compared to a reference signal obtained before administration of the sugar or referenced to signals at different time points before, during or after administration. These MR signal changes can then be used to identify regions of normal or abnormal physiologic parameters

In block 202 of FIG. 2B, changes in the recorded signals may be analyzed. The changes may result from repeatedly monitoring before and after injecting into the subject 102 a biocompatible dose of sugar that introduces MR-detectable contrast into the tissue area.

In block 203, a physiological parameter associated with said tissue area of said subject may be ascertained. Examples of physiological parameters include but are not limited to the following: tumor perfusion, tissue permeability, tissue perfusion, tissue blood flow, tissue blood volume, tissue mean transit time, tissue sugar uptake, tissue sugar metabolism, etc.

FIGS. 3A and 3B show examples of measured relative intensity and asymmetry ratio, respectively, of a phantom containing varying concentrations of glucose solutions according to some embodiments of the current invention. The data were obtained by using a Chemical Exchange Saturation Transfer (CEST) MR sequence, dubbed “glycoCEST” for glycogen and “glucoCEST” for glucose. This sequence is capable of detecting the rapidly exchanging OH groups of glycogen and glucose using CEST in phantoms and in vivo in animals and human. Such detection at first seems implausible due to the limited time available to saturate rapidly exchanging protons, but it can be understood by realizing that for CEST the only requirement is that the life time is sufficiently long to achieve partial saturation. Such partial saturation can be increased by using higher Bi or by reducing the exchange rate.

FIG. 3A illustrates the so-called Z-spectra or CEST spectra for a phantom of D-glucose in PBS buffer (pH=7.3, T=37° C.). In Z-spectra, the relative water signal intensity is displayed as a function of saturation frequency. The profile of such spectra is affected by the presence of exchangeable protons, such as in this case hydroxyl protons. The spectra were acquired using a 9.4 Tesla animal magnet, a B1 field of 3.6 μT and a saturation time of 3.5 seconds. Z-spectra can be analyzed using asymmetry analysis with respect to the water resonance frequency giving a so-called magnetization transfer ratio asymmetry (MTR_asym) spectrum.

FIG. 3B shows that the shape of the MTR_asymspectra varies with glucose concentration. It is important to realize that spectral appearance may also change with power level (higher Bi will broaden the direct saturation line shape in Z-spectra). At the power level used and under the experimental conditions in the sample, the detection sensitivity from FIG. 3B is about 0.5% of water signal per mM D-glucose. In vivo, the plasma concentration is expected to be ramped from baseline (a few mM) up to about 10-20 mM. For tumors, if total plasma and extravascular extracellular space (EES) is 20-40% of the voxel volume, this plasma concentration should give a magnitude of 2-4% in signal change (2.2-4.4M of signal).

FIG. 3C shows the appearance of the MTR_asymspectrum of glucose as a function of pH. It can be clearly seen that a reduction in pH increases the sensitivity of the OH detection. This is important as it will assist in the detection of cancer if such tumors have an EES in which the pH is low compared to physiological, which is typical for many tumors. In addition, this will allow detection of ischemic tissue, where pH is reduced if anaerobic metabolism is occurring, or any other tissue region with reduced pH.

Tumors are generally characterized by rapid glucose metabolism, increased tissue blood volume, and increased permeability to extravascular extracellular space (EES). We applied glucose infusion in vivo on xenografts in mice for two human breast cancer lines: less aggressive (MCF-7) and highly aggressive and metastatic (MDA-MB-231). Our MRI protocol used normal D-glucose (non-radioactive, non-hyperpolarized and not labeled with magnetic isotopes) and monitored this using glucose-based chemical exchange saturation transfer (glucoCEST) MRI. Images were compared with perfusion as assessed by dynamic contrast-enhanced (DCE) MRI and glucose uptake as assessed by FDG-PET. In-vivo animal experiments employed female SCID mice four to six weeks old. Tumor cells were implanted orthotopically in the mammary fat pad in an aseptic surgical procedure according to Institutional Animal Care and Use Committee (IACUC) guidelines. For cell implantation in the mammary fat pad, mice were anesthetized with ketamine and acepromazine (6.25 mg/kg and 62.5 mg/kg) injected in a volume of 0.05 ml using a sterile insulin syringe (28.5 G needle), and a cell suspension containing 1×10⁶MDA-MB-231 cells in 0.05 ml of Hanks balanced salt solution was injected into the upper right thoracic mammary fat pad using a sterile insulin syringe (28.5 G needle). For growth of the estrogen sensitive MCF-7 cells, an estrogen pellet (17β-estradiol, Innovative Research of America, Inc., Cat. No. SE-121) was inserted with a sterilized 10G trocar needle in the opposite flank at the time of the tumor cell inoculation and the opening will be closed with a single sterile suture clip. The estrogen pellet size, placed at the tip of the trocar, was ˜3 mm and the total dose was 0.18 mg/pellet released over 60 days. The procedures were performed in the sterile environment of a laminar flow cabinet. All surgical equipment, including suture clips were sterilized in an autoclave before use. The area of incision and inoculation was swabbed with 70% alcohol and iodine before and after implantation. Following tumor cell implants, animals were monitored until recovered from anesthesia and then checked after 6 and 12 h, then daily.

In vivo MR imaging of the mice was demonstrated at 9.4 T, using a company-supplied transmit and receive coil. Gas anesthesia was delivered using a commercial vaporizer through tubing connected to a nose cone. Animals were kept warm during scanning with the company-supplied heating beds. The lateral tail vein of the anesthetized mouse was cannulated prior to placing the animal on the scanner gantry for injection of glucose or contrast agents. For MR experiments, a home-built catheter was used to minimize dead volume to under 0.04 ml and yet maintain the long lines that are necessary to be able to inject the agents while the animal is in the magnet. The home-built catheter has a PE-60 tubing (0.76 mm inner diameter and 1.22 mm outer diameter, Becton Dickinson) that inserts snugly into a T-connector ( 1/16″, Cole-Parmer, 06365-77). A 4 cm length of PE-60 tubing was inserted into the top arm of the T-connector, and a 25G⅝ needle cleared of its base fits snugly into the other end of the 4 cm PE-60 piece to insert in the tail vein. The remaining two arms of the T-connector were used to attach a PE-60 tube connected to a 3 ml syringe with 0.5 M glucose solution positioned in an infusion pump (as described below) and the other arm of the T-connector was hooked to a line containing the contrast agent that can be opened or closed with a one-way stop-cock. An additional line containing saline was used to test the potency of the tail vein.

For glucose infusion, sterile glucose solution in water (0.5 M or 90 g/L) was infused to the tail vein. An initial loading bolus of 0.1 ml was followed by the continuous infusion with exponentially decreasing rates from 0.5 ml/h to 0.05 ml/h to maintain the target blood glucose concentration of 20 mM.

FIG. 4 shows preliminary glucose delivery and uptake data in a mouse inoculated with MDA-MB-231 (2.5 weeks) and MCF-7 (1 month) xenografts, with the growth period based on approximate volume matching (the malignant MDA-MB-231 grows faster). The mouse was infused with 0.5 M D-glucose in saline via the tail vein using an initial loading bolus followed by a maintenance infusion at decreasing infusion rates that provide stable steady-state blood glucose concentrations at about 20 mM (about 4-5 times the normal blood concentration). GlucoCEST MRI was performed during steady state before infusion (40-min scan) and after blood glucose stabilization (40 min scan). The GlucoCEST sequence used had a Rapid Acquisition Rapid Echo (RARE) readout, a slice thickness of 1.5 mm, an in-plane resolution of 0.4 mm×0.5 mm, Z-spectra based on 29 saturation frequencies and a total acquisition time of 40 minutes. Thereafter, a DCE-MRI with 0.2 mmole/kg GdDTPA was performed.

FIG. 4A shows a MTR asymmetry image at offset 1.2 ppm from the proton water resonance frequency for a mouse inoculated with xenograft tumor before glucose infusion.

FIG. 4B shows a MTR asymmetry image at offset 1.2 ppm for a mouse inoculated with xenograft tumor after glucose infusion.

FIG. 4C shows the difference image between FIGS. 4A and 4B. Three regions of interest (ROIs), namely, ROI 1 through ROI 3, are identified on the difference image.

FIG. 4D shows the respective Z-spectra before and after infusion.

FIG. 4E shows the respective MTR asymmetry ratio before and after infusion as well as the difference between them.

FIG. 4F shows the respective MTR asymmetry histograms before and after infusion for three different regions of interest, showing a trend towards higher glucose uptake in the malignant MDA-MB-231 tumor (ROI 1) as well as in the necrotic area in the MCF7 tumor (ROI 2). These promising preliminary data further demonstrate the feasibility of using non-labeled glucose to probe tumor metabolism and perfusion parameters.

FIG. 5A shows a reconstructed CT image of the mouse skeleton with the locations of the MDA-MB-231 tumor and the MCF7 tumor indicated. FIGS. 5B and 5C show representative ¹⁸F-FDG-PET images and post-Gd-DTPA difference images for this mouse, respectively. In FIG. 5B, the MDA-MB-231 tumor exhibits a higher glucose uptake than the MCF-7 tumor on PET at 1 hr post-injection, in line with literature. In FIG. 5C, the Gd-DTPA data demonstrates good visualization of the tumor mass relative to normal tissue. Interestingly, the Gd perfusion scan did not highlight the necrotic area in the MCF7 tumor, while the glucose infusion did. This may be due to the different plasma kinetics and indicate different transport properties for the Gd agent versus D-glucose, which will be further investigated. The Gd acquisition for this mouse did not allow quantitative analysis. Glucose uptake and metabolism are well-known to be increased in malignant tumors, a phenomenon exploited in FDG PET. Also, tumor vascularity is generally increased through angiogenesis. Thus, the preliminary data suggest that infusion of non-labeled D-glucose and detection with CEST MRI can provide combined information about glucose uptake and perfusion.

Even though the procedure of looking at steady state glucose uptake at a certain time point post infusion should already provide a viable technique (similar to clinical PET where data are taken at a single time point post-infusion), it is also possible to speed up these experiments so that one can measure active uptake and perform quantitative analysis of combined glucose delivery and utilization rate. In fact, speeding up CEST may not be difficult as in principle less irradiation frequencies, e.g. measure only differences at a single frequency of the broad glucose saturation spectrum (e.g., around 1.2 ppm) may be used during infusion. This reduction may avoid the need for CEST asymmetry analysis by measuring only difference signals. As the glucose resonance is quite broad, a small miss-setting of the frequency due to local differences in field inhomogeneity should not be a problem. In addition, before measuring the kinetics, the exact water frequency can be quickly measured using field mapping procedures available on MRI scanners, including but not limited to our water saturation shift reference (WASSR) scheme that uses the same MR scheme as CEST and water frequencies.

FIGS. 6A-6C show results for a D-glucose infusion experiment with dynamic monitoring of the MR exchange transfer signals for a mouse with two tumor xenografts (MDA-MB-231 and MCF7) at 11.7 T for which a D-glucose infusion protocol similar to the example in FIG. 4 was used. FIG. 6A shows the MTR asymmetry images for the two implanted xenografts at the time point of maximal signal change. FIG. 6B shows the MTR_asymsignal changes as a function of time during infusion for the MDA-MB-231 xenograft in the rim and tumor center. FIG. 6C shows the MTR_asymsignal changes as a function of time during infusion for the MCF7 xenograft in the rim and tumor center. It can be seen that the uptake is higher in the more malignant MDA-MB-231 tumor.

FIG. 7A compares the parameter definition for kinetic analysis for glucose uptake in tumors, for 11C-glucose PET (PET-Glc), glucoCEST (MR-CEST), FDG-PET (PET-FDG) and DCE-MRI. The envisioned kinetic analysis includes uptake, efflux, tissue permeability, and metabolism. PET-FDG constants are denoted with an asterisk as FDG uptake and metabolism differs from glucose, which is usually accounted for through a lumped constant. Capital K indicates the transfer constant into the extravascular extracellular space and small k rate constant, but the units of K1 (ml/g/min) and Ktrans (min-1) differ by the multiplication with density (g/ml) for Ktrans. Abbreviation are illustrated as: G1=glucose; CR=contrast reagent, b=blood; v=volume fraction; t=total; V=volume; Vt=Vb+Ve+Vc; Hct=hematocrit; Vp=Vb(1-Hct); n=volume fraction=Vi/Vt; i=p,e,c.

The active equations for glucoCEST (MR-CEST), ¹¹C-PET FDG-PET and DCE-MRI are very similar and are summarized for glucoCEST and DCE-MRI in FIG. 7B. To date, no universal strategy for analysis of DCE-MRI has been formulated and most groups use simple two-compartment kinetic models as indicated in FIGS. 7A and B that can provide the fractional volume of EES (v_e), the rate constant for reflux of GdDTPA from EES back to plasma (k_ep), and the transfer constant characterizing extravasation of GdDTPA from plasma (K^trans). This two-compartment model has shown potential for breast tumor grading, allowing detection of statistically significant differences between various grades of tumor. DCE-MRI relies on T₁contrast, but one of the assumptions, fast exchange of water between compartments is not really valid in the most commonly used clinical pulse sequence approaches, which affects the parameter values. Accounting for this so-called shutter-speed effect allowed improved separation of the vascular properties of benign and malignant tumors. The glucoCEST equations were derived in analogy to ¹¹C-glucose and ¹³C-glucose, assuming typical tumor microenvironment conditions: (i) negligible backflow of glucose from cell to interstitium (i.e., k˜0) and (ii) instantaneous disappearance of glucoCEST signal upon cell entry due to rapid phosphorylation and glycolytic conversion, as supported by extremely low cellular equilibrium concentrations of the glycolytic intermediates glucose-6-phosphate (˜80 μM), fructose-6-phosphate (˜14 μM), and fructose-1,6-bisphosphate (˜30 μM) that can contribute to glucoCEST via OH groups. To establish the input function, Gl_p(t), time-dependent blood glucose concentrations may be measured with a standard glucose analyzer (radiometer) in a separate group of tumor bearing animals outside the scanner.

The method described relates to administering a sufficient amount of sugar, that accomplishes either an MR detectable blood sugar concentration change or the buildup in tissue of an MR detectable amount of the sugar. The detectable range of sugar depends on the magnetic field used (smaller concentrations can be detected at the more sensitive higher fields for MRI, the radiofrequency coil used and the subject's perfusion and metabolic parameters, and the pH of the tissue. The following general guidelines apply for sugars with protons of normal MR polarization (i.e. not hyperpolarized). The detectable amount corresponds to the MR effect of the concentration of exchangeable protons when using a sugar with one hydroxyl group at a blood or tissue concentration in the range of about 50 μM or more at normal tissue pH. Lower concentrations can be detected when increasing the number of hydroxyl groups in a sugar, increasing the concentration of the sugar in the administered volume of an intravascular bolus or infusion, or increasing the volume of the administration of the intravascular bolus or infusion of the sugar, or increasing the amount of solid sugar or sugar containing food in the case of oral administration. Other ways to lower this concentration would be either by changing the pH of the infusate to enhance detectability of the sugar, monitoring tissues that have a pH that enhances detectability of the sugar, including but not limited to the extravascular extracellular space of tumors, enhancing the exchange rate of the hydroxyl group through synthetic modification of the sugar, enhancing the polarization of other protons in the sugar and transferring that to the hydroxyl protons, enhancing the polarization of other nuclei in the sugar and transferring said polarization to the hydroxyl protons in the sugar either directly or via the protons in the sugar.

The dynamic analysis may provide an alternative for dynamic and static perfusion imaging with Gd-DTPA and other paramagnetic contrast agents, which can then be used to assess for instance, tumor perfusion, blood brain barrier permeability, tissue perfusion, tissue blood flow, tissue blood volume, tissue mean transit time. In addition, glucose kinetics can relate to both perfusion and tissue metabolism. Thus the use of non-labeled glucose infusion and measurement by MRI may provide an alternative to current methods using radioactive substances, such as FDG-PET, to probe metabolism.

For PET, by contrast, values for K₁, k₂, and k₃can in principle be derived using the Sokoloff 3-compartment model. To date, only few publications on clinical PET-FDG of breast cancer employ quantitative kinetics. The majority of studies measure the dimensionless standardized uptake value (SUV=organ uptake (Bq/g)×bodyweight (g)/injected dose (Bq)), which is proportional to K₁k₃/(k₂+k₃) and can provide a semi-quantitative measure of accumulation by normalizing organ tissue radioactivity at 60 min post-injection to the injected dose and body weight. The most comprehensive study on breast cancer known to the authors was on SUV in 46 benign and 51 malignant tumors, showing more than double the SUV in malignancy. We foresee that the non-labeled glucose experiment proposed here will also allow the determination of a specific uptake value for glucose in tumors, but then based on D-glucose.

The methodology, discussed above according to some embodiments of the current invention, may be used for tumor detection, imaging tumor perfusion and metabolism, assessing tumor malignancy, and monitoring tumor treatment. Such methodology may reduce false-positive detection rates by functioning as an add-on for current high-volume screening approaches and to improve treatment monitoring.

To the extent that B₀variation across the sample can lead to systematic errors in the CEST asymmetry analysis for the steady state method, a WAter Saturation Shift Referencing (WASSR) method may be used to correct for this. For kinetics experiments in which signal differences at one frequency will be measured, as explained above, susceptibility to B₀variation will be less pronounced. A time resolution on the order of seconds (similar to fMRI and DCE-MRI) is expected. Imaging speed also depends on the spatial resolution used and can be increased using parallel imaging and EPI methods. Because ultimately the methodology according to some embodiments of the current invention is a water detection method, all commonly available fast imaging methods that remain within specific absorption ration (SAR) guidelines when combined with sugar-sensitive pulse sequences including saturation or other exchange transfer approaches can be used.

Thus, some embodiments of the current invention provide a methodology of MRI monitoring of non-labeled glucose infusion. In particular, the methodology can measure MR contrast sensitive to both metabolic and vascular properties (perfusion and permeability) of tissue. In addition, the methodology uses a biocompatible compounds (sugars, including but not limited to D-glucose). Finally, in contrast to current magnetically labeled isotopes (e.g. ¹³C-labeled) approaches, the method according to some embodiments of the current invention uses water imaging for detection. Thus, the method according to some embodiments of the current invention is directly compatible with clinical scanners.

Some embodiments of the current invention may provide a toolkit for imaging a subject in a MR system, as shown in FIG. 8. The toolkit 700 may comprise at least one of: a MR-compatible device 701 constructed to inject a biocompatible dose of glucose sufficient to generate MR contrast in the subject under observation in the MR system, a MR-compatible glucose analyzer 702 configured to measure blood glucose level in the subject before, during, and after the dose of glucose is injected into the subject. MR compatible means capable of being operative inside a magnet for MR scanning 101 or inside an MR scanner room or outside an MR scan room, but connected to the subject in the MR scanner. For example, ferromagnetic materials are not MR-compatible. The device 701 may comprise one of: an external device or an intravascular applicator. An external device may comprise, for example, an infusion pump capable of being synchronized with data acquisition on the MR system.

The toolkit 700 may further comprise a computing engine 703, in communication with the MR system and the glucose analyzer 702. The computing engine may be programmed to: receive at least one of: measured data from the MR system or measured glucose level from the glucose analyzer 702, the measured data comprising MR signal changes due to injection of the dose of glucose into the subject; and compute a physiologic parameter based on at least one of: the measured data or the measured glucose level, the physiologic parameter comprising: a perfusion parameter or a metabolism parameter; and output the physiologic parameter to a user. It may also be possible to determine a sugar input function based on specific time dependent MR data and not use the glucose analyzer, and to use this MR-determined input function for calculating physiologic parameters.

The physiologic parameter may be computed based on one of: an empirical model, or an analytic model. An empirical model is not based on an analytic model and may include, for example, an area under the curve, etc. An analytic model may include, for example, the compartmental modeling as discussed above. The physiologic parameter may comprise one of: a standard uptake value, a permeability parameter, a tissue mean transit time, or a tissue blood volume, or other related physiologic parameters as specified above.

Computing engine 703 may be a computer with at least one central processing unit (CPU) and a plurality of memories. Computing engine 703 may also be a dedicated processing machine with such devices as, for example, a field programmable gated array (FPGA), a digital signal processing (DSP) chip, a graphic processing unit (GPU), an application specific integrated circuit (ASIC), etc. Computing engine 703 may also be incorporated in the MR scanner 101.

An embodiment of the invention provides a toolkit that includes a prepared sugar solution or sugar-containing solid or liquid food in a prescribed concentration (most likely ranging from, but not limited to, 1% weight fraction to 100% weight fraction) and in the appropriate amount for MR detection such that the sugar is in ampules or solid form and can be used immediately either with an available MR-compatible injector or infusion pump or as oral contrast agent.

In describing embodiments of the invention, specific terminology is employed for the sake of clarity. However, the invention is not intended to be limited to the specific terminology so selected. The above-described embodiments of the invention may be modified or varied, without departing from the invention, as appreciated by those skilled in the art in light of the above teachings. It is therefore to be understood that, within the scope of the claims and their equivalents, the invention may be practiced otherwise than as specifically described.

	Number	Date	Country
Parent	13994600	Jun 2013	US
Child	14877754		US

USE OF NON-LABELED SUGARS AND DETECTION BY MRI FOR ASSESSING TISSUE PERFUSION AND METABOLISM

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