WO2016123446A1 - Imagerie rm sans marqueur de tumeur maligne - Google Patents

Imagerie rm sans marqueur de tumeur maligne Download PDF

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WO2016123446A1
WO2016123446A1 PCT/US2016/015577 US2016015577W WO2016123446A1 WO 2016123446 A1 WO2016123446 A1 WO 2016123446A1 US 2016015577 W US2016015577 W US 2016015577W WO 2016123446 A1 WO2016123446 A1 WO 2016123446A1
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tissue
cest
mucins
tumor
computer readable
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PCT/US2016/015577
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Jeff Bulte
Xiaolei Song
Raag Airan
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The Johns Hopkins University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/05—Detecting, measuring or recording for diagnosis by means of electric currents or magnetic fields; Measuring using microwaves or radio waves 
    • A61B5/055—Detecting, measuring or recording for diagnosis by means of electric currents or magnetic fields; Measuring using microwaves or radio waves  involving electronic [EMR] or nuclear [NMR] magnetic resonance, e.g. magnetic resonance imaging
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/48—Other medical applications
    • A61B5/4842—Monitoring progression or stage of a disease
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01R—MEASURING ELECTRIC VARIABLES; MEASURING MAGNETIC VARIABLES
    • G01R33/00—Arrangements or instruments for measuring magnetic variables
    • G01R33/20—Arrangements or instruments for measuring magnetic variables involving magnetic resonance
    • G01R33/44—Arrangements or instruments for measuring magnetic variables involving magnetic resonance using nuclear magnetic resonance [NMR]
    • G01R33/48—NMR imaging systems
    • G01R33/54—Signal processing systems, e.g. using pulse sequences ; Generation or control of pulse sequences; Operator console
    • G01R33/56—Image enhancement or correction, e.g. subtraction or averaging techniques, e.g. improvement of signal-to-noise ratio and resolution
    • G01R33/5605—Image enhancement or correction, e.g. subtraction or averaging techniques, e.g. improvement of signal-to-noise ratio and resolution by transferring coherence or polarization from a spin species to another, e.g. creating magnetization transfer contrast [MTC], polarization transfer using nuclear Overhauser enhancement [NOE]

Definitions

  • the present invention relates generally to medical imaging. More particularly the present invention relates to CEST magnetic resonance imaging.
  • Mucins a family of large molecular weight and heavily glycosylated proteins, constitute the mucous barrier at the epithelial surface, and play an important role in cell signal transduction. Alterations in mucin expression or glycosylation have long been associated with the development of cancer, as they are thought to influence cellular growth, invasion, metastasis, and immune surveillance. Mucin- 1, one of the cell-surface-associated mucins encoded by the MUC1 gene, is expressed aberrantly in -900,000 of the 1.4 million tumors diagnosed each year in the United States.
  • MUCl-overexpressing breast, colon, and thyroid cancer cells are unresponsive to chemotherapeutic agents.
  • Tumor-associated glycosylation changes have been observed for decades and are associated with tumor proliferation, metastasis, and angiogenesis.
  • Most malignant epithelial tumor cells express mucins that are heavily underglycosylated, and alterations in mucin expression or glycosylation have long been associated with the development and prognosis of cancer.
  • Cell-surface glycoproteins including mucins, in particular, Mucin-1 (MUC-1), have been used as a novel diagnostic and therapeutic target.
  • MUC-1 is a marker of epithelial cell lines that is expressed in an underglycosylated form uMUC-1 in neoplastic cells derived from both epithelial and non-epithelial cell types.
  • uMUC-1 is overexpressed in most malignant adenocarcinomas of epithelial origin (e.g. colon, breast, and ovarian cancer). The specificity of this marker for early tumorgenesis makes it a target of great interest for molecular imaging.
  • MUC-1 under glycolycosylation of mucin implies that its hydroxyl content would be much reduced compared to normally glycosylated tissue.
  • MUC-1 with a core protein mass of 120-225 kDa, increasing to 250-500 kDa with glycosylation, extends no less that 500 nm beyond the surface of the cell.
  • MUCl is often underglycosylated with fewer and truncated oligosaccharide side chains, identified as the tumor- associated underglycosylated MUCl (uMUCl) antigen (FIG. 1).
  • the reduced glycosylation of tumor cells allows exposure of a highly immunogenic core peptide epitope of the uMUCl antigen, which has been exploited for the development of immunotherapeutic vaccines and targeted radiotheraputic drugs, and is also widely used as a serum diagnostic assay to detect ovarian, breast, and colon adenocarcinomas.
  • Targeted imaging agents against the uMUCl antigen recognizing the exposed peptide sequence on the tandem repeat have been developed, including radiolabeled agents and a dual- modality probe with the near-infrared fluorescence (NIRF) dye Cy5.5 conjugated to MRI- detectable superparamagnetic iron oxide nanoparticles.
  • NIRF near-infrared fluorescence
  • these approaches may not readily be adapted for clinical tumor staging as drug development and approval is a lengthy and costly process.
  • the pharmacokinetics of the probes may be such that only a small fraction of the tumor can be targeted.
  • An imaging technique that is "label-free" i.e., that does not rely on administering an exogenous agent) and can sample the entire tumor would be extremely valuable.
  • CEST chemical exchange saturation transfer
  • MRI magnetic resonance imaging
  • FIG. 1 illustrates a schematic diagram depicting the different levels of glycosylation between normal mucin (left) and tumor-associated mucin (right).
  • the oligosaccharide side- chains consist of a variety of glycans, e.g. GalNAc (triangles, which are O-linked to the core protein and contain sialic acid terminal residues (circles).
  • FIGS. 2A-2G illustrates graphical and image views for normally glycosylated mucin, which exhibits a strong CEST signal.
  • FIG. 2 A illustrates a graphical view of a Z-spectra of 5 mg/ml mucin at different pH values.
  • FIG. 2B illustrates a graphical view of calculated MTR aS ym values.
  • FIG. 2F illustrates a graphical view of concentration- dependence of MTRasym at different offset frequencies.
  • FIG. 2G illustrates a corresponding CEST image at 1.8 ppm.
  • FIGS. 3A-3E illustrate graphical and image views of a decrease of CEST signal following deglycosylation.
  • M native (normally glycosylated) mucin
  • DM deglycosylated mucin
  • FIG. 3 A Shown are the Z-spectra in FIG. 3 A, MTRasym values in FIG. 3B, and MTRasym image at 1.8 ppm in FIG. 3C.
  • FIG. 3D illustrates an image of PAS glycoprotein staining.
  • FIG. 3E illustrates an image view of SDS-PAGE.
  • FIGS. 4A-4H illustrate in vitro imaging of encapsulated cell lines.
  • FIG. 4A illustrates a lOx bright-field image that shows individual microcapsules containing MCFIOA cells.
  • FIG. 4D illustrates an MT -weighted image showing the phantom layout.
  • FIGS. 5A-5D illustrate in vivo imaging of benign and malignant tumor xenografts.
  • FIG. 5 A illustrates a T2w image, marked with regions of U87, LS174T, and control white matter (dashed square).
  • FIG. 5C illustrates a graphical view of MTRasym curves of the 3 ROIs marked in FIG. 5 A.
  • FIGS. 6A- 6C illustrate in vitro CEST images and spectrum for encapsulated LS174T uMUC-l + and U87 uMUC-1 " .
  • FIG. 7A illustrates in vivo mouse brain MTw images.
  • FIG. 7B illustrates a CEST image at lppm and
  • FIG. 7C illustrates a CEST MTRasym spectrum.
  • FIGS. 8A-8D illustrate graphical and image views of a Zspectra and MTRasym for deglycosylated and untreated mucin.
  • FIGS. 9A-9F illustrate microscopy, MTw image, CEST spectra and images, and imunostaining of encapsulated cell lines with different MUC-1 glycosylation levels.
  • a method for magnetic resonance imaging of a subject includes using a magnetic resonance imaging machine to generate CEST contrast image data for tissue.
  • the method includes processing the CEST contrast image data to determine presence of a mucin in the tissue.
  • the method also includes processing the CEST contrast image data to differentiate glycosylated and unglycosylated mucins and generate data related to the mucins present in the tissue.
  • the method includes assessing cancer using the generated data related to the mucins present in the tissue.
  • the method includes processing the CEST contrast image data with a non-transitory computer readable medium.
  • the method also includes processing the CEST contrast image data with a computing device specifically designed for assessment of mucins in tissue. Additionally, the method includes using the data related to the mucins present in the tissue to non-invasively phenotype a tumor in the tissue, to detect early tumorgenesis, and to monitor tumor growth.
  • a system for magnetic resonance imaging of a subject includes a magnetic resonance imaging machine configured to generate CEST contrast image data for tissue.
  • the system includes a non-transitory computer readable medium programmed for processing the CEST contrast image data to determine presence of a mucin in the tissue.
  • the non-transitory computer readable medium is also programmed for processing the CEST contrast image data to differentiate glycosylated and unglycosylated mucins and generate data related to the mucins present in the tissue and assessing cancer using the generated data related to the mucins present in the tissue.
  • the system includes a computing device.
  • the computing device is specifically designed for the assessment of mucins in tissue.
  • the non-transitory computer readable medium is programmed for processing the CEST contrast image data and loading the non-transitory computer readable medium on the computing device specifically designed for assessment of mucins in tissue.
  • the non-transitory computer readable medium is programmed for using the data related to the mucins present in the tissue to non-invasively phenotype a tumor in the tissue.
  • the non-transitory computer readable medium is also programmed for using the data related to the mucins present in the tissue to detect early tumorgenesis.
  • the non-transitory computer readable medium is programmed for using the data related to the mucins present in the tissue to monitor tumor growth.
  • the non-transitory computer readable medium is programmed for using normal tissue as a reference. Additionally, the non- transitory computer readable medium is programmed for using normal tissue as a reference during longitudinal follow-up studies and for calibrating and taking a ratio using normal tissue to gauge tumor contrast changes and malignancv.
  • the magnetic resonance imaging machine and the non-transitory computer readable medium are networked together. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
  • the present invention is directed to a new MRI technique that can assess tumor malignancy non-invasively. It is based on the principle that most malignant epithelial tumor cells express mucins that are heavily underglyocsylated. This phenomenon of aberrant expression has been well-documented in the literature. When benign tumor cells having normal mucin glycosylation with CEST MRI are imaged a strong CEST MRI signal effect is seen. When tumor cells lose their glycans and become malignant the difference in MRI CEST signal is detectable. Thus, the intrinsic properties of tumor cell surface mucin glycosylation can be used as a surrogate marker for predicting tumor malignancy. This can be accomplished without the need for injecting exogenous probes or labels, which have their own problems in terms of clinical approval, cost, and pharmacokinetics of binding to all tumor cells in question.
  • mucins are natural polymers rich in glycans, it was investigated whether
  • MUCl a single core protein contains up to 120 tandem repeats, each of which has five potential sites of O-glycosylation; a single molecule can contain up to 600 oligosaccharide side chains.
  • Glycosylation is initiated by the addition of an N- acetylgalactosamine (GalNAc) residue to a serine or threonine, followed by the sequential addition of carbohydrate residues, such as N-acetylglucosamine (GlcNAc), and then terminated by sialic acid, fucose, or galactose (FIG. 1).
  • GalNAc N- acetylgalactosamine
  • oligosaccharide side-chains consist of a variety of glycans, e.g. GalNAc (triangles, which are O-linked to the core protein and contain sialic acid terminal residues (circles).
  • each chain contains 2-10 simple sugars with 4-5 -OH protons (8-50 total -OH protons per chain), it can be calculated that 1 nM of MUCl contains up to 30,000 nM
  • uMUC-l 's counterpart MUCl is a large polymer rich in glycans containing multiple exchangeable OH protons, which is readily detectable by Chemical Exchange Saturation
  • CEST Cockayne syndrome MRI.
  • BT20, HT29, and LS174T Three uMUCl+ human malignant cancer cell lines overexpressing uMUCl (BT20, HT29, and LS174T) showed a significantly lower CEST signal compared to the benign human epithelial cell line MCFIOA and the uMUCl - tumor cell line U87.
  • MCFIOA benign human epithelial cell line
  • U87 uMUCl - tumor cell line
  • LS174T and U87 were bilaterally implanted in mouse brain, CEST MRI was able to make a clear distinction between the two types of tumors.
  • mucCEST imaging can be used as a label-free surrogate marker to non-invasively assess mucin glycosylation and tumor malignancy.
  • normal tissue is used as a reference for longitudinal follow up studies. If the tumor contrast changes upon malignancy a ratio can be taken and calibrated using normal tissue (for instance breast glandular tissue).
  • a commercial mucin extract from porcine stomach (Sigma-Aldrich, M2378) was used to characterize the CEST properties.
  • This crude product contains -1% bound sialic acid, which was obtained by digestion of hog stomach with pepsin.
  • Mucin was dissolved in 0.01 M phosphate-buffered saline (PBS) at concentrations from 1.25 mg/ml to 10 mg/ml, and titrated using high concentration HCl/NaOH, to various pH values ranging from 6 to 8.
  • PBS phosphate-buffered saline
  • the solutions were placed into 1 mm glass capillaries and assembled in a holder for CEST MR imaging. The samples were kept at 37 OC during imaging.
  • the mucins were first prepared with reduced glycan chains, to mimic tumor mucins with lower glycosylation levels (FIG. l). Due to the complex O-linked glycosylation and polymerization of mucins, chemical deglycosylation is preferred over enzymatic methods.
  • the oligosaccharide chains on mucins (Sigma- Andrich, M2378) were removed using anhydrous trifluoromethanesulfonic acid (TFMS) treatment, based on the protocol of the GLYCOFREETM chemical deglycosylation kit (Glyko, GKK500).
  • the deglycosylated mucin and the untreated mucin were further analyzed by polyacrylamide gel electrophoresis (SDS-PAGE) on 4-15% polyacrylamide minigels (Bio- Rad, Gel #456-1083 S) stained with coomassie blue, with the glycosylation level was confirmed by periodic acid-Schiff (PAS) staining (Thermo Scientific, Pierce glycoprotein staining Kit, 24562).
  • MCF10A a benign human breast carcinoma and U87, a human glioblastoma cell line, were selected as uMUCl -negative cell lines.
  • MEM Eagle's minimum essential medium
  • FBS fetal bovine serum
  • penicillin and streptomycin all from Gibco, Grand Island, NY.
  • HT29 cells were cultured using ATCC-formulated McCoy's 5a Medium Modified (Catalog No. 30-2007), containing 10% FBS.
  • the control mammary epithelial cells, MCF10A were grown in a Mammary Epithelial Cell Growth Medium kit (Lonza, CC-3150), which contains mammary epithelial cell basal medium and growth factors, with the addition of 100 ng/ml cholera toxin. Cultures were maintained at 37 °C in a humidified atmosphere of 5% C02 and 95% air.
  • the cell media were changed every two to three days, and when cells were confluent, they were 1 :4 distributed to new flasks by removing cells from the surface of the culture flask gently with 0.05% trypsin EDTA and a sterile scraper.
  • the four cell lines were encapsulated in alginate-PLL-alginate microcapsules at the same density of 1000 cells/capsule. After the encapsulation, the cell-containing microcapsules were suspended in PBS, and immediately transferred to 5-mm MR tubes for CEST imaging. The empty microcapsules without cells were also imaged as controls.
  • mice 1.5x105 MCF10A, LS174T or U87 cells were bilaterally injected to the striatum of each hemisphere at a depth of 2 mm, slowly over a period of 3-4 min with the syringe removed 30 s after completion to minimize back flow. These mice were subjected to MR imaging 2-3 weeks after implantation of tumor cells. During MR imaging, mice were anesthetized using 0.5-2% isoflurane.
  • Imaging experiments were performed on a Bruker 11.7T vertical bore scanner for the in vitro experiments and on a Bruker 9.4T horizontal bore scanner for the in vivo mice experiments, both using a transmit/receive volume coil.
  • CEST images were acquired using a continuous wave (CW) saturation pulse of 3 sec. as preparation, followed by a Rapid Acquisition with Relaxation Enhancement (RARE) readout sequence.
  • the saturation field strength (B i) was varied from 1.2 ⁇ to 6.0 ⁇ for investigating the CEST properties of normal mucin phantoms, with 2.4 ⁇ and 3.6 ⁇ chosen for the cell imaging and for in vivo imaging.
  • the CEST z-spectra were acquired by incrementing the saturation frequency every 0.2 ppm from -6 to 6 ppm for phantoms, and every 0.25 ppm from -5 to 5 ppm for cells and in vivo.
  • Another set of saturation weighted images with frequency incrementing every 0.1 ppm from -1 to 1 ppm, termed as Water Saturation Shift Reference (WASSR) were also collected for Bo mapping, using a 0.5 sec saturation pulse with B i of 0.5 ⁇ .
  • MTRasym ( -AwS+Aa)/ S- ⁇ was used to increase the dynamic range.
  • FIGS. 2A-2G illustrate graphical and image views for normally glycosylated mucin, which exhibits a strong CEST signal.
  • FIG. 2A illustrates a graphical view of a Z-spectra of 5 mg/ml mucin at different pH values.
  • FIG. 2B illustrates a graphical view of calculated MTRasym values.
  • FIG. 2D illustrates a graphical view of
  • FIG. 2F illustrates a graphical view of concentration-dependence of MTRasym at different offset frequencies.
  • FIG. 2G illustrates a corresponding CEST image at 1.8 ppm.
  • the 3.6 ppm peak from the backbone amides can be clearly observed in FIG. 2D. Even at the lowest concentration of 1.25 mg/ml, the MTR aS ym peaks reach >5%, which should be easily detectable.
  • the MTRasym changes as a function of concentration for three saturation frequencies (FIG. 2F).
  • the corresponding CEST image at 1.8 ppm (FIG. 2G) demonstrates strong CEST signal changes as a function of the concentration of normal glycosylated mucin.
  • FIGS. 3 A-3E illustrate graphical and image views of a decrease of CEST signal following deglycosylation.
  • M native (normally glycosylated) mucin
  • DM deglycosylated mucin
  • FIG. 3D illustrates an image of PAS glycoprotein staining
  • FIG. 3E illustrates an image view of SDS-PAGE.
  • FIGS. 4A-4H illustrate in vitro imaging of encapsulated cell lines.
  • FIG. 4A illustrates a lOx bright-field image that shows individual microcapsules containing MCF10A cells.
  • FIG. 4D illustrates an MT-weighted image showing the phantom layout.
  • FIGS. 4B and 4C represent the average
  • FIGS. 5A-5D illustrate in vivo imaging of benign and malignant tumor xenografts.
  • FIG. 5 A illustrates a T2w image, marked with regions of U87, LS174T, and control white matter (dashed square).
  • FIG. 5B illustrates a CEST contrast map created by averaging 1.2 ppm and 0.9 ppm superimposed onto FIG. 5 A.
  • FIG. 5C illustrates a graphical view of MTRasym curves of the 3 ROIs marked in FIG. 5 A.
  • MCF10A a benign human epithelial cell line
  • U87 a tumor cell line without uMUCl expression (uMUCl-)
  • uMUCl+ malignant LS174T cells expressing uMUCl
  • mucCEST MRI has been demonstrated as able to differentiate between tumor cells that are expressing normal vs. underglycosylated MUC1.
  • these mucopolysaccharides display a broad peak from 0.5 ppm to 4 ppm, with a signal peak around ⁇ 1 ppm, owing to the abundance of glycan side chains.
  • uMUCl underglycosylated MUC1
  • mucin was deglycosylated with as result a striking difference between the treated and untreated mucin, with the former showing a >75% reduction of CEST signal from 0.5 to 2 ppm.
  • underglycosylated human malignant tumor cell lines (BT20, HT29, and LS174T) showed a significantly lower CEST signal compared to a benign normally glycosylated human epithelial cell line (MCF10A) and to another uMUCl -negative cell line (U87), was tested and found in agreement.
  • MCF10A benign normally glycosylated human epithelial cell line
  • U87 uMUCl -negative cell line
  • the homogeneous environment of brain tissue was used instead of an orthotopical tumor model, as there are still challenges associated with high-field small animal CEST imaging, including motion artifacts, field inhomogeneity corrections, and susceptibility artifacts arising from air-tissue interfaces.
  • Improved CEST imaging methods, better-equipped clinical scanners, and larger tumor volumes may allow future orthotopic imaging in patients, where longitudinal monitoring may allow for proper quantification.
  • amide proton transfer (APT) CEST imaging has already been applied to monitor the response to neoadjuvant chemotherapy in breast cancer patients.
  • APT amide proton transfer
  • mucCEST imaging represents the first approach to differentiate label-free between tumor cells expressing and not expressing a single specific molecule, which has been widely studied and shown to play a significant role in tumor malignancy.
  • CEST imaging has become an active new field, and new imaging schemes and pulse sequences are continuously being developed to improve the quantification and robustness of CEST imaging.
  • LS174T uMUC-1 positive, i.e., underglycosylated
  • U87 uMUC-1 negative, i.e., heavily glycosylated tumor cells
  • Z-spectra were calculated from sample ROIs after Bo correction for each voxel using WAS SR.
  • FIGS. 6A- 6C illustrate in vitro CEST images and spectrum for encapsulated LS174T uMUC-l + and U87 uMUC-r.
  • FIG. 7A illustrates in vivo mouse brain MTw images.
  • FIG. 7B illustrates a CEST image at lppm and
  • FIG. 7C illustrates a CEST MTR aS ym spectrum.
  • the specific reduction of CEST contrast for LS174T cells is likely due to the different uMUC-1 glycosylation levels.
  • the different chemical shifts of the maximal effects between the in vitro and in vivo preparations may be due to different pH buffering and also back-exchange effects to other protons such as amide.
  • TFMS trifluoromethanesulfonic acid
  • MCF10A non-tumorigenic human breast carcinoma
  • LS174T and HT29 both human colon carcinomas
  • MUC-1 glycosylation levels were encapsulated in alginate-PLLalginate microcapsules at 1000 cells/capsule in order to minimize cell sedimentation and variations in cell density.
  • FIGS. 8A and 8B illustrate graphical views of a Zspectra and MTRasym for deglycosylated and untreated mucin.
  • FIG.8C is a MTRasym contrast map at 1.8ppm peak.
  • FIGS.8D The deglycosylation was confirmed by SDSPAGE electrophoresis (FIG.8D), where deglycosylated mucin showed a MW of 70-100kD, whereas untreated mucin did not show any bands due to the MW being >260kD8.
  • the CEST contrast produced by 3 cell lines with different MUC-1 expression was tested: LS174T and HT29, both expressing underglycosylated MUC-1 (i.e., "uMUC-1 positive"), and MCFlOA, expressing normally glycosylated MUC-1 (i.e. "uMUC-1 negative)”.
  • FIGS. 9A-9F illustrate microscopy, MTw image, CEST spectra and images, and imunostaining of encapsulated cell lines with different MUC-1 glycosylation levels.
  • FIG. 9C MTRasym spectra
  • FIG. 9D and 9E contrast maps
  • TFMS trifluoromethanesulfonic acid
  • deglycosylated mucin could be easily differentiated in both the Z-spectra and MTR aS ym spectra with a significant reduction of CEST contrast over a broad chemical shift range, i.e. a -80% reduction from 0.5 to 2 ppm and a -50% loss from 2 to 4 ppm, respectively.
  • Deglycosylation was confirmed by SDS-PAGE, where deglycosylated mucin showed a MW of 70-100kD, in contrast to untreated mucin which has an Mw >260kD6.
  • the CEST contrast produced by LS174T and HT29 were compared, both expressing underglycosylated MUC-1 and MCF10A, expressing normally glycosylated MUC-1.
  • the sugar chains on a crude purification of porcine stomach mucin protein (Sigma-Aldrich, M-2378, St Louis, MO) by treatment with
  • TFMS trifluoromethanesulfonic acid
  • underglycosylated MUC-1 i.e. uMUC-1 -positive; LS174T and HT29 human colon cancer cell- lines
  • normally glycosylated MUC-1 i.e. uMUC-1 -negative; MCF10A non- tumorigenic human breast cells.
  • Measurements were performed in vitro by encapsulating live cells in alginate-PLL-alginate microcapsules (-1000 cells per capsule; FIG. 9A) in order to minimize variations in tumor cell density. Microcapsules were suspended in PBS and loaded into 5 mm MR tubes with their layout shown on the MT image in FIG. 9(b).
  • the MUC-1 cancer marker exhibits differential CEST contrast between 0.5 and 4 ppm depending on the glycosylation level, which is lower for the two cell lines having
  • the pulse sequences, imaging protocols, described herein can be executed with a program(s) fixed on one or more non-transitory computer readable medium.
  • the non-transitory computer readable medium can be loaded onto a computing device, server, imaging device processor, smartphone, tablet, phablet, or any other suitable device known to or conceivable by one of skill in the art.
  • the computing device can be configured especially for use with the magnetic resonance imaging machine.
  • the computing device can also be integrated into the magnetic resonance imaging machine either directly or via network. It is noted that the computing device to carry out the present invention can be a unique computing device designed especially for use with the present invention and to address specific needs of the execution of the present invention.
  • the steps of the method described can be carried out using a computer, non-transitory computer readable medium, or alternately a computing device, microprocessor, or other computer type device independent of or incorporated with an imaging or signal collection device.
  • An independent computing device can be networked together with the imaging device either with wires or wirelessly.
  • any suitable method of analysis known to or conceivable by one of skill in the art could be used.
  • any suitable method of analysis known to or conceivable by one of skill in the art could be used.
  • equations are detailed herein, variations on these equations can also be derived, and this application includes any such equation known to or conceivable by one of skill in the art.
  • the computing device is unique to this application.
  • a non-transitory computer readable medium is understood to mean any article of manufacture that can be read by a computer.
  • Such non-transitory computer readable media includes, but is not limited to, magnetic media, such as a floppy disk, flexible disk, hard disk, reel-to-reel tape, cartridge tape, cassette tape or cards, optical media such as CD-ROM, writable compact disc, magneto-optical media in disc, tape or card form, and paper media, such as punched cards and paper tape.

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Abstract

La présente invention concerne une technique d'IRM pour évaluer la malignité de tumeur de façon non invasive. Elle est basée sur le principe selon lequel la plupart des cellules tumorales épithéliales malignes expriment des mucines qui sont fortement sous-glycosylées. Ce phénomène d'expression aberrante a été correctement documenté dans la littérature. Lorsque des cellules de tumeur bénigne ayant une glycosylation de mucine normale par IRM CEST sont soumises à imagerie, un effet de signal IRM CEST est observé. Lorsque les cellules tumorales perdent leur glycanes et deviennent malignes, la différence de signal IRM CEST est détectable. Par conséquent, les propriétés intrinsèques de glycosylation de mucine de surface de cellule tumorale peuvent être utilisées en tant que marqueur de substitution pour prédire la malignité de tumeur. Cela peut être effectué sans injecter des sondes exogènes ou des marqueurs, qui ont leurs propres problèmes en termes d'approbation clinique, de coût et la pharmacocinétique de liaison à toutes les cellules tumorales en question. Un tissu normal peut être utilisé en tant que référence pour déterminer des changements de contraste associés à une tumeur maligne.
PCT/US2016/015577 2015-01-30 2016-01-29 Imagerie rm sans marqueur de tumeur maligne WO2016123446A1 (fr)

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WO2014124006A1 (fr) * 2013-02-05 2014-08-14 The Johns Hopkins University Nanoparticules pour le suivi de l'imagerie par résonance magnétique et procédés de fabrication et d'utilisation associés
WO2014165719A1 (fr) * 2013-04-05 2014-10-09 The Trustees Of The University Of Pennsylvania Imagerie non invasive de l'état redox d'un tissu par irm
WO2014172359A1 (fr) * 2013-04-19 2014-10-23 The Johns Hopkins University Irm fondée sur le transfert de saturation par échange chimique (cest) utilisant des gènes rapporteurs et des substrats et procédés associés

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