CEST Imaging Research Articles

Progress toward quantitative in vivo chemical exchange saturation transfer (CEST) MRI

AbstractChemical exchange saturation transfer (CEST) MRI provides a sensitive detection mechanism for imaging dilute labile protons, complementing the routine radiological exams. Enormous progress has been achieved in CEST MRI and image analysis, from the mathematical modeling, CEST agent design, and most importantly, increasing adoption of CEST imaging in the clinical setting. Therefore, CEST imaging represents an emerging field that involves multiple disciplines and together made a remarkable transition from the simplistic CEST‐weighted MRI to quantitative CEST (qCEST) analysis. This review focuses on the recent advancements in CEST quantification techniques and findings of in vivo CEST imaging in representative disorders of ischemia, tumor, and epilepsy. In addition, limitations of current CEST methodologies are examined that should help guide future development of more sensitive and quantitative CEST imaging techniques and ultimately, facilitate their adoption in the clinical setting.

In vivo imaging of brain glutamate defects in a knock-in mouse model of Huntington's disease

Huntington's disease (HD) is an inherited neurodegenerative disease characterized by motor, cognitive and psychiatric symptoms. Atrophy of the striatum has been proposed for several years as a biomarker to assess disease progression in HD gene carriers. However, it does not provide any information about the biological mechanisms linked to HD pathogenesis. Changes in brain metabolites have been also consistently seen in HD patients and animal models using Magnetic Resonance Spectroscopy (MRS), but metabolite measurements are generally limited to a single voxel. In this study, we used Chemical Exchange Saturation Transfer imaging of glutamate (gluCEST) in order to map glutamate distribution in the brain of a knock-in mouse model (Ki140CAG) with a precise anatomical resolution. We demonstrated that both heterozygous and homozygous mice with pathological CAG repeat expansion in gene encoding huntingtin exhibited an atrophy of the striatum and a significant alteration of their metabolic profile in the striatum as compared to wild type littermate controls. The striatal decrease was then confirmed by gluCEST imaging. Surprisingly, CEST imaging also revealed that the corpus callosum was the most affected structure in both genotype groups, suggesting that this structure could be highly vulnerable in HD. We evaluated for the first time gluCEST imaging as a potential biomarker of HD and demonstrated its potential for characterizing metabolic defects in neurodegenerative diseases in specific regions.

Insight into the quantitative metrics of chemical exchange saturation transfer (CEST) imaging.

To evaluate the reliability of four CEST imaging metrics for brain tumors, at varied saturation power levels and magnetic field strengths (3-9.4 Tesla (T)). A five-pool proton exchange model (free water, semisolid, amide, amine, and NOE-related protons) was used for the simulations. For the in vivo study, eight glioma-bearing rats were scanned at 4.7 T. The CEST ratio (CESTR), CESTR normalized with the reference value (CESTRnr ), inverse Z-spectrum-based (MTRRex ), and apparent exchange-related relaxation (AREX) were compared. The simulated CEST signal intensities using MTRRex and AREX were substantially increased at relatively high radiofrequency (RF) saturation powers at 3 T and 4.7 T, whereas CESTR and CESTRnr metrics remained relatively stable. There were tremendously high MTRRex and AREX signals around the water frequency at all field strengths because of the small denominators. In the rat tumor study at 4.7 T, both CESTR and CESTRnr showed clear contrasts in the tumor with respect to the normal tissue across all saturation power levels (0.5-3 μT), whereas the AREX showed negligible to negative insignificant contrasts. CEST metrics must be carefully selected based on the different experimental settings. CESTR and CESTRnr are more reliable at 3 T (a clinical field strength) and 4.7 T. Magn Reson Med 77:1853-1865, 2017. © 2016 International Society for Magnetic Resonance in Medicine.

Abstract 63: Novel Imaging of Protein Integrity to Better Define Ischemic Injury After Stroke

Introduction: Ischemic injury on MRI is defined acutely by measuring the diffusion of water alone. Chemical exchange saturation transfer imaging (CEST) allows measurement of intracellular pH, and changes in protein structure and mobility using Nuclear Overhauser Enhancement (NOE). When identifying treatment targets, changes to protein integrity have the potential to complement diffusion-weighted imaging (DWI) in distinguishing irreversible from reversible intracellular processes that are amenable to therapeutic intervention. Methods: Patients with non-lacunar ischemic stroke underwent serial MRI over 1 month. The imaging protocol included single slice CEST imaging. NOE was quantified using a 3-pool exchange model relative to the contralateral hemisphere (rNOE*). DWI, ASL perfusion imaging, T1-weighted and FLAIR imaging were used to define the tissue outcome: ischemic core, infarct growth, and oligemic tissue that survived. Infarct growth was classified as early or late (before or after 24 hours). Images and masks were registered to CEST native image space for voxelwise and patient-level serial analyses. Results: 30 patients were included in the analysis. Within 6 hours rNOE* in early infarct growth was significantly lower than ischemic core and late infarct growth (p<0.001 and p<0.001, see Figure). In patient-level analyses, rNOE* dropped over the initial 24 hours in the ischemic core and early infarct growth regions, but not in regions of late infarct growth or oligemia. Conclusion: rNOE* showed a discrete temporal profile in tissue that infarcted by 24 hours in contrast to that which survived or infarcted later. This is additive information to that provided by routine acute stroke MRI at presentation, and demonstrates that the imaging of intracellular protein structure and mobility may have a role alongside DWI in helping to predict tissue fate. Further work is required to understand the potential of NOE as a modifiable imaging biomarker.

Investigation of Readout RF Pulse Impact on the Chemical Exchange Saturation Transfer Spectrum.

Chemical exchange saturation transfer magnetic resonance imaging (CEST-MRI) is capable of both microenvironment and molecular imaging. The optimization of scanning parameters is important since the CEST effect is sensitive to factors such as saturation power and field homogeneity. The aim of this study was to determine if the CEST effect would be altered by changing the length of readout RF pulses. Both theoretical computer simulation and phantom experiments were performed to examine the influence of readout RF pulses. Our results showed that the length of readout RF pulses has unremarkable impact on the Z-spectrum and CEST effect in both computer simulation and phantom experiment. Moreover, we demonstrated that multiple refocusing RF pulses used in rapid acquisition with relaxation enhancement (RARE) sequence induced no obvious saturation transfer contrast. Therefore, readout RF pulse has negligible effect on CEST Z-spectrum and the optimization of readout RF pulse length can be disregarded in CEST imaging protocol.

Multi-echo length and offset VARied saturation (MeLOVARS) method for improved CEST imaging.

The aim of this study was to develop a technique for rapid collection of chemical exchange saturation transfer images with the saturation varied to modulate signal loss transfer and enhance contrast. Multi-echo Length and Offset VARied Saturation (MeLOVARS) divides the saturation pulse of length Tsat into N = 3-8 submodules, each consisting of a saturation pulse with length of Tsat /N (∼0.3-1 s), one or more low flip-angle gradient-echo readout(s) and a flip back pulse. This results in N readouts with increasing saturation time from Tsat /N to Tsat without extra scan time. For phantoms, eight images with Tsat incremented every 0.5 s from 0.5-4 s were collected simultaneously using MeLOVARS, which allows rapid determination of exchange rates for agent protons. For live mice bearing glioblastomas, the Z-spectra for five different Tsat values from 0.5 to 2.5 s were acquired in a time normally used for one Tsat . With the additional Tsat -dependence information, LOVARS phase maps were produced with a more clearly defined tumor boundary and an estimated 4.3-fold enhanced contrast-to-noise ratio (CNR). We also show that enhancing CNR is achievable by simply averaging the collected images or transforming them using the principal component analysis. MeLOVARS enables collection of multiple saturation-time-weighted images without extra time, producing a LOVARS phase map with increased CNR.

Improvement of gagCEST imaging in the human lumbar intervertebral disc by motion correction.

To investigate whether motion correction improves glycosaminoglycan chemical exchange saturation transfer imaging (gagCEST imaging) of intervertebral discs (IVDs). Magnetic resonance gagCEST imaging of 12 volunteers was obtained in lumbar IVDs at 3 T using a prototype pulse sequence. The data were motion-corrected using a prototype diffeomorphism-based motion compensation technique. For both the data with and that without motion correction (datac, datauc), CEST evaluation was performed using the magnetisation transfer ratio asymmetry (MTRasym) as a means of quantifying CEST effects. MTRasym and the signal-to-noise ratio (SNR) of the MTRasym map in the nucleus pulposus (NP) were compared for datac and datauc. A visual grading analysis was performed by a radiologist in order to subjectively quantify the quality of the MTRasym analysis (score 1: best quality, score 5: worst quality). Furthermore, a landmark analysis was performed in order to objectively quantify the motion between CEST images using the mean landmark distance dmean. MTRasym and SNR were significantly higher for the motion-corrected data than for the uncorrected CEST data (MTRasym(datac) = 3.77 % ± 0.95 %, MTRasym(datauc) = 3.41 % ± 1.54 %, p value = 0.001; SNR(datac) = 3.88 ± 2.04, SNR(datauc) = 2.77 ± 1.55, p value < 0.001, number of IVDs = 48). The visual grading analysis revealed a higher reliability for datac (maximum score = 2) compared with datauc (maximum score = 5). The landmark analysis demonstrated the superiority of the motion-corrected data (dmean(datac) = 0.08 mm ± 0.09 mm, dmean(datauc) = 0.36 mm ± 0.09 mm, p value = 0.001). Our study showed significant improvements in the ability to quantify CEST imaging in IVDs after the application of motion correction compared with uncorrected datasets.

Emerging techniques and technologies in brain tumor imaging.

The purpose of this report is to describe the state of imaging techniques and technologies for detecting response of brain tumors to treatment in the setting of multicenter clinical trials. Within currently used technologies, implementation of standardized image acquisition and the use of volumetric estimates and subtraction maps are likely to help to improve tumor visualization, delineation, and quantification. Upon further development, refinement, and standardization, imaging technologies such as diffusion and perfusion MRI and amino acid PET may contribute to the detection of tumor response to treatment, particularly in specific treatment settings. Over the next few years, new technologies such as 2(3)Na MRI and CEST imaging technologies will be explored for their use in expanding the ability to quantitatively image tumor response to therapies in a clinical trial setting.

The presence of fast-exchanging proton species in aqueous solutions of paraCEST agents can impact rate constants measured for slower exchanging species when fitting CEST spectra to the Bloch equations.

LnDOTA-tetraamide complexes typically exist in solution as a mixture of square-antiprismatic (SAP) and twisted square-antiprismatic (TSAP) coordination isomers. In most cases, the SAP isomer, which is preferred for CEST imaging, predominates, and the presence of the minor TSAP isomer is assumed to have little influence on quantitative measures of the water-exchange rate constant for the SAP isomer. Here, we sought to confirm the validity of this assumption by mixing two chelates with different SAP and TSAP isomer populations while measuring the water-exchange rate constant of the SAP isomer. The results show that an increase in the population of the TSAP isomer in solution results in as much as a 30% overestimation of the water-exchange rate constant for the SAP isomer when CEST spectra are fit to the Bloch equations. This effect was shown to be significant only when the TSAP isomer population exceeded 50%.

The pH sensitivity of –NH exchange in LnDOTA–tetraamide complexes varies with amide substituent

The amide proton exchange rates in various lanthanide(III) DOTA-tetraamide complexes were investigated by CEST as a function of variable chemical structures and charges on the amide substituents. Comparisons were made between YbDOTA-(gly)(4)(-) (Yb-1), YbDOTA-(NHCH(2)PO(3))(4 (5-) (Yb-2) and YbDOTA-(NHCH(2)PO(3)Et(2))(4)(3+) (Yb-3). The general shapes of the CEST vs pH profiles were similar for the three complexes but they showed maximum CEST intensities at different pH values, pH 8.3, 8.8 and 6.9 for Yb-1, Yb-2 and Yb-3, respectively. This indicates that a more negatively charged substituent on the amide helps stabilize the partial positive charge on the amide nitrogen and consequently more base is required to catalyze proton exchange. The chemical shifts of the -NH protons in Yb-1 and Yb-2 were similar (-17 ppm) while the -NH proton in Yb-3 was at -13 ppm. This shows that the crystal field produced by the amide oxygen donor atoms in Yb-3 is substantially weaker than that in the other two complexes. In an effort to expand the useful range of pH values that might be measured using these complexes as CEST agents, the shapes of the CEST vs pH curves were also determined for two thulium(III) complexes with much larger hyperfine shifted -NH proton resonances. The ratio of CEST from -NH exchange in Tm-1 compared with CEST from -NH exchange in Tm-3 was found to be linear over an extended pH range, from 6.3 to 7.4. This demonstrates a potential advantage of using mixtures of lanthanide(III) DOTA-tetraamides for mapping tissue pH by use of ratiometric CEST imaging.

Iron(II) PARACEST MRI Contrast Agents

The first examples of Fe(II) PARACEST magnetic resonance contrast agents are reported (PARACEST = paramagnetic chemical exchange saturation transfer). The iron(II) complexes contain a macrocyclic ligand, either 1,4,7-tris(carbamoylmethyl)-1,4,7-triazacyclononane (L1) or 1,4,7-tris[(5-amino-6-methyl-2-pyridyl)methyl]-1,4,7-triazacyclononane (L2). The macrocycles bind Fe(II) in aqueous solution with formation constants of log K = 13.5 and 19.2, respectively, and maintain the Fe(II) state in the presence of air. These complexes each contain six exchangeable protons for CEST which are amide protons in [Fe(L1)](2+) or amino protons in [Fe(L2)](2+). The CEST peak for the [Fe(L1)](2+) amide protons is at 69 ppm downfield of the bulk water resonance whereas the CEST peak for the [Fe(L2)](2+) amine protons is at 6 ppm downfield of bulk water. CEST imaging using a MRI scanner shows that the CEST effect can be observed in solutions containing low millimolar concentrations of complex at neutral pH, 100 mM NaCl, 20 mM buffer at 25 °C or 37 °C.

Methods for an improved detection of the MRI‐CEST effect

CEST imaging is a recently introduced MRI contrast modality based on the use of endogenous or exogenous molecules whose exchangeable proton pools transfer saturated magnetization to bulk water, thus creating negative contrast. One of the critical issues for further development of these agents is represented by their limited sensitivity in vivo. The aim of this work is to improve the detection of CEST agents by exploring new approaches through which the saturation transfer (ST) effect can be enhanced. The performance of the proposed methods has been tested in vitro and in vivo using highly sensitive and highly shifted lipoCEST agents, and the results were compared with the standard ST evaluation mode. The acquired Z-spectra were interpolated locally and voxel-by-voxel by smoothing splines. Besides expressing the ST in the standard mode, we explore two methods, enhanced and integral ST, which better exploit all the information contained in the Z-spectrum. By combining different modes for ST assessment a significant improvement in the detection of the lipoCEST agents, both in vitro and in vivo, has been found. The results obtained from the application of the proposed methods outline the importance of post-processing analysis for highlighting the CEST-MRI contrast.

Amplification strategies in MR imaging: Activation and accumulation of sensing contrast agents (SCAs)

We review new strategies for the development of Gd3+-based T1-relaxation agents and paramagnetic chemical exchange saturation transfer (PARACEST) "sensing" contrast agents (SCAs) designed specifically to detect small molecules or enzymatic activity in living systems. The first class of agents exhibits molecular "sensing" properties as a result of water coordination sphere effects, cleavage, or synthesis of reactive precursor compounds that recombine with macromolecules with the resultant formation of immobilized or rotationally constrained paramagnetic cations. This effect results in changes of water proton relaxation times. The second class (PARACEST) comprises a family of lanthanide-based paramagnetic compounds suitable for CEST imaging. The need for both types of MR agents is justified by efforts to utilize magnetic resonance imaging (MRI) to visualize fine structures in living tissue, and to increase the molecular specificity of MRI.