Ying Liu1,2, Botao Zhao1,2, and Xiao-Yong Zhang1,2
1Institute of Science and Technology for Brain-Inspired Intelligence, Fudan University, Shanghai, China, 2Key Laboratory of Computational Neuroscience and Brain-Inspired Intelligence (Fudan University), Ministry of Education, Shanghai, China
Synopsis
CEST-MRI
has emerged as a noninvasive pH imaging technique based on the pH-dependence of
the prototropic exchange rates. In this work, we compared several recently
published CEST pH imaging methods and proton exchange rate value in rat glioma
models at 11.7T. All of
these pH-weighted contrasts show a pH correlation and have significantly
different signal intensities between the tumor region and the surrounding
normal tissue. Our
results demonstrated that pH enhanced
method and APT imaging are not concentration-independent while the sensitivity
of AACID is affected by the weak amine signals at 2.7ppm.
Introduction
Chemical exchange
saturation transfer (CEST) MRI has emerged as a noninvasive pH imaging
technique for several kinds of diseases such as tumor, relying on the
pH-dependence of the prototropic exchange rates1,2. Recently, several endogenous CEST methods
including corrected amide proton transfer (APT) imaging3,4, amine and amide
concentration-independent detection (AACID)5,6, and pH enhanced method with the
combination of amide and guanidyl CEST (pHenh)7 have been exploited to obtain pH maps and
hold great clinical potentials. In the present work,
we aimed to compare the performance of these pH-weighted contrasts with
exchange rate value and relative proton concentration using multi-pool
Lorentzian fit8 and omega plot method9,10 in rat glioma models at 11.7T.Materials and Methods
Animal model: All experimental
protocols were approved by the Animal Ethics Committee of Fudan University. A total of 6 Male Sprague Dawley
rats (180−200 g)
were purchased from Shanghai Slac Lab Animal Ltd (Shanghai, China). C6
rat glioblastoma cells were stereotaxically injected into the right cerebral
hemisphere to establish glioma model.
Data Acquisition: MRI experiments
were performed at 11.7T BioSpec 117/16 USR MRI system with a surface coil for
receiving (Bruker BioSpin, Ettlingen, Germany). T2-weighted images of the whole brain
were acquired using a Rapid Acquisition with Relaxation Enhancement (RARE)
sequence with the following parameters: TR/TE, 3500/30 ms; RARE factor, 8;
spatial resolution, 0.125 × 0.125 × 0.8 mm3. Single-slice CEST
images were obtained by a continuous wave (CW) RF irradiation (B1=0.6,
0.8, 1.1, 1.5, 2.0μT) followed by a
RARE sequence to readout with parameters: TR/TE=5000/41ms, RARE factor, 24;
spatial resolution, 0.33 × 0.33 × 2 mm3. Saturation offsets were
distributed between –30 and 30ppm with denser sampling between –5 and 5ppm and S0
was set at 200 ppm. T1 value was measured using the same RARE
sequence with 6 different inversion times (5500ms, 3000ms, 1500ms, 800ms, 500ms,
and 300ms).
Data analysis: After normalization,
B0 correction, and adaptive denoising11, the Z- spectra were fitted as a
linear combination of multiple Lorentzian functions12. Then the exchange rate (kb)
and labile proton ratio (fb) were estimated using the omega
plot method10 with multi-pool fitted 1/MTRRex values
at 2.0 ppm. And several recently published pH-weighted contrasts including pHenh7 with B1=1.1μT at 3.6ppm and B1=0.6μT at 2ppm, AACID5 with B1=1.1μT, and APT resolved from multi-pool model4 with B1=1.1μT were also calculated for comparison.
Results and Discussion
Fig. 1a presents representative normalized Z spectra
from a normal region of interest with varied B1
from 0.6 μT to 2.0μT. Fig. 1b shows the corresponding six-pool Lorentzian fitting results and the subtracted
residual (dark stem plot) at B1=1.0μT. Individual CEST contributions
(amide, amine, NOE (-1.6), NOE (-3.5), DS, and MT) were isolated well from the
entire Z-spectrum. As shown in Fig. 1c, the omega plot analysis display the
expected linear relationship between the 1/MTRRex and 1/ ω12.
And the exchange
rate values appear lower in the tumor region (kb= 416.81±18.14) than
in the normal tissue (kb=511.25±26.90), indicating lower pH values in tumor according to the previous
reports13,14. Fig. 2 illustrates
example maps of the exchange rate (Fig. 2b), proton ratio (Fig. 2c), and
the commonly used pH-weighted MRI (Fig. 2d-e). Tumor
with
hyper-intensive signals in the T2W image (Fig.2a) was delineated by the red dotted line. Notably, signal intensity in
the tumor region shows a clear difference compared to the surrounding normal
tissue among all of these contrast maps. The variation trends of pHenh,
AACID, and APT values in these experiments agree well with the original
literature researches4,5,7.
Fig. 3 shows the scatter distributions
and Person correlation coefficients among the fb, kb with
the other three pH-weighted contrasts of all voxels from 6 mice brain. Signals
from tumor region were isolated using the red points. A significant negative correlation
was observed among pHenh with both kb and fb (Fig.
3a, d), which may reflect that pH
enhanced method is not concentration-independent for pH imaging. Fig. 3b
& 3e show that AACID approach allows the removal of the concentration but relatively
less sensitive to pH detection, which may due to the weak amine signals at
2.7ppm. As shown in the last column (Fig. 3c, f), APT effects have a strong
correlation with the proton ratio and a weaker correlation with the exchange
rate, indicating that accurate pH values cannot be calculated using this method
due to the varied concentration of labile protons in tumors.Conclusion
This
work investigated the association between several recently published endogenous
CEST pH-weighted imaging methods (corrected APT, AACID, pHenh) and direct exchange
rate value and relative proton concentration using DS-removed omega plot method in rat glioma
models at 11.7T. All of these pH-weighted contrast maps show a pH
correlation and have significantly different signal intensities between the tumor
region and the surrounding normal tissue. Our
results demonstrated that pH enhanced method and
APT imaging are not concentration-independent while the sensitivity of AACID is
affected by the weak amine signals at
2.7ppm.
Acknowledgements
This work was supported by grants from
the National Natural Science Foundation of China (81873893), Three-year action
plan for key global partners (IDH6282008/050/001/004), and Shanghai Municipal
Science and Technology Major Project (2018SHZDZX01). References
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