Dana C. Peters1, Stefan Markovic2, Qingjia Bao2, Dina Preise2, Keren Sasson2, Lilach Agemy2, Avigdor Scherz2, and Lucio Frydman2
1Radiology and Biomed Eng., Yale University, New Haven, CT, United States, 2Weizmann Institute of Science, Rehovot, Israel
Synopsis
Deuterium metabolic imaging (DMI) maps the individual in vivo fate
of 2H-enriched metabolites. Upon injecting 2H6,6’-glucose,
DMI images a 2H-water peak, and a small but diagnostic 2H3,3’-lactate
signature, highlighting tumors and their aberrant metabolism. DMI faces major sensitivity challenges, that can
be alleviated by a multi-echo balanced SSFP approach. When suitably tuned, multi-echo
bSSFP yields good spectral isolation of all metabolites, and thanks to the relatively
large T2/T1 ratios of deuterated compounds, several-fold increases in SNR vs.
chemical shift imaging are then obtained. This is demonstrated in phantoms, and in in
vivo mice studies of orthotopic pancreatic tumors.
Introduction
Deuterium
metabolic imaging (DMI) is an emerging method 1-3 for mapping in
vivo and non-invasively the uptake of deuterated metabolites, and their marking
of metabolism. So far DMI employed T1-weighted
chemical shift imaging (CSI) sequences to differentiate the spatial
distribution of the individual compounds, and image them vs time. However, even when leveraging 2H’s
short T1s, DMI’s SNR is limited.
Ongoing studies have shown that abnormal tumor metabolism4 might
be highlighted by following the small but diagnostic 2H3,3’-lactate
signature arising ca. one hour after the injection of 2H6,6’-glucose.
We hypothesized that a multi-echo balanced steady state free precession (ME-bSSFP)
approach5 would increase this the sensitivity of this diagnosis several-fold
vis-à-vis CSI, while achieving good spectral isolation of metabolites. Supporting
this is the relatively high T2/T1 ratios of the metabolites observed in vivo,
increasing bSSFP’s SNR which scales as √(T2/T1).6 Furthermore, a ME approach coupled to the IDEAL
reconstruction7, could enable bSSFP’s Dixon-like isolation of the
various chemical shifts of interest. ME-bSSFP
has in fact been applied to hyperpolarized 13C spectroscopy8-10,
with a goal to leverage it for increasing the scanning speed rather than the SNR. This study shows both theoretically and
experimentally, that substantial increases in SNR, resulting in better 2H
spectral images, can be achieved by 2H ME-bSSFP.Methods
Simulations were run to optimize the
SNR of bSSFP and CSI, based on T2/T1 values observed for 2H6,6’-glucose
(30ms/60ms), for 2H3,3’-lactate (60ms/292ms) and for 2H-water
(12ms/290ms) (1). The SNR simulations included standard signal dependency of
bSSFP and GRE on T1/T2 and T2*6,11 and assumed equal scan times, and voxel sizes. Calculations were also done to explore ME-bSSFP’s
ability to isolate these spatially overlapping metabolites, given their
respective chemical shifts (3.7ppm, 1.3ppm and 4.8ppm) at a 2H Larmor
frequency of 99.34 MHz (15.2T). These spectral
selectivity simulations included realistic field-inhomogeneity maps, multiple signal
ratios for the metabolites, and added Gaussian noise. Simulations also helped us to determine a repetition
time TR and carrier frequency combination, so that all of the metabolites were
located outside of the bSSFP “dark bands”8.
Experiments: 2H ME-bSSFP and CSI sequences were written in
Paravision 6 (Figure 1), and suitably tuned experiments were implemented at 15.2
T on a Bruker scanner utilizing a double-resonance 1H/2H
setup based on surface coils. These results were compared to optimized CSI
sequences, both in deuterated phantoms and in vivo. Scan time was 10 minutes for all experiments. CSI reconstruction used standard methods
(spatial and spectral filtering, zero-filling, 3D FFT12). ME-bSSFP reconstruction was performed using
2D FFT, then IDEAL processing using the ISMRM fat-water toolbox, and
Hierarchical IDEAL13.
Phantoms: Agar-based samples containing the targeted metabolites were made, and
the spectral isolation provided by optimized ME-bSSFP was evaluated. SNRs of CSI and ME-bSSFP at equal spatial
resolution (1x1x8mm3 with a 32x32 matrix) were compared.
Animals: A KPC mouse tumor model of pancreatic adenocarcinoma (PDAC)
was investigated, with 2H experiments performed on IACUC-approved
studies of ten mice at various stages of tumor growth. To compare SNR performance, 32x32 CSI were acquired, in consecutive randomized order, with 32x32 ME-bSSFP. SNRs
were measured in matched ROIs. However, as lactate was not visible on CSI at
this resolution except in one case, 8x8 CSI (5x5x8mm3) data sets were also acquired
and the spatial/temporal patterns of the metabolites compared. Results
Simulations
showed that for the targeted 2H compounds bSSFP could provide
several-fold increases in SNR (Figure 2), depending on T2*. Suitable settings of
the ME-bSSFP sequence –5 TEs, ΔTE=2.2ms, TR=12.2ms, θ=60° and a carrier frequency placed at 2.6ppm to
avoid dark bands– also provided well isolated metabolites after IDEAL
reconstruction even with limited homogeneities and SNR (Figure 3; smaller
echo-spacings were found useful to deal with larger field inhomogeneities). Phantom experiments executed under such
conditions showed a four-fold increase in lactate’s ME-bSSFP image, two-fold
for the glucose, and a minor improvement for water, vis-à-vis an optimized CSI collected
with TR/θ=95ms/90° (Figure 4). Figure 5 compares CSI and ME-bSSFP images from
an animal with a large tumor collected ≈2 hrs after injection of glucose, at
equal spatial resolution. The spatial
locations of glucose, water and lactate agree well, as they did in all in
vivo studies (which typically compared 8x8 CSI to 32x32 ME-bSSFP scans). At equal spatial resolution and scan time, glucose’s
in vivo ME-bSSFP SNR was 3-fold greater CSI (57±30 vs. 19±10, p<0.001),
and water’s SNR was doubled (Figure 5B).
Lactate’s SNR could not be compared, as lactate was usually invisible on
32x32 CSI studies. The sole exception to this is shown in Figure 5A, for which
ME-bSSFP’s SNR was 3-fold higher.Discussion
Simulations, phantoms and in vivo
results indicate that, for DMI at 15.2T, ME-bSSFP provides substantial boosts
in SNR per unit time compared to CSI. This can be translated into higher
resolution, faster imaging, or greater sensitivity to map lactate’s production.
Given the quadrupolar nature of 2H’s relaxation and the flexibility
afforded by the TE choices the translation of this method to lower field strengths
seems very feasible, greatly facilitating the power of DMI to image
non-invasively the Warburg effect.Acknowledgements
This work was
supported by the Kimmel and by the Clore Institutes for Magnetic Resonance
(Weizmann Institute), by the Israel Science Foundation (grant 965/18), the
Thompson Family Foundation, and the Israel Cancer Research Foundation. DCP
acknowledges the Benoziyo Endowment Fund for the Advancement of Science, for a
Visiting Faculty Fellowship to the Weizmann Institute, and thanks Yale
Radiology for sabbatical time. References
1. De Feyter HM, Behar
KL, Corbin ZA, Fulbright RK, Brown PB, McIntyre S, Nixon TW, Rothman DL, de
Graaf RA. Deuterium metabolic imaging (DMI) for MRI-based 3D mapping of
metabolism in vivo. Sci Adv 2018;4(8):eaat7314.
2. de Graaf RA,
Hendriks AD, Klomp DWJ, Kumaragamage C, Welting D, Arteaga de Castro CS, Brown
PB, McIntyre S, Nixon TW, Prompers JJ, De Feyter HM. On the magnetic field
dependence of deuterium metabolic imaging. NMR Biomed 2020;33(3):e4235.
3. Kreis F, Wright AJ,
Hesse F, Fala M, Hu DE, Brindle KM. Measuring Tumor Glycolytic Flux in Vivo by
Using Fast Deuterium MRI. Radiology 2020;294(2):289-296.
4. Warburg O, Wind F,
Negelein E. The Metabolism of Tumors in the Body. J Gen Physiol
1927;8(6):519-530.
5. Herzka DA, Kellman
P, Aletras AH, Guttman MA, McVeigh ER. Multishot EPI-SSFP in the heart. Magn
Reson Med 2002;47(4):655-664.
6. Bieri O, Scheffler
K. Fundamentals of balanced steady state free precession MRI. J Magn Reson
Imaging 2013;38(1):2-11.
7. Reeder SB, Pineda
AR, Wen Z, Shimakawa A, Yu H, Brittain JH, Gold GE, Beaulieu CH, Pelc NJ.
Iterative decomposition of water and fat with echo asymmetry and least-squares
estimation (IDEAL): application with fast spin-echo imaging. Magn Reson Med
2005;54(3):636-644.
8. Leupold J, Mansson
S, Petersson JS, Hennig J, Wieben O. Fast multiecho balanced SSFP metabolite
mapping of (1)H and hyperpolarized (13)C compounds. MAGMA 2009;22(4):251-256.
9. Leupold J, Wieben
O, Mansson S, Speck O, Scheffler K, Petersson JS, Hennig J. Fast chemical shift
mapping with multiecho balanced SSFP. MAGMA 2006;19(5):267-273.
10. Perman WH,
Bhattacharya P, Leupold J, Lin AP, Harris KC, Norton VA, Hovener JB, Ross BD.
Fast volumetric spatial-spectral MR imaging of hyperpolarized 13C-labeled
compounds using multiple echo 3D bSSFP. Magn Reson Imaging 2010;28(4):459-465.
11. Ernst RR, Anderson
WA. Application of Fourier Transform Spectroscopy to Magnetic Resonance. Rev
Sci Instrum 1966;37(1):93-+.
12 Markovic S, Fages A,
Roussel T, Hadas R, Brandis A, Neeman M, Frydman L. Placental physiology
monitored by hyperpolarized dynamic (13)C magnetic resonance. Proc Natl Acad
Sci U S A 2018;115(10):E2429-E2436.
13. Tsao J, Jiang Y.
Hierarchical IDEAL: fast, robust, and multiresolution separation of multiple
chemical species from multiple echo times. Magn Reson Med 2013;70(1):155-159.