Nicholas Drachman1, Stephen J. Kadlecek1, Mehrdad Pourfathi1,2, Yi Xin1, Harrilla Profka1, and Rahim R. Rizi1
1Radiology, University of Pennsylvania, Philadelphia, PA, United States, 2Electrical and Systems Engineering, University of Pennsylvania, Philadelphia, PA, United States
Synopsis
In this study, we investigate the possibility of simultaneously imaging pH and lactate to pyruvate ratio in vivo in the lungs. We produce hyperpolarized 13C-bicarbonate by rapidly decarboxylating hyperpolarized [1-13C]pyruvate with hydrogen peroxide. By tuning the reaction rate by altering the pH, we produce roughly equal amounts of pyruvate and bicarbonate, which allows us to image both metabolic processes simultaneously.Introduction
Many pathological processes are associated with changes
in endogenous pH, including cancer, inflammation, and ischemia [1]. In
vivo pH imaging is possible using hyperpolarized
13C-bicarbonate,
but achieving a high level of polarization in a short time remains a major
challenge. This challenge can be overcome by polarizing [1-
13C]-pyruvate
by DNP and then rapidly converting it to [
13C-]bicarbonate by
decarboxylation via hydrogen peroxide while minimizing loss of polarization.
The rate of decarboxylation can be tuned by altering the pH of the reactants,
thereby allowing us to obtain the desired ratio of pyruvate to bicarbonate. By
allowing the reaction to proceed to roughly 50% decarboxylation, both pyruvate
and bicarbonate are present in high enough concentrations to measure local pH
and lactate-to-pyruvate ratio. In this experiment, we demonstrate the
effectiveness of this method by locally obtaining these measurements in inflamed
rat lungs.
Methods
Two
Sprague Dawley rats (400g) were ventilated with tidal volume of 12mL/kg and positive
end-expiratory pressure (PEEP) of 3 cmH
2O for t = 180 minutes. After
70 minutes the lungs were injured via tracheal HCl aspiration (pH 1.25,
2.5mL/kg) to simulate gastric acid aspiration. Rats were imaged in supine position using
a
1H/
13C Quadrature birdcage coil (m2m) in a 4.7T
horizontal magnet (Varian Inc.). The
hyperpolarized agent was produced by polarizing 100.1mg of [1-
13C]pyruvate
(Cambridge Isotopes Inc.) in a Hypersense DNP system. 5mL of 280mM hydrogen
peroxide was prepared for the reaction by diluting 1.57mL of unstabilized 3%
hydrogen peroxide solution with 3.43mL of DI water. After polarizing for one
hour in the DNP system, the hyperpolarized pyruvate was diluted in 4mL of
dissolution medium, which produces a neutral, isotonic, 280mM solution. 2.5mL
of the 280mM peroxide solution and 2.5mL of the 280mM hyperpolarized pyruvate
solution were mixed together with a small amount of NaOH to raise the pH near
12 where the reaction proceeds most rapidly. The solution was given roughly 8
seconds to react before being neutralized and injected into the rat via a tail
vein catheter. The lungs were imaged using a slice selective 16x16 single-point
CSI pulse sequence with an outward spiral k-space trajectory and an in-plane field
of view 50x50 mm
2 over a 30 mm axial slice to cover the entire lungs
(TR/TE = 70/0.38 ms, SW = 6.0 kHz, α=12°). The data obtained was processed using custom
routines in MATLAB 2014b (MathWorks Inc.). The pH in each voxel was calculated
using the Henderson-Hasselbach equation, where the relative signal intensities
are used in place of the relative concentrations (the validity of this
substitution is based on the rapid interconversion of bicarbonate to CO
2 due
to the endogenous presence of carbonic anhydrase). The lactate-to-pyruvate
ratio in each voxel was similarly calculated using the integrals under each
metabolite’s respective peak. Both images were thresholded based on total
carbon signal to remove noise seen on the perimeter of and outside of the
lungs.
Results
Carbonate polarizations of over 13% were achieved using this method. A reduction in pH was observed in
nearly all regions of the inflamed lungs as compared to the value of 7.4 seen
in healthy tissue. The pH in each voxel ranges from 7.0 to 7.4 as seen in
figure
3, it is believed that these differences correspond to local variation in the
level of inflammation. The mean pH
measured over the entire volume of the lungs was 7.18±0.14, which is well
within the expected range for inflamed tissue. At this pH, the CO
2 signal
is ~10 times smaller than the bicarbonate signal, however the polarization is
high enough that the CO
2 signal is quantifiable in all voxels in the
lung. A sample voxel is shown in
figure 1. In these experiments, the
decarboxylation reaction was tuned to produce roughly equal amounts of
hyperpolarized pyruvate and bicarbonate. With these parameters, there is a high
enough concentration of pyruvate to quantify the amount of lactate produced metabolically;
the lactate-to-pyruvate map obtained is shown in
figure 4. The mean lactate-to-pyruvate
ratio over the volume of the lungs was calculated to be ~0.07, which is
consistent with other experiments done by our group.
Conclusions
In
this study we demonstrated a method to locally measure the pH in inflamed
lung tissue in vivo using
13C-bicarbonate produced via the
decarboxylation of hyperpolarized [1-
13C]-pyruvate. We have also
shown the feasibility of simultaneously imaging both pH and lactate-pyruvate
ratio in the lungs with a single injection.
Acknowledgements
No acknowledgement found.References
[1] Grinstein, et al. (1991) Regulation of cyto- plasmic
pH in phagocytic cell function and dysfunction. Clin. Biochem. 24, 241–247.