Christian Prinz1, Paula Ramos Delgado1, Thomas Wilhelm Eigentler1, Ludger Starke1, Thoralf Niendorf1,2, and Sonia Waiczies1
1Berlin Ultrahigh Field Facility, Max Delbrück Center for Molecular Medicine, Berlin, Germany, 2Experimental and Clinical Research Center, a joint cooperation between the Charité Medical Faculty and the Max Delbrück Center for Molecular Medicine, Berlin, Germany
Synopsis
Fluorine-19
(19F) magnetic resonance is a powerful tool for tracking fluorine
labelled markers, cells and drugs. Here, we studied the influence of temperature
on the 19F MR characteristics (chemical shift, T1, T2,
SNR) of four fluorinated drugs using 19F MR mapping and spectroscopy
techniques. We demonstrated the impact of temperature on the T1 relaxation
time of PFCE nanoparticles in vivo
and postmortem. Our findings open a trajectory toward 19F MR-based
thermometry and indicate the need for adapting MR sequence parameters according
to temperature induced environmental changes. This will be an essential requirement
for monitoring fluorinated compounds by 19F MR techniques in vivo.
Introduction
Fluorine-19
(19F) magnetic resonance techniques are of paramount relevance for a
multitude of biomedical applications1,2. The 19F signal of fluorinated
compounds depends on 19F MR properties and on environmental factors3. Here, we examined the influence of
temperature on the chemical shift (CS), the 19F spectrum, signal
intensity (SI), SNR, longitudinal (T1) and transversal (T2)
relaxation times of perfluoro-15-crown-5-ether (PFCE), the anesthetic
isoflurane (Iso), the anti-inflammatory drug teriflunomide (TF) and the
antipsychotic flupentixol (Flu) in phantom studies. We also investigated the impact
of temperature on the T1 relaxation of PFCE nanoparticles in vivo and postmortem.Methods
All
fluorinated compounds were prepared in 2ml syringe phantoms: 1.72M PFCE, 0.54M isoflurane
in DMSO, 100mM teriflunomide in DMSO and 169mM flupentixol in medium chain triglycerides. A water bath was used to heat the
sample (T=20-60°C in increments of 5°C). For absolute temperature measurements
and management, a fiber optic probe was used. For the in vivo and post mortem studies, a fiber optic temperature probe was
inserted under the skin in the neck region of an anesthetized C57BL/6 mouse.
PFCE nanoparticles were administered subcutaneously in the neck region.
All MR experiments
were performed on a 9.4T MR scanner (Bruker Biospec, Ettlingen, Germany) using
a dual-tunable 19F/1H mouse head RF coil4. 2D-FLASH was used for pilot scans,
3D-RARE was used for 1H and 19F imaging. Global single
pulse spectroscopy was used to detect the 19F signal and to make
frequency adjustments. T1 mapping was performed using a saturation
based RARE technique (TE=4.6ms, ETL=4, FOV=16mmx16mm, matrix size=64x64, with 9
variable repetitions times (TR=25ms-8000ms). T2 mapping was
performed using a multi-slice multi-echo technique (TR=2000ms, FOV=16mmx16mm,
matrix size=64x64) with 25 TEs (TE=40-1000ms, increment 40ms; for flupentixol
TE=8-200ms, increment 8ms).
Image
processing and spectral analysis were performed in MATLAB R2018a. We calculated CS, FIDFit (SI), SNR, T1 and T2
and all MR parameters were plotted versus the measured median temperature. Results
PFCE demonstrates a
spectrum with a single peak (Fig.1A). Upon increasing temperature, the CS
decreased (+0.0082ppm/°C)(Fig.1B+C). A negative correlation of the SI (Fig.1D) and
the SNR (Fig.1E) was observed. T1 increased with temperature (ΔT1=+16ms/°C)(Fig.1G). We also
observed a linear increase of T2 (7ms/°C)(Fig.1I).
Isoflurane
showed one major and several minor peaks (Fig.2A). Isoflurane revealed a CS dependence
versus temperature of ΔCS=+0.0025ppm/°C (Fig.2B+C). We observed a
decrease in SI (Fig.2D) and SNR (Fig.2E) with increasing temperatures. T1
increased with a linear change of ΔT1=+30ms/°C (Fig.2G), T2
with a linear increase of ΔT2=11ms/°C (Fig.2I).
Teriflunomide
showed a spectrum with a single peak (Fig.3A) and a temperature dependent CS
change of ΔCS=-0.0038ppm/°C
(Fig.3B+C). A negative correlation of SI (Fig.3D) and SNR (Fig.3E) with
increasing temperatures was found. T1 mapping revealed a linear T1
change of ΔT1=6.6ms/°C
(Fig.3G). T2 mapping revealed a T2 increase of 4.9ms/°C up
to T=35°C. For T>35°C T2 decline (Fig.3I) was observed.
Flupentixol
yielded a single-peak spectrum (Fig.4A) and, similar to teriflunomide, a negative
CS at varying temperatures of DCS= -0.0026ppm/°C (Fig.4B+C). We observed a
linear dependence of the SI (Fig.4D) and the SNR (Fig.4E) versus temperature. There
is a linear T1 increase of ΔT1=1.3ms/°C (Fig.4G) and
a linear T2 increase of ΔT2=7.4ms/°C (Fig.4I).
T1 mapping
was performed in vivo at 32.4°C in
the neck region (Fig.5B), revealing a T1 of 1325ms. At room
temperature (23.6°C) the T1 observed ex vivo was reduced by 13% (1149ms)(Fig.5C).Discussion
En route to 19F
MR-based thermometry, we investigated the influence of the temperature on the 19F
MR characteristics of several compounds with different chemical properties and configurations
of fluorine containing groups. The degree of influence varied between each
substance, such as the ranges of the CS changes and the degree of T1
and T2 change per unit temperature.
Varying temperatures lead to an increase in the molecular tumbling rate. Therefore,
temperature related changes in dipole-dipole interactions and the anisotropy of
the fluorine chemical shift and J-coupling affect the chemical shift and relaxation
times. Conclusions
Understanding the 19F MR characteristics and temperature
influence of pharmacological compounds with a low 19F content and
low availability will be crucial for in
vivo studies. Together with optimizing
scan parameters and strategies to increase the sensitivity of 19F MR, this will make 19F
MR a powerful technique for studying biological processes and monitoring drug
therapies.Acknowledgements
This work
was supported by funding from the Germany Research Council (DFG WA2804). This project was funded in part (TN) by an advanced ERC
grant (EU project ThermalMR - DLV-743077).References
1 Schmieder, A. H., Caruthers, S.
D., Keupp, J., Wickline, S. A. & Lanza, G. M. Recent Advances in 19Fluorine
Magnetic Resonance Imaging with Perfluorocarbon Emulsions. Engineering (Beijing, China) 1,
475-489, doi:10.15302/j-eng-2015103 (2015).
2 Ruiz-Cabello, J., Barnett, B. P.,
Bottomley, P. A. & Bulte, J. W. Fluorine (19F) MRS and MRI in biomedicine. NMR in biomedicine 24, 114-129, doi:10.1002/nbm.1570 (2011).
3 Colotti, R. et al. Characterization of perfluorocarbon relaxation times and
their influence on the optimization of fluorine-19 MRI at 3 tesla. Magnetic resonance in medicine 77, 2263-2271, doi:10.1002/mrm.26317
(2017).
4 Waiczies, H. et al. Visualizing brain inflammation with a shingled-leg
radio-frequency head probe for 19F/1H MRI. Scientific
reports 3, 1280,
doi:10.1038/srep01280 (2013).