On resonance variable delay multiple pulse CEST MRI detected a transient reduction in lactate levels in the cerebrospinal fluid during the disease course of a mouse model of multiple sclerosis, which was validated using magnetic resonance spectroscopy.
Purpose
The lack of specificity in diagnosing multiple sclerosis (MS) with MRI has encouraged the development of supplemental detection methods. The cerebrospinal fluid (CSF) contains certain metabolites that are known to change as the disease progresses1,2. Since on resonance variable delay multiple pulse (onVDMP) CEST MRI has previously been shown to be able to detect several of these metabolites when containing fast exchanging protons3, e.g. hydroxyl and amine protons, we explored onVDMP CEST MRI monitoring of the CSF during the disease course of experimental autoimmune encephalomyelitis (EAE), a mouse model of MS.Induction of EAE: EAE was induced in C57Bl/6 mice (n=6-8) by i.p. injection of 250 ng of pertussis toxin and s.c. injection of 300 μg of MOG35-55 in IFA supplemented with 4 mg/ml tuberculin on day 0 and day 2. Mice were monitored daily and assessed for paralysis using the following clinical scoring system: 0=healthy, 1=tail paralysis, 2=mild hind limb paralysis, 3=total hind limb paralysis, 4=forelimb paralysis, and 5=moribund/death. Naïve mice were used as a control (n=4-5).
MRI/MRS: Mice were imaged weekly using a horizontal bore Biospec 11.7 T scanner. For transmission, a 72-mm volume resonator was used. For acquisition, 2x2 and 4x1 phased array coils were used for the brain and spinal cord, respectively. Slice thickness was 2 mm for all images except those used for lesion detection (0.7 mm for T1w/FLASH and for T2w images). Other parameters are:
· T2w imaging: TE=20 ms, TR=2 s, RARE factor=8, number of averages (NA)=4, and FOV=128x128.
· T1w/FLASH imaging: TE=6 ms and TR=300 ms, NA=2, and FOV=128x128.
· onVDMP CEST imaging: TE=5 ms, TR=5 s, Rare factor=10, NA=1, repetitions=9, B1=46.8 μT, 32 pulse-exchange modules with a 2 ms pulse width, eight delays (mixing times exponentially distributed from 1.14 to 100 ms), and FOV=128x128. Variation of onVDMP CEST signal over time (%) was defined as 100 * (onVDMPAFTER – onVDMPBEFORE) / onVDMPBEFORE.
· STEAM spectroscopy: TE=3 ms, TR=2.5 s, NA=128, and voxel size = 4x2x2 mm, TARQUIN4 was used to determine metabolite concentrations (mM) of peaks fitted with the standard 1H brain library. Metabolite content analyzed in these studies was normalized to total creatine content.
Statistics: Significance was defined at p<0.05 using an ANOVA with Tukey post-hoc, Student t-test or Pearson correlation, as appropriate.
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