The bSSFP sequence provides high spatial and temporal resolution capabilities, but has a difficult to manage frequency response at 3T with regards to hyperpolarized [1-13C]pyruvate and its products. The purpose of this project was to integrate a spectral suppression pulse, designed to suppress alanine and pyruvate-hydrate, with the bSSFP sequence to image [1-13C]pyruvate and its conversion to [1-13C]lactate. The results showed no significant effect on quantitative analysis of lactate-to-pyruvate ratios or kpl after suppression of alanine and pyruvate-hydrate. Subsequently, dynamic imaging of [1-13C]pyruvate and [1-13C]lactate at high in-plane spatial resolution was achieved with the bSSFP sequence.
A 20ms SLR maximum-phase suppression pulse6 with an excitation bandwidth of 150Hz was initially tested in a slab-selective chemical shift imaging (CSI) sequence. The in vivo parameters were: two 12cm axial slabs, one localized on liver and one on kidney, 8cm FOV, progressive flip angle scheme,8 double-spin echo, 120ms TE, 25kHz spectral width, 2048 spectral points, 3s temporal resolution, 10 time-points, with one suppression pulse (centered ~158Hz upfield of lactate, between the alanine and pyruvate-hydrate resonances) played out prior to each time-point, ending 1ms before slab excitation. Each Sprague-Dawley rat was subjected to one scan with spectral suppression and one without spectral suppression. Thermal 13C phantom parameters were similar, except the suppression pulse center frequency was centered on lactate and moved downfield in 10Hz increments up to 180Hz.
Subsequent bSSFP imaging, acquired as a coronal projection, utilized the following parameters: 12x6 cm2 FOV, 60x30 matrix size for 2x2 mm2 in-plane resolution (or 40x20 for 3x3 mm2), progressive flip angle scheme, 6 time-points and 5s temporal resolution for each metabolite, three suppression pulses played before each time-point as described above. The RF pulse width and TR/TE were simulated and chosen to selectively excite either pyruvate or lactate, whereby a 6.8ms TBW2 sinc pulse led to a TR/TE of 15.3/7.65ms. An alternating center frequency scheme was utilized, with lactate acquired first, followed by pyruvate 2.5s later. [13C]Urea phantom experiments were acquired for one time-point in a similar manner, with 10x10 mm2 spatial resolution, and alternating between on-resonance and 390Hz off-resonance.
A dynamic 2D CSI sequence was used for comparison of lactate-to-pyruvate ratios and had the following parameters: 8cm FOV, axial 8cm slab, 8x8 matrix size leading to 10x10 mm2 in-plane resolution, 5kHz spectral width, 256 spectral points, TR of 76ms, progressive flip angle scheme, 6 time-points. All data was analyzed in Matlab, except the 2D CSI, which was analyzed in SIVIC.9 The experiments were conducted on a 3T MR scanner and DNP experiments used a HyperSense polarizer. All scans started at 20s after beginning of injection and 3mL of 80mM [1-13C]pyruvate was injected over 12s via tail vein catheters in six different rats.
As seen in the phantom results, the spectral suppression pulse showed no effect on resonances 100+Hz away (Fig. 1). Within the kidneys and liver, the pulse was able to successfully suppress both alanine and pyruvate-hydrate without having an effect on the lactate or pyruvate resonances (Fig. 2). Calculations of lactate-to-pyruvate ratios from the integration of summed spectra and calculations of kpl showed no statistically significant difference between spectral suppression and no spectral suppression in either kidney (p=0.499, p=0.416) or liver (p=0.400, p=0.057), indicating no significant effect on quantitative analysis.
Simulations and phantom tests demonstrated individual selectivity of pyruvate and lactate with the bSSFP sequence after spectral suppression (Fig. 3). While part a shows the drawback to having a long TR with the bSSFP sequence, the pulse width and TR used here can be shortened by a factor of 2-3 using an optimally designed RF spectrally selective pulse,7 which also has improved off-resonance insensitivity. Figure 4 shows successful acquisition of each metabolite at both 2x2 mm2 and 3x3 mm2, while Figure 5 indicates correlation in dynamic lactate-to-pyruvate ratios between the bSSFP acquisitions and the 2D CSI acquisitions.
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