PH is a tightly regulated physiological parameter that is often altered in diseased states like cancer. The development of biosensors that can be used to non-invasively image pH with hyperpolarized (HP) magnetic resonance spectroscopic imaging has therefore recently gained tremendous interest. However, most of the known HP-sensors have only individually and not comprehensively been analyzed for their biocompatibility, their pH sensitivity under physiological conditions, and the effects of chemical derivatization on their logarithmic acid dissociation constant (pK_a). Proteinogenic amino acids are biocompatible, can be hyperpolarized and have at least two pH sensitive moieties. However, they do not exhibit a pH sensitivity in the physiologically relevant pH range. Here, we developed a systematic approach to tailor the pK_a of molecules using modifications of carbon chain length and derivatization rendering these molecules interesting for pH biosensing. Notably, we identified several derivatives such as [1-¹³C]serine amide and [1-¹³C]-2,3-diaminopropionic acid as novel pH sensors. They bear several spin-1/2 nuclei (¹³C, ¹⁵N, ³¹P) with high sensitivity up to 4.8 ppm/pH and we show that ¹³C spins can be hyperpolarized with dissolution dynamic polarization (DNP). Our findings elucidate the molecular mechanisms of chemical shift pH sensors that might help to design tailored probes for specific pH in vivo imaging applications.

Increased extracellular sodium activates Th17 cells, which provide protection from bacterial and fungal infections. Whilst high salt diets have been shown to worsen autoimmune disease, the immunological consequences of clinical salt depletion are unknown. Here, we investigate immunity in patients with inherited salt-losing tubulopathies (SLT). Forty-seven genotyped SLT patients (with Bartter, Gitelman or EAST Syndromes) are recruited. Clinical features of dysregulated immunity are recorded with a standardised questionnaire and immunological investigations of IL-17 responsiveness undertaken. The effects of altering extracellular ionic concentrations on immune responses are then assessed. Patients are hypokalaemic and hypomagnesaemic, with reduced interstitial sodium stores determined by ²³Na-magnetic resonance imaging. SLT patients report increased mucosal infections and allergic disease compared to age-matched controls. Aligned with their clinical phenotype, SLT patients have an increased ratio of Th2:Th17 cells. SLT Th17 and Tc17 polarisation is reduced in vitro, yet STAT1 and STAT3 phosphorylation and calcium flux following T cell activation are unaffected. In control cells, the addition of extracellular sodium (+40 mM), potassium (+2 mM), or magnesium (+1 mM) reduces Th2:Th17 ratio and augments Th17 polarisation. Our results thus show that the ionic environment typical in SLT impairs IL-17 immunity, but the intracellular pathways that mediate salt-driven Th17 polarisation are intact and in vitro IL-17 responses can be reinvigorated by increasing extracellular sodium concentration. Whether better correction of extracellular ions can rescue the immunophenotype in vivo in SLT patients remains unknown.

Background: Three‐dimensional (3D) multiecho balanced steady‐state free precession (ME‐bSSFP) has previously been demonstrated in preclinical hyperpolarized (HP) ¹³C‐MRI in vivo experiments, and it may be suitable for clinical metabolic imaging of prostate cancer (PCa).Purpose: To validate a signal simulation framework for the use of sequence parameter optimization. To demonstrate the feasibility of ME‐bSSFP for HP ¹³C‐MRI in patients. To evaluate the metabolism in PCa measured by ME‐bSSFP.Study Type: Retrospective single‐center cohort study.Phantoms/Population: Phantoms containing aqueous solutions of [1‐¹³C] lactate (2.3 M) and [¹³C] urea (8 M). Eight patients (mean age 67 ± 6 years) with biopsy‐confirmed Gleason 3 + 4 (n = 7) and 4 + 3 (n = 1) PCa.Field Strength/Sequences: ¹H MRI at 3 T with T₂‐weighted turbo spin‐echo sequence used for spatial localization and spoiled dual gradient‐echo sequence used for B₀‐field measurement. ME‐bSSFP sequence for ¹³C MR spectroscopic imaging with retrospective multipoint IDEAL metabolite separation.Assessment: The primary endpoint was the analysis of pyruvate‐to‐lactate conversion in PCa and healthy prostate regions of interest (ROIs) using model‐free area under the curve (AUC) ratios and a one‐directional kinetic model (k_P). The secondary objectives were to investigate the correlation between simulated and experimental ME‐bSSFP metabolite signals for HP ¹³C‐MRI parameter optimization.Statistical Tests: Pearson correlation coefficients with 95% confidence intervals and paired t‐tests. The level of statistical significance was set at P < 0.05.Results: Strong correlations between simulated and empirical ME‐bSSFP signals were found (r > 0.96). Therefore, the simulation framework was used for sequence optimization. Whole prostate metabolic HP ¹³C‐MRI, observing the conversion of pyruvate into lactate, with a temporal resolution of 6 seconds was demonstrated using ME‐bSSFP. Both assessed metrics resulted in significant differences between PCa (mean ± SD) (AUC = 0.33 ± 012, k_P = 0.038 ± 0.014) and healthy (AUC = 0.15 ± 0.10, k_P = 0.011 ± 0.007) ROIs.Data Conclusion: Metabolic HP ¹³C‐MRI in the prostate using ME‐bSSFP allows for differentiation between aggressive PCa and healthy tissue.Evidence Level: 2Technical Efficacy: Stage 1

Hyperpolarized contrast agents (HyCAs) have enabled unprecedented magnetic resonance imaging (MRI) of metabolism and pH in vivo. Producing HyCAs with currently available methods, however, is typically time and cost intensive. Here, we show virtually-continuous production of HyCAs using parahydrogen-induced polarization (PHIP), without stand-alone polarizer, but using a system integrated in an MRI instead. Polarization of ≈2% for [1-¹³C]succinate-d₂ or ≈19% for hydroxyethyl-[1-¹³C]propionate-d₃ was created every 15 s, for which fast, effective, and well-synchronized cycling of chemicals and reactions in conjunction with efficient spin-order transfer was key. We addressed these challenges using a dedicated, high-pressure, high-temperature reactor with integrated water-based heating and a setup operated via the MRI pulse program. As PHIP of several biologically relevant HyCAs has recently been described, this Rapid-PHIP technique promises fast preclinical studies, repeated administration or continuous infusion within a single lifetime of the agent, as well as a prolonged window for observation with signal averaging and dynamic monitoring of metabolic alterations.

The aim of this study was to acquire the transient MRI signal of hyperpolarized tracers and their metabolites efficiently, for which specialized imaging sequences are required. In this work, a multi‐echo balanced steady‐state free precession (me‐bSSFP) sequence with Iterative Decomposition with Echo Asymmetry and Least squares estimation (IDEAL) reconstruction was implemented on a clinical 3 T positron‐emission tomography/MRI system for fast 2D and 3D metabolic imaging. Simulations were conducted to obtain signal‐efficient sequence protocols for the metabolic imaging of hyperpolarized biomolecules. The sequence was applied in vitro and in vivo for probing the enzymatic exchange of hyperpolarized [1–¹³C]pyruvate and [1–¹³C]lactate. Chemical shift resolution was achieved using a least‐square, iterative chemical species separation algorithm in the reconstruction. In vitro, metabolic conversion rate measurements from me‐bSSFP were compared with NMR spectroscopy and free induction decay‐chemical shift imaging (FID‐CSI). In vivo, a rat MAT‐B‐III tumor model was imaged with me‐bSSFP and FID‐CSI. 2D metabolite maps of [1–¹³C]pyruvate and [1–¹³C]lactate acquired with me‐bSSFP showed the same spatial distributions as FID‐CSI. The pyruvate‐lactate conversion kinetics measured with me‐bSSFP and NMR corresponded well. Dynamic 2D metabolite mapping with me‐bSSFP enabled the acquisition of up to 420 time frames (scan time: 180‐350 ms/frame) before the hyperpolarized [1–¹³C]pyruvate was relaxed below noise level. 3D metabolite mapping with a large field of view (180 × 180 × 48 mm³) and high spatial resolution (5.6 × 5.6 × 2 mm³) was conducted with me‐bSSFP in a scan time of 8.2 seconds. It was concluded that Me‐bSSFP improves the spatial and temporal resolution for metabolic imaging of hyperpolarized [1–¹³C]pyruvate and [1–¹³C]lactate compared with either of the FID‐CSI or EPSI methods reported at 3 T, providing new possibilities for clinical and preclinical applications.