Linewidth-related bias in modelled concentration estimates from GABA-edited 1H-MRS

J-difference-edited MRS is widely used to study GABA in the human brain. Editing for low-concentration target molecules (such as GABA) typically exhibits lower signal-to-noise ratio (SNR) than conventional non-edited MRS, varying with acquisition region, volume and duration. Moreover, spectral lineshape may be influenced by age-, pathology-, or brain-region-specific effects of metabolite T2, or by task-related blood-oxygen level dependent (BOLD) changes in functional MRS contexts. Differences in both SNR and lineshape may have systematic effects on concentration estimates derived from spectral modelling. The present study characterises the impact of lineshape and SNR on GABA+ estimates from different modelling algorithms: FSL-MRS, Gannet, LCModel, Osprey, spant and Tarquin. Publicly available multi-site GABA-edited data (222 healthy subjects from 20 sites; conventional MEGA-PRESS editing; TE = 68 ms) were pre-processed with a standardised pipeline, then filtered to apply controlled levels of Lorentzian and Gaussian linebroadening and SNR reduction. Increased Lorentzian linewidth was associated with a 2-5% decrease in GABA+ estimates per Hz, observed consistently (albeit to varying degrees) across datasets and most algorithms. Weaker, often opposing effects were observed for Gaussian linebroadening. Variations are likely caused by differing baseline parametrization and lineshape constraints between models. Effects of linewidth on other metabolites (e.g., Glx and tCr) varied, suggesting that a linewidth confound may persist after scaling to an internal reference. These findings indicate a potentially significant confound for studies where linewidth may differ systematically between groups or experimental conditions, e.g. due to T2 differences between brain regions, age, or pathology, or varying T2* due to BOLD-related changes. We conclude that linewidth effects need to be rigorously considered during experimental design and data processing, for example by incorporating linewidth into statistical analysis of modelling outcomes or development of appropriate lineshape matching algorithms. Highlights In-vivo GABA-edited 1H-MRS data from 222 subjects were filtered to simulate varying linewidth and SNR conditions Filtered datasets were quantified with six different modelling algorithms to assess the impact of linewidth and SNR on the metabolite level estimates. Synthetic spectra with controlled GABA+ levels and in-vivo-like background signals (applied incrementally) were also assessed. For both in-vivo and synthetic datasets, GABA+ estimates showed a significant association with Lorentzian linewidth across most algorithms, even for small changes in linewidth. Weaker, often opposing associations were observed for Gaussian linebroadening. This indicates a potentially significant confound for studies where linewidth or lineshape may be expected to differ, even slightly, between groups. The need for appropriate strategies to account for lineshape differences is highlighted. Graphical Abstract To assess the degree to which aspects of linewidth, lineshape and SNR may confound GABA+ estimates, a collection of in-vivo datasets were quantified with six modelling algorithms, with linebroadening and SNR varied experimentally. Most algorithms showed a strong association between GABA+ estimate and Lorentzian linebroadening (2-5% decrease per Hz), with weaker effects for Gaussian broadening. This indicates a potentially significant confound in cases of differing relaxation parameters between groups or experimental conditions.