Journal of Cystic Fibrosis

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Journal of Cystic Fibrosis Pilot study to test inhaled nitric oxide in cystic fibrosis patients with refractory Mycobacterium abscessus lung infection Lea Bentur, Michal Gur, Moshe Ashkenazi, Galit Livnat-Levanon, Marko Mizrahi, Asher Tal, Abdi Ghaffari, Yuval Geffen, Micha Aviram, Ori Efrati Journal of Cystic Fibrosis DOI: 10.1016/j.jcf.2019.05.002 Copyright © 2019 The Authors Terms and Conditions

Fig. 1 Methemoglobin and NO2 remained within accepted range during intermittent iNO treatment. Panel A: Pooled MetHb levels from before (baseline) and after NO inhalation (NO) are shown in a Box-Whisker plot. The dotted line marks the 7% MetHb safety threshold set in this study. Panel B: Inspired nitrogen dioxide (NO2) levels in delivery line were continuously monitored by sampling from the mouthpiece sample port with an AeroNOx electrochemical analyzer. Pooled inspired NO2 levels from start to end of NO inhalation are shown. The dotted line marks the 5% NO2 safety threshold set in the study. Box and Whisker plot description: boxes extend from the 25th to 75th percentile, middle line represents the median, and + symbol shows the mean value. Whiskers extend to minimum and maximum values. Journal of Cystic Fibrosis DOI: (10.1016/j.jcf.2019.05.002) Copyright © 2019 The Authors Terms and Conditions

Fig. 2 The effect of iNO treatment on lung function and 6MWD. Mean changes from baseline (day 1) in FEV1 (panel A) and FVC (panel B) measured by spirometry in all patients (n = 9) during iNO treatment (iNO Therapy: week 1–3) and at follow up (Off Therapy: weeks 7 and 11). CI indicates 95% confidence intervals. Matched non-parametric ANOVA (Friedman test) did not reveal a significant difference (FEV1 p = .220; FVC p = .41). 6MWD in meters (panel C) was assessed each morning before NO inhalation (iNO Therapy) and at follow up (Off Therapy). Matched non-parametric ANOVA (Friedman test) did not reveal a significant difference (p = .71). Values above each bar represent means change of all patients. Journal of Cystic Fibrosis DOI: (10.1016/j.jcf.2019.05.002) Copyright © 2019 The Authors Terms and Conditions

Fig. 3 iNO treatment increases time to positivity in mycobacterium culture. Panels A: time to positivity of mycobacteria culture was assessed in sputum samples of all patients at screening, during NO treatment (NO Therapy), and at follow up (Off Therapy). Median values plus 95% confidence intervals (CI) are presented here. No significant difference was found from baseline (Wilcoxon matched-pairs signed rank test). Panel B: assessment of uncultured sputum samples for M. abscessus DNA by qPCR using M. abscessus-specific rpoB gene. Data generated in qPCR was converted to CFU/ml by generating a CFU/qPCR standard curve. Mean values (plus standard deviation – SD) from all 9 patients are represented here. No significant difference was observed between baseline and time points (Wilcoxon matched-pairs signed rank test). Journal of Cystic Fibrosis DOI: (10.1016/j.jcf.2019.05.002) Copyright © 2019 The Authors Terms and Conditions