ENDOCANNABINOID CEREBROSPINAL FLUID LEVELS IN MIGRAINE 81 4 individuals there was not enough CSF sample available (< 1.5 mL). Next, one population outlier with migraine with aura was excluded based on the PCA plot, leaving 194 people with migraine and 94 controls for the statistical analysis. After inclusion, n = 5 healthy controls were found to have a first-degree relative with migraine. However, their results were kept in the study, because CSF from healthy volunteers is very difficult to obtain. The clinical characteristics are shown in Table 1. There were no differences between the migraine groups, except for a higher monthly attack frequency in migraine with aura (1.5 IQR 1.0 - 3.0) versus migraine without aura (2.0 IQR 1.0 - 3.5). However, there was a difference between the three groups in, stress during LP (P = 0.024), all HADS related scores (p < 0.001), total CES-D score (p < 0.001) and a lower weekly alcohol consumption (P = 0.009), the latter three being in line with prior results on migraine comorbidities and lifestyle characteristics (Table 1).2, 3, 34 Data was assumed to be missing at random for weekly alcohol consumption (n = 6), lifetime depression (n = 45), HADS scores (n = 9), total CES-D (n = 48) and drug use (n = 1). The missing values used for the regression analysis were imputed. No group differences were observed for fasting time (in hours) and time of CSF collection (data not shown). CSF levels of AEA, 2-AG and DHEA in migraine patients and controls Multivariate linear regression analyses showed that there were no differences in concentrations in migraine with aura and migraine without aura versus control for each of the three eCBs (Figure 1A-C) P = 0.46 and P = 0.07 for AEA, P = 0.40 and P = 1.00 for 2-AG and P = 0.65 and P = 0.79 for DHEA, respectively. Some of the covariates, however, did have a significant effect on eCB concentration in the regression model. AEA concentration was associated with sex (β = -0.077; 95% CI: -0.12 to -0.04, P < 0.001) and number of cigarettes per day (β = 0.007; 95% CI: 0.00 to 0.01, P = 0.008). Female sex was associated with a lower AEA concentration (Figure 2A) and the number of cigarettes per day was positively correlated with the AEA concentration (Figure 2B). 2-AG concentration was negatively correlated with female sex (β = -0.083; 95% CI: -0.12 to -0.05, P < 0.001) (Figure 3A) and positively correlated with lifetime depression (β = 0.060; 95% CI: 0.020 to 0.10, P = 0.003) (Figure 3B and 3C). DHEA concentration was positively correlated with age (β = 0.003; 95% CI: 0.00 to 0.00, P < 0.001) and BMI (β = 0.010; 95% CI: 0.00 to 0.02, P = 0.010) (Figure 4A-B).
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