Extended Data Fig. 1: Analysis of miRNAs that increase during ALS course.
From: Circulating miR-181 is a prognostic biomarker for amyotrophic lateral sclerosis
(a) Plasma levels of four miRNAs of the 129 miRNAs analyzed in main Fig. 1a displayed low inter-individual variability, but increased with the disease course, suggesting that, although they are not suited for prognostic use, they could potentially monitor disease progression. (miR-423/484/92a/b, t4/t1 > 1.5 SD, X-axis) (B-D) MA plots of differential miRNA expression upon repeated sampling relative to the first phlebotomy. Red features denote miRNAs with statistically significant change in levels. Temporal changes in the levels of (E) miR-423-5p, (F) miR-484, (G) miR-92a-3p, or (H) miR-92b-3p revealed correlation with time passing from enrollment (in months). Spaghetti plots of individual patient trajectories (t1-t4) denoted for (I) miR-423-5p, t21 = 4.52, p = 0.0002; (J) miR-484, t21 = 4.05,p = 0.0006; (K) miR-92a-3p, t21 = 3.85, p = 0.0009 or (L) miR-92b-3p, t21 = 4.77, p = 0.0001. Time intervals: t1-t2 6.3 ± 0.3 m.; t1-t3 13.0 ± 0.3 m.; t1-t4 32.7 ± 3 m. Disease duration: t1 28.8 ± 3 m.; t4 61.5 ± 3 m. Validation of changes to miRNA levels in an independent replication cohort (N = 26 individuals, Table 2). Spaghetti plots of individual patient trajectories (t1-t2 13.7 ± 1.6 months) in a replication cohort, for (M) miR-423-5p, t24 = 0.98, p = 0.17 (N) miR-484, t25 = 2.08, p = 0.02; (O) miR-92a-3p, t25 = 2.13, p = 0.02; or (P) miR-92b-3p, t23 = 1.55, p = 0.067. Together, miR-484 and miR-92a/b may be considered as candidate molecular biomarkers of functional decline over the course of disease. Data presented as Mean ± SEM. *p < 0.05, ***p < 0.001, paired t-test. Analysis of a single miR-423-5p sample and two miR-92b-3p samples in the replication cohort deviated from the mean according to Grubbs test and these were excluded as outliers. Correlation between the relative disease covered (rD50) in longitudinal plasma collections (X-axis) and levels of (Q) miR-423-5p, (R) miR-484, (S) miR-92a-3p, and (T) miR-92b-3p (Y-axis). The relative D50 (rD50) is a derivative of ALS Functional Rating Scale-Revised (ALSFRS-R) decay that reveals the disease covered by individual patients independent of the rate of progression24,39. For example, an rD50 of 0.0 signifies ALS onset, and 0.5 signifies the time-point where functionality is reduced by half. Longitudinal miR-484/92a/b levels in blood correlated with rD50 at the time of sampling (R-T). All statistical tests were two-sided, except for panels M-P.
