Plasma microRNA interpersonal variability in healthy individuals, pregnant women, and an individual with a stably-altered neuroendocrine phenotype

Large cohort: exRNA composition of platelet-depleted plasma from a cohort of 173 female and 63 male healthy volunteers of diverse races and ages, collected at two visits spaced two weeks apart; 4 formerly-collected aliquots from study subject P12, who displayed a stable neuroendocrine miRNA signature in plasma; 20 plasma samples of pregnant women spanning all trimesters T1 to T3 of pregnancy, with an expected placenta miRNA contribution. P12 Family: exRNA from two replicates of platelet-depleted plasma from three unstudied P12 family members and a sample from P12 collected four years after the initial discovery of the phenotype, along with samples of two healthy volunteers. Plasma subfractions: exRNA of samples from P12 and his family, two healthy male volunteers, two non-pregnant, and nine pregnant women, separated by differential ultracentrifugation into 10,000 × g and 100,000 × g pellets, and corresponding 10,000 × g and 100,000 × g supernatants. sRNA-derived cDNA profiling of exRNA isolated from platelet-depleted EDTA plasma and 10,000 x g and 100,000 x g plasma subfractions. Pellet and supernatant fractions were generated by differential ultracentrifugation. Automated RNA isolation and multiplexed library preparation were done using propriatory protocols (KEA Max et al., PNAS, 2018).