Low CLOCK and CRY2 in 2nd trimester human maternal blood and risk of preterm birth: a nested case-control study†

Biol Reprod. 2021 Oct 11;105(4):827-836. doi: 10.1093/biolre/ioab119.


Previous studies have observed an association between maternal circadian rhythm disruption and preterm birth (PTB). However, the underlying molecular mechanisms and the potential of circadian clock genes to serve as predictors of PTB remain unexplored. We examined the association of 10 core circadian transcripts in maternal blood with spontaneous PTB (sPTB) vs term births using a nested case-control study design. We used a public gene expression dataset (GSE59491), which was nested within the All Our Babies (AOB) study cohort in Canada. Maternal blood was sampled in Trimesters 2-3 from women with sPTB (n = 51) and term births (n = 106), matched for five demographic variables. In 2nd trimester maternal blood, only CLOCK and CRY2 transcripts were significantly lower in sPTB vs term (P = 0.02-0.03, false discovery rate (FDR) < 0.20). A change of PER3 mRNA from trimesters 2-3 was significantly associated with sPTB (decline in sPTB, P = 0.02, FDR < 0.20). When CLOCK and CRY2 were modeled together in 2nd trimester blood, the odds of being in the low level of both circadian gene transcripts was greater in sPTB vs term (OR = 4.86, 95%CI = (1.75,13.51), P < 0.01). Using GSVA and Pearson correlation, we identified 98 common pathways that were negatively or positively correlated with CLOCK and CRY2 expression (all P < 0.05, FDR < 0.10). The top three identified pathways were amyotrophic lateral sclerosis, degradation of extracellular matrix, and inwardly rectifying potassium channels. These three processes have previously been shown to be involved in neuron death, parturition, and uterine excitability during pregnancy, respectively.

Keywords: CLOCK; CRY2; circadian clock genes; gene set variation analysis; human; logistic regression; pregnancy; spontaneous preterm birth.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Alberta / epidemiology
  • CLOCK Proteins / blood
  • CLOCK Proteins / deficiency*
  • Case-Control Studies
  • Cryptochromes / blood
  • Cryptochromes / deficiency*
  • Female
  • Humans
  • Mothers
  • Pregnancy
  • Pregnancy Trimester, Second
  • Premature Birth / epidemiology*
  • Premature Birth / etiology
  • Young Adult


  • CRY2 protein, human
  • Cryptochromes
  • CLOCK Proteins
  • CLOCK protein, human