Co-alterations of circadian clock gene transcripts in human placenta in preeclampsia.

Pre-eclampsia (PE) is a hypertensive condition that occurs during pregnancy and complicates up to 4% of pregnancies. PE exhibits several circadian-related characteristics, and the placenta possesses a functioning molecular clock. We examined the associations of 17 core circadian gene transcripts in placenta with PE vs. non-PE (a mixture of pregnant women with term, preterm, small-for-gestational-age, or chorioamnionitis) using two independent gene expression datasets: GSE75010-157 (80 PE vs. 77 non-PE) and GSE75010-173 (77 PE and 96 non-PE). We found a robust difference in circadian gene expression between PE and non-PE across the two datasets, where CRY1 mRNA increases and NR1D2 and PER3 transcripts decrease in PE placenta. Gene set variation analysis revealed an interplay between co-alterations of circadian clock genes and PE with altered hypoxia, cell migration/invasion, autophagy, and membrane trafficking pathways. Using human placental trophoblast HTR-8 cells, we show that CRY1/2 and NR1D1/2 regulate trophoblast migration. A subgroup study including only term samples demonstrated that CLOCK, NR1D2, and PER3 transcripts were simultaneously decreased in PE placenta, a finding supported by CLOCK protein downregulation in an independent cohort of human term PE placenta samples. These findings provide novel insights into the roles of the molecular clock in the pathogenesis of PE.

Quercetin supplementation to the thawing and incubation media of boar sperm improves post-thaw sperm characteristics and the in vitro production of pig embryos.

Elevated levels of reactive oxygen species can cause oxidative stress, which could lead to membrane damage, decreased fertility, and spermatozoan morphological deformities. Antioxidants can be supplemented to reduce the impacts of oxidative stress. The objective of this study was to determine the effects of supplementing quercetin (0.25, 0.50, 0.75 mM) during the thawing and incubation of frozen-thawed boar semen on spermatozoan characteristics, IVF kinetics (n = 400) and subsequent embryonic development (n = 1340). Spermatozoa were evaluated for motility, viability, and membrane lipid peroxidation levels at 0, 2, 4, 6, 8, and 10 h after thawing. Embryos were evaluated for IVF kinetics 12 h after IVF (penetration, polyspermy, male pronucleus formation, IVF efficiency) and cleavage and blastocyst formation at 48 h and 144 h after IVF, respectively. Spermatozoa supplemented with 0.25 mM quercetin had significantly higher (P < 0.05) motility (51.67±8.50 %) and percent of viable cells (61.21 ± 2.44 %) compared to all other treatments at 10 h after thawing, in addition to having significantly (P < 0.05) lower levels of hydroperoxide (3.38 ± 0.88 µM/107cells). There were no differences in penetration rates and male pronucleus formation between treatment groups. Supplementation of quercetin significantly decreased (P < 0.05) polyspermy and significantly increased (P < 0.05) the percentage of embryos reaching blastocyst stage of development by 144 h after IVF compared to no supplementation. Results indicated that supplementing frozen-thawed boar semen with 0.25 mM quercetin improves sperm characteristics up to 10 h after thawing and decreases polyspermy while improving early embryonic development in pigs.

Using antagonistic pleiotropy to design a chemotherapy-induced evolutionary trap to target drug resistance in cancer.

Local adaptation directs populations towards environment-specific fitness maxima through acquisition of positively selected traits. However, rapid environmental changes can identify hidden fitness trade-offs that turn adaptation into maladaptation, resulting in evolutionary traps. Cancer, a disease that is prone to drug resistance, is in principle susceptible to such traps. We therefore performed pooled CRISPR-Cas9 knockout screens in acute myeloid leukemia (AML) cells treated with various chemotherapies to map the drug-dependent genetic basis of fitness trade-offs, a concept known as antagonistic pleiotropy (AP). We identified a PRC2-NSD2/3-mediated MYC regulatory axis as a drug-induced AP pathway whose ability to confer resistance to bromodomain inhibition and sensitivity to BCL-2 inhibition templates an evolutionary trap. Across diverse AML cell-line and patient-derived xenograft models, we find that acquisition of resistance to bromodomain inhibition through this pathway exposes coincident hypersensitivity to BCL-2 inhibition. Thus, drug-induced AP can be leveraged to design evolutionary traps that selectively target drug resistance in cancer.