Multi-omics analysis of primary cytotrophoblasts from second trimester and term placentas — ASN Events
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  2. Lunch/Poster Session Two
  3. Multi-omics analysis of primary cytotrophoblasts from second trimester and term placentas

Multi-omics analysis of primary cytotrophoblasts from second trimester and term placentas (#221)

Christie Hunter 1 , Katy Williams 2 , Andrew Olson 3
  1. SCIEX, Redwood City, CA, United States
  2. University of California, San Francisco, CA, United States
  3. Advaita Biosciences, Plymouth, MI, United States

During human pregnancy, a subset of placental cytotrophoblasts (CTBs) differentiates into cells that aggressively invade the uterus and its vasculature, anchoring the progeny and rerouting maternal blood to the placenta. Defects in this process are the hallmark of the pregnancy complication preeclampsia. While disease-associated genes or transcripts may serve as useful biomarkers, they are not necessarily predictive of disease mechanisms. Thus, we performed global proteomic and transcriptional profiling to measure expression patterns of CTBs from second trimester and term normal placentas to gain further understanding of CTB differentiation in healthy pregnancy.

Primary CTBs were isolated using collagenase and trypsin digestion and Percoll gradient centrifugation. For proteomics analysis, cells were lysed and digested with trypsin. Variable window SWATH® Acquisition data were acquired using the TripleTOF 6600 System (SCIEX). A 180 min gradient using a nanoLC™ 425 with a cHiPLC® system (SCIEX) was performed for separation. SWATH data were processed using OneOmics™ applications in BaseSpace (Illumina). For transcriptomics, a second set of CTBs were analyzed using RNAseq (Kundaje, Nature 2015). iPathwayGuide (Advaita) was used to compare protein and RNA levels for pathway and gene ontology analyses.

Approximately 3000 proteins were quantified and ~400 showed differential expression in second trimester CTBs vs. term. Proteins known to function in CTB differentiation processes, e.g. angiogenesis and hypoxia response, as well as previously uncharacterized processes, e.g., NF-kappaB signaling, metal ion transport and muscle contraction, were altered. Integration with RNAseq data showed variations at the molecular level but concordance among biological processes. Multi-omics data sets comprised of SWATH MS protein quantification and RNAseq expression results showed gestation age differences in healthy CTB populations corresponding to known and novel processes. In future, these methods will be used to study CTBs from patients with preeclampsia is likely to identify aberrations that could contribute to disease and/or serve as diagnostic markers.