DESCRIPTION OF DIFFERENTIALLY EXPRESSED GENES BETWEEN HEALTHY NON‐PREGNANT AND PREGNANT WOMEN IN AN RNAseq TRANSCRIPTOME STUDY
Authors: de Lima Hirata Andrea Harumy , de Jesus Rocha Luiz Antonio , Silva Valdelena Alessandra , de Almeida Robson José , Rodrigues Albertina Gomes , Delle Humberto , Camacho Cleber P. 
Published: June 2021

Circulating RNA is detected in the blood, making it an excellent ally for the discovery of new pathophysiological mechanisms and new biomarkers. The biomarkers not only give the diagnosis but also can provide a prognosis in a non‐invasive way and they are already well established in some diseases. The purpose of this study was to establish the differential gene expression (DGE) between healthy non‐pregnant and pregnant women since homeostasis during pregnancy is critical to avoid future problems for the mother and her offspring.METHODSThree non‐pregnant and three pregnant women in the third trimester were selected, both healthy, all of whom signed a free and informed consent form. The project was approved by the Institutional Review Board (IRB) numbers 665.331 and 679.727. For the RNA extraction, blood samples were collected from the PAXgene Blood RNA tube (QIAGEN, NL, DE) and the PAXgene Blood RNA kit (Qiagen) was subsequently used. It was used Qubit fluorometer 2.0 (Thermo Fisher Scientific, MA, USA) for the quantification and SuperScript VILO cDNA Synthesis Kit (Thermo Fisher Scientific) for cDNA synthesis. The AmpliSeq Ion Transcriptome Human Gene Expression Kit (Thermo Fisher Scientific) and the Proton Ion platform (Thermo Fisher Scientific) were used to constructing the transcriptome library. Software R (version 3.4.1) was used with the edgeR package (version 3.16.5) for analyses, and Gene Ontology was used to analyze the biological processes (geneontology.org/).RESULTSWe were able to find 713 genes that were differentially expressed between the non‐pregnant and pregnant women. 2 were antisense RNA, 2 long intergenic noncoding RNA (lincRNA), 5 processed pseudogene, 2 processed transcript, 595 protein coding, 2 small cajalbody‐specific RNA (scaRNA), 6 small nucleolar RNA (snoRNA), 3 transcribed processed pseudogene, 3 transcribed unprocessed pseudogene and 2 unprocessed pseudogenes. Regarding molecular function, 20.9% of the genes are part of the catalytic activity, 21.8% binding, 12.1% receptor activity, 7.7% signal transducer activity, 5.4% transporter activity, 1.1% structural molecule activity, 0.4% translation and channel regulator activity. In the biological process, we found 211 genes related to cellular communication and signal transduction in cascade MAPK, 111 response to stimulus genes and 20 immune system process. The most abundant protein class found was the receptor type (7.8%). We found 67 pathways altered, among these, 13 genes were related to the Wnt signaling pathway where CSNK1A1L was upregulated with logFC 2.98, and NFATC2 downregulated with logFC − 3.26; 11 genes with the apoptosis signaling pathway being HSPA6 upregulated with logFC 2.23 and GZMH downregulated with logFC −1.74; 11 genes with the CCKR signaling map pathway (upregulated: MMP9logFC 3.44, downregulated: NFATC2 logFC −3.26); and 11 genes with a pathway Gonadotropin‐releasing hormone receptor (upregulated: MAPK14 logFC 2.21, downregulated: NFATC2 logFC −3.26).CONCLUSIONWe were able to determine the expression profile in the peripheral blood during gestation, which may be useful for future work with biomarkers.Support or Funding InformationThe authors' research grant was supported by the São Paulo State Research Foundation ‐ FAPESP (2014/04193‐0 to C.P.C.).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.