{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T08:04:54Z","timestamp":1768464294458,"version":"3.49.0"},"reference-count":25,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2021,11,28]],"date-time":"2021-11-28T00:00:00Z","timestamp":1638057600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000071","name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development","doi-asserted-by":"publisher","award":["5R01 HD088386"],"award-info":[{"award-number":["5R01 HD088386"]}],"id":[{"id":"10.13039\/100000071","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Neutrophils are activated and extensively infiltrate blood vessels in preeclamptic women. To identify genes that contribute to neutrophil activation and infiltration, we analyzed the transcriptomes of circulating neutrophils from normal pregnant and preeclamptic women. Neutrophils were collected at 30 weeks\u2019 gestation and RNA and DNA were isolated for RNA sequencing and 5-hydroxy-methylcytosine (5-hmC) sequencing as an index of dynamic changes in neutrophil DNA methylation. Women with normal pregnancy who went on to develop mild preeclampsia at term had the most uniquely expressed genes (697) with 325 gene ontology pathways upregulated, many related to neutrophil activation and function. Women with severe preeclampsia who delivered prematurely had few pathways up- or downregulated. Cluster analysis revealed that gene expression in women with severe preeclampsia was an inverse mirror image of gene expression in normal pregnancy, while gene expression in women who developed mild preeclampsia was remarkably different from both. DNA methylation marks, key regulators of gene expression, are removed by the action of ten-eleven translocation (TET) enzymes, which oxidize 5-methylcytosines (5mCs), resulting in locus-specific reversal of DNA methylation. DNA sequencing for 5-hmC revealed no differences among the three groups. Genome-wide DNA methylation revealed extremely low levels in circulating neutrophils suggesting they are de-methylated. Collectively, these data demonstrate that neutrophil gene expression profiles can distinguish different preeclampsia phenotypes, and in the case of mild preeclampsia, alterations in gene expression occur well before clinical symptoms emerge. These findings serve as a foundation for further evaluation of neutrophil transcriptomes as biomarkers of preeclampsia phenotypes. Changes in DNA methylation in circulating neutrophils do not appear to mediate differential patterns of gene expression in either mild or severe preeclampsia.<\/jats:p>","DOI":"10.3390\/ijms222312876","type":"journal-article","created":{"date-parts":[[2021,11,30]],"date-time":"2021-11-30T23:22:28Z","timestamp":1638314548000},"page":"12876","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Patterns of Maternal Neutrophil Gene Expression at 30 Weeks of Gestation, but Not DNA Methylation, Distinguish Mild from Severe Preeclampsia"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8234-025X","authenticated-orcid":false,"given":"Scott W.","family":"Walsh","sequence":"first","affiliation":[{"name":"Department of Obstetrics and Gynecology, Virginia Commonwealth University, Richmond, VA 23298-0034, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marwah","family":"Al Dulaimi","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Virginia Commonwealth University, Richmond, VA 23298-0034, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kellie J.","family":"Archer","sequence":"additional","affiliation":[{"name":"Division of Biostatistics, College of Public Health, The Ohio State University, Columbus, OH 43210, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6199-0480","authenticated-orcid":false,"suffix":"III","given":"Jerome F.","family":"Strauss","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Virginia Commonwealth University, Richmond, VA 23298-0034, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,28]]},"reference":[{"key":"ref_1","unstructured":"Cunningham, F.G., Gant, N.F., Leveno, K.J., Gilstrap, L.C., Hauth, J.C., and Wenstrom, K.D. (2001). Williams Obstetrics, McGraw-Hill. [21st ed.]."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1161\/01.HYP.0000130483.83154.37","article-title":"Neutrophils infiltrate resistance-sized vessels of subcutaneous fat in women with preeclampsia","volume":"44","author":"Leik","year":"2004","journal-title":"Hypertension"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"867","DOI":"10.1161\/HYPERTENSIONAHA.111.176602","article-title":"Mechanisms of enhanced vascular reactivity in preeclampsia","volume":"58","author":"Mishra","year":"2011","journal-title":"Hypertension"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"48.e1","DOI":"10.1016\/j.ajog.2006.08.038","article-title":"Activation of NF-kappaB and expression of COX-2 in association with neutrophil infiltration in systemic vascular tissue of women with preeclampsia","volume":"196","author":"Shah","year":"2007","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1332","DOI":"10.1177\/1933719112450336","article-title":"DNA Methylation is Altered in Maternal Blood Vessels of Women With Preeclampsia","volume":"19","author":"Mousa","year":"2012","journal-title":"Reprod. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.1016\/j.ajpath.2012.06.019","article-title":"Preeclampsia is associated with alterations in DNA methylation of genes involved in collagen metabolism","volume":"181","author":"Mousa","year":"2012","journal-title":"Am. J. Pathol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1249","DOI":"10.1161\/HYPERTENSIONAHA.111.188730","article-title":"Reduced methylation of the thromboxane synthase gene is correlated with its increased vascular expression in preeclampsia","volume":"59","author":"Mousa","year":"2012","journal-title":"Hypertension"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1303","DOI":"10.1126\/science.1210944","article-title":"Tet-mediated formation of 5-carboxylcytosine and its excision by TDG in mammalian DNA","volume":"333","author":"He","year":"2011","journal-title":"Science"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1300","DOI":"10.1126\/science.1210597","article-title":"Tet proteins can convert 5-methylcytosine to 5-formylcytosine and 5-carboxylcytosine","volume":"333","author":"Ito","year":"2011","journal-title":"Science"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1126\/science.1170116","article-title":"Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1","volume":"324","author":"Tahiliani","year":"2009","journal-title":"Science"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"10396","DOI":"10.1021\/ja4028346","article-title":"Ascorbic Acid enhances tet-mediated 5-methylcytosine oxidation and promotes DNA demethylation in mammals","volume":"135","author":"Yin","year":"2013","journal-title":"J. Am. Chem. Soc."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1229","DOI":"10.1126\/science.1211917","article-title":"Molecular biology. Demystifying DNA demethylation","volume":"333","author":"Nabel","year":"2011","journal-title":"Science"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"R46","DOI":"10.1186\/gb-2013-14-5-r46","article-title":"5-Hydroxymethylcytosine is an essential intermediate of active DNA demethylation processes in primary human monocytes","volume":"14","author":"Klug","year":"2013","journal-title":"Genome Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2436","DOI":"10.1101\/gad.179184.111","article-title":"Mechanisms and functions of Tet protein-mediated 5-methylcytosine oxidation","volume":"25","author":"Wu","year":"2011","journal-title":"Genes Dev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"R63","DOI":"10.1186\/gb-2010-11-6-r63","article-title":"Active DNA demethylation in human postmitotic cells correlates with activating histone modifications, but not transcription levels","volume":"11","author":"Klug","year":"2010","journal-title":"Genome Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2115","DOI":"10.1007\/s43032-020-00232-4","article-title":"Proteases Activate Pregnancy Neutrophils by a Protease-Activated Receptor 1 Pathway: Epigenetic Implications for Preeclampsia","volume":"27","author":"Walsh","year":"2020","journal-title":"Reprod. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Munchel, S., Rohrback, S., Randise-Hinchliff, C., Kinnings, S., Deshmukh, S., Alla, N., Tan, C., Kia, A., Greene, G., and Leety, L. (2020). Circulating transcripts in maternal blood reflect a molecular signature of early-onset preeclampsia. Sci. Transl. Med., 12.","DOI":"10.1126\/scitranslmed.aaz0131"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2158","DOI":"10.1007\/s43032-020-00234-2","article-title":"Placental Production of Eicosanoids and Sphingolipids in Women Who Developed Preeclampsia on Low-Dose Aspirin","volume":"27","author":"Walsh","year":"2020","journal-title":"Reprod. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1189\/jlb.0903412","article-title":"Gene expression in mature neutrophils: Early responses to inflammatory stimuli","volume":"75","author":"Zhang","year":"2004","journal-title":"J. Leukoc. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"817038","DOI":"10.1155\/2015\/817038","article-title":"Regulation of Neutrophil Degranulation and Cytokine Secretion: A Novel Model Approach Based on Linear Fitting","volume":"2015","author":"Naegelen","year":"2015","journal-title":"J. Immunol. Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1800","DOI":"10.1111\/j.1538-7836.2005.01377.x","article-title":"Protease-activated receptors in hemostasis, thrombosis and vascular biology","volume":"3","author":"Coughlin","year":"2005","journal-title":"J. Thromb. Haemost."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1016\/j.ajpath.2010.11.003","article-title":"Increased expression of matrix metalloproteinase-1 in systemic vessels of preeclamptic women a critical mediator of vascular dysfunction","volume":"178","author":"Cappello","year":"2011","journal-title":"Am. J. Pathol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1415","DOI":"10.1016\/j.ajog.2006.02.053","article-title":"Thromboxane mediates neutrophil superoxide production in pregnancy","volume":"195","author":"Vaughan","year":"2006","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2183","DOI":"10.1093\/bioinformatics\/btm311","article-title":"beadarray: R classes and methods for Illumina bead-based data","volume":"23","author":"Dunning","year":"2007","journal-title":"Bioinformatics"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"9440","DOI":"10.1073\/pnas.1530509100","article-title":"Statistical significance for genomewide studies","volume":"100","author":"Storey","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/23\/12876\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:37:03Z","timestamp":1760168223000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/23\/12876"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,28]]},"references-count":25,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["ijms222312876"],"URL":"https:\/\/doi.org\/10.3390\/ijms222312876","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,28]]}}}