{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T03:30:44Z","timestamp":1785123044871,"version":"3.55.0"},"reference-count":49,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2023,1,21]],"date-time":"2023-01-21T00:00:00Z","timestamp":1674259200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Institutes of Health","award":["R01HL049277"],"award-info":[{"award-number":["R01HL049277"]}]},{"name":"National Institutes of Health","award":["R01HD101485"],"award-info":[{"award-number":["R01HD101485"]}]},{"name":"National Institutes of Health","award":["R03HD080788"],"award-info":[{"award-number":["R03HD080788"]}]},{"name":"National Institutes of Health","award":["K23HD088742"],"award-info":[{"award-number":["K23HD088742"]}]},{"name":"UNC Lineberger Comprehensive Cancer Center","award":["R01HL049277"],"award-info":[{"award-number":["R01HL049277"]}]},{"name":"UNC Lineberger Comprehensive Cancer Center","award":["R01HD101485"],"award-info":[{"award-number":["R01HD101485"]}]},{"name":"UNC Lineberger Comprehensive Cancer Center","award":["R03HD080788"],"award-info":[{"award-number":["R03HD080788"]}]},{"name":"UNC Lineberger Comprehensive Cancer Center","award":["K23HD088742"],"award-info":[{"award-number":["K23HD088742"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Obesity is a risk factor for preeclampsia. We investigated how obesity influences preeclampsia in mice lacking ankyrin-repeat-and-SOCS-box-containing-protein 4 (ASB4), which promotes trophoblast differentiation via degrading the inhibitor of DNA-binding protein 2 (ID2). Asb4\u2212\/\u2212 mice on normal chow (NC) develop mild preeclampsia-like phenotypes during pregnancy, including hypertension, proteinuria, and reduced litter size. Wild-type (WT) and Asb4\u2212\/\u2212 females were placed on a high-fat diet (HFD) starting at weaning. At the age of 8\u20139 weeks, they were mated with WT or Asb4\u2212\/\u2212 males, and preeclamptic phenotypes were assessed. HFD-WT dams had no obvious adverse outcomes of pregnancy. In contrast, HFD-Asb4\u2212\/\u2212 dams had significantly more severe preeclampsia-like phenotypes compared to NC-Asb4\u2212\/\u2212 dams. The HFD increased white fat weights and plasma leptin and insulin levels in Asb4\u2212\/\u2212 females. In the HFD-Asb4\u2212\/\u2212 placenta, ID2 amounts doubled without changing the transcript levels, indicating that insulin likely increases ID2 at a level of post-transcription. In human first-trimester trophoblast HTR8\/SVneo cells, exposure to insulin, but not to leptin, led to a significant increase in ID2. HFD-induced obesity markedly worsens the preeclampsia-like phenotypes in the absence of ASB4. Our data indicate that hyperinsulinemia perturbs the timely removal of ID2 and interferes with proper trophoblast differentiation, contributing to enhanced preeclampsia.<\/jats:p>","DOI":"10.3390\/ijms24032149","type":"journal-article","created":{"date-parts":[[2023,1,23]],"date-time":"2023-01-23T02:27:45Z","timestamp":1674440865000},"page":"2149","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Insulin Elevates ID2 Expression in Trophoblasts and Aggravates Preeclampsia in Obese ASB4-Null Mice"],"prefix":"10.3390","volume":"24","author":[{"given":"Yukako","family":"Kayashima","sequence":"first","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, The University of North Carolina, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W. H. Davin","family":"Townley-Tilson","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, The University of North Carolina, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Neeta L.","family":"Vora","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Division of Maternal Fetal Medicine, The University of North Carolina, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim","family":"Boggess","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Division of Maternal Fetal Medicine, The University of North Carolina, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathon W.","family":"Homeister","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, The University of North Carolina, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nobuyo","family":"Maeda-Smithies","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, The University of North Carolina, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6779-5077","authenticated-orcid":false,"given":"Feng","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine, The University of North Carolina, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1016\/S0140-6736(05)17987-2","article-title":"Pre-eclampsia","volume":"365","author":"Sibai","year":"2005","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"S907","DOI":"10.1016\/j.ajog.2020.09.047","article-title":"Syncytiotrophoblast stress in preeclampsia: The convergence point for multiple pathways","volume":"226","author":"Redman","year":"2022","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"f6564","DOI":"10.1136\/bmj.f6564","article-title":"Pre-eclampsia rates in the United States, 1980\u20132010: Age-period-cohort analysis","volume":"347","author":"Ananth","year":"2013","journal-title":"BMJ"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1016\/j.annepidem.2004.12.008","article-title":"The risk of preeclampsia rises with increasing prepregnancy body mass index","volume":"15","author":"Bodnar","year":"2005","journal-title":"Ann. Epidemiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e05079","DOI":"10.1016\/j.heliyon.2020.e05079","article-title":"Risk factors and fetal outcomes for preeclampsia in a Colombian cohort","volume":"6","author":"Serrano","year":"2020","journal-title":"Heliyon"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e218401","DOI":"10.1001\/jamanetworkopen.2021.8401","article-title":"Preeclampsia Prevalence, Risk Factors, and Pregnancy Outcomes in Sweden and China","volume":"4","author":"Yang","year":"2021","journal-title":"JAMA Netw. Open"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1016\/j.preghy.2010.10.013","article-title":"The Role of Obesity in Preeclampsia","volume":"1","author":"Roberts","year":"2011","journal-title":"Pregnancy Hypertens."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Townley-Tilson, W.H., Wu, Y., Ferguson, J.E., and Patterson, C. (2014). The ubiquitin ligase ASB4 promotes trophoblast differentiation through the degradation of ID2. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0089451"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"6407","DOI":"10.1128\/MCB.00511-07","article-title":"ASB4 is a hydroxylation substrate of FIH and promotes vascular differentiation via an oxygen-dependent mechanism","volume":"27","author":"Ferguson","year":"2007","journal-title":"Mol. Cell. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1016\/B978-0-12-405943-6.00005-1","article-title":"Id proteins: Small molecules, mighty regulators","volume":"110","author":"Ling","year":"2014","journal-title":"Curr. Top. Dev. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.mce.2004.04.016","article-title":"Id2 is a primary partner for the E2-2 basic helix-loop-helix transcription factor in the human placenta","volume":"222","author":"Liu","year":"2004","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"13450","DOI":"10.1073\/pnas.1614947113","article-title":"Nicotinamide benefits both mothers and pups in two contrasting mouse models of preeclampsia","volume":"113","author":"Li","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1536","DOI":"10.1161\/HYPERTENSIONAHA.121.18832","article-title":"Midgestation Leptin Infusion Induces Characteristics of Clinical Preeclampsia in Mice, Which Is Ablated by Endothelial Mineralocorticoid Receptor Deletion","volume":"79","author":"Faulkner","year":"2022","journal-title":"Hypertension"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Alston, M.C., Redman, L.M., and Sones, J.L. (2022). An Overview of Obesity, Cholesterol, and Systemic Inflammation in Preeclampsia. Nutrients, 14.","DOI":"10.3390\/nu14102087"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.placenta.2004.03.009","article-title":"Leptin expression in the fetus and placenta during mouse pregnancy","volume":"26","author":"Malik","year":"2005","journal-title":"Placenta"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"301","DOI":"10.7150\/ijms.11644","article-title":"Fetal\/Placental weight ratio in term Japanese pregnancy: Its difference among gender, parity, and infant growth","volume":"12","author":"Matsuda","year":"2015","journal-title":"Int. J. Med. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Sui, X., Zhang, L., Zhang, X.F., and Zhang, Y. (2021). TRIB3-Regulated Akt Signal Pathway Affects Trophoblast Invasion in the Development of Preeclampsia. Am. J. Perinatol.","DOI":"10.1055\/s-0041-1735872"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"894","DOI":"10.1161\/HYPERTENSIONAHA.117.10849","article-title":"Causative Effects of Genetically Determined High Maternal\/Fetal Endothelin-1 on Preeclampsia-Like Conditions in Mice","volume":"71","author":"Li","year":"2018","journal-title":"Hypertension"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1161\/HYPERTENSIONAHA.110.160242","article-title":"Adenoviral delivery of VEGF121 early in pregnancy prevents spontaneous development of preeclampsia in BPH\/5 mice","volume":"57","author":"Woods","year":"2011","journal-title":"Hypertension"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1529","DOI":"10.1042\/CS20160832","article-title":"Visceral adipose tissue activated macrophage content and inflammatory adipokine secretion is higher in pre-eclampsia than in healthy pregnancys","volume":"131","author":"Huda","year":"2017","journal-title":"Clin. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2038","DOI":"10.1210\/me.2007-0454","article-title":"Inhibitor of DNA binding 2 is a small molecule-inducible modulator of peroxisome proliferator-activated receptor-gamma expression and adipocyte differentiation","volume":"22","author":"Park","year":"2008","journal-title":"Mol. Endocrinol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Mathew, D., Zhou, P., Pywell, C.M., van der Veen, D.R., Shao, J., Xi, Y., Bonar, N.A., Hummel, A.D., Chapman, S., and Leevy, W.M. (2013). Ablation of the ID2 gene results in altered circadian feeding behavior, and sex-specific enhancement of insulin sensitivity and elevated glucose uptake in skeletal muscle and brown adipose tissue. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0073064"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1445","DOI":"10.1093\/ajh\/hpt137","article-title":"Euglycemic hyperinsulinemia increases blood pressure in pregnant rats independent of placental antiangiogenic and inflammatory factors","volume":"26","author":"Palei","year":"2013","journal-title":"Am. J. Hypertens."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Takao, T., Asanoma, K., Kato, K., Fukushima, K., Tsunematsu, R., Hirakawa, T., Matsumura, S., Seki, H., Takeda, S., and Wake, N. (2011). Isolation and characterization of human trophoblast side-population (SP) cells in primary villous cytotrophoblasts and HTR-8\/SVneo cell line. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0021990"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1016\/j.fertnstert.2018.11.032","article-title":"Direct toxicity of insulin on the human placenta and protection by metformin","volume":"111","author":"Vega","year":"2019","journal-title":"Fertil. Steril."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1530\/JOE-19-0127","article-title":"Hyperinsulinemia restrains endometrial angiogenesis during decidualization in early pregnancy","volume":"243","author":"Chen","year":"2019","journal-title":"J. Endocrinol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"F182","DOI":"10.1152\/ajprenal.00548.2012","article-title":"Insulin directly stimulates VEGF-A production in the glomerular podocyte","volume":"305","author":"Hale","year":"2013","journal-title":"Am. J. Physiol. Renal. Physiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1016\/j.placenta.2014.07.002","article-title":"Placental miR-1301 is dysregulated in early-onset preeclampsia and inversely correlated with maternal circulating leptin","volume":"35","author":"Sheng","year":"2014","journal-title":"Placenta"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1016\/j.mce.2011.07.047","article-title":"Treatment with a constitutive androstane receptor ligand ameliorates the signs of preeclampsia in high-fat diet-induced obese pregnant mice","volume":"348","author":"Masuyama","year":"2012","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1093\/ajh\/hpw153","article-title":"Role of Nitric Oxide Synthase on Blood Pressure Regulation and Vascular Function in Pregnant Rats on a High-Fat Diet","volume":"30","author":"Palei","year":"2017","journal-title":"Am. J. Hypertens."},{"key":"ref_31","first-page":"3532","article-title":"Effect of high-fat diet on liver and placenta fatty infiltration in early onset preeclampsia-like mouse model","volume":"125","author":"Sun","year":"2012","journal-title":"Chin. Med. J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1242\/dev.127.3.549","article-title":"Id-2 regulates critical aspects of human cytotrophoblast differentiation, invasion and migration","volume":"127","author":"Janatpour","year":"2000","journal-title":"Development"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1089\/scd.2016.0010","article-title":"Id2 Mediates Differentiation of Labyrinthine Placental Progenitor Cell Line, SM10","volume":"25","author":"Selesniemi","year":"2016","journal-title":"Stem Cells Dev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1006\/mgme.1997.2658","article-title":"Insulin sensitivity indices calculated from basal and OGTT-induced insulin, glucose, and FFA levels","volume":"63","author":"Belfiore","year":"1998","journal-title":"Mol. Genet. Metab."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Jeyarajah, M.J., Jaju Bhattad, G., Hillier, D.M., and Renaud, S.J. (2020). The Transcription Factor OVOL2 Represses ID2 and Drives Differentiation of Trophoblast Stem Cells and Placental Development in Mice. Cells, 9.","DOI":"10.3390\/cells9040840"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1042\/bj3280329","article-title":"Molecular mechanisms for the control of translation by insulin","volume":"328","author":"Proud","year":"1997","journal-title":"Biochem. J."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"582","DOI":"10.1177\/1933719116667220","article-title":"Insulin Exhibits an Antiproliferative and Hypertrophic Effect in First Trimester Human Extravillous Trophoblasts","volume":"24","author":"Silva","year":"2017","journal-title":"Reprod. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"8977","DOI":"10.1038\/s41598-018-27119-4","article-title":"Human trophoblast stem cell self-renewal and differentiation: Role of decorin","volume":"8","author":"Nandi","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Li, J.Y., Chai, B., Zhang, W., Wu, X., Zhang, C., Fritze, D., Xia, Z., Patterson, C., and Mulholland, M.W. (2011). Ankyrin repeat and SOCS box containing protein 4 (Asb-4) colocalizes with insulin receptor substrate 4 (IRS4) in the hypothalamic neurons and mediates IRS4 degradation. BMC Neurosci., 12.","DOI":"10.1186\/1471-2202-12-95"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1210\/en.2009-0560","article-title":"Expression of ankyrin repeat and suppressor of cytokine signaling box protein 4 (Asb-4) in proopiomelanocortin neurons of the arcuate nucleus of mice produces a hyperphagic, lean phenotype","volume":"151","author":"Li","year":"2010","journal-title":"Endocrinology"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"eabj8204","DOI":"10.1126\/scisignal.abj8204","article-title":"ASB4 modulates central melanocortinergic neurons and calcitonin signaling to control satiety and glucose homeostasis","volume":"15","author":"Vagena","year":"2022","journal-title":"Sci. Signal"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1038\/s41574-022-00704-4","article-title":"Obesity-associated ASB4 controls satiety","volume":"18","author":"Kriebs","year":"2022","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"101504","DOI":"10.1016\/j.redox.2020.101504","article-title":"Oral high dose vitamin B12 decreases renal superoxide and post-ischemia\/reperfusion injury in mice","volume":"32","author":"Li","year":"2020","journal-title":"Redox Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2122","DOI":"10.1111\/j.1538-7836.2010.03976.x","article-title":"Elevated tissue factor expression contributes to exacerbated diabetic nephropathy in mice lacking eNOS fed a high fat diet","volume":"8","author":"Li","year":"2010","journal-title":"J. Thromb. Haemost."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1006\/excr.1993.1139","article-title":"Establishment and characterization of first trimester human trophoblast cells with extended lifespan","volume":"206","author":"Graham","year":"1993","journal-title":"Exp. Cell Res."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"773","DOI":"10.7150\/ijms.11610","article-title":"Progesterone Inhibits Leptin-Induced Invasiveness of BeWo Cells","volume":"12","author":"Jo","year":"2015","journal-title":"Int. J. Med. Sci."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Toro, A.R., Maymo, J.L., Ibarbalz, F.M., Perez-Perez, A., Maskin, B., Faletti, A.G., Sanchez-Margalet, V., and Varone, C.L. (2014). Leptin is an anti-apoptotic effector in placental cells involving p53 downregulation. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0099187"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"506","DOI":"10.1016\/j.cmet.2007.10.011","article-title":"Increased energy expenditure, dietary fat wasting, and resistance to diet-induced obesity in mice lacking renin","volume":"6","author":"Takahashi","year":"2007","journal-title":"Cell Metab."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1002\/(SICI)1097-0177(199603)205:3<281::AID-AJA8>3.0.CO;2-E","article-title":"Cardiac neural crest is essential for the persistence rather than the formation of an arch artery","volume":"205","author":"Waldo","year":"1996","journal-title":"Dev. Dyn."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/3\/2149\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:12:51Z","timestamp":1760119971000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/3\/2149"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,21]]},"references-count":49,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2023,2]]}},"alternative-id":["ijms24032149"],"URL":"https:\/\/doi.org\/10.3390\/ijms24032149","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,21]]}}}