{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T21:27:38Z","timestamp":1783718858201,"version":"3.55.0"},"reference-count":45,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,3,12]],"date-time":"2025-03-12T00:00:00Z","timestamp":1741737600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["1R15HL152257"],"award-info":[{"award-number":["1R15HL152257"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Gestational diabetes mellitus (GDM) affects placental metabolism, influencing both maternal and fetal outcomes. This study investigated the expression of metabolic regulators\u2014Pyruvate Kinase M2 (PKM2), AMP-activated protein kinase (AMPK), and mTOR pathway components\u2014in placental tissues from GDM pregnancies managed with either insulin (GDM-I) or dietary interventions (GDM-D). We hypothesize that metabolic adaptation in GDM is differentially regulated by treatment modality. This study analyzed 30 cases, including 10 control pregnancies,10 GDM-D cases, and 10 GDM-I cases. Analytical methods included immunofluorescence and immunoblotting. We observed an upregulation of PKM2 in both GDM-I and GDM-D placentas, suggesting enhanced glycolytic adaptation under GDM-induced metabolic stress. AMPK expression was significantly elevated in GDM-I and moderately increased in GDM-D placentas, potentially compensating for insulin resistance by promoting glucose uptake and energy homeostasis. Furthermore, mTOR pathway activation differed by treatment type, suggesting a treatment-specific mTOR response. The metabolic changes observed suggest that treatment modality in GDM may have direct implications for maternal and fetal health. Our findings indicate that while insulin and dietary management support metabolic adaptation in GDM, they do so through distinct mechanisms. These findings support a personalized approach in GDM treatment, where patient-specific metabolic responses should guide therapeutic decisions.<\/jats:p>","DOI":"10.3390\/cells14060416","type":"journal-article","created":{"date-parts":[[2025,3,12]],"date-time":"2025-03-12T07:31:42Z","timestamp":1741764702000},"page":"416","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Differential Regulation of PKM2, AMPK, and mTOR in Response to Insulin and Dietary Management"],"prefix":"10.3390","volume":"14","author":[{"given":"Emily","family":"Broberg","sequence":"first","affiliation":[{"name":"Department of Cell Biology and Physiology, Brigham Young University, Provo, UT 84602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jillise","family":"English","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, Brigham Young University, Provo, UT 84602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Derek M.","family":"Clarke","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, Brigham Young University, Provo, UT 84602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marley J.","family":"Shin","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, Brigham Young University, Provo, UT 84602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin T.","family":"Bikman","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, Brigham Young University, Provo, UT 84602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0931-3025","authenticated-orcid":false,"given":"Paul R.","family":"Reynolds","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, Brigham Young University, Provo, UT 84602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8818-8786","authenticated-orcid":false,"given":"Juan A.","family":"Arroyo","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, Brigham Young University, Provo, UT 84602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"338","DOI":"10.4239\/wjd.v6.i2.338","article-title":"Effects of maternal diabetes on trophoblast cells","volume":"6","author":"Aires","year":"2015","journal-title":"World J. Diabetes"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1016\/j.jdiacomp.2014.03.010","article-title":"Gestational diabetes mellitus alters apoptotic and inflammatory gene expression of trophobasts from human term placenta","volume":"28","author":"Magee","year":"2014","journal-title":"J. Diabetes Complicat."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1007\/s00441-016-2496-5","article-title":"Decreased activation of placental mTOR family members is associated with the induction of intrauterine growth restriction by secondhand smoke in the mouse","volume":"367","author":"Mejia","year":"2017","journal-title":"Cell Tissue Res."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Plows, J.F., Stanley, J.L., Baker, P.N., Reynolds, C.M., and Vickers, M.H. (2018). The Pathophysiology of Gestational Diabetes Mellitus. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19113342"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Modzelewski, R., Stefanowicz-Rutkowska, M.M., Matuszewski, W., and Bandurska-Stankiewicz, E.M. (2022). Gestational Diabetes Mellitus-Recent Literature Review. J. Clin. Med., 11.","DOI":"10.3390\/jcm11195736"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Zhang, L., Yu, X., Wu, Y., Fu, H., Xu, P., Zheng, Y., Wen, L., Yang, X., Zhang, F., and Hu, M. (2021). Gestational Diabetes Mellitus-Associated Hyperglycemia Impairs Glucose Transporter 3 Trafficking in Trophoblasts Through the Downregulation of AMP-Activated Protein Kinase. Front. Cell Dev. Biol., 9.","DOI":"10.3389\/fcell.2021.722024"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1038\/nature06734","article-title":"The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth","volume":"452","author":"Christofk","year":"2008","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"221","DOI":"10.3233\/CBM-210111","article-title":"Targeting PKM2 promotes chemosensitivity of breast cancer cells in vitro and in vivo","volume":"32","author":"Wang","year":"2021","journal-title":"Cancer Biomark."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Zahra, K., Dey, T., Mishra, S.P., and Pandey, U. (2020). Pyruvate Kinase M2 and Cancer: The Role of PKM2 in Promoting Tumorigenesis. Front. Oncol., 10.","DOI":"10.3389\/fonc.2020.00159"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"969","DOI":"10.1016\/j.biocel.2010.02.005","article-title":"Pyruvate kinase type M2: A key regulator of the metabolic budget system in tumor cells","volume":"43","author":"Mazurek","year":"2011","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1016\/j.canlet.2020.11.018","article-title":"Pyruvate kinase M2 (PKM2) in cancer and cancer therapeutics","volume":"503","author":"Zhu","year":"2021","journal-title":"Cancer Lett."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Tsai, K., Tullis, B., Jensen, T., Graff, T., Reynolds, P., and Arroyo, J. (2021). Differential expression of mTOR related molecules in the placenta from gestational diabetes mellitus (GDM), intrauterine growth restriction (IUGR) and preeclampsia patients. Reprod. Biol., 21.","DOI":"10.1016\/j.repbio.2021.100503"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Li, N., Meng, D., Xu, Y., Gao, L., Shen, F., Tie, X., Zhang, Y., Yi, Z., Shen, W., and Liu, Z. (2020). Pyruvate Kinase M2 Knockdown Suppresses Migration, Invasion, and Epithelial-Mesenchymal Transition of Gastric Carcinoma via Hypoxia-Inducible Factor Alpha\/B-Cell Lymphoma 6 Pathway. Biomed. Res. Int., 2020.","DOI":"10.1155\/2020\/7467104"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"9417","DOI":"10.1021\/bi0474923","article-title":"Structural basis for tumor pyruvate kinase M2 allosteric regulation and catalysis","volume":"44","author":"Dombrauckas","year":"2005","journal-title":"Biochemistry"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/S0969-2126(98)00021-5","article-title":"The allosteric regulation of pyruvate kinase by fructose-1,6-bisphosphate","volume":"6","author":"Jurica","year":"1998","journal-title":"Structure"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4941","DOI":"10.1074\/jbc.272.8.4941","article-title":"Effect of extracellular AMP on cell proliferation and metabolism of breast cancer cell lines with high and low glycolytic rates","volume":"272","author":"Mazurek","year":"1997","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"864","DOI":"10.1016\/j.molcel.2011.10.015","article-title":"Tyrosine phosphorylation of mitochondrial pyruvate dehydrogenase kinase 1 is important for cancer metabolism","volume":"44","author":"Hitosugi","year":"2011","journal-title":"Mol. Cell"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e245","DOI":"10.1038\/emm.2016.81","article-title":"Regulation and function of AMPK in physiology and diseases","volume":"48","author":"Jeon","year":"2016","journal-title":"Exp. Mol. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1038\/nrm.2017.95","article-title":"AMPK: Guardian of metabolism and mitochondrial homeostasis","volume":"19","author":"Herzig","year":"2018","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"9075","DOI":"10.1007\/s11033-020-05900-x","article-title":"The role of AMPK in metabolism and its influence on DNA damage repair","volume":"47","author":"Szewczuk","year":"2020","journal-title":"Mol. Biol. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Xiao, Z., Liu, X., Luan, X., Duan, R., Peng, W., Tong, C., Qiao, J., and Qi, H. (2024). Glucose uptake in trophoblasts of GDM mice is regulated by the AMPK-CLUT3 signaling pathway. Sci. Rep., 14.","DOI":"10.1038\/s41598-024-61719-7"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Kumagai, A., Itakura, A., Koya, D., and Kanasaki, K. (2018). AMP-Activated Protein (AMPK) in Pathophysiology of Pregnancy Complications. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19103076"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Hung, T.H., Wu, C.P., and Chen, S.F. (2021). Differential Changes in Akt and AMPK Phosphorylation Regulating mTOR Activity in the Placentas of Pregnancies Complicated by Fetal Growth Restriction and Gestational Diabetes Mellitus with Large-For-Gestational Age Infants. Front. Med., 8.","DOI":"10.3389\/fmed.2021.788969"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Torres-Torres, J., Monroy-Munoz, I.E., Perez-Duran, J., Solis-Paredes, J.M., Camacho-Martinez, Z.A., Baca, D., Espino, Y.S.S., Martinez-Portilla, R., Rojas-Zepeda, L., and Borboa-Olivares, H. (2024). Cellular and Molecular Pathophysiology of Gestational Diabetes. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms252111641"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1007\/s40200-024-01420-8","article-title":"AMPK pathway: An emerging target to control diabetes mellitus and its related complications","volume":"23","author":"Kakoti","year":"2024","journal-title":"J. Diabetes Metab. Disord."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"bqaa076","DOI":"10.1210\/endocr\/bqaa076","article-title":"Placental Regulation of Energy Homeostasis During Human Pregnancy","volume":"161","author":"Armistead","year":"2020","journal-title":"Endocrinology"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"dev202088","DOI":"10.1242\/dev.202088","article-title":"Maternal-fetal cross-talk via the placenta: Influence on offspring development and metabolism","volume":"150","author":"Kramer","year":"2023","journal-title":"Development"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Dong, J., Xu, Q., Qian, C., Wang, L., DiSciullo, A., Lei, J., Lei, H., Yan, S., Wang, J., and Jin, N. (2024). Fetal growth restriction exhibits various mTOR signaling in different regions of mouse placentas with altered lipid metabolism. Cell Biol. Toxicol., 40.","DOI":"10.1007\/s10565-024-09855-8"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Arroyo, J.A., Garcia-Jones, P., Graham, A., Teng, C.C., Battaglia, F.C., and Galan, H.L. (2012). Placental TonEBP\/NFAT5 osmolyte regulation in an ovine model of intrauterine growth restriction. Biol. Reprod., 86.","DOI":"10.1095\/biolreprod.111.094797"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"14791641231173627","DOI":"10.1177\/14791641231173627","article-title":"Comparative study of microvascular structural changes in the gestational diabetic placenta","volume":"20","author":"Liang","year":"2023","journal-title":"Diabetes Vasc. Dis. Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/s40170-021-00239-8","article-title":"Pharmacological activation of pyruvate kinase M2 reprograms glycolysis leading to TXNIP depletion and AMPK activation in breast cancer cells","volume":"9","author":"Almouhanna","year":"2021","journal-title":"Cancer Metab."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1038\/ncb1547","article-title":"Insulin signalling to mTOR mediated by the Akt\/PKB substrate PRAS40","volume":"9","author":"Lee","year":"2007","journal-title":"Nat. Cell Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3113","DOI":"10.7150\/ijbs.89942","article-title":"The PI3K\/Akt Pathway and Glucose Metabolism: A Dangerous Liaison in Cancer","volume":"20","author":"Fontana","year":"2024","journal-title":"Int. J. Biol. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4225","DOI":"10.1210\/jc.2018-02778","article-title":"Insulin Stimulates GLUT4 Trafficking to the Syncytiotrophoblast Basal Plasma Membrane in the Human Placenta","volume":"104","author":"Arbet","year":"2019","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1016\/j.placenta.2015.04.006","article-title":"Inflammation in maternal obesity and gestational diabetes mellitus","volume":"36","author":"Pantham","year":"2015","journal-title":"Placenta"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Liu, Z., Le, Y., Chen, H., Zhu, J., and Lu, D. (2021). Role of PKM2-Mediated Immunometabolic Reprogramming on Development of Cytokine Storm. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.748573"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1016\/j.freeradbiomed.2019.08.007","article-title":"Pyruvate kinase M2: A simple molecule with complex functions","volume":"143","author":"Alquraishi","year":"2019","journal-title":"Free Radic. Biol. Med."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1151","DOI":"10.3892\/ol.2012.928","article-title":"Energy metabolism of cancer: Glycolysis versus oxidative phosphorylation (Review)","volume":"4","author":"Zheng","year":"2012","journal-title":"Oncol. Lett."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"4816","DOI":"10.1128\/JB.00705-13","article-title":"Depletion of glycolytic intermediates plays a key role in glucose-phosphate stress in Escherichia coli","volume":"195","author":"Richards","year":"2013","journal-title":"J. Bacteriol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1016\/j.tem.2012.06.010","article-title":"Emerging roles of PKM2 in cell metabolism and cancer progression","volume":"23","author":"Luo","year":"2012","journal-title":"Trends Endocrinol. Metab."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Puckett, D.L., Alquraishi, M., Chowanadisai, W., and Bettaieb, A. (2021). The Role of PKM2 in Metabolic Reprogramming: Insights into the Regulatory Roles of Non-Coding RNAs. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22031171"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1295","DOI":"10.1038\/ncb2629","article-title":"ERK1\/2-dependent phosphorylation and nuclear translocation of PKM2 promotes the Warburg effect","volume":"14","author":"Yang","year":"2012","journal-title":"Nat. Cell Biol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.diabres.2018.10.017","article-title":"Increased placental IGF-1\/mTOR activity in macrosomia born to women with gestational diabetes","volume":"146","author":"Shang","year":"2018","journal-title":"Diabetes Res. Clin. Pract."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"961","DOI":"10.1042\/CS20190266","article-title":"Placental function in maternal obesity","volume":"134","author":"Kelly","year":"2020","journal-title":"Clin. Sci."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"839","DOI":"10.1038\/nchembio.1060","article-title":"Pyruvate kinase M2 activators promote tetramer formation and suppress tumorigenesis","volume":"8","author":"Anastasiou","year":"2012","journal-title":"Nat. Chem. Biol."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/6\/416\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:51:15Z","timestamp":1760028675000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/6\/416"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,12]]},"references-count":45,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["cells14060416"],"URL":"https:\/\/doi.org\/10.3390\/cells14060416","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,12]]}}}