An in vitro Model to Study Placental Functions in Gestational Diabetes Mellitus
DOI:
https://doi.org/10.48048/tis.2024.7427Keywords:
Gestational diabetes mellitus, Biomarker, Trophoblast, Insulin resistance, BeWo cells, VEGF, Flow cytometryAbstract
Background: Gestational diabetes mellitus (GDM) is a serious pregnancy complication that affects around 14 % of pregnancies worldwide. It not only causes short-term complications in the mother and child but also significantly increases the risks of several chronic diseases, including cardiovascular diseases and diabetes. An excess production of the soluble vascular endothelial growth factor (VEGF) receptor 1 in the placenta, which is a natural inhibitor of VEGF, has been linked to various pregnancy complications. However, placentas affected by GDM often exhibit increased vascularization, suggesting that the role of VEGF in GDM differs from its impact on typical pregnancy complications. Unfortunately, limited studies on placental functions in GDM have been conducted due to a lack of reliable culture models. Methods: In this study, we developed an in vitro model of GDM using the human choriocarcinoma trophoblast cell line BeWo and examined the effects of VEGF on glucose uptake. The BeWo cells were treated with a high glucose-containing (25 mM) syncytialization differentiation medium to produce insulin-resistant cells. Insulin resistance was assessed by a glucose uptake assay using the fluorescent glucose analog 2-NBDG and flow cytometry analysis. Results: The cells treated with high glucose exhibited a significant decrease in glucose uptake, suggesting the successful development of glucose resistance in these cells. However, VEGF treatment did not show a significant impact on glucose uptake in high glucose-treated cells. Conclusions: It is important to note that glucose homeostasis is a complex process involving multiple cell types, and further studies are necessary to fully understand the functions of VEGF on GDM placentas. Nevertheless, the in vitro model we have developed and validated will be a valuable tool for studying placental pathophysiology in GDM and evaluating the effectiveness of potential therapeutic agents.
HIGHLIGHTS
- BeWo cells are the choriocarcinoma cell line which were cultured in the in vitro condition
- BeWo cells were differentiated using forskolin and 8-Br Camp
- Insulin resistant model of BeWo cells were prepared by high glucose (25 mM) treatment
- The effect of VEGF on glucose homeostasis in GDM model of BeWo cells was observed
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H Wang, N Li, T Chivese, M Werfalli, H Sun, L Yuen, CA Hoegfeldt, CE Powe, J Immanuel, S Karuranga, H Divakar, N Levitt, C Li, D Simmons, X Yang and IDF Diabetes Atlas Committee Hyperglycaemia in Pregnancy Special Interest Group. IDF diabetes atlas: Estimation of global and regional gestational diabetes mellitus prevalence for 2021 by International Association of Diabetes in Pregnancy Study Group's Criteria. Diabetes Res. Clin. Pract. 2022; 183, 109050.
LA Barbour, CE McCurdy, TL Hernandez, JP Kirwan, PM Catalano and JE Friedman. Cellular mechanisms for insulin resistance in normal pregnancy and gestational diabetes. Diabetes Care 2007; 30, S112-S119.
L Martinez-Fernandez, E Gonzalez-Rodriguez, I Gomez-Hurtado, A Jimenez-Gomez, M Lopez-Miranda, J M Morales, J M Moreno-Otero, J M Quesada-Gomez, J M Ramirez-Jimenez, J M Ruiz-Ruiz, J M Sanchez-Margallo, J M Torres-Cantero. Hyperglycemia-induced oxidative stress and inflammation in the placenta: a potential factor involved in the pathophysiology of gestational diabetes mellitus. Placenta 2021; 105, 119-27.
X Chen, TO Scholl and TP Stein. Maternal glucose concentration during pregnancy is associated with the umbilical artery flow velocity waveform: The healthy start study. Diabetes Care 2020; 43, 1923-9.
PM Catalano, HD McIntyre, JK Cruickshank, DR McCance, AR Dyer, BE Metzger, LP Lowe, ER Trimble, DR Coustan, DR Hadden, B Persson, M Hod and JJN Oats. The hyperglycemia and adverse pregnancy outcome study: Associations of GDM and obesity with pregnancy outcomes. Diabetes Care 2012; 35, 780-6.
American Diabetes Association. Classification and diagnosis of diabetes: Standards of medical care in diabetes - 2020. Diabetes Care 2020; 43, S14-S31.
S Balci, A Gorur, DD Yıldırım, F Cayan and L Tamer. Expression level of miRNAs in patients with gestational diabetes. Turkish J. Biochem. 2020; 45, 825-31.
V Seshiah, V Balaji, MS Balaji, CB Sanjeevi and A Green. Gestational diabetes mellitus in India. J. Assoc. Phys. India 2004; 52, 707-11.
M Leoni, Padilla, A Fabbri, D Della-Morte, C Ricordi and M Infante. Mechanisms of Insulin Resistance during Pregnancy. IntechOpen, 2022.
N Jayabalan, A Lai, S Nair, D Guanzon, K Scholz-Romero, C Palma, HD McIntyre, M Lappas and C Salomon. Quantitative proteomics by SWATH-MS suggest an association between circulating exosomes and maternal metabolic changes in gestational diabetes mellitus. Proteomics 2019; 19, 1800164.
SH Jung, CH Jung, MJ Lee, JH Choi, OS Kwon, SW Kang and WS Kim. Vascular endothelial growth factor receptor 2 signaling affects glucose uptake and proliferation in human adipose tissue-derived mesenchymal stem cells. J. Clin. Endocrinol. Metab. 2014; 99, E686-E695.
F Troncoso, J Acurio, K Herlitz, C Aguayo, P Bertoglia, E Guzman-Gutierrez, M Loyola, M Gonzalez, M Rezgaoui, G Desoye and C Escudero. Gestational diabetes mellitus is associated with increased pro-migratory activation of vascular endothelial growth factor receptor 2 and reduced expression of vascular endothelial growth factor receptor 1. PLoS One 2017; 12, e0182509.
NP Joshi, AR Mane, AS Sahay, DP Sundrani, SR Joshi and CS Yajnik. Role of placental glucose transporters in determining fetal growth. Reprod. Sci. 2022; 29, 2744-59.
WL Lowe Jr and DM Scholtens. Pathophysiology of gestational diabetes mellitus: The past, present and future. Diabetic Med. 2020; 37, 812-20.
T Radaelli, A Varastehpour, P Catalano and S Hauguel-de Mouzon. Gestational diabetes induces placental genes for chronic stress and inflammatory pathways. Diabetes 2009; 58, 760-8.
CH Hulme, A Stevens, W Dunn, AEP Heazell, K Hollywood, P Begley, M Westwood and JE Myers. Identification of the functional pathways altered by placental cell exposure to high glucose: Lessons from the transcript and metabolite interactome. Sci. Rep. 2018; 8, 5270.
ZJW Easton, X Luo, L Li and TRH Regnault. The impact of hyperglycemia upon BeWo trophoblast cell metabolic function: A multi-OMICS and functional metabolic analysis. bioRxiv 2022, https://doi.org/10.1101/2022.02.15.480604
I Arutyunyan, T Fatkhudinov, E Kananykhina, N Usman, A Elchaninov, A Makarov, G Bolshakova, D Goldshtein and G Sukhikh. Role of VEGF-A in angiogenesis promoted by umbilical cord-derived mesenchymal stromal/stem cells: in vitro study. Stem Cell Res. Ther. 2016; 7, 46.
SK Saxena, AK Singh and MM Ansari. Development of new diagnostic methods and therapeutic compounds against gestational diabetes mellitus using cell culture technology. Expert Opin. Drug Dis. 2016; 21, 1213-27.
C Nguyen-Ngo, N Jayabalan, C Salomon and M Lappas. Molecular pathways disrupted by gestational diabetes mellitus. J. Mol. Endocrinol. 2019; 63, R51-R72.
M Lappas. Activation of inflammasomes in adipose tissue of women with gestational diabetes. Mol. Cell. Endocrinol. 2014; 382, 74-83.
LA Barbour, CE Mccurdy, TL Hernan-Dez, BDL Houssaye, B Draznin and JE Friedman. Reduced IRS-1 and increased serine IRS-1 phosphorylation skeletal muscle from women with GDM. Diabetes 2006; 55, A39.
V Aguirre, ED Werner, J Giraud, YH Lee, SE Shoelson and MF White. Phosphorylation of Ser307 in insulin receptor substrate-1 blocks interactions with the insulin receptor and inhibits insulin action. J. Biol. Chem. 2002; 277, 1531-7.
K Moeschel, A Beck, C Weigert, R Lammers, H Kalbacher, W Voelter, ED Schleicher, HU Häring and R Lehmann. Protein kinase C-ζ-induced phosphorylation of Ser318 in insulin receptor substrate-1 (IRS-1) attenuates the interaction with the insulin receptor and the tyrosine phosphorylation of IRS-1. J. Biol. Chem. 2004; 279, 25157-63.
PM Catalano, SE Nizielski, J Shao, L Preston, L Qiao and JE Friedman. Downregulated IRS-1 and PPARγ in obese women with gestational diabetes: Relationship to FFA during pregnancy. Am. J. Physiol. Endocrinol. Metabol. 2002; 282, E522-E533.
N Chandrasekhar and R Deepa. Gestational diabetes mellitus in India: A comprehensive review. Indian J. Med. Res. 2018; 147, 17-27.
CR Kahn and SE Kahn. The physiology of insulin resistance: Insights from humans and animals. J. Clin. Investigat. 2006; 116, 1496-502.
The HAPO Study Cooperative Research Group. Hyperglycemia and adverse pregnancy outcomes. New Engl. J. Med. 2008; 358, 1991-2002.
KY Lain, SC Robson, JC Kingdom and GC Smith. Maternal metabolic syndrome risk factors and ultrasound-determined measures of fetal growth in early pregnancy. American J. Obstet. Gynecol. 2008; 198, 1-7.
X Luo, RR Li, YQ Li, HP Yu, HN Yu, WG Jiang and YN Li. Reducing VEGFB expression regulates the balance of glucose and lipid metabolism in mice via VEGFR1. Mol. Med. Rep. 2022; 26, 285.
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