Metabolic signatures suggest o-phosphocholine to UDP-N-acetylglucosamine ratio as a potential biomarker for high-glucose and/or palmitate exposure in pancreatic β-cells
- 200 Downloads
Chronic exposure to high-glucose and free fatty acids (FFA) alone/or in combination; and the resulting gluco-, lipo- and glucolipo-toxic conditions, respectively, have been known to induce dysfunction and apoptosis of β-cells in Diabetes. The molecular mechanisms and the development of biomarkers that can be used to predict similarities and differences behind these conditions would help in easier and earlier diagnosis of Diabetes.
This study aims to use metabolomics to gain insight into the mechanisms by which β-cells respond to excess-nutrient stress and identify associated biomarkers.
INS-1E cells were cultured in high-glucose, palmitate alone/or in combination for 24 h to mimic gluco-, lipo- and glucolipo-toxic conditions, respectively. Biochemical and cellular experiments were performed to confirm the establishment of these conditions. To gain molecular insights, abundant metabolites were identified and quantified using 1H-NMR.
No loss of cellular viability was observed in high-glucose while exposure to FFA alone/in combination with high-glucose was associated with increased ROS levels, membrane damage, lipid accumulation, and DNA double-strand breaks. Forty-nine abundant metabolites were identified and quantified using 1H-NMR. Chemometric pair-wise analysis in glucotoxic and lipotoxic conditions, when compared with glucolipotoxic conditions, revealed partial overlap in the dysregulated metabolites; however, the dysregulation was more significant under glucolipotoxic conditions.
The current study compared gluco-, lipo- and glucolipotoxic conditions in parallel and elucidated differences in metabolic pathways that play major roles in Diabetes. o-phosphocholine and UDP-N-acetylglucosamine were identified as common dysregulated metabolites and their ratio was proposed as a potential biomarker for these conditions.
KeywordsGlucotoxicity Glucolipotoxicity Lipotoxicity Metabolic markers Pancreatic beta cell Type 2 diabetes mellitus
The authors acknowledge HF-NMR facility at IISER-Pune (co-funded by DST-FIST and IISER Pune) and the flow cytometer facility at Institute for Applied Biological Research and Development, Pune. SY is thankful for the financial assistance from UGC-JRF, Government of India. JC acknowledges the funding from IISER Pune, Government of India. SS acknowledges the funding from Ramalingaswami fellowship (BT/RLF/Re-entry/11/2012; Department of Biotechnology — DBT, Government of India); University Grants Commission (UGC, Government of India F.4-5(18-FRP)(IV-Cycle)/2017(BSR)) and Board of College and University Development (BCUD) grant (SPPU). SS lab has been supported by Research and Development grant and DST-Purse grant to the Department of Biotechnology, SPPU; and UPE Phase II grant to SPPU. DMS is thankful for the financial assistance from DBT-JRF program and ABM acknowledges DBT, GOI for her Masters in Biotechnology fellowship. JDA acknowledges the funding from Start-up research grant by Science and Engineering Research board (SB/YS/LS-23/2014; SERB), Government of India. INS-1E cells were obtained as a kind gift from Prof. Claes Wollheim and Prof. Pierre Maechler, University Medical Centre, Geneva, Switzerland.
Conceived and designed the experiments: SS, JC. Performed the experiments: SY, DMS, ABM, RK, JDA, SS. Analysis and Interpretation of data: SY, JDA, SS, JC. Contributed reagents/materials/analysis tools: JDA, SS, JC. Compilation of data: SY, JDA, SS. Preparation of the manuscript: SS, JC. All authors have read and approved the final manuscript.
This study was funded by Ramalingaswami fellowship (BT/RLF/Re-entry/11/2012; Department of Biotechnology — DBT, Government of India) to SS; University Grants Commission (UGC, Government of India F.4-5(18-FRP)(IV-Cycle)/2017(BSR)) to SS; Board of College and University Development (BCUD) grant (SPPU) to SS; Start-up research grant by Science and Engineering Research board (SB/YS/LS-23/2014; SERB) to JDA; and funding from IISER Pune, Government of India to JC.
Compliance with ethical standards
Conflict of interest
All authors declare no conflict of interest.
This article does not contain any studies with human and/or animal participants performed by any of the authors.
- Basu, A., Basu, R., Shah, P., Vella, A., Johnson, C. M., Jensen, M., et al. (2001). Type 2 diabetes impairs splanchnic uptake of glucose but does not alter intestinal glucose absorption during enteral glucose feeding: Additional evidence for a defect in hepatic glucokinase activity. Diabetes, 50(6), 1351–1362.CrossRefGoogle Scholar
- Bhakkiyalakshmi, E., Shalini, D., Sekar, T. V., Rajaguru, P., Paulmurugan, R., & Ramkumar, K. M. (2014). Therapeutic potential of pterostilbene against pancreatic beta-cell apoptosis mediated through Nrf2. British Journal of Pharmacology, 171(7), 1747–1757. https://doi.org/10.1111/bph.12577.CrossRefPubMedPubMedCentralGoogle Scholar
- Boslem, E., MacIntosh, G., Preston, A. M., Bartley, C., Busch, A. K., Fuller, M., et al. (2011). A lipidomic screen of palmitate-treated MIN6 beta-cells links sphingolipid metabolites with endoplasmic reticulum (ER) stress and impaired protein trafficking. Biochemical Journal, 435(1), 267–276. https://doi.org/10.1042/BJ20101867.CrossRefPubMedGoogle Scholar
- Carlson, O. D., David, J. D., Schrieder, J. M., Muller, D. C., Jang, H. J., Kim, B. J., et al. (2007). Contribution of nonesterified fatty acids to insulin resistance in the elderly with normal fasting but diabetic 2-h postchallenge plasma glucose levels: The Baltimore Longitudinal Study of Aging. Metabolism, 56(10), 1444–1451. https://doi.org/10.1016/j.metabol.2007.06.009.CrossRefPubMedPubMedCentralGoogle Scholar
- Coute, Y., Brunner, Y., Schvartz, D., Hernandez, C., Masselot, A., Lisacek, F., et al. (2010). Early activation of the fatty acid metabolism pathway by chronic high glucose exposure in rat insulin secretory beta-cells. Proteomics, 10(1), 59–71. https://doi.org/10.1002/pmic.200900080.CrossRefPubMedGoogle Scholar
- Croze, M. L., Geloen, A., & Soulage, C. O. (2015). Abnormalities in myo-inositol metabolism associated with type 2 diabetes in mice fed a high-fat diet: Benefits of a dietary myo-inositol supplementation. British Journal of Nutrition, 113(12), 1862–1875. https://doi.org/10.1017/S000711451500121X.CrossRefPubMedGoogle Scholar
- Cunha, D. A., Hekerman, P., Ladriere, L., Bazarra-Castro, A., Ortis, F., Wakeham, M. C., et al. (2008). Initiation and execution of lipotoxic ER stress in pancreatic beta-cells. Journal of Cell Science, 121(Pt 14), 2308–2318. https://doi.org/10.1242/jcs.026062.CrossRefPubMedPubMedCentralGoogle Scholar
- da Silva, R. P., Leonard, K. A., & Jacobs, R. L. (2017). Dietary creatine supplementation lowers hepatic triacylglycerol by increasing lipoprotein secretion in rats fed high-fat diet. Journal of Nutritional Biochemistry, 50, 46–53. https://doi.org/10.1016/j.jnutbio.2017.08.010.CrossRefPubMedGoogle Scholar
- Deminice, R., da Silva, R. P., Lamarre, S. G., Brown, C., Furey, G. N., McCarter, S. A., et al. (2011). Creatine supplementation prevents the accumulation of fat in the livers of rats fed a high-fat diet. Journal of Nutrition, 141(10), 1799–1804. https://doi.org/10.3945/jn.111.144857.CrossRefPubMedGoogle Scholar
- Dubey, R., Minj, P., Malik, N., Sardesai, D. M., Kulkarni, S. H., Acharya, J. D., et al. (2017). Recombinant human islet amyloid polypeptide forms shorter fibrils and mediates beta-cell apoptosis via generation of oxidative stress. Biochemical Journal, 474(23), 3915–3934. https://doi.org/10.1042/BCJ20170323.CrossRefPubMedGoogle Scholar
- Eccleston, H. B., Andringa, K. K., Betancourt, A. M., King, A. L., Mantena, S. K., Swain, T. M., et al. (2011). Chronic exposure to a high-fat diet induces hepatic steatosis, impairs nitric oxide bioavailability, and modifies the mitochondrial proteome in mice. Antioxidants & Redox Signaling, 15(2), 447–459. https://doi.org/10.1089/ars.2010.3395.CrossRefGoogle Scholar
- El-Azzouny, M., Evans, C. R., Treutelaar, M. K., Kennedy, R. T., & Burant, C. F. (2014). Increased glucose metabolism and glycerolipid formation by fatty acids and GPR40 receptor signaling underlies the fatty acid potentiation of insulin secretion. Journal of Biological Chemistry, 289(19), 13575–13588. https://doi.org/10.1074/jbc.M113.531970.CrossRefPubMedGoogle Scholar
- El Mesallamy, H. O., El-Demerdash, E., Hammad, L. N., & El Magdoub, H. M. (2010). Effect of taurine supplementation on hyperhomocysteinemia and markers of oxidative stress in high fructose diet induced insulin resistance. Diabetology & Metabolic Syndrome, 2, 46. https://doi.org/10.1186/1758-5996-2-46.CrossRefGoogle Scholar
- Erion, K. A., Berdan, C. A., Burritt, N. E., Corkey, B. E., & Deeney, J. T. (2015). Chronic exposure to excess nutrients left-shifts the concentration dependence of glucose-stimulated insulin secretion in pancreatic beta-cells. Journal of Biological Chemistry, 290(26), 16191–16201. https://doi.org/10.1074/jbc.M114.620351.CrossRefPubMedGoogle Scholar
- Franconi, F., Bennardini, F., Mattana, A., Miceli, M., Ciuti, M., Mian, M., et al. (1995). Plasma and platelet taurine are reduced in subjects with insulin-dependent diabetes mellitus: Effects of taurine supplementation. American Journal of Clinical Nutrition, 61(5), 1115–1119. https://doi.org/10.1093/ajcn/61.4.1115.CrossRefPubMedGoogle Scholar
- Goehring, I., Sauter, N. S., Catchpole, G., Assmann, A., Shu, L., Zien, K. S., et al. (2011). Identification of an intracellular metabolic signature impairing beta cell function in the rat beta cell line INS-1E and human islets. Diabetologia, 54(10), 2584–2594. https://doi.org/10.1007/s00125-011-2249-7.CrossRefPubMedGoogle Scholar
- Gu, W., Rebsdorf, A., Hermansen, K., Gregersen, S., & Jeppesen, P. B. (2018). The dynamic effects of isosteviol on insulin secretion and its inability to counteract the impaired beta-cell function during gluco-, lipo-, and aminoacidotoxicity: Studies in vitro. Nutrients, 10(2), 127. https://doi.org/10.3390/nu10020127.CrossRefPubMedCentralGoogle Scholar
- Gualano, B., Painneli, V. D. E. S., Roschel, H., Artioli, G. G., Neves, M., Jr., De Sa Pinto, A. L., et al. (2011). Creatine in type 2 diabetes: A randomized, double-blind, placebo-controlled trial. Medicine and Science in Sports and Exercise, 43(5), 770–778. https://doi.org/10.1249/MSS.0b013e3181fcee7d.CrossRefPubMedGoogle Scholar
- Guan, M., Xie, L., Diao, C., Wang, N., Hu, W., Zheng, Y., et al. (2013). Systemic perturbations of key metabolites in diabetic rats during the evolution of diabetes studied by urine metabonomics. PLoS ONE, 8(4), e60409. https://doi.org/10.1371/journal.pone.0060409.CrossRefPubMedPubMedCentralGoogle Scholar
- Haber, C. A., Lam, T. K., Yu, Z., Gupta, N., Goh, T., Bogdanovic, E., et al. (2003). N-acetylcysteine and taurine prevent hyperglycemia-induced insulin resistance in vivo: Possible role of oxidative stress. American Journal of Physiology. Endocrinology and Metabolism, 285(4), E744–753. https://doi.org/10.1152/ajpendo.00355.2002.CrossRefPubMedGoogle Scholar
- Jansen, S. M., Groener, J. E., Bax, W., Suter, A., Saftig, P., Somerharju, P., et al. (2001). Biosynthesis of phosphatidylcholine from a phosphocholine precursor pool derived from the late endosomal/lysosomal degradation of sphingomyelin. Journal of Biological Chemistry, 276(22), 18722–18727. https://doi.org/10.1074/jbc.M101817200.CrossRefPubMedGoogle Scholar
- Kim, K. S., Oh, D. H., Kim, J. Y., Lee, B. G., You, J. S., Chang, K. J., et al. (2012). Taurine ameliorates hyperglycemia and dyslipidemia by reducing insulin resistance and leptin level in Otsuka Long-Evans Tokushima fatty (OLETF) rats with long-term diabetes. Experimental & Molecular Medicine, 44(11), 665–673. https://doi.org/10.3858/emm.2012.44.11.075.CrossRefGoogle Scholar
- Kim, S. H., Yang, S. O., Kim, H. S., Kim, Y., Park, T., & Choi, H. K. (2009). 1H-nuclear magnetic resonance spectroscopy-based metabolic assessment in a rat model of obesity induced by a high-fat diet. Analytical and Bioanalytical Chemistry, 395(4), 1117–1124. https://doi.org/10.1007/s00216-009-3054-8.CrossRefPubMedGoogle Scholar
- Merglen, A., Theander, S., Rubi, B., Chaffard, G., Wollheim, C. B., & Maechler, P. (2004). Glucose sensitivity and metabolism-secretion coupling studied during two-year continuous culture in INS-1E insulinoma cells. Endocrinology, 145(2), 667–678. https://doi.org/10.1210/en.2003-1099.CrossRefPubMedGoogle Scholar
- Mugabo, Y., Zhao, S., Lamontagne, J., Al-Mass, A., Peyot, M. L., Corkey, B. E., et al. (2017). Metabolic fate of glucose and candidate signaling and excess-fuel detoxification pathways in pancreatic beta-cells. Journal of Biological Chemistry, 292(18), 7407–7422. https://doi.org/10.1074/jbc.M116.763060.CrossRefPubMedGoogle Scholar
- Ogurtsova, K., da Rocha Fernandes, J. D., Huang, Y., Linnenkamp, U., Guariguata, L., Cho, N. H., et al. (2017). IDF Diabetes Atlas: Global estimates for the prevalence of diabetes for 2015 and 2040. Diabetes Research and Clinical Practice, 128, 40–50. https://doi.org/10.1016/j.diabres.2017.03.024.CrossRefPubMedGoogle Scholar
- Pouwels, M. J., Tack, C. J., Span, P. N., Olthaar, A. J., Sweep, C. G., Huvers, F. C., et al. (2004). Role of hexosamines in insulin resistance and nutrient sensing in human adipose and muscle tissue. Journal of Clinical Endocrinology and Metabolism, 89(10), 5132–5137. https://doi.org/10.1210/jc.2004-0291.CrossRefPubMedGoogle Scholar
- Somesh, B. P., Verma, M. K., Sadasivuni, M. K., Mammen-Oommen, A., Biswas, S., Shilpa, P. C., et al. (2013). Chronic glucolipotoxic conditions in pancreatic islets impair insulin secretion due to dysregulated calcium dynamics, glucose responsiveness and mitochondrial activity. BMC Cell Biol, 14, 31. https://doi.org/10.1186/1471-2121-14-31.CrossRefPubMedPubMedCentralGoogle Scholar
- Spegel, P., Andersson, L. E., Storm, P., Sharoyko, V., Gohring, I., Rosengren, A. H., et al. (2015). Unique and shared metabolic regulation in clonal beta-cells and primary islets derived from rat revealed by metabolomics analysis. Endocrinology, 156(6), 1995–2005. https://doi.org/10.1210/en.2014-1391.CrossRefPubMedGoogle Scholar
- Sumner, L. W., Amberg, A., Barrett, D., Beale, M. H., Beger, R., Daykin, C. A., et al. (2007). Proposed minimum reporting standards for chemical analysis Chemical Analysis Working Group (CAWG) Metabolomics Standards Initiative (MSI). Metabolomics, 3(3), 211–221. https://doi.org/10.1007/s11306-007-0082-2.CrossRefPubMedPubMedCentralGoogle Scholar
- Tam, Z. Y., Ng, S. P., Tan, L. Q., Lin, C. H., Rothenbacher, D., Klenk, J., et al. (2017). Metabolite profiling in identifying metabolic biomarkers in older people with late-onset type 2 diabetes mellitus. Scientific Reports, 7(1), 4392. https://doi.org/10.1038/s41598-017-01735-y.CrossRefPubMedPubMedCentralGoogle Scholar
- Testa, R., Vanhooren, V., Bonfigli, A. R., Boemi, M., Olivieri, F., Ceriello, A., et al. (2015). N-glycomic changes in serum proteins in type 2 diabetes mellitus correlate with complications and with metabolic syndrome parameters. PLoS ONE, 10(3), e0119983. https://doi.org/10.1371/journal.pone.0119983.CrossRefPubMedPubMedCentralGoogle Scholar
- Trachtman, H., Futterweit, S., Maesaka, J., Ma, C., Valderrama, E., Fuchs, A., et al. (1995). Taurine ameliorates chronic streptozocin-induced diabetic nephropathy in rats. American Journal of Physiology, 269(3 Pt 2), F429–438. https://doi.org/10.1152/ajprenal.1995.269.3.F429.CrossRefPubMedGoogle Scholar
- Williams, R. E., Lenz, E. M., Rantalainen, M., & Wilson, I. D. (2006). The comparative metabonomics of age-related changes in the urinary composition of male Wistar-derived and Zucker (fa/fa) obese rats. Molecular BioSystems, 2(3–4), 193–202. https://doi.org/10.1039/b517195d.CrossRefPubMedGoogle Scholar
- Yuzefovych, L., Wilson, G., & Rachek, L. (2010). Different effects of oleate vs. palmitate on mitochondrial function, apoptosis, and insulin signaling in L6 skeletal muscle cells: Role of oxidative stress. American Journal of Physiology Heart and Circulatory Physiology, 299(6), 1096–1105. https://doi.org/10.1152/ajpendo.00238.2010.CrossRefGoogle Scholar