Metadichol-Induced Differentiation of Pancreatic Ductal Cells (PANC-1) into Insulin-Producing Cells
Main Article Content
Abstract
Endocrine gene expression in PANC-1, a type of pancreatic cancer cell, has been studied in the context of their potential to be reprogrammed toward a normal, differentiated state. Alkaline phosphatase activity has also been shown in pluripotent stem cells to differentiate between feeder and parental cells in reprogramming experiments. Metadichol®-based cell programming holds promise as a versatile and potentially safer approach for manipulating cellular behavior without the use of viral vectors, gating, or CRISPR. Using qRT‒PCR the results show multifold increase in the gene expression of CA9, GCG, INS MAFA, NEUROD1, NGN3, NKX2-2, PAX6: PDX1, SLC2A2, FOXO1, and SIRT1. ALP levels increased and this activity is often used to distinguish stem cells from feeder cells as well as from parental cells in reprogramming experiments. Pluripotency was confirmed by the presence of islet-like structures on day eight. Metadichol exhibits anticancer activity with a CC50 value of 5.50 µg/ml compared to standard doxorubicin with a CC50 value of 10.28 µg/ml. At 100 ug/ml Metadichol is 82% cytotoxic.in a MTT assay Anti-tumor gene Klotho’s expression was increased 70fold on day eight. All the genes seen expressed regulate endocrine cell development in the pancreas and are involved in insulin and glucagon secretion. Gene network analysis is presented to show how Metadichol induced expression leads to a closed loop feedback network and biological process that would help in mitigating diabetes and other related disorders.
Keywords:
Metadichol, PANC-1, Gene expression, beta cells, Glut2, Nuclear receptors, reprograming cancer cells, Diabetes, Insulin, Glucagon, Alkaline phosphatase, Klotho
Article Details
How to Cite
RAGHAVAN, Palayakotai R.
Metadichol-Induced Differentiation of Pancreatic Ductal Cells (PANC-1) into Insulin-Producing Cells.
Medical Research Archives, [S.l.], v. 11, n. 11, nov. 2023.
ISSN 2375-1924.
Available at: <https://esmed.org/MRA/mra/article/view/4654>. Date accessed: 08 may 2024.
doi: https://doi.org/10.18103/mra.v11i11.4654.
Section
Research Articles
The Medical Research Archives grants authors the right to publish and reproduce the unrevised contribution in whole or in part at any time and in any form for any scholarly non-commercial purpose with the condition that all publications of the contribution include a full citation to the journal as published by the Medical Research Archives.
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3. Donadel G, Pastore D, Della-Morte D, et al. FGF-2b and h-PL transform duct and nonendocrine human pancreatic cells into endocrine insulin secreting cells by modulating differentiating genes. Int J Mol Sci. 2017;18(11):2234. doi:10.3390/ijms18112234.
4. Zhang S, Huang F, Tian W, et al. Andrographolide promotes pancreatic duct cells differentiation into insulin-producing cells by targeting PDX-1. Biochem Pharmacol. 2020;174:113785. doi:10.1016/j.bcp.2019.113785.
5. Damame H, Rooge S, Patil R, Garad C, Arvindekar A. In vitro differentiation of human pancreatic duct–derived PANC-1 cells into β-cell phenotype using Tinospora cordifolia. In Vitro Cell Dev Biol Anim. 2022;58(5):376-383. doi:10.1007/s11626-022-00690-x.
6. Heydari M, Yazdanparast R. Differentiation of PANC-1 ductal cells to β-like cells via cellular GABA modulation by magainin and CPF-7 peptides. Biochem Biophys Res Commun. 2022;597:128-133. doi:10.1016/j.bbrc.2022.01.126.
7. Schmidtlein PM, Volz C, Hackel A, et al. Activation of a ductal-to-endocrine transdifferentiation transcriptional program in the pancreatic cancer cell line PANC-1 is controlled by RAC1 and RAC1b through antagonistic regulation of stemness factors. Cancers. 2021;13(21):5541. doi:10.3390/cancers13215541.
8. Fomina-Yadlin D, Kubicek S, Walpita D, et al. Small-molecule inducers of insulin expression in pancreatic α-cells. Proc Natl Acad Sci U S A. 2010;107(34):15099-15104. doi:10.1073/pnas.1010018107.
9. Thakur G, Lee HJ, Jeon RH, Lee SL, Rho GJ. Small molecule-induced pancreatic β-like cell development: mechanistic approaches and available strategies. Int J Mol Sci. 2020;21(7):2388. doi:10.3390/ijms21072388.
10. Fu Z, Gilbert ER, Liu D. Regulation of insulin synthesis and secretion and pancreatic Beta-cell dysfunction in diabetes. Curr Diabetes Rev. 2013;9(1):25-53. doi:10.2174/157339913804143225.
11. Rorsman P, Braun M. Regulation of insulin secretion in human pancreatic islets. Annu Rev Physiol. 2013;75:155-179. doi:10.1146/annurev-physiol-030212-183754.
12. Dong S, Li W, Li X, et al. Glucose metabolism and tumor microenvironment in pancreatic cancer: A key link in cancer progression. Front Immunol. 2022;13:1038650. doi:10.3389/fimmu.2022.1038650.
13. Hidalgo M, Cascinu S, Kleeff J, et al. Addressing the challenges of pancreatic cancer: future directions for improving outcomes. Pancreatology. 2015;15(1):8-18. doi:10.1016/j.pan.2014.10.001.
14. Ghani MW, Bin L, Jie Y, et al. Differentiation of rat pancreatic duct stem cells into insulin-secreting islet-like cell clusters through BMP7 inducement. Tissue Cell. 2020;67:101439. doi:10.1016/j.tice.2020.101439.
15. Pedica F, Beccari S, Pedron S, et al. PDX-1 (pancreatic/duodenal homeobox-1 protein 1). Pathologica. 2014;106(4):315-321.
16. Vinogradova TV, Sverdlov ED. PDX1: a Unique Pancreatic Master Regulator Constantly Changes Its Functions during Embryonic Development and Progression of Pancreatic Cancer. Biochemistry (Mosc). 2017;82(8):887-893. doi:10.1134/S000629791708003X.
17. Ebrahim N, Shakirova K, Dashinimaev E. PDX1 is the cornerstone of pancreatic β-cell functions and identity. Front Mol Biosci. 2022;9:1091757. doi:10.3389/fmolb.2022.1091757.
18. Nishimura W, Iwasa H, Tumurkhuu M. Role of the transcription factor MAFA in the maintenance of pancreatic β-cells. Int J Mol Sci. 2022;23(9):4478. doi:10.3390/ijms23094478.
19. Raghavan PR, US Patents 8 722 093; vol 2014(9),034:383 (2015) ; 9,006,292; 2015.
20. Berger C, Zdzieblo D. Glucose transporters in pancreatic islets. Pflugers Arch. 2020;472(9):1249-1272. doi:10.1007/s00424-020-02383-4.
21. Yang M, Darwish T, Larraufie P, et al. Inhibition of mitochondrial function by metformin increases glucose uptake, glycolysis and GDF-15 release from intestinal cells. Sci Rep. 2021;11(1):2529. doi:10.1038/s41598-021-81349-7.
22. Afshar N, Safaei S, Nickerson DP, Hunter PJ, Suresh V. Computational modeling of glucose uptake by SGLT1 and Apical GLUT2 in the enterocyte. Front Physiol. 2021;12:699152. doi:10.3389/fphys.2021.699152.
23. Párrizas M, Maestro MA, Boj SF, et al. Hepatic nuclear factor 1-alpha directs nucleosomal hyperacetylation to its tissue-specific transcriptional targets. Mol Cell Biol. 2001;21(9):3234-3243. doi:10.1128/MCB.21.9.3234-3243.2001.
24. Ono Y, Kataoka K. MafA, NeuroD1, and HNF1β synergistically activate the Slc2a2 (Glut2) gene in β-cells. J Mol Endocrinol. 2021;67(3):71-82. doi:10.1530/JME-20-0339.
25. Yang MX, Coates RF, Ambaye A, et al. NKX2.2, PDX-1 and CDX-2 as potential biomarkers to differentiate well-differentiated neuroendocrine tumors. Biomark Res. 2018;6:15. doi:10.1186/s40364-018-0129-8.
26. Anderson KR, Torres CA, Solomon K, et al. Cooperative transcriptional regulation of the essential pancreatic islet gene NeuroD1 (beta2) by Nkx2.2 and neurogenin 3. J Biol Chem. 2009;284(45):31236-31248. doi:10.1074/jbc.M109.048694.
27. Heremans Y, Van De Casteele M, in’t Veld P, et al. Recapitulation of embryonic neuroendocrine differentientiation in adult human pancreatic duct cells expressing neurogenin 3. J Cell Biol. 2002;159(2):303-312. doi:10.1083/jcb.200203074.
28. Rukstalis JM, Habener JF. Neurogenin3: A master regulator of pancreatic islet differentiation and regeneration. Islets. 2009;1(3):177-184. doi:10.4161/isl.1.3.9877.
29. Zhu Y, Liu Q, Zhou Z, Ikeda Y. PDX1, neurogenin-3, and MAFA: critical transcription regulators for beta cell development and regeneration. Stem Cell Res Ther. 2017;8(1):240. doi:10.1186/s13287-017-0694-z.
30. Ouyang J, et al 2006. Pax6 overexpression suppresses cell proliferation and retards the cell cycle in corneal epithelial cells. Invest Ophthalmol Vis Sci. 47(6):2397-407. doi: 10.1167/iovs.05-1083.
31. Ochi S, Manabe S, Kikkawa T, Osumi N. Thirty years’ history since the discovery of Pax6: from central nervous system development to neurodevelopmental disorders. Int J Mol Sci. 2022;23(11):6115. doi:10.3390/ijms23116115.
32. Matsuoka TA, Kaneto H, Miyatsuka T, et al. Regulation of MafA expression in pancreatic β-cells in db/db mice with diabetes. Diabetes. 2010;59(7):1709-1720. doi:10.2337/db08-0693.
33. Liang J, Chirikjian M, Pajvani UB, Bartolomé A. MafA regulation in β-cells: from transcriptional to Post Translational mechanisms. Biomolecules. 2022;12(4):535. doi:10.3390/biom12040535.
34. Fujimoto K, Polonsky KS. Pdx1 and other factors that regulate pancreatic beta-cell survival. Diabetes Obes Metab Suppl. 2009;4(suppl 4):30-37. doi:10.1111/j.1463-1326.2009.01121.x.
35. Puthanveetil P, Wan A, Rodrigues B. FoxO1 is crucial for sustaining cardiomyocyte metabolism and cell survival. Cardiovasc Res. 2013;97(3):393-403. doi:10.1093/cvr/cvs426.
36. Martinez SC, Cras-Méneur C, Bernal-Mizrachi E, Permutt MA. Glucose regulates FoxO1 through insulin receptor signaling in the pancreatic islet beta-cell. Diabetes. 2006;55(6):1581-1591. doi:10.2337/db05-0678.
37. Singh U, Quintanilla RH, Grecian S, Gee KR, Rao MS, Lakshmipathy U. Novel live alkaline phosphatase substrate for identification of pluripotent stem cells. Stem Cell Rev Rep. 2012;8(3):1021-1029. doi:10.1007/s12015-012-9359-6.
38. Raghavan PR. Metadichol® Is a Nano Lipid Emulsion That Expresses All 48 Nuclear Receptors in Stem and Somatic Cells T; 2022. doi:10.21203/rs.3.rs-1797646/v4.
39. Miyazaki S, Taniguchi H, Moritoh Y, et al. nuclear hormone retinoid X receptor (RXR) negatively regulates the glucose-stimulated insulin secretion of pancreatic β-cells. Diabetes. 2010;59(11):2854-2861. doi:10.2337/db09-1897.
40. Ghyselinck NB, Gregg Duester G. Retinoic acid signaling pathways. Development. 2019;46:dev167502. doi:10.1242/dev.167502.
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