Sat, Aug 23, 2025
[Archive]
Volume 17, Issue 1 (March-2025 2025)
Iranian Journal of Blood and Cancer 2025, 17(1): 20-28 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Razavi Babaheidari S R, Salahi-Niri A, Mohammadi M H, Hamidpour M, Esmaeili S. DNA Methylation-Dependent Regulation of Lipoprotein Lipase Expression During Human Mesenchymal Stem Cell Differentiation into Adipocytes. Iranian Journal of Blood and Cancer 2025; 17 (1) :20-28
URL: http://ijbc.ir/article-1-1647-en.html
URL: http://ijbc.ir/article-1-1647-en.html
Seied Rasoul Razavi Babaheidari1 
, Aryan Salahi-Niri2 , Mohammad Hossein Mohammadi3 , Mohsen Hamidpour3 , Shadi Esmaeili *3
, Aryan Salahi-Niri2 , Mohammad Hossein Mohammadi3 , Mohsen Hamidpour3 , Shadi Esmaeili *3
1- Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
2- Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
3- Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
2- Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
3- Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Abstract: (524 Views)
Background: Lipoprotein lipase (LPL) is a critical enzyme in lipid metabolism that hydrolyzes triglyceride-rich lipoproteins. While its role in mature adipose tissue is well understood, the epigenetic regulation of LPL during mesenchymal stem cell (MSC) differentiation into adipocytes remains poorly characterized. This study aimed to investigate the temporal expression pattern of LPL and its relationship with DNA methylation during adipogenic differentiation of human bone marrow-derived MSCs.
Methods: Human bone marrow MSCs were isolated and characterized using flow cytometry for specific surface markers (CD34, CD31, CD90, and CD105). Cells were differentiated into adipocytes over 14 days and osteoblasts over 21 days using specific differentiation media. Differentiation was confirmed through Oil Red O and Alizarin Red staining respectively. LPL gene expression was analyzed using both qualitative RT-PCR and quantitative real-time PCR at day 0 (undifferentiated) and day 14 (differentiated) timepoints. DNA methylation patterns were assessed using methylation-specific PCR (MSP) following bisulfite conversion, with collagen gene serving as an internal control.
Results: Flow cytometry confirmed MSC identity through positive expression of CD166, CD13, CD105, and CD44. Successful adipogenic differentiation was demonstrated by Oil Red O-positive lipid droplet accumulation, while osteogenic differentiation was confirmed by Alizarin Red S staining of calcium deposits. LPL gene expression was absent in undifferentiated MSCs but showed significant expression in differentiated adipocytes at day 14, coinciding with morphological changes and lipid accumulation.
Conclusion: This study demonstrates that LPL expression is epigenetically regulated during MSC differentiation into adipocytes, with significant changes in both gene expression and DNA methylation patterns. The temporal correlation between LPL expression, methylation status, and adipogenic differentiation suggests that LPL serves as a key molecular switch in this process.
Methods: Human bone marrow MSCs were isolated and characterized using flow cytometry for specific surface markers (CD34, CD31, CD90, and CD105). Cells were differentiated into adipocytes over 14 days and osteoblasts over 21 days using specific differentiation media. Differentiation was confirmed through Oil Red O and Alizarin Red staining respectively. LPL gene expression was analyzed using both qualitative RT-PCR and quantitative real-time PCR at day 0 (undifferentiated) and day 14 (differentiated) timepoints. DNA methylation patterns were assessed using methylation-specific PCR (MSP) following bisulfite conversion, with collagen gene serving as an internal control.
Results: Flow cytometry confirmed MSC identity through positive expression of CD166, CD13, CD105, and CD44. Successful adipogenic differentiation was demonstrated by Oil Red O-positive lipid droplet accumulation, while osteogenic differentiation was confirmed by Alizarin Red S staining of calcium deposits. LPL gene expression was absent in undifferentiated MSCs but showed significant expression in differentiated adipocytes at day 14, coinciding with morphological changes and lipid accumulation.
Conclusion: This study demonstrates that LPL expression is epigenetically regulated during MSC differentiation into adipocytes, with significant changes in both gene expression and DNA methylation patterns. The temporal correlation between LPL expression, methylation status, and adipogenic differentiation suggests that LPL serves as a key molecular switch in this process.
Keywords: Lipoprotein lipase, DNA methylation, Mesenchymal stem cells, Adipogenic differentiation, Epigenetic regulation, Gene expression
: Original Article |
Subject:
Genetics
Received: 2025/01/19 | Accepted: 2025/03/17 | Published: 2025/03/30
Received: 2025/01/19 | Accepted: 2025/03/17 | Published: 2025/03/30
References
1. Geldenhuys WJ, Lin L, Darvesh AS, Sadana P. Emerging strategies of targeting lipoprotein lipase for metabolic and cardiovascular diseases. Drug Discov Today. 2017;22(2):352-65. [DOI:10.1016/j.drudis.2016.10.007]
2. Wang H, Eckel RH. Lipoprotein lipase: from gene to obesity. American Journal of Physiology-Endocrinology and Metabolism. 2009;297(2):E271-E88. [DOI:10.1152/ajpendo.90920.2008]
3. Kockx M, Kritharides L. Triglyceride-Rich Lipoproteins. Cardiology Clinics. 2018;36(2):265-75. [DOI:10.1016/j.ccl.2017.12.008]
4. Liu SS, Fang X, Wen X, Liu JS, Alip M, Sun T, et al. How mesenchymal stem cells transform into adipocytes: Overview of the current understanding of adipogenic differentiation. World J Stem Cells. 2024;16(3):245-56. [DOI:10.4252/wjsc.v16.i3.245]
5. Pamplona JH, Zoehler B, Shigunov P, Barisón MJ, Severo VR, Erich NM, et al. Alternative Methods as Tools for Obesity Research: In Vitro and In Silico Approaches. Life (Basel). 2022;13(1). [DOI:10.3390/life13010108]
6. Pittenger MF, Discher DE, Péault BM, Phinney DG, Hare JM, Caplan AI. Mesenchymal stem cell perspective: cell biology to clinical progress. npj Regenerative Medicine. 2019;4(1):22. [DOI:10.1038/s41536-019-0083-6]
7. Kolf CM, Cho E, Tuan RS. Mesenchymal stromal cells: Biology of adult mesenchymal stem cells: regulation of niche, self-renewal and differentiation. Arthritis Research & Therapy. 2007;9(1):204. [DOI:10.1186/ar2116]
8. Ahmad B, Serpell CJ, Fong IL, Wong EH. Molecular Mechanisms of Adipogenesis: The Anti-adipogenic Role of AMP-Activated Protein Kinase. Frontiers in Molecular Biosciences. 2020;7. [DOI:10.3389/fmolb.2020.00076]
9. Feng P, Pang P, Sun Z, Xie Z, Chen T, Wang S, et al. Enhancer-mediated FOXO3 expression promotes MSC adipogenic differentiation by activating autophagy. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease. 2024;1870(2):166975. [DOI:10.1016/j.bbadis.2023.166975]
10. Gonzales AM, Orlando RA. Role of adipocyte-derived lipoprotein lipase in adipocyte hypertrophy. Nutr Metab (Lond). 2007;4:22. [DOI:10.1186/1743-7075-4-22]
11. Enerbäck S, Ohlsson BG, Samuelsson L, Bjursell G. Characterization of the human lipoprotein lipase (LPL) promoter: evidence of two cis-regulatory regions, LP-alpha and LP-beta, of importance for the differentiation-linked induction of the LPL gene during adipogenesis. Mol Cell Biol. 1992;12(10):4622-33. [DOI:10.1128/MCB.12.10.4622]
12. Kuryłowicz A. Estrogens in Adipose Tissue Physiology and Obesity-Related Dysfunction. Biomedicines [Internet]. 2023; 11(3). [DOI:10.3390/biomedicines11030690]
13. Lorzadeh A, Romero-Wolf M, Goel A, Jadhav U. Epigenetic Regulation of Intestinal Stem Cells and Disease: A Balancing Act of DNA and Histone Methylation. Gastroenterology. 2021;160(7):2267-82. [DOI:10.1053/j.gastro.2021.03.036]
14. Fagnocchi L, Mazzoleni S, Zippo A. Integration of Signaling Pathways with the Epigenetic Machinery in the Maintenance of Stem Cells. Stem Cells Int. 2016;2016:8652748. [DOI:10.1155/2016/8652748]
15. Li X, Jiang O, Wang S. Molecular mechanisms of cellular metabolic homeostasis in stem cells. International Journal of Oral Science. 2023;15(1):52. [DOI:10.1038/s41368-023-00262-z]
16. Pereira B, Correia FP, Alves IA, Costa M, Gameiro M, Martins AP, et al. Epigenetic reprogramming as a key to reverse ageing and increase longevity. Ageing Research Reviews. 2024;95:102204. [DOI:10.1016/j.arr.2024.102204]
17. Castellano-Castillo D, Moreno-Indias I, Fernández-García JC, Alcaide-Torres J, Moreno-Santos I, Ocaña L, et al. Adipose Tissue LPL Methylation is Associated with Triglyceride Concentrations in the Metabolic Syndrome. Clin Chem. 2018;64(1):210-8. [DOI:10.1373/clinchem.2017.277921]
18. Heindel JJ, Blumberg B, Cave M, Machtinger R, Mantovani A, Mendez MA, et al. Metabolism disrupting chemicals and metabolic disorders. Reproductive Toxicology. 2017;68:3-33. [DOI:10.1016/j.reprotox.2016.10.001]
19. Choi MR, In Y-H, Park J, Park T, Jung KH, Chai JC, et al. Genome-scale DNA methylation pattern profiling of human bone marrow mesenchymal stem cells in long-term culture. Experimental & Molecular Medicine. 2012;44(8):503-12. [DOI:10.3858/emm.2012.44.8.057]
20. Chinn CA, Ren H, Morival JLP, Nie Q, Wood MA, Downing TL. Examining age-dependent DNA methylation patterns and gene expression in the male and female mouse hippocampus. Neurobiol Aging. 2021;108:223-35. [DOI:10.1016/j.neurobiolaging.2021.08.006]
21. Guay SP, Brisson D, Lamarche B, Marceau P, Vohl MC, Gaudet D, et al. DNA methylation variations at CETP and LPL gene promoter loci: new molecular biomarkers associated with blood lipid profile variability. Atherosclerosis. 2013;228(2):413-20. [DOI:10.1016/j.atherosclerosis.2013.03.033]
22. Shi Y, Zhang H, Huang S, Yin L, Wang F, Luo P, et al. Epigenetic regulation in cardiovascular disease: mechanisms and advances in clinical trials. Signal Transduction and Targeted Therapy. 2022;7(1):200. [DOI:10.1038/s41392-022-01055-2]
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
- Iranian Journal of Blood and Cancer
- Journal Tel: +989369936116
- Publisher Tel: +982122275917
- Mail: Info@ijbc.ir
