Metadichol, a Natural Ligand for the Expression of Yamanaka Reprogramming Factors in Human Cardiac, Fibroblast, and Cancer Cell Lines
Main Article Content
Abstract
The reprogramming of somatic cells into embryonic-like stem cells requires the activation of four essential transcription factors, referred to as the Yamanaka factors: Sox2, Oct4, Klf4, and c-Myc. However, the utilization of these genes and viral vectors for their delivery presents a risk of carcinogenesis, which renders induced pluripotent stem cell (iPSC) lines unsuitable for clinical applications. Typically, direct reprogramming involves the use of viral vectors to induce the expression of these factors. In contrast, metadichol, a novel approach, enhances the expression of Yamanaka factors in cells using a dose ranging from 1 pg to 100 ng, thereby preventing the need for viral vectors. This non-viral method renders cellular reprogramming safer and more clinically viable. Treatment with metadichol within the specific dose range resulted in a substantial augmentation of the fibroblast expression of OCT4, SOX2, KLF4, KLF2, and Nanog, which was confirmed through qRT-PCR and western blot analyses. The expression levels of OCT4, KLF4, Nanog, and Sox2, exhibited an increase of 4.01-, 3.51-, 1.26-, and 2.5-fold, respectively, compared to the controls. Notably, A549 and Colo-205 cancer cells demonstrated a marked elevation in expression levels. The reprogramming of primary human cancer cells presents considerable challenges. However, in triple-negative primary breast cancer cells, metadichol treatment led to the substantial upregulation of the expression levels of OCT4, KLF4, Nanog, and Sox2 by 19.6-, 8.07-, 2.45-, and 6.91-fold, respectively, across the dose range of 1 pg to 100 ng. Klotho, an antiaging gene that is modulated by metadichol, down regulates TP53, a crucial factor for the production of somatic cell iPSCs. Additionally, metadichol enhances the availability of vitamin C, which is essential for generating iPSCs from somatic cells. Increased levels of alkaline phosphatase, indicative of cellular differentiation, were observed in the treated group compared to the controls. Metadichol is a nontoxic nanoemulsion that is derived from long-chain C26–C28 alcohols from food sources and activates Yamanaka factors across various cell types without requiring the use of viral or CRISPR-based methodologies, thus substantially advancing somatic cell reprogramming.
Keywords:
Metadichol, Yamanaka Factors, Cellular Reprogramming, Induced Pluripotent Stem Cells (iPSCs), Non-Viral Reprogramming, Fibroblast Differentiation, Cancer Cell Lines, OCT4, Sox2, KLF4
Article Details
How to Cite
RAGHAVAN, Palayakotai R.
Metadichol, a Natural Ligand for the Expression of Yamanaka Reprogramming Factors in Human Cardiac, Fibroblast, and Cancer Cell Lines.
Medical Research Archives, [S.l.], v. 12, n. 6, july 2024.
ISSN 2375-1924.
Available at: <https://esmed.org/MRA/mra/article/view/5323>. Date accessed: 02 july 2024.
doi: https://doi.org/10.18103/mra.v12i6.5323.
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.
References
1. Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006;126(4):663-676.
2. Wang L, Su Y, Huang C, et al. NANOG and LIN28 dramatically improve human cell reprogramming by modulating LIN41 and canonical WNT activities. Biol Open. 2019;8(1 2):bio047225.
3. Lapasset L, Milhavet O, Prieur A, et al. Rejuvenating senescent and centenarian human cells by reprogramming through the pluripotent state. Genes Dev. 2011;25(21) :2248-2253.
4. López-Otín C, Blasco MA, Partridge L, Serrano M, Kroemer G. The hallmarks of aging. Cell. 2013;153(6):1194-1217.
5. Abad M, Mosteiro L, Pantoja C, et al. Reprogramming in vivo produces teratomas and iPS cells with totipotency features. Nature. 2013;502(7471):340-345.
6. Ocampo A, Reddy P, Martinez-Redondo P, et al. In vivo amelioration of age-associated hallmarks by partial reprogramming. Cell. 2016;167(7):1719-1733.e12.
7. Lu Y, Brommer B, Tian X, David.A. Sinclair et al. Reprogramming to recover youthful epigenetic information and restore vision. Nature. 2020;588(7836):124-129.
8. Gill D, Parry A, Santos F, et al. Multi-omic rejuvenation of human cells by maturation phase transient reprogramming. eLife. 2022; 11:e71624.
9. Tan Y, Ooi S, Wang L. Immunogenicity and tumorigenicity of pluripotent stem cells and their derivatives: genetic and epigenetic perspectives. Curr Stem Cell Res Ther. 2014;9(1):63-72.
10. Deng XY, Wang H, Wang T, et al. Non-viral methods for generating integration-free, induced pluripotent stem cells. Curr Stem Cell Res Ther. 2015;10(2):153-158.
11. Plotnikov A, Kozer N, Cohen G, et al. PRMT1 inhibition induces differentiation of colon cancer cells. Sci Rep. 2020;10(1):20030.
12. Sato M, Saitoh I, Kiyokawa Y, et al. Tissue-nonspecific alkaline phosphatase, a possible mediator of cell maturation: towards a new paradigm. Cells. 2021;10(12):3338.
13. Fedde KN, Whyte MP. Alkaline phosphatase (tissue-nonspecific isoenzyme) is a phosphoethanolamine and pyridoxal-5’-phosphate ectophosphatase: normal and hypophosphatasia fibroblast study. Am J Hum Genet. 1990;47(5):767-775.
14. Raghavan PR. Policosanol nanoparticles. US patent,2014, 8722,093
15. Raghavan PR. Metadichol, a natural ligand for the expression of yamanaka reprogramming factors in somatic and primary cancer cell lines. 28 June 2022, PREPRINT (Version 4) available at Research Square. https://doi.org/10.21203/rs.3.rs-1727437/v4.
16. Raghavan PR. Metadichol treatment of fibroblasts and embryonic stem cells regulates key cardiac progenitors. Cardiol Cardiovasc Med. 2023;7(4):302-310.
17. Hu K, Yu J, Suknuntha K, et al. Efficient generation of transgene-free induced pluripotent stem cells from normal and neoplastic bone marrow and cord blood mononuclear cells. Blood. 2011;117(14):e109-e119.
18. Yeo JC, Jiang J, Tan ZY, et al. Klf2 is an essential factor that sustains ground state pluripotency. Cell Stem Cell. 2014;14(6):864-872.
19. Bialkowska AB, Yang VW, Mallipattu SK. Krüppel-like factors in mammalian stem cells and development. Development. 2017;144(5) :737-754.
20. Qiu D, Ye S, Ruiz B, et al. Klf2 and Tfcp2l1, two Wnt/β-catenin targets, act synergistically to induce and maintain naive pluripotency. Stem Cell Rep. 2015;5(3):314-322.
21. Raghavan PR. Metadichol: an agonist that expresses the anti-aging gene klotho in various cell lines. Fortune J Health Sci. 2023;6(4):357-362.
22. Shmulevich R, Nissim TB, Wolf I, et al. Klotho rewires cellular metabolism of breast cancer cells through alteration of calcium shuttling and mitochondrial activity. Oncogene. 2020;39(24):4636-4649.
23. Fan J, Sun Z. The antiaging gene klotho regulates proliferation and differentiation of adipose-derived stem cells. Stem Cells. 2016;34(6):1615-1625.
24. Trivanović D, Jauković A, Popović B, et al. Mesenchymal stem cells of different origin: comparative evaluation of proliferative capacity, telomere length and pluripotency marker expression. Life Sci. 2015;141:61-73.
25. Raghavan PR. Metadichol® A Nano lipid emulsion that expresses all 49 nuclear receptors in stem and somatic cells. Arch Clin Biomed Res. 2023;7(5):524-536.
26. Morohashi KI, Inoue M, Baba T. Coordination of Multiple Cellular Processes by NR5A1/Nr5a1. Endocrinol Metab (Seoul). 2020 Dec;35(4):756-764.
27. Kim, KP., Han, D.W., Kim, J. et al. Biological importance of OCT transcription factors in reprogramming and development. Exp Mol Med 2021; 53, 1018–1028 .
28. Wang T, Chen K, Zeng X, et al. The histone demethylases Jhdm1a/1b enhance somatic cell reprogramming in a vitamin-C-dependent manner. Cell Stem Cell. 2011;9 (6):575-587.
29. Stadtfeld M, Apostolou E, Ferrari F, et al. Ascorbic acid prevents loss of Dlk1-Dio3 imprinting and facilitates generation of all-iPS cell mice from terminally differentiated B cells. Nat Genet. 2012;44(4):398-405.
30. Raghavan PR. Metadichol®, vitamin C and GULO gene expression in mouse adipocytes. Biol Med. 2017;10(1):426.
31. Raghavan PR. Metadichol® and vitamin C increase in vivo, an open-label study. Vitam Miner. 2017;6:163.
32. Raghavan PR. Metadichol®-induced high levels of vitamin C: case studies. Vitam Miner. 2017;6:169.
33. Banito A, Rashid ST, Acosta JC, et al. Senescence impairs successful reprogramming to pluripotent stem cells. Genes Dev. 2009; 23(18):2134-2139.
34. Wang S, Tian X, et al. Intracellular Reactive Oxygen Species Mediate the Therapeutic Effect of Induced Pluripotent Stem Cells for Acute Kidney Injury. Oxid Med Cell Longev. 2020 Mar 26;2020:1609638
35. Esteban MA, Wang T, Qin B, et al. Vitamin C enhances the generation of mouse and human induced pluripotent stem cells. Cell Stem Cell. 2010;6(1):71-79.
36. Lee Chong T, Ahearn EL, Cimmino L. Reprogramming the epigenome with vitamin C. Front Cell Dev Biol. 2019;7:128.
37. Cloos PA, Christensen J, Agger K, Helin K. Erasing the methyl mark: histone demethylases at the center of cellular differentiation and disease. Genes Dev. 2008;22(9):1115-1140.
38. Liang J, Wang N, He J, et al. Induction of Sertoli-like cells from human fibroblasts by NR5A1 and GATA4. eLife. 2019;8:e48767.
39. Kato T, Esaki M, Matsuzawa A, Ikeda Y. NR5A1 is required for functional maturation of Sertoli cells during postnatal development. Reproduction. 2012;143(5):663-672.
40. Heng JC, Feng B, Han J, et al. The nuclear receptor Nr5a2 can replace Oct4 in the reprogramming of murine somatic cells to pluripotent cells. Cell Stem Cell. 2010;6 (2):167-174.
41. Washburn RL, Hibler T, Kaur G, Dufour JM. Sertoli cell immune regulation: a double-edged sword. Front Immunol. 2022;13:913502.
42. Kaur G, Thompson LA, Dufour J. Therapeutic potential of immune privileged Sertoli cells. Anim Reprod. 2015;12:105-117.
43. Mital P, Kaur G, Dufour JM. Immunoprotective Sertoli cells: making allogeneic and xenogeneic transplantation feasible. Reproduction. 2010;139(3):495-504.
44. Koc G, Ozdemir AA, Girgin G, et al. Male infertility in Sertoli cell-only syndrome: an investigation of autosomal gene defects. Int J Urol. 2019;26(2):292-298.
45. Simandi Z, Horvath A, Wright LC, et al. OCT4 acts as an integrator of pluripotency and signal-induced differentiation. Mol Cell. 2016;63(4):647-661.
46. Patra SK. Roles of OCT4 in pathways of embryonic development and cancer progression. Mech Ageing Dev. 2020;189:111286.
47. Heng JC, Ng HH. Transcriptional regulation in embryonic stem cells. Adv Exp Med Biol. 2010;695:76-91.
48. Hu H, Qian K, Ho MC, Zheng YG. Small Molecule Inhibitors of Protein Arginine Methyltransferases. Expert Opin Investig Drugs. 2016;25(3):335-58.
49. Rao SR, Snaith AE, Marino D, et al. Tumor-derived alkaline phosphatase regulates tumour growth, epithelial plasticity and disease-free survival in metastatic prostate cancer. Br J Cancer. 2017;116(2):227-236.
50. Cook B, Rafiq R, Lee H, et al. Discovery of a small molecule promoting mouse and human osteoblast differentiation via activation of p38 MAPK-β. Cell Chem Biol. 2019;26(7):926-935.e6.
51. Haydon RC, Zhou L, Feng T, et al. Nuclear receptor agonists as potential differentiation therapy agents for human osteosarcoma. Clin Cancer Res. 2002;8(5):1288-1294.
52. He BC, Chen L, et al. Synergistic antitumor effect of the activated PPARgamma and retinoid receptors on human osteosarcoma. Clin Cancer Res. 2010 Apr 15;16(8):2235-45.
53. Colapietro F, Gershwin ME, Lleo A. PPAR agonists for the treatment of primary biliary cholangitis: Old and new tales. J Transl Autoimmun. 2023 Jan 5;6:100188.
54. Shi Z, Shen T, Liu Y, Huang Y, Jiao J. Retinoic acid receptor γ (Rarg) and nuclear receptor subfamily 5, group A, member 2 (Nr5a2) promote conversion of fibroblasts to functional neurons. J Biol Chem. 2014;289(10) :6415-6428.
55. Jiang T, Zeng Q, He J. Do alkaline phosphatases have great potential in the diagnosis, prognosis, and treatment of tumors? Transl Cancer Res. 2023 Oct 31;12(10 ):2932-2945
56. Nikitin A, Egorov S, Daraselia N, Mazo I. Pathway studio--the analysis and navigation of molecular networks. Bioinformatics. 2003;19( 16):2155-2157.
57. Sivachenko AY, Yuryev A, Daraselia N, Mazo I. Molecular networks in microarray analysis. J Bioinform Comput Biol. 2007;5(2B) :429-456.
58. Kenakin T. Inverse, protean, and ligand-selective agonism: matters of receptor conformation. FASEB J. 2001;15(3):598-611.
59. De Oliveira RM. Klotho RNAi induces premature senescence of human cells via a p53/p21 dependent pathway. FEBS Lett. 2006;580(24):5753-5758.
60. Hong H, Takahashi K, Ichisaka T, et al. Suppression of induced pluripotent stem cell generation by the p53–p21 pathway. Nature. 2009;460(7259):1132-1135.
61. Kawamura T, Suzuki J, Wang YV, et al. Linking the p53 tumour suppressor pathway to somatic cell reprogramming. Nature. 2009;460(7259):1140-1144.
62. Forster RE, Jurutka PW, Hsieh JC, et al. Vitamin D receptor controls expression of the anti-aging klotho gene in mouse and human renal cells. Biochem Biophys Res Commun. 2011;414(3):557-562.
63. Salehi-Tabar R, Nguyen-Yamamoto L, Tavera-Mendoza LE, etal. Vitamin D receptor as a master regulator of the c-MYC/MXD1 network. Proc Natl Acad Sci U S A. 2012;109 (46):18827-18832.
64. Raghavan PR. The quest for immortality: introducing metadichol® a novel telomerase activator. Stem Cell Res Ther. 2019;9:446.
65. Tomczak A, Mortensen JM, Winnenburg R, et al. Interpretation of biological experiments changes with evolution of the gene ontology and its annotations. Sci Rep. 2018;8(1):5115.
66. Alemán CL, Más R, Hernández C, et al. A 12-month study of policosanol oral toxicity in Sprague Dawley rats. Toxicol Lett. 1994;70 (1):77-87.
67. Alemán CL, Más Ferreiro R, Noa Puig M, Rodeiro Guerra I, Hernández Ortega C, Capote A. Carcinogenicity of policosanol in Sprague Dawley rats: a 24 month study. Teratog Carcinog Mutagen. 1994;14(5):239-249.
68. Alemán CL, Puig MN, Elías EC, et al. Carcinogenicity of policosanol in mice: an 18-month study. Food Chem Toxicol. 1995;33(7):573-578.
69. Raghavan PR. Metadichol® and healthy skin: one approach many possible cures. J Clin Exp Dermatol Res. 2018;09(2):1.
70. Raghavan PR. Metadichol, A novel ROR gamma inverse agonist and its applications in psoriasis. J Clin Exp Dermatol Res. 2017;8:433.
2. Wang L, Su Y, Huang C, et al. NANOG and LIN28 dramatically improve human cell reprogramming by modulating LIN41 and canonical WNT activities. Biol Open. 2019;8(1 2):bio047225.
3. Lapasset L, Milhavet O, Prieur A, et al. Rejuvenating senescent and centenarian human cells by reprogramming through the pluripotent state. Genes Dev. 2011;25(21) :2248-2253.
4. López-Otín C, Blasco MA, Partridge L, Serrano M, Kroemer G. The hallmarks of aging. Cell. 2013;153(6):1194-1217.
5. Abad M, Mosteiro L, Pantoja C, et al. Reprogramming in vivo produces teratomas and iPS cells with totipotency features. Nature. 2013;502(7471):340-345.
6. Ocampo A, Reddy P, Martinez-Redondo P, et al. In vivo amelioration of age-associated hallmarks by partial reprogramming. Cell. 2016;167(7):1719-1733.e12.
7. Lu Y, Brommer B, Tian X, David.A. Sinclair et al. Reprogramming to recover youthful epigenetic information and restore vision. Nature. 2020;588(7836):124-129.
8. Gill D, Parry A, Santos F, et al. Multi-omic rejuvenation of human cells by maturation phase transient reprogramming. eLife. 2022; 11:e71624.
9. Tan Y, Ooi S, Wang L. Immunogenicity and tumorigenicity of pluripotent stem cells and their derivatives: genetic and epigenetic perspectives. Curr Stem Cell Res Ther. 2014;9(1):63-72.
10. Deng XY, Wang H, Wang T, et al. Non-viral methods for generating integration-free, induced pluripotent stem cells. Curr Stem Cell Res Ther. 2015;10(2):153-158.
11. Plotnikov A, Kozer N, Cohen G, et al. PRMT1 inhibition induces differentiation of colon cancer cells. Sci Rep. 2020;10(1):20030.
12. Sato M, Saitoh I, Kiyokawa Y, et al. Tissue-nonspecific alkaline phosphatase, a possible mediator of cell maturation: towards a new paradigm. Cells. 2021;10(12):3338.
13. Fedde KN, Whyte MP. Alkaline phosphatase (tissue-nonspecific isoenzyme) is a phosphoethanolamine and pyridoxal-5’-phosphate ectophosphatase: normal and hypophosphatasia fibroblast study. Am J Hum Genet. 1990;47(5):767-775.
14. Raghavan PR. Policosanol nanoparticles. US patent,2014, 8722,093
15. Raghavan PR. Metadichol, a natural ligand for the expression of yamanaka reprogramming factors in somatic and primary cancer cell lines. 28 June 2022, PREPRINT (Version 4) available at Research Square. https://doi.org/10.21203/rs.3.rs-1727437/v4.
16. Raghavan PR. Metadichol treatment of fibroblasts and embryonic stem cells regulates key cardiac progenitors. Cardiol Cardiovasc Med. 2023;7(4):302-310.
17. Hu K, Yu J, Suknuntha K, et al. Efficient generation of transgene-free induced pluripotent stem cells from normal and neoplastic bone marrow and cord blood mononuclear cells. Blood. 2011;117(14):e109-e119.
18. Yeo JC, Jiang J, Tan ZY, et al. Klf2 is an essential factor that sustains ground state pluripotency. Cell Stem Cell. 2014;14(6):864-872.
19. Bialkowska AB, Yang VW, Mallipattu SK. Krüppel-like factors in mammalian stem cells and development. Development. 2017;144(5) :737-754.
20. Qiu D, Ye S, Ruiz B, et al. Klf2 and Tfcp2l1, two Wnt/β-catenin targets, act synergistically to induce and maintain naive pluripotency. Stem Cell Rep. 2015;5(3):314-322.
21. Raghavan PR. Metadichol: an agonist that expresses the anti-aging gene klotho in various cell lines. Fortune J Health Sci. 2023;6(4):357-362.
22. Shmulevich R, Nissim TB, Wolf I, et al. Klotho rewires cellular metabolism of breast cancer cells through alteration of calcium shuttling and mitochondrial activity. Oncogene. 2020;39(24):4636-4649.
23. Fan J, Sun Z. The antiaging gene klotho regulates proliferation and differentiation of adipose-derived stem cells. Stem Cells. 2016;34(6):1615-1625.
24. Trivanović D, Jauković A, Popović B, et al. Mesenchymal stem cells of different origin: comparative evaluation of proliferative capacity, telomere length and pluripotency marker expression. Life Sci. 2015;141:61-73.
25. Raghavan PR. Metadichol® A Nano lipid emulsion that expresses all 49 nuclear receptors in stem and somatic cells. Arch Clin Biomed Res. 2023;7(5):524-536.
26. Morohashi KI, Inoue M, Baba T. Coordination of Multiple Cellular Processes by NR5A1/Nr5a1. Endocrinol Metab (Seoul). 2020 Dec;35(4):756-764.
27. Kim, KP., Han, D.W., Kim, J. et al. Biological importance of OCT transcription factors in reprogramming and development. Exp Mol Med 2021; 53, 1018–1028 .
28. Wang T, Chen K, Zeng X, et al. The histone demethylases Jhdm1a/1b enhance somatic cell reprogramming in a vitamin-C-dependent manner. Cell Stem Cell. 2011;9 (6):575-587.
29. Stadtfeld M, Apostolou E, Ferrari F, et al. Ascorbic acid prevents loss of Dlk1-Dio3 imprinting and facilitates generation of all-iPS cell mice from terminally differentiated B cells. Nat Genet. 2012;44(4):398-405.
30. Raghavan PR. Metadichol®, vitamin C and GULO gene expression in mouse adipocytes. Biol Med. 2017;10(1):426.
31. Raghavan PR. Metadichol® and vitamin C increase in vivo, an open-label study. Vitam Miner. 2017;6:163.
32. Raghavan PR. Metadichol®-induced high levels of vitamin C: case studies. Vitam Miner. 2017;6:169.
33. Banito A, Rashid ST, Acosta JC, et al. Senescence impairs successful reprogramming to pluripotent stem cells. Genes Dev. 2009; 23(18):2134-2139.
34. Wang S, Tian X, et al. Intracellular Reactive Oxygen Species Mediate the Therapeutic Effect of Induced Pluripotent Stem Cells for Acute Kidney Injury. Oxid Med Cell Longev. 2020 Mar 26;2020:1609638
35. Esteban MA, Wang T, Qin B, et al. Vitamin C enhances the generation of mouse and human induced pluripotent stem cells. Cell Stem Cell. 2010;6(1):71-79.
36. Lee Chong T, Ahearn EL, Cimmino L. Reprogramming the epigenome with vitamin C. Front Cell Dev Biol. 2019;7:128.
37. Cloos PA, Christensen J, Agger K, Helin K. Erasing the methyl mark: histone demethylases at the center of cellular differentiation and disease. Genes Dev. 2008;22(9):1115-1140.
38. Liang J, Wang N, He J, et al. Induction of Sertoli-like cells from human fibroblasts by NR5A1 and GATA4. eLife. 2019;8:e48767.
39. Kato T, Esaki M, Matsuzawa A, Ikeda Y. NR5A1 is required for functional maturation of Sertoli cells during postnatal development. Reproduction. 2012;143(5):663-672.
40. Heng JC, Feng B, Han J, et al. The nuclear receptor Nr5a2 can replace Oct4 in the reprogramming of murine somatic cells to pluripotent cells. Cell Stem Cell. 2010;6 (2):167-174.
41. Washburn RL, Hibler T, Kaur G, Dufour JM. Sertoli cell immune regulation: a double-edged sword. Front Immunol. 2022;13:913502.
42. Kaur G, Thompson LA, Dufour J. Therapeutic potential of immune privileged Sertoli cells. Anim Reprod. 2015;12:105-117.
43. Mital P, Kaur G, Dufour JM. Immunoprotective Sertoli cells: making allogeneic and xenogeneic transplantation feasible. Reproduction. 2010;139(3):495-504.
44. Koc G, Ozdemir AA, Girgin G, et al. Male infertility in Sertoli cell-only syndrome: an investigation of autosomal gene defects. Int J Urol. 2019;26(2):292-298.
45. Simandi Z, Horvath A, Wright LC, et al. OCT4 acts as an integrator of pluripotency and signal-induced differentiation. Mol Cell. 2016;63(4):647-661.
46. Patra SK. Roles of OCT4 in pathways of embryonic development and cancer progression. Mech Ageing Dev. 2020;189:111286.
47. Heng JC, Ng HH. Transcriptional regulation in embryonic stem cells. Adv Exp Med Biol. 2010;695:76-91.
48. Hu H, Qian K, Ho MC, Zheng YG. Small Molecule Inhibitors of Protein Arginine Methyltransferases. Expert Opin Investig Drugs. 2016;25(3):335-58.
49. Rao SR, Snaith AE, Marino D, et al. Tumor-derived alkaline phosphatase regulates tumour growth, epithelial plasticity and disease-free survival in metastatic prostate cancer. Br J Cancer. 2017;116(2):227-236.
50. Cook B, Rafiq R, Lee H, et al. Discovery of a small molecule promoting mouse and human osteoblast differentiation via activation of p38 MAPK-β. Cell Chem Biol. 2019;26(7):926-935.e6.
51. Haydon RC, Zhou L, Feng T, et al. Nuclear receptor agonists as potential differentiation therapy agents for human osteosarcoma. Clin Cancer Res. 2002;8(5):1288-1294.
52. He BC, Chen L, et al. Synergistic antitumor effect of the activated PPARgamma and retinoid receptors on human osteosarcoma. Clin Cancer Res. 2010 Apr 15;16(8):2235-45.
53. Colapietro F, Gershwin ME, Lleo A. PPAR agonists for the treatment of primary biliary cholangitis: Old and new tales. J Transl Autoimmun. 2023 Jan 5;6:100188.
54. Shi Z, Shen T, Liu Y, Huang Y, Jiao J. Retinoic acid receptor γ (Rarg) and nuclear receptor subfamily 5, group A, member 2 (Nr5a2) promote conversion of fibroblasts to functional neurons. J Biol Chem. 2014;289(10) :6415-6428.
55. Jiang T, Zeng Q, He J. Do alkaline phosphatases have great potential in the diagnosis, prognosis, and treatment of tumors? Transl Cancer Res. 2023 Oct 31;12(10 ):2932-2945
56. Nikitin A, Egorov S, Daraselia N, Mazo I. Pathway studio--the analysis and navigation of molecular networks. Bioinformatics. 2003;19( 16):2155-2157.
57. Sivachenko AY, Yuryev A, Daraselia N, Mazo I. Molecular networks in microarray analysis. J Bioinform Comput Biol. 2007;5(2B) :429-456.
58. Kenakin T. Inverse, protean, and ligand-selective agonism: matters of receptor conformation. FASEB J. 2001;15(3):598-611.
59. De Oliveira RM. Klotho RNAi induces premature senescence of human cells via a p53/p21 dependent pathway. FEBS Lett. 2006;580(24):5753-5758.
60. Hong H, Takahashi K, Ichisaka T, et al. Suppression of induced pluripotent stem cell generation by the p53–p21 pathway. Nature. 2009;460(7259):1132-1135.
61. Kawamura T, Suzuki J, Wang YV, et al. Linking the p53 tumour suppressor pathway to somatic cell reprogramming. Nature. 2009;460(7259):1140-1144.
62. Forster RE, Jurutka PW, Hsieh JC, et al. Vitamin D receptor controls expression of the anti-aging klotho gene in mouse and human renal cells. Biochem Biophys Res Commun. 2011;414(3):557-562.
63. Salehi-Tabar R, Nguyen-Yamamoto L, Tavera-Mendoza LE, etal. Vitamin D receptor as a master regulator of the c-MYC/MXD1 network. Proc Natl Acad Sci U S A. 2012;109 (46):18827-18832.
64. Raghavan PR. The quest for immortality: introducing metadichol® a novel telomerase activator. Stem Cell Res Ther. 2019;9:446.
65. Tomczak A, Mortensen JM, Winnenburg R, et al. Interpretation of biological experiments changes with evolution of the gene ontology and its annotations. Sci Rep. 2018;8(1):5115.
66. Alemán CL, Más R, Hernández C, et al. A 12-month study of policosanol oral toxicity in Sprague Dawley rats. Toxicol Lett. 1994;70 (1):77-87.
67. Alemán CL, Más Ferreiro R, Noa Puig M, Rodeiro Guerra I, Hernández Ortega C, Capote A. Carcinogenicity of policosanol in Sprague Dawley rats: a 24 month study. Teratog Carcinog Mutagen. 1994;14(5):239-249.
68. Alemán CL, Puig MN, Elías EC, et al. Carcinogenicity of policosanol in mice: an 18-month study. Food Chem Toxicol. 1995;33(7):573-578.
69. Raghavan PR. Metadichol® and healthy skin: one approach many possible cures. J Clin Exp Dermatol Res. 2018;09(2):1.
70. Raghavan PR. Metadichol, A novel ROR gamma inverse agonist and its applications in psoriasis. J Clin Exp Dermatol Res. 2017;8:433.