Thyroid Hormone Synergizes with PPARγ and cAMP to Drive UCP1 Transcription and Brown-like Adipocyte Phenotype in 3T3-L1 Cells

Manal Malibary

Abstract


Background: Brown adipose tissue expresses uncoupling protein 1(Ucp1), a mitochondrial protein essential for energy balance and non-shivering thermogenesis. This study aimed to show how cyclic adenosine monophosphate (cAMP), peroxisome proliferator-activated receptor γ (Pparγ), and triiodothyronine (T3) pathways together stimulate Ucp1 expression in white adipose tissue.

Methods: Differentiated 3T3-L1 cells, both transiently transfected and stably transduced with a UCP1 vector, were used to assess the effects of  T3, Pparγ, cAMP agonists, and their combinations on Ucp1 expression.

Results: The results showed that treatment with T3, Pparγ agonists, and cAMP agonists significantly increased Ucp1 promoter activity in both undifferentiated and differentiated 3T3-L1 cells. In differentiated cells, combined treatment with rosiglitazone, T3, and forskolin led to a time-dependent increase in Ucp1 expression: 5-fold on Day 4, 7.5-fold on Day 8, and 10.5-fold on Day 12 (P < 0.001). Prdm16 mRNA increased 1.5-fold on Days 4 and 8 (P < 0.001), and 3-fold on Day 12 (P < 0.01) after T3 and rosiglitazone treatment, with forskolin added in the final 12 hours. Pgc1α expression peaked at a 2-fold increase on Day 12 (P < 0.05). Cidea expression was markedly upregulated, showing a maximum increase approximately 3 fold on Day 12 (P < 0.001). Elovl3 doubled on Days 4 and 8, and increased approximately 3-fold by Day 12 (P < 0.001).

Conclusion: This study suggests that activating PPARγ, cAMP, and T3 pathways can induce browning of white adipose tissue, offering potential therapeutic strategies for obesity management.

Keywords: Brown Adipose Tissue; cAMP; PPARγ; Thyroid Hormones; Ucp1; White Adipose Tissue; Transdifferentiation, Brown-Like Adipocyte


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Choe SS, Huh JY, Hwang IJ, Kim JI, Kim JB. Adipose Tissue Remodeling: Its Role in Energy Metabolism and Metabolic Disorders. Frontiers in Endocrinology, (2016); 7: 30.

Yau WW, Singh BK, Lesmana R, Zhou J, Sinha RA, Wong KA, et al. Thyroid hormone (T(3)) stimulates brown adipose tissue activation via mitochondrial biogenesis and MTOR-mediated mitophagy. Autophagy, (2019); 15(1): 131-150.

Kim S, Li A, Monti S, Schlezinger JJ. Tributyltin induces a transcriptional response without a brite adipocyte signature in adipocyte models. Archives of Toxicology, (2018); 92(9): 2859-2874.

Sharp LZ, Shinoda K, Ohno H, Scheel DW, Tomoda E, Ruiz L, et al. Human BAT possesses molecular signatures that resemble beige/brite cells. PLoS ONE, (2012); 7(11): e49452.

Coulter AA, Bearden CM, Liu X, Koza RA, Kozak LP. Dietary fat interacts with QTLs controlling induction of Pgc-1 alpha and Ucp1 during conversion of white to brown fat. Physiological Genomics, (2003); 14(2): 139-147.

Iida S, Chen W, Nakadai T, Ohkuma Y, Roeder RG. PRDM16 enhances nuclear receptor-dependent transcription of the brown fat-specific Ucp1 gene through interactions with Mediator subunit MED1. Genes & Development, (2015); 29(3): 308-321.

Finlin BS, Memetimin H, Confides AL, Kasza I, Zhu B, Vekaria HJ, et al. Human adipose beiging in response to cold and mirabegron. JCI Insight, (2018); 3(15): e121510.

Villarroya F, Peyrou M, Giralt M. Transcriptional regulation of the uncoupling protein-1 gene. Biochimie, (2017); 134: 86-92.

Tabuchi C, Sul HS. Signaling Pathways Regulating Thermogenesis. Frontiers in Endocrinology, (2021); 12: 595020.

Yamamoto T, Diao Z, Murakami M, Shimokawa F, Matsui T, Funaba M. Factors affecting the induction of uncoupling protein 1 in C2C12 myogenic cells. Cytokine, (2022); 157: 155936.

Hamaoka T, Fu X, Tomonaga S, Hashimoto O, Murakami M, Funaba M. Stimulation of uncoupling protein 1 expression by beta-alanine in brown adipocytes. Archives of Biochemistry and Biophysics, (2022); 727: 109341.

McAninch EA, Bianco AC. Thyroid hormone signaling in energy homeostasis and energy metabolism. Annals of the New York Academy of Sciences, (2014); 1311: 77-87.

Wu Y, Xu B, Koenig RJ. Thyroid hormone response element sequence and the recruitment of retinoid X receptors for thyroid hormone responsiveness. Journal of Biological Chemistry, (2001); 276(6): 3929-3936.

Malibary MA. Vitamin A: A Key Inhibitor of Adipocyte Differentiation. PPAR Research, (2023); 2023: 7405954.

Song Y, Yao X, Ying H. Thyroid hormone action in metabolic regulation. Protein & Cell, (2011); 2(5): 358-368.

Ribeiro MO, Bianco SD, Kaneshige M, Schultz JJ, Cheng SY, Bianco AC, et al. Expression of uncoupling protein 1 in mouse brown adipose tissue is thyroid hormone receptor-beta isoform specific and required for adaptive thermogenesis. Endocrinology, (2010); 151(1): 432-440.

Chen W, Yang Q, Roeder RG. Dynamic interactions and cooperative functions of PGC-1alpha and MED1 in TRalpha-mediated activation of the brown-fat-specific UCP-1 gene. Molecular Cell, (2009); 35(6): 755-768.

Rim JS, Kozak LP. Regulatory motifs for CREB-binding protein and Nfe2l2 transcription factors in the upstream enhancer of the mitochondrial uncoupling protein 1 gene. Journal of Biological Chemistry, (2002); 277(37): 34589-34600.

Liu Q, Hill PJ, Karamitri A, Ryan KJ, Chen HY, Lomax MA. Construction of a doxycycline inducible adipogenic lentiviral expression system. Plasmid, (2013); 69(1): 96-103.

Tews D, Pula T, Funcke JB, Jastroch M, Keuper M, Debatin KM, et al. Elevated UCP1 levels are sufficient to improve glucose uptake in human white adipocytes. Redox Biology, (2019); 26: 101286.

So J, Taleb S, Wann J, Strobel O, Kim K, Roh HC. Chronic cAMP activation induces adipocyte browning through discordant biphasic remodeling of transcriptome and chromatin accessibility. Molecular Metabolism, (2022); 66: 101619.

Desai A, Loureiro ZY, DeSouza T, Yang Q, Solivan-Rivera J, Corvera S. cAMP driven UCP1 induction in human adipocytes requires ATGL-catalyzed lipolysis. Molecular Metabolism, (2024); 90: 102051.

Castillo M, Hall JA, Correa-Medina M, Ueta C, Kang HW, Cohen DE, et al. Disruption of thyroid hormone activation in type 2 deiodinase knockout mice causes obesity with glucose intolerance and liver steatosis only at thermoneutrality. Diabetes, (2011); 60(4): 1082-1089.

Thomas C, Auwerx J, Schoonjans K. Bile acids and the membrane bile acid receptor TGR5--connecting nutrition and metabolism. Thyroid, (2008); 18(2): 167-174.

Lee JY, Takahashi N, Yasubuchi M, Kim YI, Hashizaki H, Kim MJ, et al. Triiodothyronine induces UCP-1 expression and mitochondrial biogenesis in human adipocytes. American Journal of Physiology-Cell Physiology, (2012); 302(2): C463-C472.

Szychowski KA, Skóra B, Kryshchyshyn-Dylevych A, Kaminskyy D, Tobiasz J, Lesyk RB, et al. 4-Thiazolidinone-based derivatives do not affect differentiation of mouse embryo fibroblasts (3T3-L1 cell line) into adipocytes. Chemico-Biological Interactions, (2021); 345: 109538.

Fayyad AM, Khan AA, Abdallah SH, Alomran SS, Bajou K, Khattak MNK. Rosiglitazone enhances browning adipocytes in association with MAPK and PI3-K pathways during the differentiation of telomerase-transformed mesenchymal stromal cells into adipocytes. International Journal of Molecular Sciences, (2019); 20(7): 1618.

Kurylowicz A, Puzianowska-Kuznicka M. Induction of adipose tissue browning as a strategy to combat obesity. International Journal of Molecular Sciences, (2020); 21(17): 6241.

Sohn JH, Kim JI, Jeon YG, Park J, Kim JB. Effects of Three Thiazolidinediones on Metabolic Regulation and cold-induced thermogenesis. Molecules and Cells, (2018); 41(10): 900-908.

Bogacka I, Ukropcova B, McNeil M, Gimble JM, Smith SR. Structural and functional consequences of mitochondrial biogenesis in human adipocytes in vitro. The Journal of Clinical Endocrinology & Metabolism, (2005); 90(12): 6650-6656.

Asano H, Kanamori Y, Higurashi S, Nara T, Kato K, Matsui T, et al. Induction of beige-like adipocytes in 3T3-L1 cells. Journal of Veterinary Medical Science, (2014); 76(1): 57-64.

Fogli S, Pellegrini S, Adinolfi B, Mariotti V, Melissari E, Betti L, et al. Rosiglitazone reverses salbutamol-induced beta(2) -adrenoceptor tolerance in airway smooth muscle. British Journal of Pharmacology, (2011); 162(2): 378-391.

Kovaničová Z, Kurdiová T, Baláž M, Štefanička P, Varga L, Kulterer OC, et al. Cold exposure distinctively modulates parathyroid and thyroid hormones in cold-acclimatized and non-acclimatized humans. Endocrinology, (2020); 161(7): bqaa051.

Karamitri A, Shore AM, Docherty K, Speakman JR, Lomax MA. Combinatorial transcription factor regulation of the cyclic AMP-response element on the Pgc-1alpha promoter in white 3T3-L1 and brown HIB-1B preadipocytes. Journal of Biological Chemistry, (2009); 284(31): 20738-20752.

Abe Y, Fujiwara Y, Takahashi H, Matsumura Y, Sawada T, Jiang S, et al. Histone demethylase JMJD1A coordinates acute and chronic adaptation to cold stress via thermogenic phospho-switch. Nature Communications, (2018); 9(1): 1566.

Leiva M, Matesanz N, Pulgarín-Alfaro M, Nikolic I, Sabio G. Uncovering the Role of p38 Family Members in Adipose Tissue Physiology. Frontiers in Endocrinology, (2020); 11: 572089.

Wang CZ, Wei D, Guan MP, Xue YM. Triiodothyronine regulates distribution of thyroid hormone receptors by activating AMP-activated protein kinase in 3T3-L1 adipocytes and induces uncoupling protein-1 expression. Molecular and Cellular Biochemistry, (2014); 393(1-2): 247-254.

Puigserver P, Wu Z, Park CW, Graves R, Wright M, Spiegelman BM. A cold-inducible coactivator of nuclear receptors linked to adaptive thermogenesis. Cell, (1998); 92(6): 829-839.

Obregon MJ. Thyroid hormone and adipocyte differentiation. Thyroid, (2008); 18(2): 185-195.

Fukano K, Okamatsu-Ogura Y, Tsubota A, Nio-Kobayashi J, Kimura K. Cold Exposure Induces Proliferation of Mature Brown Adipocyte in a ß3-Adrenergic Receptor-Mediated Pathway. PLoS ONE, (2016); 11(11): e0166579.

Westerberg R, Månsson JE, Golozoubova V, Shabalina IG, Backlund EC, Tvrdik P, et al. ELOVL3 is an important component for early onset of lipid recruitment in brown adipose tissue. Journal of Biological Chemistry, (2006); 281(8): 4958-4968.

Qin X, Park HG, Zhang JY, Lawrence P, Liu GW, Subramanian N, et al. Brown but not white adipose cells synthesize omega-3 docosahexaenoic acid in culture. Prostaglandins, Leukotrienes and Essential Fatty Acids, (2016); 104: 19-24.

Jash S, Banerjee S, Lee MJ, Farmer SR, Puri V. CIDEA Transcriptionally Regulates UCP1 for Britening and Thermogenesis in Human Fat Cells. iScience, (2019); 20: 73-89.

Egusa G, Ohno H, Nagano G, Sagawa J, Shinjo H, Yamamoto Y, et al. Selective activation of PPARα maintains thermogenic capacity of beige adipocytes. iScience, (2023); 26(7): 107143.




DOI: http://dx.doi.org/10.62940/als.v12i4.3714

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