NSC-640358 acts as RXRα ligand to promote TNFα-mediated apoptosis of cancer cell

Abstract

Retinoid X receptor α (RXRα) and its N-terminally truncated version tRXRα play important roles in tumorigenesis, while some RXRα ligands possess potent anticancer activities by targeting and modulating the tumorigenic effects of RXRα and tRXRα. Here we describe NSC-640358 (N-6), a thiazolyl-pyrazole derived compound, acts as a selective RXRα ligand to promote TNFα-mediated apoptosis of cancer cell. N-6 binds to RXRα and inhibits the transactivation of RXRα homodimer and RXRα/TR3 heterodimer. Using mutational analysis and computational study, we determine that Arg316 in RXRα, essential for 9-cis-retinoic acid binding and activating RXRα transactivation, is not required for antagonist effects of N-6, whereas Trp305 and Phe313 are crucial for N-6 binding to RXRα by forming extra π–π stacking interactions with N-6, indicating a distinct RXRα binding mode of N-6. N-6 inhibits TR3-stimulated transactivation of Gal4-DBD-RXRα-LBD by binding to the ligand binding pocket of RXRα-LBD, suggesting a strategy to regulate TR3 activity indirectly by using small molecules to target its interacting partner RXRα. For its physiological activities, we show that N-6 strongly inhibits tumor necrosis factor α (TNFα)-induced AKT activation and stimulates TNFα-mediated apoptosis in cancer cells in an RXRα/tRXRα dependent manner. The inhibition of TNFα-induced tRXRα/p85α complex formation by N-6 implies that N-6 targets tRXRα to inhibit TNFα-induced AKT activation and to induce cancer cell apoptosis. Together, our data illustrate a new RXRα ligand with a unique RXRα binding mode and the abilities to regulate TR3 activity indirectly and to induce TNFα-mediated cancer cell apoptosis by targeting RXRα/tRXRα.

Key words： NSC-640358; ligand; RXRα; tRXRα; TNFα; apoptosis

References

1 Altucci L, Leibowitz MD, Ogilvie KM, de Lera AR, Gronemeyer H (2007) RAR and RXR modulation in cancer and metabolic disease. Nat Rev Drug Discov 6: 793―810

2 Balkwill F (2009) Tumour necrosis factor and cancer. Nat Rev Cancer 9: 361―371

3 Cao X, Liu W, Lin F, Li H, Kolluri SK, Lin B, Han YH, Dawson MI, Zhang XK (2004) Retinoid X receptor regulates Nur77/TR3- dependent apoptosis [corrected] by modulating its nuclear export and mitochondrial targeting. Mol Cell Biol 24: 9705―9725

4 Casas F, Daury L, Grandemange S, Busson M, Seyer P, Hatier R, Carazo A, Cabello G, Wrutniak-Cabello C (2003) Endocrine regulation of mitochondrial activity: involvement of truncated RXRalpha and c-Erb Aalpha1 proteins. FASEB J 17: 426―436

5 Dawson MI, Xia Z (2012) The retinoid X receptors and their ligands. Biochim Biophys Acta 1821: 21―56

6 Dawson MI, Hobbs PD, Peterson VJ, Leid M, Lange CW, Feng KC, Chen G, Gu J, Li H, Kolluri SK (2001) Apoptosis induction in cancer cells by a novel analogue of 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalenecarboxylic acid lacking retinoid receptor transcriptional activation activity. Cancer Res 61: 4723―4730

7 de Lera AR, Bourguet W, Altucci L, Gronemeyer H (2007) Design of selective nuclear receptor modulators: RAR and RXR as a case study. Nat Rev Drug Discov 6: 811―820

8 Egea PF, Mitschler A, Rochel N, Ruff M, Chambon P, Moras D (2000) Crystal structure of the human RXRalpha ligand-binding domain bound to its natural ligand: 9-cis retinoic acid. EMBO J 19: 2592―2601

9 Egea PF, Mitschler A, Moras D (2002) Molecular recognition of agonist ligands by RXRs. Mol Endocrinol 16: 987―997

10 Evans RM, Mangelsdorf DJ (2014) Nuclear receptors, RXR, and the big bang. Cell 157: 255―266

11 Gao W, Liu J, Hu M, Huang M, Cai S, Zeng Z, Lin B, Cao X, Chen J, Zeng JZ (2013) Regulation of proteolytic cleavage of retinoid X receptor-alpha by GSK-3beta. Carcinogenesis 34: 1208―1215

12 Ghose R, Zimmerman TL, Thevananther S, Karpen SJ (2004) Endotoxin leads to rapid subcellular re-localization of hepatic RXRalpha: a novel mechanism for reduced hepatic gene expression in inflammation. Nucl Recept 2: 4

13 Gronemeyer H, Gustafsson JA, Laudet V (2004) Principles for modulation of the nuclear receptor superfamily. Nat Rev Drug Discov 3: 950―964

14 Huang J, Powell WC, Khodavirdi AC, Wu J, Makita T, Cardiff RD, Cohen MB, Sucov HM, Roy-Burman P (2002) Prostatic intraepithelial neoplasia in mice with conditional disruption of the retinoid X receptor alpha allele in the prostate epithelium. Cancer Res 62: 4812―4819

15 Jiang SY, Shen SR, Shyu RY, Yu JC, Harn HJ, Yeh MY, Lee MM, Chang YC (1999) Expression of nuclear retinoid receptors in normal, premalignant and malignant gastric tissues determined by in situ hybridization. Br J Cancer 80: 206―214

16 Jorgensen WL, Maxwell DS, TiradoRives J (1996) Development and testing of the OPLS all-atom force field on conformational energetics and properties of organic liquids. J Am Chem Soc 118: 11225―11236

17 Katagiri Y, Takeda K, Yu ZX, Ferrans VJ, Ozato K, Guroff G (2000) Modulation of retinoid signalling through NGF-induced nuclear export of NGFI-B. Nat Cell Biol 2: 435―440

18 Kolluri SK, Bruey-Sedano N, Cao X, Lin B, Lin F, Han YH, Dawson MI, Zhang XK (2003) Mitogenic effect of orphan receptor TR3 and its regulation by MEKK1 in lung cancer cells. Mol Cell Biol 23: 8651―8667

19 Lee SO, Li X, Khan S, Safe S (2011) Targeting NR4A1 (TR3) in cancer cells and tumors. Expert Opin Ther Targets 15: 195―206

20 Lefebvre P, Benomar Y, Staels B (2010) Retinoid X receptors: common heterodimerization partners with distinct functions. Trends Endocrinol Metab 21: 676―683

21 Li H, Kolluri SK, Gu J, Dawson MI, Cao X, Hobbs PD, Lin B, Chen G, Lu J, Lin F (2000a) Cytochrome c release and apoptosis induced by mitochondrial targeting of nuclear orphan receptor TR3. Science 289: 1159―1164

22 Li M, Indra AK, Warot X, Brocard J, Messaddeq N, Kato S, Metzger D, Chambon P (2000b) Skin abnormalities generated by temporally controlled RXRalpha mutations in mouse epidermis. Nature 407: 633―636

23 Lin B, Kolluri SK, Lin F, Liu W, Han YH, Cao X, Dawson MI, Reed JC, Zhang XK (2004) Conversion of Bcl-2 from protector to killer by interaction with nuclear orphan receptor Nur77/TR3. Cell 116: 527―540

24 Lotan Y, Xu XC, Shalev M, Lotan R, Williams R, Wheeler TM, Thompson TC, Kadmon D (2000) Differential expression of nuclear retinoid receptors in normal and malignant prostates. J Clin Oncol 18: 116―121

25 Matsushima-Nishiwaki R, Okuno M, Adachi S, Sano T, Akita K, Moriwaki H, Friedman SL, Kojima S (2001) Phosphorylation of retinoid X receptor alpha at serine 260 impairs its metabolism and function in human hepatocellular carcinoma. Cancer Res 61: 7675―7682

26 Mocellin S, Nitti D (2008) TNF and cancer: the two sides of the coin. Front Biosci 13: 2774―2783

27 Moll UM, Marchenko N, Zhang XK (2006) p53 and Nur77/TR3: transcription factors that directly target mitochondria for cell death induction. Oncogene 25: 4725―4743

28 Nagaya T, Murata Y, Yamaguchi S, Nomura Y, Ohmori S, Fujieda M, Katunuma N, Yen PM, Chin WW, Seo H (1998) Intracellular proteolytic cleavage of 9-cis-retinoic acid receptor alpha by cathepsin L-type protease is a potential mechanism for modulating thyroid hormone action. J Biol Chem 273: 33166―33173

29 Nomura Y, Nagaya T, Yamaguchi S, Katunuma N, Seo H (1999) Cleavage of RXRalpha by a lysosomal enzyme, cathepsin L-type protease. Biochem Biophys Res Commun 254: 388―394

30 Perez E, Bourguet W, Gronemeyer H, de Lera AR (2012) Modulation of RXR function through ligand design. Biochim Biophys Acta 1821: 57―69

31 Remacle AG, Golubkov VS, Shiryaev SA, Dahl R, Stebbins JL, Chernov AV, Cheltsov AV, Pellecchia M, Strongin AY (2012) Novel MT1-MMP small-molecule inhibitors based on insights into hemopexin domain function in tumor growth. Cancer Res 72: 2339―2349

32 Sato Y, Ramalanjaona N, Huet T, Potier N, Osz J, Antony P, Peluso-Iltis C, Poussin-Courmontagne P, Ennifar E, Mely Y (2010) The “Phantom Effect” of the Rexinoid LG100754: structural and functional insights. PLoS One 5: e15119

33 Shiryaev SA, Cheltsov AV, Gawlik K, Ratnikov BI, Strongin AY (2011) Virtual ligand screening of the National Cancer Institute (NCI) compound library leads to the allosteric inhibitory scaffolds of the West Nile Virus NS3 proteinase. Assay Drug Dev Technol 9: 69―78

34 Shiryaev SA, Cheltsov AV, Strongin AY (2012) Probing of exosites leads to novel inhibitor scaffolds of HCV NS3/4A proteinase. PLoS One 7: e40029

35 Shulman AI, Mangelsdorf DJ (2005) Retinoid X receptor heterodimers in the metabolic syndrome. N Engl J Med 353: 604―615

36 Szanto A, Narkar V, Shen Q, Uray IP, Davies PJ, Nagy L (2004) Retinoid X receptors: X-ploring their (patho)physiological functions. Cell Death Differ 11(Suppl 2): S126―S143

37 Takiyama Y, Miyokawa N, Sugawara A, Kato S, Ito K, Sato K, Oikawa K, Kobayashi H, Kimura S, Tateno M (2004) Decreased expression of retinoid X receptor isoforms in human thyroid carcinomas. J Clin Endocrinol Metab 89: 5851―5861

38 Tang XH, Gudas LJ (2011) Retinoids, retinoic acid receptors, and cancer. Annu Rev Pathol 6: 345―364

39 Thomas M, Sukhai MA, Kamel-Reid S (2012) An emerging role for retinoid X receptor alpha in malignant hematopoiesis. Leuk Res 36: 1075―1081

40 Wang Z, Benoit G, Liu J, Prasad S, Aarnisalo P, Liu X, Xu H, Walker NP, Perlmann T (2003) Structure and function of Nurr1 identifies a class of ligand-independent nuclear receptors. Nature 423: 555―560

41 Wang GH, Jiang FQ, Duan YH, Zeng ZP, Chen F, Dai Y, Chen JB, Liu JX, Liu J, Zhou H (2013) Targeting truncated retinoid X receptor-alpha by CF31 induces TNF-alpha-dependent apoptosis. Cancer Res 73: 307―318

42 Wansa KD, Harris JM, Muscat GE (2002) The activation function-1 domain of Nur77/NR4A1 mediates trans-activation, cell specificity, and coactivator recruitment. J Biol Chem 277: 33001―33011

43 Waters JP, Pober JS, Bradley JR (2013) Tumour necrosis factor and cancer. J Pathol 230: 241―248

44 Zhang XK, Hoffmann B, Tran PB, Graupner G, Pfahl M (1992a) Retinoid X receptor is an auxiliary protein for thyroid hormone and retinoic acid receptors. Nature 355: 441―446

45 Zhang XK, Lehmann J, Hoffmann B, Dawson MI, Cameron J, Graupner G, Hermann T, Tran P, Pfahl M (1992b) Homodimer formation of retinoid X receptor induced by 9-cis retinoic acid. Nature 358: 587―591

46 Zhang H, Li L, Chen L, Hu L, Jiang H, Shen X (2011a) Structure basis of bigelovin as a selective RXR agonist with a distinct binding mode. J Mol Biol 407: 13―20

47 Zhang H, Xu X, Chen L, Chen J, Hu L, Jiang H, Shen X (2011b) Molecular determinants of magnolol targeting both RXRalpha and PPARgamma. PLoS One 6: e28253

48 Zhang H, Zhou R, Li L, Chen J, Chen L, Li C, Ding H, Yu L, Hu L, Jiang H (2011c) Danthron functions as a retinoic X receptor antagonist by stabilizing tetramers of the receptor. J Biol Chem 286: 1868―1875

49 Zhou H, Liu W, Su Y, Wei Z, Liu J, Kolluri SK, Wu H, Cao Y, Chen J, Wu Y (2010) NSAID sulindac and its analog bind RXRalpha and inhibit RXRalpha-dependent AKT signaling. Cancer Cell 17: 560―573

50 Zimmerman TL, Thevananther S, Ghose R, Burns AR, Karpen SJ (2006) Nuclear export of retinoid X receptor alpha in response to interleukin-1beta-mediated cell signaling: roles for JNK and SER260. J Biol Chem 281: 15434―15440