nach oben

Erschienen in:

01.10.2014 | Original Article

Anticancer activity of VDR-coregulator inhibitor PS121912

verfasst von: Preetpal S. Sidhu, Kelly Teske, Belaynesh Feleke, Nina Y. Yuan, Margaret L. Guthrie, Grant B. Fernstrum, Nishita D. Vyas, Lanlan Han, Joshua Preston, Jonathan W. Bogart, Nicholas R. Silvaggi, James M. Cook, Rakesh K. Singh, Daniel D. Bikle, Leggy A. Arnold

Erschienen in: Cancer Chemotherapy and Pharmacology | Ausgabe 4/2014

Einloggen, um Zugang zu erhalten

Abstract

Purpose

PS121912 has been developed as selective vitamin D receptor (VDR)-coregulator inhibitor starting from a high throughput screening campaign to identify new agents that modulate VDR without causing hypercalcemia. Initial antiproliferative effects of PS121912 were observed that are characterized herein to enable future in vivo investigation with this molecule.

Methods

Antiproliferation and apoptosis were determined using four different cancer cell lines (DU145, Caco2, HL-60 and SKOV3) in the presence of PS121912, 1,25-(OH)₂D₃, or a combination of 1,25-(OH)₂D₃ and PS121912. VDR si-RNA was used to identify the role of VDR during this process. The application of ChIP enabled us to determine the involvement of coregulator recruitment during transcription, which was investigated by RT-PCR with VDR target genes and those affiliated with cell cycle progression. Translational changes of apoptotic proteins were determined with an antibody array. The preclinical characterization of PS121912 includes the determination of metabolic stability and CYP3A4 inhibition.

Results

PS121912 induced apoptosis in all four cancer cells, with HL-60 cells being the most sensitive. At sub-micromolar concentrations, PS121912 amplified the growth inhibition of cancer cells caused by 1,25-(OH)₂D₃ without being antiproliferative by itself. A knockout study with VDR si-RNA confirmed the mediating role of VDR. VDR target genes induced by 1,25-(OH)₂D₃ were down-regulated with the co-treatment of PS121912. This process was highly dependent on the recruitment of coregulators that in case of CYP24A1 was SRC2. The combination of PS121912 and 1,25-(OH)₂D₃ reduced the presence of SRC2 and enriched the occupancy of corepressor NCoR at the promoter site. E2F transcription factors 1 and 4 were down-regulated in the presence of PS121912 and 1,25-(OH)₂D₃ that in turn reduced the transcription levels of cyclin A and D, thus arresting HL-60 cells in the S or G2/M phase. In addition, proteins with hematopoietic functions such as cyclin-dependent kinase 6, histone deacetylase 9 and transforming growth factor beta 2 and 3 were down-regulated as well. Elevated levels of P21 and GADD45, in concert with cyclin D1, also mediated the antiproliferative response of HL-60 in the presence of 1,25-(OH)₂D₃ and PS121912. Studies at higher concentration of P121912 identified a VDR-independent pathway of antiproliferation that included the enzymatic and transcriptional activation of caspase 3/7.

Conclusion

Overall, we conclude that PS121912 behaves like a VDR antagonist at low concentrations but interacts with more targets at higher concentrations leading to apoptosis mediated by caspase 3/7 activation. In addition, PS121912 showed an acceptable metabolic stability to enable in vivo cancer studies.

Nur mit Berechtigung zugänglich

Eisman JA, Martin TJ, MacIntyre I, Moseley JM (1979) 1,25-dihydroxyvitamin-D-receptor in breast cancer cells. Lancet 2:1335–1336PubMedCrossRef

Colston K, Colston MJ, Feldman D (1981) 1,25-dihydroxyvitamin D3 and malignant melanoma: the presence of receptors and inhibition of cell growth in culture. Endocrinology 108:1083–1086PubMedCrossRef

Savli H, Aalto Y, Nagy B, Knuutila S, Pakkala S (2002) Gene expression analysis of 1,25(OH)2D3-dependent differentiation of HL-60 cells: a cDNA array study. Br J Haematol 118:1065–1070PubMedCrossRef

Swami S, Raghavachari N, Muller UR, Bao YP, Feldman D (2003) Vitamin D growth inhibition of breast cancer cells: gene expression patterns assessed by cDNA microarray. Breast Cancer Res Treat 80:49–62PubMedCrossRef

Krishnan AV, Shinghal R, Raghavachari N, Brooks JD, Peehl DM, Feldman D (2004) Analysis of vitamin D-regulated gene expression in LNCaP human prostate cancer cells using cDNA microarrays. Prostate 59:243–251PubMedCrossRef

Wood RJ, Tchack L, Angelo G, Pratt RE, Sonna LA (2004) DNA microarray analysis of vitamin D-induced gene expression in a human colon carcinoma cell line. Physiol Genomics 17:122–129PubMedCrossRef

Zhang X, Li P, Bao J, Nicosia SV, Wang H, Enkemann SA, Bai W (2005) Suppression of death receptor-mediated apoptosis by 1,25-dihydroxyvitamin D3 revealed by microarray analysis. J Biol Chem 280:35458–35468PubMedCrossRefPubMedCentral

Akutsu N, Lin R, Bastien Y, Bestawros A, Enepekides DJ, Black MJ, White JH (2001) Regulation of gene Expression by 1alpha,25-dihydroxyvitamin D3 and its analog EB1089 under growth-inhibitory conditions in squamous carcinoma Cells. Mol Endocrinol 15:1127–1139PubMed

Brumbaugh PF, Haussler MR (1974) 1 Alpha, 25-dihydroxycholecalciferol receptors in intestine. II. Temperature-dependent transfer of the hormone to chromatin via a specific cytosol receptor. J Biol Chem 249:1258–1262PubMed

10.

Mellon WS, DeLuca HF (1979) An equilibrium and kinetic study of 1,25-dihydroxyvitamin D3 binding to chicken intestinal cytosol employing high specific activity 1,25-dehydroxy[3H-26, 27] vitamin D3. Arch Biochem Biophys 197:90–95PubMedCrossRef

11.

Gross C, Stamey T, Hancock S, Feldman D (1998) Treatment of early recurrent prostate cancer with 1, 25-dihydroxyvitamin D3 (calcitriol). J Urol 159:2035–2039 discussion 2039-2040PubMedCrossRef

12.

Beer TM, Myrthue A (2004) Calcitriol in cancer treatment: from the lab to the clinic. Mol Cancer Ther 3:373–381PubMed

13.

Gulliford T, English J, Colston KW, Menday P, Moller S, Coombes RC (1998) A phase I study of the vitamin D analogue EB 1089 in patients with advanced breast and colorectal cancer. Br J Cancer 78:6–13PubMedCrossRefPubMedCentral

14.

Jain RK, Trump DL, Egorin MJ, Fernandez M, Johnson CS, Ramanathan RK (2011) A phase I study of the vitamin D3 analogue ILX23-7553 administered orally to patients with advanced solid tumors. Invest New Drugs 29:1420–1425PubMedCrossRef

15.

McKenna NJ, Lanz RB, O’Malley BW (1999) Nuclear receptor coregulators: cellular and molecular biology. Endocr Rev 20:321–344PubMed

16.

Tagami T, Lutz WH, Kumar R, Jameson JL (1998) The interaction of the vitamin D receptor with nuclear receptor corepressors and coactivators. Biochem Biophys Res Commun 253:358–363PubMedCrossRef

17.

Masuyama H, Brownfield CM, St-Arnaud R, MacDonald PN (1997) Evidence for ligand-dependent intramolecular folding of the AF-2 domain in vitamin D receptor-activated transcription and coactivator interaction. Mol Endocrinol 11:1507–1517PubMedCrossRef

18.

Hong H, Kohli K, Garabedian MJ, Stallcup MR (1997) GRIP1, a transcriptional coactivator for the AF-2 transactivation domain of steroid, thyroid, retinoid, and vitamin D receptors. Mol Cell Biol 17:2735–2744PubMedPubMedCentral

19.

Li H, Gomes PJ, Chen JD (1997) RAC3, a steroid/nuclear receptor-associated coactivator that is related to SRC-1 and TIF2. Proc Natl Acad Sci U S A 94:8479–8484PubMedCrossRefPubMedCentral

20.

Nandhikonda P, Lynt WZ, McCallum MM, Ara T, Baranowski AM, Yuan NY, Pearson D, Bikle DD, Guy RK, Arnold LA (2012) Discovery of the first irreversible small molecule inhibitors of the interaction between the vitamin D receptor and coactivators. J Med Chem 55:4640–4651PubMedCrossRefPubMedCentral

21.

Mita Y, Dodo K, Noguchi-Yachide T, Miyachi H, Makishima M, Hashimoto Y, Ishikawa M (2010) LXXLL peptide mimetics as inhibitors of the interaction of vitamin D receptor with coactivators. Bioorg Med Chem Lett 20:1712–1717PubMedCrossRef

22.

Mita Y, Dodo K, Noguchi-Yachide T, Hashimoto Y, Ishikawa M (2013) Structure-activity relationship of benzodiazepine derivatives as LXXLL peptide mimetics that inhibit the interaction of vitamin D receptor with coactivators. Bioorg Med Chem 21:993–1005PubMedCrossRef

23.

Sidhu PS, Nassif N, McCallum MM, Teske K, Feleke B, Yuan NY, Nandhikonda P, Cook JM, Singh RK, Bikle DD, Arnold LA (2014) Development of Novel Vitamin D Receptor–Coactivator Inhibitors. ACS Med Chem Lett 5(2):199–204PubMedCrossRef

24.

Miura D, Manabe K, Ozono K, Saito M, Gao Q, Norman AW, Ishizuka S (1999) Antagonistic action of novel 1alpha,25-dihydroxyvitamin D3-26, 23-lactone analogs on differentiation of human leukemia cells (HL-60) induced by 1alpha,25-dihydroxyvitamin D3. J Biol Chem 274:16392–16399PubMedCrossRef

25.

Bury Y, Steinmeyer A, Carlberg C (2000) Structure activity relationship of carboxylic ester antagonists of the vitamin D(3) receptor. Mol Pharmacol 58:1067–1074PubMed

26.

Igarashi M, Yoshimoto N, Yamamoto K, Shimizu M, Ishizawa M, Makishima M, DeLuca HF, Yamada S (2007) Identification of a highly potent vitamin D receptor antagonist: (25S)-26-adamantyl-25-hydroxy-2-methylene-22,23-didehydro-19,27-dinor-20-epi-vita min D3 (ADMI3). Arch Biochem Biophys 460:240–253PubMedCrossRef

27.

Inaba Y, Yoshimoto N, Sakamaki Y, Nakabayashi M, Ikura T, Tamamura H, Ito N, Shimizu M, Yamamoto K (2009) A new class of vitamin D analogues that induce structural rearrangement of the ligand-binding pocket of the receptor. J Med Chem 52:1438–1449PubMedCrossRef

28.

Teske K, Nandhikonda P, Bogart JW, Feleke B, Sidhu PS, Yuan NY, Prestron J, Goy R, Singh RK, Bikle DD, Cook JM, Arnold LA (2014) Identification of VDR antagonists among nuclear receptor ligands using virtual screening. Nucl Receptor Res 1:1–8CrossRef

29.

Nandhikonda P, Yasgar A, Baranowski AM, Sidhu PS, McCallum MM, Pawlak AJ, Teske K, Feleke B, Yuan NY, Kevin C, Bikle DD, Ayers SD, Webb P, Rai G, Simeonov A, Jadhav A, Maloney D, Arnold LA (2013) Peroxisome Proliferation-Activated Receptor delta Agonist GW0742 Interacts Weakly with Multiple Nuclear Receptors, Including the Vitamin D Receptor. Biochemistry-Us 52:4193–4203CrossRef

30.

Thornberry NA, Lazebnik Y (1998) Caspases: enemies within. Science 281:1312–1316PubMedCrossRef

31.

Meyer MB, Goetsch PD, Pike JW (2010) A downstream intergenic cluster of regulatory enhancers contributes to the induction of CYP24A1 expression by 1alpha,25-dihydroxyvitamin D3. J Biol Chem 285:15599–15610PubMedCrossRefPubMedCentral

32.

Meyer MB, Watanuki M, Kim S, Shevde NK, Pike JW (2006) The human transient receptor potential vanilloid type 6 distal promoter contains multiple vitamin D receptor binding sites that mediate activation by 1,25-dihydroxyvitamin D3 in intestinal cells. Mol Endocrinol 20:1447–1461PubMedCrossRef

33.

Peng L, Malloy PJ, Feldman D (2004) Identification of a functional vitamin D response element in the human insulin-like growth factor binding protein-3 promoter. Mol Endocrinol 18:1109–1119PubMedCrossRef

34.

Zou A, Elgort MG, Allegretto EA (1997) Retinoid X receptor (RXR) ligands activate the human 25-hydroxyvitamin D3-24-hydroxylase promoter via RXR heterodimer binding to two vitamin D-responsive elements and elicit additive effects with 1,25-dihydroxyvitamin D3. J Biol Chem 272:19027–19034PubMedCrossRef

35.

Liu M, Lee MH, Cohen M, Bommakanti M, Freedman LP (1996) Transcriptional activation of the Cdk inhibitor p21 by vitamin D3 leads to the induced differentiation of the myelomonocytic cell line U937. Genes Dev 10:142–153PubMedCrossRef

36.

Jiang F, Li P, Fornace AJ Jr, Nicosia SV, Bai W (2003) G2/M arrest by 1,25-dihydroxyvitamin D3 in ovarian cancer cells mediated through the induction of GADD45 via an exonic enhancer. J Biol Chem 278:48030–48040PubMedCrossRef

37.

Kriebitzsch C, Verlinden L, Eelen G, Tan BK, Van Camp M, Bouillon R, Verstuyf A (2009) The impact of 1,25(OH)2D3 and its structural analogs on gene expression in cancer cells–a microarray approach. Anticancer Res 29:3471–3483PubMed

38.

Anderson MG, Nakane M, Ruan X, Kroeger PE, Wu-Wong JR (2006) Expression of VDR and CYP24A1 mRNA in human tumors. Cancer Chemother Pharmacol 57:234–240PubMedCrossRef

39.

Chung HK, Yi YW, Jung NC, Kim D, Suh JM, Kim H, Park KC, Song JH, Kim DW, Hwang ES, Yoon SH, Bae YS, Kim JM, Bae I, Shong M (2003) CR6-interacting factor 1 interacts with Gadd45 family proteins and modulates the cell cycle. J Biol Chem 278:28079–28088PubMedCrossRef

40.

Kearsey JM, Coates PJ, Prescott AR, Warbrick E, Hall PA (1995) Gadd45 is a nuclear cell cycle regulated protein which interacts with p21Cip1. Oncogene 11:1675–1683PubMed

41.

Jiang H, Lin J, Su ZZ, Collart FR, Huberman E, Fisher PB (1994) Induction of differentiation in human promyelocytic HL-60 leukemia cells activates p21, WAF1/CIP1, expression in the absence of p53. Oncogene 9:3397–3406PubMed

42.

LaBaer J, Garrett MD, Stevenson LF, Slingerland JM, Sandhu C, Chou HS, Fattaey A, Harlow E (1997) New functional activities for the p21 family of CDK inhibitors. Genes Dev 11:847–862PubMedCrossRef

43.

Yang R, Nakamaki T, Lubbert M, Said J, Sakashita A, Freyaldenhoven BS, Spira S, Huynh V, Muller C, Koeffler HP (1999) Cyclin A1 expression in leukemia and normal hematopoietic cells. Blood 93:2067–2074PubMed

44.

Yang R, Muller C, Huynh V, Fung YK, Yee AS, Koeffler HP (1999) Functions of cyclin A1 in the cell cycle and its interactions with transcription factor E2F-1 and the Rb family of proteins. Mol Cell Biol 19:2400–2407PubMedPubMedCentral

45.

Meyerson M, Harlow E (1994) Identification of G1 kinase activity for cdk6, a novel cyclin D partner. Mol Cell Biol 14:2077–2086PubMedPubMedCentral

46.

Kozar K, Sicinski P (2005) Cell cycle progression without cyclin D-CDK4 and cyclin D-CDK6 complexes. Cell Cycle 4:388–391PubMedCrossRef

47.

Munger JS, Harpel JG, Gleizes PE, Mazzieri R, Nunes I, Rifkin DB (1997) Latent transforming growth factor-beta: structural features and mechanisms of activation. Kidney Int 51:1376–1382PubMedCrossRef

48.

Wang Z, Smith KS, Murphy M, Piloto O, Somervaille TC, Cleary ML (2008) Glycogen synthase kinase 3 in MLL leukaemia maintenance and targeted therapy. Nature 455:1205–1209PubMedCrossRefPubMedCentral

49.

Antony R, Sheng X, Ehsanipour EA, Ng E, Pramanik R, Klemm L, Ichihara B, Mittelman SD (2012) Vitamin D protects acute lymphoblastic leukemia cells from dexamethasone. Leuk Res 36:591–593PubMedCrossRefPubMedCentral

Titel: Anticancer activity of VDR-coregulator inhibitor PS121912
verfasst von: Preetpal S. Sidhu
Kelly Teske
Belaynesh Feleke
Nina Y. Yuan
Margaret L. Guthrie
Grant B. Fernstrum
Nishita D. Vyas
Lanlan Han
Joshua Preston
Jonathan W. Bogart
Nicholas R. Silvaggi
James M. Cook
Rakesh K. Singh
Daniel D. Bikle
Leggy A. Arnold
Publikationsdatum: 01.10.2014
Verlag: Springer Berlin Heidelberg
Erschienen in: Cancer Chemotherapy and Pharmacology / Ausgabe 4/2014
Print ISSN: 0344-5704
Elektronische ISSN: 1432-0843
DOI: https://doi.org/10.1007/s00280-014-2549-y

Springer Medizin

Anticancer activity of VDR-coregulator inhibitor PS121912