Abstract
Fifty years ago, prescription of the sedative thalidomide caused a worldwide epidemic of multiple birth defects. The drug is now used in the treatment of leprosy and multiple myeloma. However, its use is limited due to its potent teratogenic activity. The mechanism by which thalidomide causes limb malformations and other developmental defects is a long-standing question. Multiple hypotheses exist to explain the molecular mechanism of thalidomide action. Among them, theories involving oxidative stress and anti-angiogenesis have been widely supported. Nevertheless, until recently, the direct target of thalidomide remained elusive. We identified a thalidomide-binding protein, cereblon (CRBN), as a primary target for thalidomide teratogenicity. Our data suggest that thalidomide initiates its teratogenic effects by binding to CRBN and inhibiting its ubiquitin ligase activity. In this review, we summarize the biology of thalidomide, focusing on the molecular mechanisms of its teratogenic effects. In addition, we discuss the questions still to be addressed.
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Franks ME, Macpherson GR, Figg WD (2004) Thalidomide. Lancet 363:1802–1811
Bartlett JB, Dredge K, Dalgleish AG (2004) The evolution of thalidomide and its IMiD derivatives as anticancer agents. Nat Rev Cancer 4:314–322
Melchert M, List A (2007) The thalidomide saga. Int J Biochem Cell Biol 39:1489–1499
Knobloch J, Ruther U (2008) Shedding light on an old mystery: thalidomide suppresses survival pathways to induce limb defects. Cell Cycle 7:1121–1127
Vargesson N (2009) Thalidomide-induced limb defects: resolving a 50-year-old puzzle. Bioessays 31:1327–1336
McBride WG (1977) Thalidomide embryopathy. Teratology 16:79–82
Lenz W (1988) A short history of thalidomide embryopathy. Teratology 38:203–215
Sheskin J (1965) Thalidomide in the treatment of lepra reactions. Clin Pharmacol Ther 6:303–306
Gutierrez-Rodriguez O (1984) Thalidomide. A promising new treatment for rheumatoid arthritis. Arthritis Rheum 27:1118–1121
Hamza MH (1986) Treatment of Behcet’s disease with thalidomide. Clin Rheumatol 5:365–371
McCarthy DM, Kanfer EJ, Barrett AJ (1989) Thalidomide for the therapy of graft-versus-host disease following allogeneic bone marrow transplantation. Biomed Pharmacother 43:693–697
Vogelsang GB, Farmer ER, Hess AD, Altamonte V, Beschorner WE, Jabs DA, Corio RL, Levin LS, Colvin OM, Wingard JR, Santos GW (1992) Thalidomide for the treatment of chronic graft-versus-host disease. N Engl J Med 326:1055–1058
Atra E, Sato EI (1993) Treatment of the cutaneous lesions of systemic lupus erythematosus with thalidomide. Clin Exp Rheumatol 11:487–493
Makonkawkeyoon S, Limson-Pobre RN, Moreira AL, Schauf V, Kaplan G (1993) Thalidomide inhibits the replication of human immunodeficiency virus type 1. Proc Natl Acad Sci USA 90:5974–5978
Sampaio EP, Sarno EN, Galilly R, Cohn ZA, Kaplan G (1991) Thalidomide selectively inhibits tumor necrosis factor alpha production by stimulated human monocytes. J Exp Med 173:699–703
Moreira AL, Sampaio EP, Zmuidzinas A, Frindt P, Smith KA, Kaplan G (1993) Thalidomide exerts its inhibitory action on tumor necrosis factor alpha by enhancing mRNA degradation. J Exp Med 177:1675–1680
D’Amato RJ, Loughnan MS, Flynn E, Folkman J (1994) Thalidomide is an inhibitor of angiogenesis. Proc Natl Acad Sci USA 91:4082–4085
Singhal S, Mehta J, Desikan R, Ayers D, Roberson P, Eddlemon P, Munshi N, Anaissie E, Wilson C, Dhodapkar M, Zeddis J, Barlogie B (1999) Antitumor activity of thalidomide in refractory multiple myeloma. N Engl J Med 341:1565–1571
Zeldis JB, Williams BA, Thomas SD, Elsayed ME (1999) S.T.E.P.S.: a comprehensive program for controlling and monitoring access to thalidomide. Clin Ther 21:319–330
Castilla EE, Ashton-Prolla P, Barreda-Mejia E, Brunoni D, Cavalcanti DP, Correa-Neto J, Delgadillo JL, Dutra MG, Felix T, Giraldo A, Juarez N, Lopez-Camelo JS, Nazer J, Orioli IM, Paz JE, Pessoto MA, Pina-Neto JM, Quadrelli R, Rittler M, Rueda S, Saltos M, Sanchez O, Schuler L (1996) Thalidomide, a current teratogen in South America. Teratology 54:273–277
Schuler-Faccini L, Soares RC, de Sousa AC, Maximino C, Luna E, Schwartz IV, Waldman C, Castilla EE (2007) New cases of thalidomide embryopathy in Brazil. Birth Defects Res A Clin Mol Teratol 79:671–672
Hansen JM, Harris C (2004) A novel hypothesis for thalidomide-induced limb teratogenesis: redox misregulation of the NF-kappaB pathway. Antioxid Redox Signal 6:1–14
Parman T, Wiley MJ, Wells PG (1999) Free radical-mediated oxidative DNA damage in the mechanism of thalidomide teratogenicity. Nat Med 5:582–585
Therapontos C, Erskine L, Gardner ER, Figg WD, Vargesson N (2009) Thalidomide induces limb defects by preventing angiogenic outgrowth during early limb formation. Proc Natl Acad Sci USA 106:8573–8578
Ito T, Ando H, Suzuki T, Ogura T, Hotta K, Imamura Y, Yamaguchi Y, Handa H (2010) Identification of a primary target of thalidomide teratogenicity. Science 327:1345–1350
Eriksson T, Bjorkman S, Roth B, Hoglund P (2000) Intravenous formulations of the enantiomers of thalidomide: pharmacokinetic and initial pharmacodynamic characterization in man. J Pharm Pharmacol 52:807–817
Braun AG, Harding FA, Weinreb SL (1986) Teratogen metabolism: thalidomide activation is mediated by cytochrome P-450. Toxicol Appl Pharmacol 82:175–179
Ando Y, Fuse E, Figg WD (2002) Thalidomide metabolism by the CYP2C subfamily. Clin Cancer Res 8:1964–1973
Chen TL, Vogelsang GB, Petty BG, Brundrett RB, Noe DA, Santos GW, Colvin OM (1989) Plasma pharmacokinetics and urinary excretion of thalidomide after oral dosing in healthy male volunteers. Drug Metab Dispos 17:402–405
Chung F, Lu J, Palmer BD, Kestell P, Browett P, Baguley BC, Tingle M, Ching LM (2004) Thalidomide pharmacokinetics and metabolite formation in mice, rabbits, and multiple myeloma patients. Clin Cancer Res 10:5949–5956
Miller MT, Stromland K (1999) Teratogen update: thalidomide: a review, with a focus on ocular findings and new potential uses. Teratology 60:306–321
Mellin GW, Katzenstein M (1962) The saga of thalidomide. Neuropathy to embryopathy, with case reports of congenital anomalies. N Engl J Med 267:1184–1192
Spouge D, Baird PA (1986) Imperforate anus in 700,000 consecutive liveborn infants. Am J Med Genet Suppl 2:151–161
Davis MC, Dahn RD, Shubin NH (2007) An autopodial-like pattern of Hox expression in the fins of a basal actinopterygian fish. Nature 447:473–476
Tanaka M, Munsterberg A, Anderson WG, Prescott AR, Hazon N, Tickle C (2002) Fin development in a cartilaginous fish and the origin of vertebrate limbs. Nature 416:527–531
Moon AM, Capecchi MR (2000) Fgf8 is required for outgrowth and patterning of the limbs. Nat Genet 26:455–459
Lewandoski M, Sun X, Martin GR (2000) Fgf8 signalling from the AER is essential for normal limb development. Nat Genet 26:460–463
Hansen JM, Gong SG, Philbert M, Harris C (2002) Misregulation of gene expression in the redox-sensitive NF-kappab-dependent limb outgrowth pathway by thalidomide. Dev Dyn 225:186–194
Brent RL (1964) Drug testing in animals for teratogenic effects. Thalidomide in the pregnant rat. J Pediatr 64:762–770
Kenyon BM, Browne F, D’Amato RJ (1997) Effects of thalidomide and related metabolites in a mouse corneal model of neovascularization. Exp Eye Res 64:971–978
Fratta ID, Sigg EB, Maiorana K (1965) Teratogenic effects of thalidomide in rabbits, rats, hamsters, and mice. Toxicol Appl Pharmacol 7:268–286
Stephens TD (1988) Proposed mechanisms of action in thalidomide embryopathy. Teratology 38:229–239
Stephens TD, Fillmore BJ (2000) Hypothesis: thalidomide embryopathy-proposed mechanism of action. Teratology 61:189–195
Stephens TD, Bunde CJ, Fillmore BJ (2000) Mechanism of action in thalidomide teratogenesis. Biochem Pharmacol 59:1489–1499
Parman T, Chen G, Wells PG (1998) Free radical intermediates of phenytoin and related teratogens. Prostaglandin H synthase-catalyzed bioactivation, electron paramagnetic resonance spectrometry, and photochemical product analysis. J Biol Chem 273:25079–25088
Wells PG, Winn LM (1996) Biochemical toxicology of chemical teratogenesis. Crit Rev Biochem Mol Biol 31:1–40
Ohuchi H, Nakagawa T, Yamamoto A, Araga A, Ohata T, Ishimaru Y, Yoshioka H, Kuwana T, Nohno T, Yamasaki M, Itoh N, Noji S (1997) The mesenchymal factor, FGF10, initiates and maintains the outgrowth of the chick limb bud through interaction with FGF8, an apical ectodermal factor. Development 124:2235–2244
Knobloch J, Shaughnessy JD Jr, Ruther U (2007) Thalidomide induces limb deformities by perturbing the Bmp/Dkk1/Wnt signaling pathway. FASEB J 21:1410–1421
Knobloch J, Schmitz I, Gotz K, Schulze-Osthoff K, Ruther U (2008) Thalidomide induces limb anomalies by PTEN stabilization, Akt suppression, and stimulation of caspase-dependent cell death. Mol Cell Biol 28:529–538
Pizette S, Niswander L (1999) BMPs negatively regulate structure and function of the limb apical ectodermal ridge. Development 126:883–894
Scherz PJ, Harfe BD, McMahon AP, Tabin CJ (2004) The limb bud Shh-Fgf feedback loop is terminated by expansion of former ZPA cells. Science 305:396–399
Leslie NR, Downes CP (2004) PTEN function: how normal cells control it and tumour cells lose it. Biochem J 382:1–11
Pajni-Underwood S, Wilson CP, Elder C, Mishina Y, Lewandoski M (2007) BMP signals control limb bud interdigital programmed cell death by regulating FGF signaling. Development 134:2359–2368
Andela VB, Sheu TJ, Puzas EJ, Schwarz EM, O’Keefe RJ, Rosier RN (2002) Malignant reversion of a human osteosarcoma cell line, Saos-2, by inhibition of NFkappaB. Biochem Biophys Res Commun 297:237–241
Grotewold L, Ruther U (2002) The Wnt antagonist Dickkopf-1 is regulated by Bmp signaling and c-Jun and modulates programmed cell death. EMBO J 21:966–975
Beurel E, Jope RS (2006) The paradoxical pro- and anti-apoptotic actions of GSK3 in the intrinsic and extrinsic apoptosis signaling pathways. Prog Neurobiol 79:173–189
Sauer H, Gunther J, Hescheler J, Wartenberg M (2000) Thalidomide inhibits angiogenesis in embryoid bodies by the generation of hydroxyl radicals. Am J Pathol 156:151–158
Yabu T, Tomimoto H, Taguchi Y, Yamaoka S, Igarashi Y, Okazaki T (2005) Thalidomide-induced antiangiogenic action is mediated by ceramide through depletion of VEGF receptors, and is antagonized by sphingosine-1-phosphate. Blood 106:125–134
Bauer KS, Dixon SC, Figg WD (1998) Inhibition of angiogenesis by thalidomide requires metabolic activation, which is species-dependent. Biochem Pharmacol 55:1827–1834
Lu J, Palmer BD, Kestell P, Browett P, Baguley BC, Muller G, Ching LM (2003) Thalidomide metabolites in mice and patients with multiple myeloma. Clin Cancer Res 9:1680–1688
Ng SS, Gutschow M, Weiss M, Hauschildt S, Teubert U, Hecker TK, Luzzio FA, Kruger EA, Eger K, Figg WD (2003) Antiangiogenic activity of N-substituted and tetrafluorinated thalidomide analogues. Cancer Res 63:3189–3194
Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TF (1995) Stages of embryonic development of the zebrafish. Dev Dyn 203:253–310
Grandel H, Schulte-Merker S (1998) The development of the paired fins in the zebrafish (Danio rerio). Mech Dev 79:99–120
Lebrin F, Srun S, Raymond K, Martin S, van den Brink S, Freitas C, Breant C, Mathivet T, Larrivee B, Thomas JL, Arthur HM, Westermann CJ, Disch F, Mager JJ, Snijder RJ, Eichmann A, Mummery CL (2010) Thalidomide stimulates vessel maturation and reduces epistaxis in individuals with hereditary hemorrhagic telangiectasia. Nat Med 16:420–428
Sakamoto S, Kabe Y, Hatakeyama M, Yamaguchi Y, Handa H (2009) Development and application of high-performance affinity beads: toward chemical biology and drug discovery. Chem Rec 9:66–85
Shimizu N, Sugimoto K, Tang J, Nishi T, Sato I, Hiramoto M, Aizawa S, Hatakeyama M, Ohba R, Hatori H, Yoshikawa T, Suzuki F, Oomori A, Tanaka H, Kawaguchi H, Watanabe H, Handa H (2000) High-performance affinity beads for identifying drug receptors. Nat Biotechnol 18:877–881
Nishio K, Masaike Y, Ikeda M, Narimatsu H, Gokon N, Tsubouchi S, Hatakeyama M, Sakamoto S, Hanyu N, Sandhu A, Kawaguchi H, Abe M, Handa H (2008) Development of novel magnetic nano-carriers for high-performance affinity purification. Colloids Surf B Biointerfaces 64:162–169
Higgins JJ, Pucilowska J, Lombardi RQ, Rooney JP (2004) A mutation in a novel ATP-dependent Lon protease gene in a kindred with mild mental retardation. Neurology 63:1927–1931
Angers S, Li T, Yi X, MacCoss MJ, Moon RT, Zheng N (2006) Molecular architecture and assembly of the DDB1-CUL4A ubiquitin ligase machinery. Nature 443:590–593
Jo S, Lee KH, Song S, Jung YK, Park CS (2005) Identification and functional characterization of cereblon as a binding protein for large-conductance calcium-activated potassium channel in rat brain. J Neurochem 94:1212–1224
Hohberger B, Enz R (2009) Cereblon is expressed in the retina and binds to voltage-gated chloride channels. FEBS Lett 583:633–637
Wittschieben BO, Wood RD (2003) DDB complexities. DNA Repair (Amst) 2:1065–1069
Petroski MD, Deshaies RJ (2005) Function and regulation of cullin-RING ubiquitin ligases. Nat Rev Mol Cell Biol 6:9–20
Groisman R, Polanowska J, Kuraoka I, Sawada J, Saijo M, Drapkin R, Kisselev AF, Tanaka K, Nakatani Y (2003) The ubiquitin ligase activity in the DDB2 and CSA complexes is differentially regulated by the COP9 signalosome in response to DNA damage. Cell 113:357–367
Lee J, Zhou P (2007) DCAFs, the missing link of the CUL4-DDB1 ubiquitin ligase. Mol Cell 26:775–780
Pickart CM (2004) Back to the future with ubiquitin. Cell 116:181–190
Sugasawa K, Okuda Y, Saijo M, Nishi R, Matsuda N, Chu G, Mori T, Iwai S, Tanaka K, Hanaoka F (2005) UV-induced ubiquitylation of XPC protein mediated by UV-DDB–ubiquitin ligase complex. Cell 121:387–400
Groisman R, Kuraoka I, Chevallier O, Gaye N, Magnaldo T, Tanaka K, Kisselev AF, Harel-Bellan A, Nakatani Y (2006) CSA-dependent degradation of CSB by the ubiquitin–proteasome pathway establishes a link between complementation factors of the Cockayne syndrome. Genes Dev 20:1429–1434
Lieschke GJ, Currie PD (2007) Animal models of human disease: zebrafish swim into view. Nat Rev Genet 8:353–367
Nasevicius A, Ekker SC (2000) Effective targeted gene ‘knockdown’ in zebrafish. Nat Genet 26:216–220
Lee KJ, Lee KM, Jo S, Kang KW, Park CS (2010) Induction of cereblon by NF-E2-related factor 2 in neuroblastoma cells exposed to hypoxia-reoxygenation. Biochem Biophys Res Commun 399:711–715
Kang KW, Lee SJ, Kim SG (2005) Molecular mechanism of nrf2 activation by oxidative stress. Antioxid Redox Signal 7:1664–1673
Su AI, Wiltshire T, Batalov S, Lapp H, Ching KA, Block D, Zhang J, Soden R, Hayakawa M, Kreiman G, Cooke MP, Walker JR, Hogenesch JB (2004) A gene atlas of the mouse and human protein-encoding transcriptomes. Proc Natl Acad Sci USA 101:6062–6067
Field HA, Dong PD, Beis D, Stainier DY (2003) Formation of the digestive system in zebrafish. II. Pancreas morphogenesis. Dev Biol 261:197–208
Knobloch J, Reimann K, Klotz LO, Ruther U (2008) Thalidomide resistance is based on the capacity of the glutathione-dependent antioxidant defense. Mol Pharm 5:1138–1144
Janer G, Verhoef A, Gilsing HD, Piersma AH (2008) Use of the rat postimplantation embryo culture to assess the embryotoxic potency within a chemical category and to identify toxic metabolites. Toxicol In Vitro 22:1797–1805
Uga H, Kuramori C, Ohta A, Tsuboi Y, Tanaka H, Hatakeyama M, Yamaguchi Y, Takahashi T, Kizaki M, Handa H (2006) A new mechanism of methotrexate action revealed by target screening with affinity beads. Mol Pharmacol 70:1832–1839
List A, Kurtin S, Roe DJ, Buresh A, Mahadevan D, Fuchs D, Rimsza L, Heaton R, Knight R, Zeldis JB (2005) Efficacy of lenalidomide in myelodysplastic syndromes. N Engl J Med 352:549–557
Andritsos LA, Johnson AJ, Lozanski G, Blum W, Kefauver C, Awan F, Smith LL, Lapalombella R, May SE, Raymond CA, Wang DS, Knight RD, Ruppert AS, Lehman A, Jarjoura D, Chen CS, Byrd JC (2008) Higher doses of lenalidomide are associated with unacceptable toxicity including life-threatening tumor flare in patients with chronic lymphocytic leukemia. J Clin Oncol 26:2519–2525
Chanan-Khan A, Miller KC, Musial L, Lawrence D, Padmanabhan S, Takeshita K, Porter CW, Goodrich DW, Bernstein ZP, Wallace P, Spaner D, Mohr A, Byrne C, Hernandez-Ilizaliturri F, Chrystal C, Starostik P, Czuczman MS (2006) Clinical efficacy of lenalidomide in patients with relapsed or refractory chronic lymphocytic leukemia: results of a phase II study. J Clin Oncol 24:5343–5349
Chanan-Khan AA, Cheson BD (2008) Lenalidomide for the treatment of B-cell malignancies. J Clin Oncol 26:1544–1552
Xu Y, Li J, Ferguson GD, Mercurio F, Khambatta G, Morrison L, Lopez-Girona A, Corral LG, Webb DR, Bennett BL, Xie W (2009) Immunomodulatory drugs reorganize cytoskeleton by modulating Rho GTPases. Blood 114:338–345
Aragon-Ching JB, Li H, Gardner ER, Figg WD (2007) Thalidomide analogues as anticancer drugs. Recent Pat Anticancer Drug Discov 2:167–174
Acknowledgments
We thank Drs. Yuki Yamaguchi, Toshihiko Ogura and Takayuki Suzuki for aiding us in our research. Our research was supported by the Global COE (Center of Excellence) Program from the Japan Ministry of Education, Culture, Sports, Science, and Technology; and by a grant for Research and Development Projects in Cooperation with Academic Institutions from the New Energy and Technology Development Organization; and by Special Coordination Funds for Promoting Science and Technology from the Japan Science and Technology Agency (JST).
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Ito, T., Ando, H. & Handa, H. Teratogenic effects of thalidomide: molecular mechanisms. Cell. Mol. Life Sci. 68, 1569–1579 (2011). https://doi.org/10.1007/s00018-010-0619-9
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DOI: https://doi.org/10.1007/s00018-010-0619-9