The patients with mantle cell lymphoma (MCL) have translocation t(11;14) associated with cyclin D1 overexpression. We observed that iron (an essential cofactor of dioxygenases including prolyl hydroxylases [PHDs]) depletion by deferoxamine blocked MCL cells' proliferation, increased expression of DNA damage marker γH2AX, induced cell cycle arrest and decreased cyclin D1 level. Treatment of MCL cell lines with dimethyloxalylglycine, which blocks dioxygenases involving PHDs by competing with their substrate 2-oxoglutarate, leads to their decreased proliferation and the decrease of cyclin D1 level. We then postulated that loss of EGLN2/PHD1 in MCL cells may lead to down-regulation of cyclin D1 by blocking the degradation of FOXO3A, a cyclin D1 suppressor. However, the CRISPR/Cas9-based loss-of-function of EGLN2/PHD1 did not affect cyclin D1 expression and the loss of FOXO3A did not restore cyclin D1 levels after iron chelation. These data suggest that expression of cyclin D1 in MCL is not controlled by ENGL2/PHD1-FOXO3A pathway and that chelation- and 2-oxoglutarate competition-mediated down-regulation of cyclin D1 in MCL cells is driven by yet unknown mechanism involving iron- and 2-oxoglutarate-dependent dioxygenases other than PHD1. These data support further exploration of the use of iron chelation and 2-oxoglutarate-dependent dioxygenase inhibitors as a novel therapy of MCL.

Department of Biology Faculty of Medicine and Dentistry Palacky University Olomouc Olomouc Czech Republic

Department of Cell and Developmental Biology Institute of Molecular Genetics Academy of Sciences of the Czech Republic Prague Czech Republic

Division of Hematology and Hematologic Malignancies Department of Internal Medicine University of Utah School of Medicine and VAH Salt Lake City Utah

See more in PubMed

Schieber M, Gordon L, Karmali R. Current overview and treatment of mantle cell lymphoma. F1000Research. 2018;7:1136. PubMed PMC

Maddocks K. Update on mantle cell lymphoma. Blood. 2018;132:1647‐1656. PubMed

Ladha A, Zhao J, Epner EM, Pu JJ. Mantle cell lymphoma and its management: where are we now? Exp Hematol Oncol. 2019;8:2. PubMed PMC

Gilbert H, Cumming J, Prchal J. Cell cycle regulation by iron: novel approaches to mantle cell lymphoma therapy. Blood. 2009;114:2515. PubMed

Vazana‐Barad L, Granot G, Mor‐Tzuntz R, et al. Mechanism of the antitumoral activity of deferasirox, an iron chelation agent, on mantle cell lymphoma. Leuk Lymphoma. 2013;54:851‐859. PubMed

Richardson DR, Kalinowski DS, Lau S, Jansson PJ, Lovejoy DB. Cancer cell iron metabolism and the development of potent iron chelators as anti‐tumour agents. Biochim Biophys Acta. 2009;1790:702‐717. PubMed

Corcé V, Gouin SG, Renaud S, Gaboriau F, Deniaud D. Recent advances in cancer treatment by iron chelators. Bioorg Med Chem Lett. 2016;26:251‐256. PubMed

Le N, Richardson D. The role of iron in cell cycle progression and the proliferation of neoplastic cells. Biochim Biophys Acta. 2002;1603:31‐46. PubMed

Zhang C. Essential functions of iron‐requiring proteins in DNA replication, repair and cell cycle control. Protein Cell. 2014;5:750‐760. PubMed PMC

Furukawa T, Naitoh Y, Kohno H, et al. Iron deprivation decreases ribonucleotide reductase and DNA synthesis. Life Sci. 1992;50:2059‐2065. PubMed

Shao J, Zhou B, Chu B, et al. Ribonucleotide reductase inhibitors and future drug design. Curr Cancer Drug Targets. 2006;6:409‐431. PubMed

Nurtjahja‐Tjendraputra E, Fu D, Phang JM, Richardson DR. Iron chelation regulates cyclin D1 expression via the proteasome: a link to iron deficiency‐mediated growth suppression. Blood. 2007;109:4054‐4154. PubMed

Zhang Q, Gu J, Li L, et al. Control of cyclin D1 and breast tumorigenesis by the EglN2 prolyl hydroxylase. Cancer Cell. 2014;16:413‐424. PubMed PMC

Zheng X, Zhai B, Koivunen P, et al. Prolyl hydroxylation by EglN2 destabilizes FOXO3a by blocking its interaction with the USP9x deubiquitinase. Genes Dev. 2014;28:1429‐1444. PubMed PMC

Metzen E, Berchner‐Pfannschmidt U, Stengel P, et al. Intracellular localisation of human HIF‐1 alpha hydroxylases: implications for oxygen sensing. J Cell Sci. 2003;116:1319‐1326. PubMed

Yasumoto K‐I, Kowata Y, Yoshida A, Torii S, Sogawa K. Role of the intracellular localization of HIF‐prolyl hydroxylases. Biochim Biophys Acta. 2009;1793:792‐797. PubMed

Erez N, Milyavsky M, Goldfinger N, Peles E, Gudkov AV, Rotter V. Falkor, a novel cell growth regulator isolated by a functional genetic screen. Oncogene. 2002;21:6713‐6721. PubMed

Rodriguez J, Herrero A, Li S, et al. PHD3 regulates p53 protein stability by hydroxylating proline 359. Cell Rep. 2018;24:1316‐1329. PubMed PMC

Xie X, Xiao H, Ding F, et al. Over‐expression of prolyl hydroxylase‐1 blocks NF‐κB‐mediated cyclin D1 expression and proliferation in lung carcinoma cells. Cancer Genet. 2014;207:188‐194. PubMed

Scholz C, Cavadas M, Tambuwala M, et al. Regulation of IL‐1β‐induced NF‐κB by hydroxylases links key hypoxic and inflammatory signaling pathways. Proc Natl Acad Sci USA. 2013;110:18490‐18495. PubMed PMC

Frei C, Edgar B. Drosophila cyclin D/Cdk4 requires Hif‐1 prolyl hydroxylase to drive cell growth. Dev Cell. 2004;6:241‐251. PubMed

Erez N, Milyavsky M, Eilam R, et al. Expression of prolyl‐hydroxylase‐1 (PHD1/EGLN2) suppresses hypoxia inducible factor‐1alpha activation and inhibits tumor growth. Cancer Res. 2003;63:8777‐8783. PubMed

Takeda K, Aguila HL, Parikh NS, et al. Regulation of adult erythropoiesis by prolyl hydroxylase domain proteins. Blood. 2008;111:3229‐3235. PubMed PMC

Seth P, Krop I, Porter D, Polyak K. Novel estrogen and tamoxifen induced genes identified by SAGE (Serial Analysis of Gene Expression). Oncogene. 2002;21:836‐843. PubMed

Flamme I, Oehme F, Ellinghaus P, Jeske M, Keldenich J, Thuss U. Mimicking hypoxia to treat anemia: HIF‐stabilizer BAY 85‐3934 (Molidustat) stimulates erythropoietin production without hypertensive effects. PLoS One. 2014;9:e111838. PubMed PMC

Holdstock L, Meadowcroft AM, Maier R, et al. Four‐week studies of oral hypoxia‐inducible factor‐prolyl hydroxylase inhibitor GSK1278863 for treatment of anemia. J Am Soc Nephrol. 2016;27:1234‐1244. PubMed PMC

Eltzschig H, Bratton D, Colgan S. Targeting hypoxia signalling for the treatment of ischaemic and inflammatory diseases. Nat Rev Drug Discov. 2014;13:852‐869. PubMed PMC

Koury M, Haase V. Anaemia in kidney disease: harnessing hypoxia responses for therapy. Nat Rev Nephrol. 2015;11:394‐410. PubMed PMC

Daschner H, Lehner K, Rechl H, Heuck A. Die Darstellung der Epiphyseolysis capitis femoris (ECF) im Magnetresonanztomogramm (MRT). RöFo: Fortschritte auf dem Gebiete der Röntgenstrahlen und der Nuklearmedizin. 1990;152:583‐586. PubMed

Burroughs SK, Kaluz S, Wang D, Wang KE, Van Meir EG, Wang B. Hypoxia inducible factor pathway inhibitors as anticancer therapeutics. Future Med Chem. 2013;5:553‐572. PubMed PMC

Yeoh KK, Chan MC, Thalhammer A, et al. Dual‐action inhibitors of HIF prolyl hydroxylases that induce binding of a second iron ion. Org Biomol Chem. 2013;11:732‐745. PubMed PMC

Yeh T‐L, Leissing T, Abboud MI, et al. Molecular and cellular mechanisms of HIF prolyl hydroxylase inhibitors in clinical trials. Chem Sci. 2017;8:7651‐7668. PubMed PMC

Klier M, Anastasov N, Hermann A, et al. Specific lentiviral shRNA‐mediated knockdown of cyclin D1 in mantle cell lymphoma has minimal effects on cell survival and reveals a regulatory circuit with cyclin D2. Leukemia. 2008;22:2097‐2105. PubMed

Erez N, Stambolsky P, Shats I, Milyavsky M, Kachko T, Rotter V. Hypoxia‐dependent regulation of PHD1: cloning and characterization of the human PHD1/EGLN2 gene promoter. FEBS Lett. 2004;567:311‐315. PubMed

Bakker W, Harris I, Duncan G. FOXO3a is activated in response to hypoxic stress and inhibits HIF1‐induced apoptosis via regulation of CITED2. Mol Cell. 2008;28:941‐953. PubMed

Lin ZP, Belcourt MF, Carbone R, et al. Excess ribonucleotide reductase R2 subunits coordinate the S phase checkpoint to facilitate DNA damage repair and recovery from replication stress. Biochem Pharmacol. 2007;73:760‐772. PubMed

Mannava S, Moparthy KC, Wheeler LJ, et al. Depletion of deoxyribonucleotide pools is an endogenous source of DNA damage in cells undergoing oncogene‐induced senescence. Am J Pathol. 2012;182:142‐151. PubMed PMC

Aye Y, Li M, Long M, Weiss RS. Ribonucleotide reductase and cancer: biological mechanisms and targeted therapies. Oncogene. 2015;34:2011‐2021. PubMed

Turinetto V, Orlando L, Sanchez‐Ripoll Y, et al. High basal γH2AX levels sustain self‐renewal of mouse embryonic and induced pluripotent stem cells. Stem Cells. 2012;30:1414‐1423. PubMed

Raskova Kafkova L, Krupkova L, Divoky V. Aberrant regulation of checkpoint response in mouse embryonic stem cells treated with iron chelator deferoxamine mesylate. Am J Hematol. 2013;88:e184.

Kunos CA, Ferris G, Pyatka N, Pink J, Radivoyevitch T. Deoxynucleoside salvage facilitates DNA repair during ribonucleotide reductase blockade in human cervical cancers. Radiat Res. 2011;176:425‐433. PubMed PMC

Lui G, Obeidy P, Ford SJ, et al. The iron chelator, deferasirox, as a novel strategy for cancer treatment: oral activity against human lung tumor xenografts and molecular mechanism of action. Mol Pharmacol. 2012;83:179‐190. PubMed

Beltran E, Fresquet V, Martinez‐Useros J, et al. A cyclin‐D1 interaction with BAX underlies its oncogenic role and potential as a therapeutic target in mantle cell lymphoma. Proc Natl Acad Sci USA. 2011;108:12461‐12466. PubMed PMC

Porter J. Deferoxamine pharmacokinetics. Semin Hematol. 2001;38:63‐68. PubMed

Tuderman L, MyllylÄ R, Kivirikko K. Mechanism of the prolyl hydroxylase reaction. 1. Role of co‐substrates. Eur J Biochem. 2008;80:341‐348. PubMed

Allinne J, Pichugin A, Iarovaia O, et al. Perinucleolar relocalization and nucleolin as crucial events in the transcriptional activation of key genes in mantle cell lymphoma. Blood. 2014;123:2044‐2053. PubMed

Woo KJ, Lee T‐J, Park J‐W, Kwon TK. Desferrioxamine, an iron chelator, enhances HIF‐1α accumulation via cyclooxygenase‐2 signaling pathway. Biochem Biophys Res Commun. 2006;343:8‐14. PubMed

Brunet A, Bonni A, Zigmond MJ, et al. Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell. 1999;96:857‐868. PubMed

Stahl M, Dijkers P, Kops G, et al. The forkhead transcription factor FoxO regulates transcription of p27Kip1 and Bim in response to IL‐2. J Immunol. 2002;168:5024‐5031. PubMed

Ekoff M, Kaufmann T, Engstrom M, et al. The BH3‐only protein Puma plays an essential role in cytokine deprivation induced apoptosis of mast cells. Blood. 2007;110:3209‐3217. PubMed PMC

Nho R, Hergert P. FoxO3a and disease progression. World J Biol Chem. 2014;5:346‐354. PubMed PMC

Zhang X, Tang N, Hadden TJ, Rishi AK. Akt, FoxO and regulation of apoptosis. Biochim Biophys Acta – Mol Cell Res. 2011;1813:1978‐1986. PubMed

Obrador‐Hevia A, Serra‐Sitjar M, Rodríguez J, Villalonga P, de Mattos SF. The tumour suppressor FOXO3 is a key regulator of mantle cell lymphoma proliferation and survival. Br J Haematol. 2011;156:334‐345. PubMed

Hopkinson RJ, Tumber A, Yapp C, et al. 5‐Carboxy‐8‐hydroxyquinoline is a broad spectrum 2‐oxoglutarate oxygenase inhibitor which causes iron translocation. Chem Sci. 2013;4:3110‐3117. PubMed PMC