Abstract
Cancer chemotherapy with monospecific agents is often hampered by the rapid development of tumor resistance to the drug used. Therefore, combination treatments aiming at several different targets are sought. Viral regulatory proteins, modified or not, appear ideal for this purpose because of their multimodal killing action against neoplastically transformed cells. The large nonstructural protein NS1of rodent parvoviruses is an excellent candidate for an anticancer agent, shown to interfere specifically with cancer cell growth and survival. The present review describes the structure, functions, and regulation of the multifunctional protein NS1, its specific interference with cell processes and cell protein activities, and what is known so far about the mechanisms underlying NS1 interference with cancer growth. It further outlines prospects for the development of new, multimodal cancer toxins and their potential applications.
Keywords
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Geletneky K, Hartkopf AD, Krempien R, Rommelaere J, Schlehofer JR (2010) Improved killing of human high-grade glioma cells by combining ionizing radiation with oncolytic parvovirus H-1 infection. J Biomed Biotechnol 2010:350748
Geletneky K, Hartkopf AD, Krempien R, Rommelaere J, Schlehofer JR (2010) Therapeutic implications of the enhanced short and long-term cytotoxicity of radiation treatment followed by oncolytic parvovirus H-1 infection in high-grade glioma cells. Bioeng Bugs 1:429–433
Geletneky K, Kiprianova I, Ayache A, Koch R, Herrero YCM, Deleu L et al (2010) Regression of advanced rat and human gliomas by local or systemic treatment with oncolytic parvovirus H-1 in rat models. Neuro Oncol 12:804–814
Rommelaere J, Cornelis JJ (1991) Antineoplastic activity of parvoviruses. J Virol Methods 33:233–251
Rommelaere J, Geletneky K, Angelova AL, Daeffler L, Dinsart C, Kiprianova I et al (2010) Oncolytic parvoviruses as cancer therapeutics. Cytokine Growth Factor Rev 21:185–195
Geletneky K, Huesing J, Rommelaere J, Schlehofer JR, Leuchs B, Dahm M et al (2012) Phase I/IIa study of intratumoral/intracerebral or intravenous/intracerebral administration of Parvovirus H-1 (ParvOryx) in patients with progressive primary or recurrent glioblastoma multiforme: ParvOryx01 protocol. BMC Cancer 12:99
Cotmore SF, Tattersall P (1987) The autonomously replicating parvoviruses of vertebrates. Adv Virus Res 33:91–174
Nüesch JPF (2006) Regulation of non-structural protein functions by differential synthesis, modification and trafficking. In: Kerr CSB, Linden ME, Parrish CR, Cotmore SF (eds) Parvoviruses. Edward Arnold, Ltd, London, pp 275–290
Corbau R, Salom N, Rommelaere J, Nuesch JP (1999) Phosphorylation of the viral nonstructural protein NS1 during MVMp infection of A9 cells. Virology 259:402–415
Corbau R, Duverger V, Rommelaere J, Nuesch JP (2000) Regulation of MVM NS1 by protein kinase C: impact of mutagenesis at consensus phosphorylation sites on replicative functions and cytopathic effects. Virology 278:151–167
Nuesch JP, Corbau R, Tattersall P, Rommelaere J (1998) Biochemical activities of minute virus of mice nonstructural protein NS1 are modulated In vitro by the phosphorylation state of the polypeptide. J Virol 72:8002–8012
Nuesch JP, Christensen J, Rommelaere J (2001) Initiation of minute virus of mice DNA replication is regulated at the level of origin unwinding by atypical protein kinase C phosphorylation of NS1. J Virol 75:5730–5739
Nuesch JP, Lachmann S, Corbau R, Rommelaere J (2003) Regulation of minute virus of mice NS1 replicative functions by atypical PKClambda in vivo. J Virol 77:433–442
Dettwiler S, Rommelaere J, Nuesch JP (1999) DNA unwinding functions of minute virus of mice NS1 protein are modulated specifically by the lambda isoform of protein kinase C. J Virol 73:7410–7420
Lachmann S, Rommeleare J, Nuesch JP (2003) Novel PKCeta is required to activate replicative functions of the major nonstructural protein NS1 of minute virus of mice. J Virol 77:8048–8060
Raimondi C, Falasca M (2011) Targeting PDK1 in cancer. Curr Med Chem 18:2763–2769
Nuesch JP, Lacroix J, Marchini A, Rommelaere J (2012) Molecular pathways: rodent parvoviruses–mechanisms of oncolysis and prospects for clinical cancer treatment. Clin Cancer Res 18:3516–3523
Hristov G, Kramer M, Li J, El-Andaloussi N, Mora R, Daeffler L et al (2010) Through its nonstructural protein NS1, parvovirus H-1 induces apoptosis via accumulation of reactive oxygen species. J Virol 84:5909–5922
Moehler M, Blechacz B, Weiskopf N, Zeidler M, Stremmel W, Rommelaere J et al (2001) Effective infection, apoptotic cell killing and gene transfer of human hepatoma cells but not primary hepatocytes by parvovirus H1 and derived vectors. Cancer Gene Ther 8:158–167
Ran Z, Rayet B, Rommelaere J, Faisst S (1999) Parvovirus H-1-induced cell death: influence of intracellular NAD consumption on the regulation of necrosis and apoptosis. Virus Res 65:161–174
Rayet B, Lopez-Guerrero JA, Rommelaere J, Dinsart C (1998) Induction of programmed cell death by parvovirus H-1 in U937 cells: connection with the tumor necrosis factor alpha signalling pathway. J Virol 72:8893–8903
Sieben M, Herzer K, Zeidler M, Heinrichs V, Leuchs B, Schuler M et al (2008) Killing of p53-deficient hepatoma cells by parvovirus H-1 and chemotherapeutics requires promyelocytic leukemia protein. World J Gastroenterol 14:3819–3828
Di Piazza M, Mader C, Geletneky K, Herrero YCM, Weber E, Schlehofer J et al (2007) Cytosolic activation of cathepsins mediates parvovirus H-1-induced killing of cisplatin and TRAIL-resistant glioma cells. J Virol 81:4186–4198
Bar S, Rommelaere J, Nuesch JP (2013) Vesicular transport of progeny parvovirus particles through ER and Golgi regulates maturation and cytolysis. PLoS Pathog 9:e1003605
Caillet-Fauquet P, Perros M, Brandenburger A, Spegelaere P, Rommelaere J (1990) Programmed killing of human cells by means of an inducible clone of parvoviral genes encoding non-structural proteins. EMBO J 9:2989–2995
Mousset S, Ouadrhiri Y, Caillet-Fauquet P, Rommelaere J (1994) The cytotoxicity of the autonomous parvovirus minute virus of mice nonstructural proteins in FR3T3 rat cells depends on oncogene expression. J Virol 68:6446–6453
Christensen J, Cotmore SF, Tattersall P (1997) Parvovirus initiation factor PIF: a novel human DNA-binding factor which coordinately recognizes two ACGT motifs. J Virol 71:5733–5741
Christensen J, Cotmore SF, Tattersall P (1997) A novel cellular site-specific DNA-binding protein cooperates with the viral NS1 polypeptide to initiate parvovirus DNA replication. J Virol 71:1405–1416
Cotmore SF, Nuesch JP, Tattersall P (1992) In vitro excision and replication of 5′ telomeres of minute virus of mice DNA from cloned palindromic concatemer junctions. Virology 190:365–377
Cotmore SF, Nuesch JP, Tattersall P (1993) Asymmetric resolution of a parvovirus palindrome in vitro. J Virol 67:1579–1589
Cotmore SF, Tattersall P (1994) An asymmetric nucleotide in the parvoviral 3′ hairpin directs segregation of a single active origin of DNA replication. EMBO J 13:4145–4152
Cotmore SF, Christensen J, Tattersall P (2000) Two widely spaced initiator binding sites create an HMG1-dependent parvovirus rolling-hairpin replication origin. J Virol 74:1332–1341
Legendre D, Rommelaere J (1992) Terminal regions of the NS-1 protein of the parvovirus minute virus of mice are involved in cytotoxicity and promoter trans inhibition. J Virol 66:5705–5713
Li X, Rhode SL 3rd (1990) Mutation of lysine 405 to serine in the parvovirus H-1 NS1 abolishes its functions for viral DNA replication, late promoter trans activation, and cytotoxicity. J Virol 64:4654–4660
Rhode SL 3rd (1985) trans-Activation of parvovirus P38 promoter by the 76 K noncapsid protein. J Virol 55:886–889
Brandenburger A, Legendre D, Avalosse B, Rommelaere J (1990) NS-1 and NS-2 proteins may act synergistically in the cytopathogenicity of parvovirus MVMp. Virology 174:576–584
Nuesch JP, Lachmann S, Rommelaere J (2005) Selective alterations of the host cell architecture upon infection with parvovirus minute virus of mice. Virology 331:159–174
Daeffler L, Horlein R, Rommelaere J, Nuesch JP (2003) Modulation of minute virus of mice cytotoxic activities through site-directed mutagenesis within the NS coding region. J Virol 77:12466–12478
Christensen J, Tattersall P (2002) Parvovirus initiator protein NS1 and RPA coordinate replication fork progression in a reconstituted DNA replication system. J Virol 76:6518–6531
Cziepluch C, Kordes E, Poirey R, Grewenig A, Rommelaere J, Jauniaux JC (1998) Identification of a novel cellular TPR-containing protein, SGT, that interacts with the nonstructural protein NS1 of parvovirus H-1. J Virol 72:4149–4156
Harris CE, Boden RA, Astell CR (1999) A novel heterogeneous nuclear ribonucleoprotein-like protein interacts with NS1 of the minute virus of mice. J Virol 73:72–80
Krady JK, Ward DC (1995) Transcriptional activation by the parvoviral nonstructural protein NS-1 is mediated via a direct interaction with Sp1. Mol Cell Biol 15:524–533
Lorson C, Pearson J, Burger L, Pintel DJ (1998) An Sp1-binding site and TATA element are sufficient to support full transactivation by proximally bound NS1 protein of minute virus of mice. Virology 240:326–337
Nuesch JP, Rommelaere J (2006) NS1 interaction with CKII alpha: novel protein complex mediating parvovirus-induced cytotoxicity. J Virol 80:4729–4739
Nuesch JP, Rommelaere J (2007) A viral adaptor protein modulating casein kinase II activity induces cytopathic effects in permissive cells. Proc Natl Acad Sci U S A 104:12482–12487
Li J, Werner E, Hergenhahn M, Poirey R, Luo Z, Rommelaere J et al (2005) Expression profiling of human hepatoma cells reveals global repression of genes involved in cell proliferation, growth, and apoptosis upon infection with parvovirus H-1. J Virol 79:2274–2286
Adeyemi RO, Landry S, Davis ME, Weitzman MD, Pintel DJ (2010) Parvovirus minute virus of mice induces a DNA damage response that facilitates viral replication. PLoS Pathog 6:e1001141
Bashir T, Rommelaere J, Cziepluch C (2001) In vivo accumulation of cyclin A and cellular replication factors in autonomous parvovirus minute virus of mice-associated replication bodies. J Virol 75:4394–4398
Ruiz Z, Mihaylov IS, Cotmore SF, Tattersall P (2011) Recruitment of DNA replication and damage response proteins to viral replication centers during infection with NS2 mutants of Minute Virus of Mice (MVM). Virology 410:375–384
Adeyemi RO, Pintel DJ (2012) Replication of minute virus of mice in murine cells is facilitated by virally induced depletion of p21. J Virol 86:8328–8332
Op De Beeck A, Anouja F, Mousset S, Rommelaere J, Caillet-Fauquet P (1995) The nonstructural proteins of the autonomous parvovirus minute virus of mice interfere with the cell cycle, inducing accumulation in G2. Cell Growth Differ 6:781–787
Op De Beeck A, Sobczak-Thepot J, Sirma H, Bourgain F, Brechot C, Caillet-Fauquet P (2001) NS1- and minute virus of mice-induced cell cycle arrest: involvement of p53 and p21(cip1). J Virol 75:11071–11078
Op De Beeck A, Caillet-Fauquet P (1997) Viruses and the cell cycle. Prog Cell Cycle Res 3:1–19
Doerig C, Hirt B, Beard P, Antonietti JP (1988) Minute virus of mice non-structural protein NS-1 is necessary and sufficient for trans-activation of the viral P39 promoter. J Gen Virol 69(Pt 10):2563–2573
Doerig C, Hirt B, Antonietti JP, Beard P (1990) Nonstructural protein of parvoviruses B19 and minute virus of mice controls transcription. J Virol 64:387–396
Nuesch JP, Cotmore SF, Tattersall P (1992) Expression of functional parvoviral NS1 from recombinant vaccinia virus: effects of mutations in the nucleotide-binding motif. Virology 191:406–416
Best SM, Wolfinbarger JB, Bloom ME (2002) Caspase activation is required for permissive replication of Aleutian mink disease parvovirus in vitro. Virology 292:224–234
Best SM, Bloom ME (2004) Caspase activation during virus infection: more than just the kiss of death? Virology 320:191–194
Lachmann S, Bar S, Rommelaere J, Nuesch JP (2008) Parvovirus interference with intracellular signalling: mechanism of PKCeta activation in MVM-infected A9 fibroblasts. Cell Microbiol 10:755–769
Salome N, van Hille B, Duponchel N, Meneguzzi G, Cuzin F, Rommelaere J et al (1990) Sensitization of transformed rat cells to parvovirus MVMp is restricted to specific oncogenes. Oncogene 5:123–130
Bar S, Daeffler L, Rommelaere J, Nuesch JP (2008) Vesicular egress of non-enveloped lytic parvoviruses depends on gelsolin functioning. PLoS Pathog 4:e1000126
Nuesch JP, Bar S, Lachmann S, Rommelaere J (2009) Ezrin-radixin-moesin family proteins are involved in parvovirus replication and spreading. J Virol 83:5854–5863
Nuesch JP, Bar S, Rommelaere J (2008) Viral proteins killing tumor cells: new weapons in the fight against cancer. Cancer Biol Ther 7:1374–1376
Christensen J, Cotmore SF, Tattersall P (1995) Minute virus of mice transcriptional activator protein NS1 binds directly to the transactivation region of the viral P38 promoter in a strictly ATP-dependent manner. J Virol 69:5422–5430
Jindal HK, Yong CB, Wilson GM, Tam P, Astell CR (1994) Mutations in the NTP-binding motif of minute virus of mice (MVM) NS-1 protein uncouple ATPase and DNA helicase functions. J Biol Chem 269:3283–3289
Mouw M, Pintel DJ (1998) Amino acids 16–275 of minute virus of mice NS1 include a domain that specifically binds (ACCA)2-3-containing DNA. Virology 251:123–131
Nuesch JP, Tattersall P (1993) Nuclear targeting of the parvoviral replicator molecule NS1: evidence for self-association prior to nuclear transport. Virology 196:637–651
Nuesch JP, Cotmore SF, Tattersall P (1995) Sequence motifs in the replicator protein of parvovirus MVM essential for nicking and covalent attachment to the viral origin: identification of the linking tyrosine. Virology 209:122–135
Pujol A, Deleu L, Nuesch JP, Cziepluch C, Jauniaux JC, Rommelaere J (1997) Inhibition of parvovirus minute virus of mice replication by a peptide involved in the oligomerization of nonstructural protein NS1. J Virol 71:7393–7403
Wilson GM, Jindal HK, Yeung DE, Chen W, Astell CR (1991) Expression of minute virus of mice major nonstructural protein in insect cells: purification and identification of ATPase and helicase activities. Virology 185:90–98
Legendre D, Rommelaere J (1994) Targeting of promoters for trans activation by a carboxy-terminal domain of the NS-1 protein of the parvovirus minute virus of mice. J Virol 68:7974–7985
Ilyina TV, Koonin EV (1992) Conserved sequence motifs in the initiator proteins for rolling circle DNA replication encoded by diverse replicons from eubacteria, eucaryotes and archaebacteria. Nucleic Acids Res 20:3279–3285
Gorbalenya AE, Koonin EV, Wolf YI (1990) A new superfamily of putative NTP-binding domains encoded by genomes of small DNA and RNA viruses. FEBS Lett 262:145–148
Hickman AB, Ronning DR, Kotin RM, Dyda F (2002) Structural unity among viral origin binding proteins: crystal structure of the nuclease domain of adeno-associated virus Rep. Mol Cell 10:327–337
Tewary SK, Zhao H, Shen W, Qiu J, Tang L (2013) Structure of the NS1 protein N-terminal origin-recognition/nickase domain from the emerging human bocavirus. J Virol 87(21):11487–11493
Cotmore SF, Christensen J, Nuesch JP, Tattersall P (1995) The NS1 polypeptide of the murine parvovirus minute virus of mice binds to DNA sequences containing the motif [ACCA]2–3. J Virol 69:1652–1660
Cotmore SF, Tattersall P (1998) High-mobility group 1/2 proteins are essential for initiating rolling-circle-type DNA replication at a parvovirus hairpin origin. J Virol 72:8477–8484
Mastrangelo IA, Hough PV, Wall JS, Dodson M, Dean FB, Hurwitz J (1989) ATP-dependent assembly of double hexamers of SV40 T antigen at the viral origin of DNA replication. Nature 338:658–662
Gu ML, Chen FX, Rhode SL (1992) Parvovirus H-1 P38 promoter requires the trans-activation region (tar), an SP1 site, and a TATA box for full activity. Virology 187:10–17
Gu ML, Rhode SL (1992) Trans-activation of H-1 parvovirus P38 promoter is correlated with increased binding of cellular protein(s) to the trans-activation responsive element (tar). Virology 190:116–123
Nuesch JP, Dettwiler S, Corbau R, Rommelaere J (1998) Replicative functions of minute virus of mice NS1 protein are regulated in vitro by phosphorylation through protein kinase C. J Virol 72:9966–9977
Li J, Bonifati S, Hristov G, Marttila T, Valmary-Degano S, Stanzel S et al (2013) Synergistic combination of valproic acid and oncolytic parvovirus H-1PV as a potential therapy against cervical and pancreatic carcinomas. EMBO Mol Med 5:1537–1555
Bayascas JR (2008) Dissecting the role of the 3-phosphoinositide-dependent protein kinase-1 (PDK1) signalling pathways. Cell Cycle 7:2978–2982
Raimondi C, Chikh A, Wheeler AP, Maffucci T, Falasca M (2012) A novel regulatory mechanism links PLCgamma1 to PDK1. J Cell Sci 125:3153–3163
Vasudevan KM, Barbie DA, Davies MA, Rabinovsky R, McNear CJ, Kim JJ et al (2009) AKT-independent signaling downstream of oncogenic PIK3CA mutations in human cancer. Cancer Cell 16:21–32
Wollmann G, Tattersall P, van den Pol AN (2005) Targeting human glioblastoma cells: comparison of nine viruses with oncolytic potential. J Virol 79:6005–6022
Chakravarti A, Chakladar A, Delaney MA, Latham DE, Loeffler JS (2002) The epidermal growth factor receptor pathway mediates resistance to sequential administration of radiation and chemotherapy in primary human glioblastoma cells in a RAS-dependent manner. Cancer Res 62:4307–4315
Lefranc F, Brotchi J, Kiss R (2005) Possible future issues in the treatment of glioblastomas: special emphasis on cell migration and the resistance of migrating glioblastoma cells to apoptosis. J Clin Oncol 23:2411–2422
Hafsi S, Pezzino FM, Candido S, Ligresti G, Spandidos DA, Soua Z et al (2012) Gene alterations in the PI3K/PTEN/AKT pathway as a mechanism of drug-resistance (review). Int J Oncol 40:639–644
Wilson JL, Hemann MT, Fraenkel E, Lauffenburger DA (2013) Integrated network analyses for functional genomic studies in cancer. Semin Cancer Biol 23:213–218
Raykov Z, Grekova S, Galabov AS, Balboni G, Koch U, Aprahamian M et al (2007) Combined oncolytic and vaccination activities of parvovirus H-1 in a metastatic tumor model. Oncol Rep 17:1493–1499
Raykov Z, Grekova S, Leuchs B, Aprahamian M, Rommelaere J (2008) Arming parvoviruses with CpG motifs to improve their oncosuppressive capacity. Int J Cancer 122:2880–2884
Bhat R, Dempe S, Dinsart C, Rommelaere J (2011) Enhancement of NK cell antitumor responses using an oncolytic parvovirus. Int J Cancer 128:908–919
Bhat R, Rommelaere J (2013) NK-cell-dependent killing of colon carcinoma cells is mediated by natural cytotoxicity receptors (NCRs) and stimulated by parvovirus infection of target cells. BMC Cancer 13:367
Moehler MH, Zeidler M, Wilsberg V, Cornelis JJ, Woelfel T, Rommelaere J et al (2005) Parvovirus H-1-induced tumor cell death enhances human immune response in vitro via increased phagocytosis, maturation, and cross-presentation by dendritic cells. Hum Gene Ther 16:996–1005
Dempe S, Stroh-Dege AY, Schwarz E, Rommelaere J, Dinsart C (2010) SMAD4: a predictive marker of PDAC cell permissiveness for oncolytic infection with parvovirus H-1PV. Int J Cancer 126:2914–2927
Lavie M, Struyf S, Stroh-Dege A, Rommelaere J, Van Damme J, Dinsart C (2013) Capacity of wild-type and chemokine-armed parvovirus H-1PV for inhibiting neo-angiogenesis. Virology 447:221–232
Vollmers EM, Tattersall P (2013) Distinct host cell fates for human malignant melanoma targeted by oncolytic rodent parvoviruses. Virology 446:37–48
Weiss N, Stroh-Dege A, Rommelaere J, Dinsart C, Salome N (2012) An in-frame deletion in the NS protein-coding sequence of parvovirus H-1PV efficiently stimulates export and infectivity of progeny virions. J Virol 86:7554–7564
El-Andaloussi N, Bonifati S, Kaufmann JK, Mailly L, Daeffler L, Deryckere F et al (2012) Generation of an adenovirus-parvovirus chimera with enhanced oncolytic potential. J Virol 86:10418–10431
Bashir T, Horlein R, Rommelaere J, Willwand K (2000) Cyclin A activates the DNA polymerase delta -dependent elongation machinery in vitro: A parvovirus DNA replication model. Proc Natl Acad Sci U S A 97:5522–5527
Deleu L, Pujol A, Faisst S, Rommelaere J (1999) Activation of promoter P4 of the autonomous parvovirus minute virus of mice at early S phase is required for productive infection. J Virol 73:3877–3885
Breitbach CJ, Thorne SH, Bell JC, Kirn DH (2012) Targeted and armed oncolytic poxviruses for cancer: the lead example of JX-594. Curr Pharm Biotechnol 13:1768–1772
Donnelly OG, Errington-Mais F, Prestwich R, Harrington K, Pandha H, Vile R et al (2012) Recent clinical experience with oncolytic viruses. Curr Pharm Biotechnol 13:1834–1841
Nyberg KA et al (2002) Toward maintaining the genome: DNA damage and replication checkpoints. Annu Rev Genet 36:617–656
Bhattacharya B et al (1990) Tropomyosins of human mammary epithelial cells: consistent defects of expression in mammary carcinoma cell lines. Cancer Res 50(7):2105–2112
Fay N, Pante N (2013) The intermediate filament network protein, vimentin, is required for parvoviral infection. Virology 444(1–2):181–190
Cornelis JJ et al (1988) Transformation of human fibroblasts by ionizing radiation, a chemical carcinogen, or simian virus 40 correlates with an increase in susceptibility to the autonomous parvoviruses H-1 virus and minute virus of mice. J Virol 62(5):1679–1686
Grekova SP et al (2011) Interferon gamma improves the vaccination potential of oncolytic parvovirus H-1PV for the treatment of peritoneal carcinomatosis in pancreatic cancer. Cancer Biol Ther 12(10):888–895
Bar S, Rommelaere J, Nüesch JPF. PKCeta/Rdx-driven phosphorylation of PDK1: a novel mechanism promoting survival of cancer cells and permissiveness for parvovirus-induced lysis. Cell Host Microbe, submitted
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Nüesch, J.P.F., Rommelaere, J. (2014). Tumor Suppressing Properties of Rodent Parvovirus NS1 Proteins and Their Derivatives. In: Grimm, S. (eds) Anticancer Genes. Advances in Experimental Medicine and Biology, vol 818. Springer, London. https://doi.org/10.1007/978-1-4471-6458-6_5
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