[en] H-1 parvovirus (H-1PV) is a promising anticancer therapy. However, in-depth understanding of its life cycle, including the host cell factors needed for infectivity and oncolysis, is lacking. This understanding may guide the rational design of combination strategies, aid development of more effective viruses, and help identify biomarkers of susceptibility to H-1PV treatment. To identify the host cell factors involved, we carry out siRNA library screening using a druggable genome library. We identify one crucial modulator of H-1PV infection: laminin γ1 (LAMC1). Using loss- and gain-of-function studies, competition experiments, and ELISA, we validate LAMC1 and laminin family members as being essential to H-1PV cell attachment and entry. H-1PV binding to laminins is dependent on their sialic acid moieties and is inhibited by heparin. We show that laminins are differentially expressed in various tumour entities, including glioblastoma. We confirm the expression pattern of laminin γ1 in glioblastoma biopsies by immunohistochemistry. We also provide evidence of a direct correlation between LAMC1 expression levels and H-1PV oncolytic activity in 59 cancer cell lines and in 3D organotypic spheroid cultures with different sensitivities to H-1PV infection. These results support the idea that tumours with elevated levels of γ1 containing laminins are more susceptible to H-1PV-based therapies.
Disciplines :
Oncology
Author, co-author :
Kulkarni, Amit ; Laboratory of Oncolytic Virus Immuno-Therapeutics, German Cancer Research Center, Heidelberg, Germany. ; Laboratory of Oncolytic Virus Immuno-Therapeutics, Luxembourg Institute of Health, Luxembourg, Luxembourg.
Ferreira, Tiago ; Laboratory of Oncolytic Virus Immuno-Therapeutics, German Cancer Research Center, Heidelberg, Germany.
Bretscher, Clemens; Laboratory of Oncolytic Virus Immuno-Therapeutics, German Cancer Research Center, Heidelberg, Germany.
Grewenig, Annabel; Laboratory of Oncolytic Virus Immuno-Therapeutics, German Cancer Research Center, Heidelberg, Germany.
El-Andaloussi, Nazim; Laboratory of Oncolytic Virus Immuno-Therapeutics, German Cancer Research Center, Heidelberg, Germany. ; Lonza Cologne GmbH, Köln, Germany.
Bonifati, Serena; Laboratory of Oncolytic Virus Immuno-Therapeutics, German Cancer Research Center, Heidelberg, Germany. ; Center for Retrovirus Research, Department of Veterinary Biosciences, The Ohio State University, Columbus, OH, USA.
Marttila, Tiina; Laboratory of Oncolytic Virus Immuno-Therapeutics, German Cancer Research Center, Heidelberg, Germany. ; Laboratory of Oncolytic Virus Immuno-Therapeutics, Luxembourg Institute of Health, Luxembourg, Luxembourg.
PALISSOT, Valerie ; Laboratory of Oncolytic Virus Immuno-Therapeutics, Luxembourg Institute of Health, Luxembourg, Luxembourg.
Hossain, Jubayer A; Laboratory of Oncolytic Virus Immuno-Therapeutics, Luxembourg Institute of Health, Luxembourg, Luxembourg. ; Department of Biomedicine, University of Bergen, Bergen, Norway. ; Department of Pathology, Haukeland University Hospital, Bergen, Norway.
AZUAJE, Francisco ; Quantitative Biology Unit, Luxembourg Institute of Health, Luxembourg, Luxembourg. ; Genomics England, London, United Kingdom.
Miletic, Hrvoje; Department of Biomedicine, University of Bergen, Bergen, Norway. ; Department of Pathology, Haukeland University Hospital, Bergen, Norway.
Ystaas, Lars A R; Department of Biomedicine, University of Bergen, Bergen, Norway.
GOLEBIEWSKA, Anna ; NorLux Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg.
NICLOU, Simone P. ; NorLux Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg.
Roeth, Ralf; nCounter Core Facility, Institute of Human Genetics, University of Heidelberg, Heidelberg, Germany. ; Department of Human Molecular Genetics, University of Heidelberg, Heidelberg, Germany.
Niesler, Beate ; nCounter Core Facility, Institute of Human Genetics, University of Heidelberg, Heidelberg, Germany. ; Department of Human Molecular Genetics, University of Heidelberg, Heidelberg, Germany.
Weiss, Amélie; Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Illkirch, France.
Brino, Laurent; Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Illkirch, France.
MARCHINI, Antonio ; Laboratory of Oncolytic Virus Immuno-Therapeutics, German Cancer Research Center, Heidelberg, Germany. antonio.marchini@lih.lu. ; Laboratory of Oncolytic Virus Immuno-Therapeutics, Luxembourg Institute of Health, Luxembourg, Luxembourg. antonio.marchini@lih.lu.
Lichty, B. D., Breitbach, C. J., Stojdl, D. F. & Bell, J. C. Going viral with cancer immunotherapy. Nat. Rev. Cancer 14, 559–567 (2014). DOI: 10.1038/nrc3770
Lemay, C. G., Keller, B. A., Edge, R. E., Abei, M. & Bell, J. C. Oncolytic viruses: the best is yet to come. Curr. Cancer Drug Targets 18, 109–123 (2018). DOI: 10.2174/1568009617666170206111609
Achard, C. et al. Lighting a fire in the tumor microenvironment using oncolytic immunotherapy. EBioMedicine 31, 17–24 (2018). DOI: 10.1016/j.ebiom.2018.04.020
Marchini, A., Daeffler, L., Pozdeev, V. I., Angelova, A. & Rommelaere, J. Immune conversion of tumor microenvironment by oncolytic viruses: the protoparvovirus H-1PV case study. Front. Immunol. 10, 1848 (2019). DOI: 10.3389/fimmu.2019.01848
Andtbacka, R. H. et al. Talimogene laherparepvec improves durable response rate in patients with advanced melanoma. J. Clin. Oncol. 33, 2780–2788 (2015). DOI: 10.1200/JCO.2014.58.3377
Fountzilas, C., Patel, S. & Mahalingam, D. Review: oncolytic virotherapy, updates and future directions. Oncotarget 8, 102617–102639 (2017). DOI: 10.18632/oncotarget.18309
Twumasi-Boateng, K., Pettigrew, J. L., Kwok, Y. Y. E., Bell, J. C. & Nelson, B. H. Oncolytic viruses as engineering platforms for combination immunotherapy. Nat. Rev. Cancer 18, 419–432 (2018). DOI: 10.1038/s41568-018-0009-4
LaRocca, C. J. & Warner, S. G. Oncolytic viruses and checkpoint inhibitors: combination therapy in clinical trials. Clin. Transl. Med. 7, 35 (2018). DOI: 10.1186/s40169-018-0214-5
Guedan, S. & Alemany, R. CAR-T cells and oncolytic viruses: joining forces to overcome the solid tumor challenge. Front. Immunol. 9, 2460 (2018). DOI: 10.3389/fimmu.2018.02460
Bommareddy, P. K., Shettigar, M. & Kaufman, H. L. Integrating oncolytic viruses in combination cancer immunotherapy. Nat. Rev. Immunol. 18, 498–513 (2018). DOI: 10.1038/s41577-018-0014-6
Marchini, A., Scott, E. M. & Rommelaere, J. Overcoming barriers in oncolytic virotherapy with HDAC Inhibitors and immune checkpoint blockade. Viruses 8, https://doi.org/10.3390/v8010009 (2016).
Marchini, A., Bonifati, S., Scott, E. M., Angelova, A. L. & Rommelaere, J. Oncolytic parvoviruses: from basic virology to clinical applications. Virol. J. 12, 6 (2015). DOI: 10.1186/s12985-014-0223-y
Bretscher, C. Marchini, A. H-1 parvovirus as a cancer-killing agent: past, present, and future. Viruses 11 https://doi.org/10.3390/v11060562 (2019).
Geletneky, K. et al. Oncolytic H-1 parvovirus shows safety and signs of immunogenic activity in a first phase I/IIa glioblastoma trial. Mol. Ther. 25, 2620–2634 (2017). DOI: 10.1016/j.ymthe.2017.08.016
Ungerechts, G. et al. Virotherapy research in Germany: from engineering to translation. Hum. Gene Ther. 28, 800–819 (2017). DOI: 10.1089/hum.2017.138
Hartley, A., Kavishwar, G., Salvato, I. & Marchini, A. A roadmap for the success of oncolytic parvovirus-based anticancer therapies. Annu Rev. Virol. 7, 537–557 (2020). DOI: 10.1146/annurev-virology-012220-023606
Nuesch, J. P., Lacroix, J., Marchini, A. & Rommelaere, J. Molecular pathways: rodent parvoviruses—mechanisms of oncolysis and prospects for clinical cancer treatment. Clin. Cancer Res. 18, 3516–3523 (2012). DOI: 10.1158/1078-0432.CCR-11-2325
Hristov, G. et al. Through its nonstructural protein NS1, parvovirus H-1 induces apoptosis via accumulation of reactive oxygen species. J. Virol. 84, 5909–5922 (2010). DOI: 10.1128/JVI.01797-09
Li, J. et al. 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 (2013). DOI: 10.1002/emmm.201302796
Bar, S., Rommelaere, J. & Nuesch, J. P. PKCeta/Rdx-driven phosphorylation of PDK1: a novel mechanism promoting cancer cell survival and permissiveness for parvovirus-induced lysis. PLoS Pathog. 11, e1004703 (2015). DOI: 10.1371/journal.ppat.1004703
Singh, B., Fleury, C., Jalalvand, F. & Riesbeck, K. Human pathogens utilize host extracellular matrix proteins laminin and collagen for adhesion and invasion of the host. FEMS Microbiol. Rev. 36, 1122–1180 (2012). DOI: 10.1111/j.1574-6976.2012.00340.x
Baum, L. G., Garner, O. B., Schaefer, K. & Lee, B. Microbe-host interactions are positively and negatively regulated by galectin-glycan interactions. Front. Immunol. 5, 284 (2014). DOI: 10.3389/fimmu.2014.00284
Di Pasquale, G. et al. Identification of PDGFR as a receptor for AAV-5 transduction. Nat. Med. 9, 1306–1312 (2003). DOI: 10.1038/nm929
Pillay, S. et al. An essential receptor for adeno-associated virus infection. Nature 530, 108–112 (2016). DOI: 10.1038/nature16465
Pillay, S. & Carette, J. E. Host determinants of adeno-associated viral vector entry. Curr. Opin. Virol. 24, 124–131 (2017). DOI: 10.1016/j.coviro.2017.06.003
Weller, M. L. et al. Epidermal growth factor receptor is a co-receptor for adeno-associated virus serotype 6. Nat. Med 16, 662–664 (2010). DOI: 10.1038/nm.2145
Allaume, X. et al. Retargeting of rat parvovirus H-1PV to cancer cells through genetic engineering of the viral capsid. J. Virol. 86, 3452–3465 (2012). DOI: 10.1128/JVI.06208-11
Halder, S. et al. Structural characterization of H-1 parvovirus: comparison of infectious virions to empty capsids. J. Virol. 87, 5128–5140 (2013). DOI: 10.1128/JVI.03416-12
Lopez-Bueno, A. et al. Host-selected amino acid changes at the sialic acid binding pocket of the parvovirus capsid modulate cell binding affinity and determine virulence. J. Virol. 80, 1563–1573 (2006). DOI: 10.1128/JVI.80.3.1563-1573.2006
Boisvert, M., Fernandes, S. & Tijssen, P. Multiple pathways involved in porcine parvovirus cellular entry and trafficking toward the nucleus. J. Virol. 84, 7782–7792 (2010). DOI: 10.1128/JVI.00479-10
Ferreira, T. et al. Oncolytic H-1 parvovirus enters cancer cells through clathrin-mediated endocytosis. Viruses 12, 1199 (2020). DOI: 10.3390/v12101199
Ros, C., Bayat, N., Wolfisberg, R. & Almendral, J. M. Protoparvovirus cell entry. Viruses 9, 313 (2017). DOI: 10.3390/v9110313
Houzet, L. & Jeang, K. T. Genome-wide screening using RNA interference to study host factors in viral replication and pathogenesis. Exp. Biol. Med. 236, 962–967 (2011). DOI: 10.1258/ebm.2010.010272
Sieben, M., Schafer, P., Dinsart, C., Galle, P. R. & Moehler, M. Activation of the human immune system via toll-like receptors by the oncolytic parvovirus H-1. Int J. Cancer 132, 2548–2556 (2013). DOI: 10.1002/ijc.27938
Sakashita, S., Engvall, E. & Ruoslahti, E. Basement membrane glycoprotein laminin binds to heparin. FEBS Lett. 116, 243–246 (1980). DOI: 10.1016/0014-5793(80)80654-5
Kouzi-Koliakos, K., Koliakos, G. G., Tsilibary, E. C., Furcht, L. T. & Charonis, A. S. Mapping of three major heparin-binding sites on laminin and identification of a novel heparin-binding site on the B1 chain. J. Biol. Chem. 264, 17971–17978 (1989). DOI: 10.1016/S0021-9258(19)84667-7
Di Piazza, M. et al. Cytosolic activation of cathepsins mediates parvovirus H-1-induced killing of cisplatin and TRAIL-resistant glioma cells. J. Virol. 81, 4186–4198 (2007). DOI: 10.1128/JVI.02601-06
Lacroix, J. et al. Parvovirus H1 selectively induces cytotoxic effects on human neuroblastoma cells. Int. J. Cancer 127, 1230–1239 (2010). DOI: 10.1002/ijc.25168
Bougnaud, S. et al. Molecular crosstalk between tumour and brain parenchyma instructs histopathological features in glioblastoma. Oncotarget 7, 31955–31971 (2016). DOI: 10.18632/oncotarget.7454
Malinda, K. M. & Kleinman, H. K. The laminins. Int. J. Biochem. Cell Biol. 28, 957–959 (1996). DOI: 10.1016/1357-2725(96)00042-8
Pupa, S. M., Menard, S., Forti, S. & Tagliabue, E. New insights into the role of extracellular matrix during tumor onset and progression. J. Cell Physiol. 192, 259–267 (2002). DOI: 10.1002/jcp.10142
Cousin, J. M. Cloninger, M. J. The role of galectin-1 in cancer progression, and synthetic multivalent systems for the study of galectin-1. Int. J. Mol. Sci. 17, https://doi.org/10.3390/ijms17091566 (2016).
Robinson, B. S., Arthur, C., Kamili, N. A. & Stowell, S. Galectin regulation of host microbial interactions. Trends Glycosci. Glycotechnol. 30, SE185-SE198(2018).
Garcin, P. O., Nabi, I. R. & Pante, N. Galectin-3 plays a role in minute virus of mice infection. Virology 481, 63–72 (2015). DOI: 10.1016/j.virol.2015.02.019
Schafer, G., Blumenthal, M. J. & Katz, A. A. Interaction of human tumor viruses with host cell surface receptors and cell entry. Viruses 7, 2592–2617 (2015). DOI: 10.3390/v7052592
Shieh, M. T., WuDunn, D., Montgomery, R. I., Esko, J. D. & Spear, P. G. Cell surface receptors for herpes simplex virus are heparan sulfate proteoglycans. J. Cell Biol. 116, 1273–1281 (1992). DOI: 10.1083/jcb.116.5.1273
Dechecchi, M. C., Tamanini, A., Bonizzato, A. & Cabrini, G. Heparan sulfate glycosaminoglycans are involved in adenovirus type 5 and 2-host cell interactions. Virology 268, 382–390 (2000). DOI: 10.1006/viro.1999.0171
Qin, Y., Rodin, S., Simonson, O. E. & Hollande, F. Laminins and cancer stem cells: partners in crime? Semin. Cancer Biol. 45, 3–12 (2017). DOI: 10.1016/j.semcancer.2016.07.004
Patarroyo, M., Tryggvason, K. & Virtanen, I. Laminin isoforms in tumor invasion, angiogenesis and metastasis. Semin. Cancer Biol. 12, 197–207 (2002). DOI: 10.1016/S1044-579X(02)00023-8
Zhang, Y. et al. Overexpression of LAMC1 predicts poor prognosis and enhances tumor cell invasion and migration in hepatocellular carcinoma. J. Cancer 8, 2992–3000 (2017). DOI: 10.7150/jca.21038
Kashima, H. et al. Laminin C1 expression by uterine carcinoma cells is associated with tumor progression. Gynecol. Oncol. 139, 338–344 (2015). DOI: 10.1016/j.ygyno.2015.08.025
Huang, S. X. et al. The correlation of microRNA-181a and target genes with poor prognosis of glioblastoma patients. Int. J. Oncol. 49, 217–224 (2016). DOI: 10.3892/ijo.2016.3511
Angelova, A. L., Geletneky, K., Nuesch, J. P. & Rommelaere, J. Tumor selectivity of oncolytic parvoviruses: from in vitro and animal models to cancer patients. Front. Bioeng. Biotechnol. 3, 55 (2015). DOI: 10.3389/fbioe.2015.00055
Golebiewska, A. et al. Patient-derived organoids and orthotopic xenografts of primary and recurrent gliomas represent relevant patient avatars for precision oncology. Acta Neuropathol. 140, 919–949 (2020). DOI: 10.1007/s00401-020-02226-7
El-Andaloussi, N. et al. Novel adenovirus-based helper system to support production of recombinant parvovirus. Cancer Gene Ther. 18, 240–249 (2011). DOI: 10.1038/cgt.2010.73
Raffelsberger, W. et al. RReportGenerator: automatic reports from routine statistical analysis using R. Bioinformatics 24, 276–278 (2008). DOI: 10.1093/bioinformatics/btm556
Kramer, A., Green, J., Pollard, J. Jr. & Tugendreich, S. Causal analysis approaches in Ingenuity Pathway Analysis. Bioinformatics 30, 523–530 (2014). DOI: 10.1093/bioinformatics/btt703
Mi, H., Muruganujan, A. & Thomas, P. D. PANTHER in 2013: modeling the evolution of gene function, and other gene attributes, in the context of phylogenetic trees. Nucleic Acids Res. 41, D377–D386 (2013). DOI: 10.1093/nar/gks1118
El-Andaloussi, N., Leuchs, B., Bonifati, S., Rommelaere, J. & Marchini, A. Efficient recombinant parvovirus production with the help of adenovirus-derived systems. J. Vis. Exp. 10.3791/3518 (2012). DOI: 10.3791/3518
Shalem, O. et al. Genome-scale CRISPR-Cas9 knockout screening in human cells. Science 343, 84–87 (2014). DOI: 10.1126/science.1247005
Ishihara, J. et al. Laminin heparin-binding peptides bind to several growth factors and enhance diabetic wound healing. Nat. Commun. 9, 2163 (2018). DOI: 10.1038/s41467-018-04525-w
Leuchs, B., Roscher, M., Muller, M., Kurschner, K. & Rommelaere, J. Standardized large-scale H-1PV production process with efficient quality and quantity monitoring. J. Virol. Methods 229, 48–59 (2016). DOI: 10.1016/j.jviromet.2015.11.022
Geiss, G. K. et al. Direct multiplexed measurement of gene expression with color-coded probe pairs. Nat. Biotechnol. 26, 317–325 (2008). DOI: 10.1038/nbt1385
Andersen, C. L., Jensen, J. L. & Orntoft, T. F. Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res. 64, 5245–5250 (2004). DOI: 10.1158/0008-5472.CAN-04-0496
Hossain, J. A., Riecken, K., Miletic, H. & Fehse, B. Cancer suicide gene therapy with TK.007. Methods Mol. Biol. 1895, 11–26 (2019). DOI: 10.1007/978-1-4939-8922-5_2
Hossain, J. A. et al. Long-term treatment with valganciclovir improves lentiviral suicide gene therapy of glioblastoma. Neuro Oncol. 21, 890–900 (2019). DOI: 10.1093/neuonc/noz060
Kinsner-Ovaskainen, A., Prieto, P., Stanzel, S. & Kopp-Schneider, A. Selection of test methods to be included in a testing strategy to predict acute oral toxicity: an approach based on statistical analysis of data collected in phase 1 of the ACuteTox project. Toxicol. Vitr. 27, 1377–1394 (2013). DOI: 10.1016/j.tiv.2012.11.010
Barretina, J. et al. The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature 483, 603–607 (2012). DOI: 10.1038/nature11003
Anaya, J. OncoLnc: linking TCGA survival data to mRNAs, miRNAs, and lncRNAs. PeerJ Comput. Sci. 2, e67 (2016). DOI: 10.7717/peerj-cs.67
Bowman, R. L., Wang, Q., Carro, A., Verhaak, R. G. & Squatrito, M. GlioVis data portal for visualization and analysis of brain tumor expression datasets. Neuro Oncol. 19, 139–141 (2017). DOI: 10.1093/neuonc/now247
