BioKnife, a uPA activity-dependent oncolytic Sendai virus, eliminates pleural spread of malignant mesothelioma via simultaneous stimulation of uPA expression

Our plain-English summary of BioKnife, a uPA activity-dependent oncolytic Sendai virus, eliminates pleural spread of malignant mesothelioma via simultaneous stimulation of uPA expression, by Y. Morodomi, T. Yano, H. Kinoh, et al (Molecular Therapy, 2012).

The study

• Authors: Y. Morodomi, T. Yano, H. Kinoh, Y. Harada, S. Saito, R. Kyuragi, K. Yoshida, M. Onimaru, F. Shoji, T. Yoshida, K. Ito, Y. Shikada, R. Maruyama, M. Hasegawa, Y. Maehara, Y. Yonemitsu
• Journal: Molecular Therapy
• Published: 2012;20(4):769-77
• Cited by: 20 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Malignant pleural mesothelioma (MPM) is highly intractable and readily spreads throughout the surface of the pleural cavity, and these cells have been shown to express urokinase-type plasminogen activator receptor (uPAR). We here examined the potential of our new and powerful recombinant Sendai virus (rSeV), which shows uPAR-specific cell-to-cell fusion activity (rSeV/dMFct14 (uPA2), named "BioKnife"), for tumor cell killing in two independent orthotopic xenograft models of human. Multicycle treatment using BioKnife resulted in the efficient rescue of these models, in association with tumor-specific fusion and apoptosis. Such an effect was also seen on both MSTO-211H and H226 cells in vitro; however, we confirmed that the latter expressed uPAR but not uPA. Of interest, infection with BioKnife strongly facilitated the uPA release from H226 cells, and this effect was completely abolished by use of either pyrrolidine dithiocarbamate (PDTC) or BioKnife expressing the C-terminus-deleted dominant negative inhibitor for retinoic acid-inducible gene-I (RIG-IC), indicating that BioKnife-dependent expression of uPA was mediated by the RIG-I/nuclear factor-κB (NF-κB) axis, detecting RNA viral genome replication. Therefore, these results suggest a proof of concept that the tumor cell-killing mechanism via BioKnife may have significant potential to treat patients with MPM that is characterized by frequent uPAR expression in a clinical setting.

In plain English

This laboratory and animal study tested BioKnife (rSeV/dMFct14 (uPA2)), a recombinant Sendai virus engineered to kill cells through urokinase-type plasminogen activator (uPA) activity-dependent cell-to-cell fusion, against human malignant pleural mesothelioma (MPM). Asbestos is mentioned only in the introduction as the background cause of MPM; it was not studied here. The authors established orthotopic models in nude mice using the human MPM lines H226 (epithelioid) and MSTO-211H (biphasic), and reported that six intrathoracic injections of BioKnife-GFP (1 x 10(7) cell-infectious units per dose) gave the optimal survival effect in both models (P less than 0.01). In a separate H226 experiment, tumours examined two days after a single injection of BioKnife-GFP had a larger TUNEL-positive (apoptotic) area than tumours given control virus; in H226-luc tumour mice imaged on days 0, 7 and 14 after vector injection, luciferase signal fell with BioKnife-GFP while it rose with control virus. In cell culture, caspase 3/7 activity rose only in the MPM lines, and killing of H226 was reduced by plasminogen activator inhibitor-1, a pan-caspase inhibitor and uPAR knockdown. uPAR staining was found in all nine surgical MPM specimens. H226 cells made no uPA, but virus infection induced uPA, an effect abolished by the NF-kappaB inhibitor PDTC or by RIG-IC. The authors concluded these findings suggest a proof of concept for treating MPM.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Preclinical laboratory study: in vitro cell experiments plus orthotopic xenograft experiments in nude mice, with immunohistochemistry of nine human surgical specimens
• Who was studied: Human MPM cell lines H226 (epithelioid) and MSTO-211H (biphasic), control cells AoSMC and Met5A; 4-week-old male balb/c nu/nu mice bearing orthotopic pleural tumours; and surgical specimens from nine MPM patients operated at Kyushu University Hospital between June 2001 and October 2009
• Exposure: Intrapleural administration of the uPA activity-dependent oncolytic Sendai virus BioKnife-GFP (1 x 10(7) ciu/dose, once, three or six times), compared with control virus rSeV/dM-GFP or PBS; in vitro infection at MOI 5, 20 or 50, with PAI-1, Z-VAD-FMK, PDTC, uPAR siRNA or RIG-IC
• Main results:
  • Model establishment: inoculation of 5 x 10(6) cells killed all mice, within 128 days for H226 and within 38 days for MSTO-211H
  • Repeated dosing: six injections of BioKnife-GFP gave the optimal survival effect in both tumour models (H226 and MSTO-211H)
  • The apoptosis experiment used H226 tumour-bearing mice examined two days after a single intrapleural injection given on day 7 after tumour cell inoculation
  • In those H226 tumours, BioKnife-GFP produced a significantly larger TUNEL-positive (apoptotic) area than the control virus rSeV/dM-GFP
  • Luciferase imaging of H226-luc tumour-bearing mice was done on days 0, 7 and 14 after vector injection
  • In that imaging experiment, luciferase activity rose over time with control virus but fell in mice given BioKnife-GFP
  • In vitro: caspase 3/7 activity rose markedly only in the MPM cells (H226, MSTO-211H), not in control AoSMC or Met5A cells, with corresponding cytotoxicity in the MPM cells
  • PAI-1 and the pan-caspase inhibitor Z-VAD-FMK significantly reduced BioKnife-GFP-mediated killing of H226 cells
  • uPAR expression was found in all nine human MPM surgical specimens; western blotting showed uPAR (~55 kDa) strongly in H226 and MSTO-211H, a small amount in AoSMC, and none in Met5A
  • siRNA knockdown of uPAR in H226 cells reduced the cytotoxic effect of BioKnife-GFP but not of control virus
  • H226 cells released no detectable uPA, yet infection with rSeV/dM-GFP or BioKnife-GFP induced uPA mRNA and protein; PDTC or BioKnife expressing RIG-IC almost completely abolished this induction
  • In vivo, uPA protein in H226 tumour nodules was raised by control virus and significantly more so by BioKnife-GFP
  • Non-malignant Met5A cells, which express uPA but not uPAR, nonetheless showed about 30% cytotoxicity
• The authors' conclusion: The authors concluded that BioKnife had antitumour activity and survival benefit in these MPM models, that its effect did not depend on tumour release of uPA because infection itself induced uPA via RIG-I and NF-kappaB, and that this suggests potential utility for treating MPM, with further study needed
• Limitations the authors state:
  • The authors state BioKnife theoretically cannot reach distant metastases
  • The authors state that uPAR-negative tumours may not always be resistant to BioKnife
  • The authors state that more basic research is needed to predict the efficacy of BioKnife
  • The authors state further preclinical studies in large animals are warranted before clinical use
  • Several dose-response results are reported only as 'data not shown'

Topics

The National Library of Medicine indexes this study under subjects our site covers: Mesothelioma.

Medical terms in this study, explained in our Lexicon: Malignant Mesothelioma, Pleural Mesothelioma, Pleural Cavity, Apoptosis.

Related studies

• Synergistic effect of non-transmissible Sendai virus vector encoding the c-myc suppressor FUSE-binding protein-interacting repressor plus cisplatin in the treatment of malignant pleural mesothelioma — A. Kitamura, K. Matsushita, et al. (2011)
• Frequent Homozygous Deletions of Type I Interferon Genes in Pleural Mesothelioma Confer Sensitivity to Oncolytic Measles Virus — T. Delaunay, C. Achard, et al. (2020)
• Expression and activity of urokinase and its receptor in endothelial and pulmonary epithelial cells exposed to asbestos — A. Barchowsky, R.R. Roussel, et al. (1998)
• Enhanced antitumor efficacy of fiber-modified, midkine promoter-regulated oncolytic adenovirus in human malignant mesothelioma — M. Takagi-Kimura, T. Yamano, et al. (2013)
• Antitumor-specific T-cell responses induced by oncolytic adenovirus ONCOS-102 (AdV5/3-D24-GM-CSF) in peritoneal mesothelioma mouse model — L. Kuryk, A.W. Møller, et al. (2018)
• Oncolytic Viral Therapy for Malignant Pleural Mesothelioma — H. Yang, D. Xu, et al. (2020)

Browse return to medical research.