Induction of apoptosis in vitro and in vivo by H-1 parvovirus infection.

Ohshima, Takayuki; Iwama, Mizuho; Ueno, Yutaka; Sugiyama, Fumihiro; Nakajima, Takeshi; Fukamizu, Akiyoshi; Yagami, Ken‐ichi

doi:10.1099/0022-1317-79-12-3067

Cited by 45 publications

(41 citation statements)

References 23 publications

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“…H1 virus was also able to induce programmed cell death in human hepatoma cells, as it was shown for other cell types and by other hepatotropic viruses. 18,19,35,37 In several cases, tumor cells were more sensitive to H1 -induced toxicity than their nontransformed parental cells. 12,22,23 Thus, human hepatoma cells constitute an additional system in which a selective killing by H1 virus could be achieved in the transformed, but not in the primary, hepatocytes.…”

Section: Discussionmentioning

confidence: 99%

Effective infection, apoptotic cell killing and gene transfer of human hepatoma cells but not primary hepatocytes by parvovirus H1 and derived vectors

Moehler¹,

Blechacz

Weiskopf³

et al. 2001

Cancer Gene Ther

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Autonomous parvoviruses preferentially replicate in and kill in vitro ± transformed cells and reduce the incidence of spontaneous and implanted tumors in animals. Because of these natural oncotropic and oncolytic properties, parvoviruses deserve to be considered as potential antitumor vectors. Here, we assessed whether parvovirus H1 is able to kill human hepatoma cells by induction of apoptosis but spares primary human liver cells, and whether the former cells can efficiently be transduced by H1 virus ± based vectors. Cell death, infectivity, and transgene transduction were investigated in Hep3B, HepG2, and Huh7 cells and in primary human hepatocytes with natural and recombinant H1 virus. All hepatoma cells were susceptible to H1 virus ± induced cytolyis. Cell death correlated with H1 virus DNA replication, nonstructural protein expression, and with morphological features of apoptosis. H1 virus ± induced apoptosis was more pronounced in p53 -deleted Hep3B and p53 -mutated Huh7 cells than in HepG2 cells which express wild -type p53. In Hep3B cells, apoptosis was partially inhibited by DEVD -CHO, a caspase -3 inhibitor. In contrast, H1 virus ± infected primary hepatocytes were neither positive for nonstructural protein expression nor susceptible to H1 virus ± induced killing. Infection with a recombinant parvovirus vector carrying the luciferase gene under control of parvovirus promoter P38 led to higher transgene activities in hepatoma cells than in the hepatocytes. Taken together, H1 virus kills human hepatoma cells at low virus multiplicity but not primary hepatocytes. Thus, recombinant H1 viruses carrying antitumor transgenes may be considered as potential therapeutic options for the treatment of hepatocellular carcinomas. Cancer Gene Therapy ( 2001 ) 8, 158 ± 167

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Section: Discussionmentioning

confidence: 99%

Effective infection, apoptotic cell killing and gene transfer of human hepatoma cells but not primary hepatocytes by parvovirus H1 and derived vectors

Moehler¹,

Blechacz

Weiskopf³

et al. 2001

Cancer Gene Ther

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“…This study focuses on rat H-1PV, which infects and kills human tumor cell lines of various origins (e.g., of brain [23], pancreas [4,14], blood [3], colon [38], cervix [20], and breast [66,67]) and which is currently under evaluation in a phase I/IIa clinical trial for the treatment of patients with recurrent glioblastoma multiforme (57). H-1PV has the ability to induce different cell death pathways in cancer cells, including necrosis (53), apoptosis (28,46,54,65), and lysosome-dependent cell death (16), while sparing nontransformed cells. Recently, we have reported the capacity of the virus to induce oxidative stress in cancer cells leading to DNA damage, cell cycle arrest, and apoptosis.…”

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confidence: 99%

Retargeting of Rat Parvovirus H-1PV to Cancer Cells through Genetic Engineering of the Viral Capsid

Allaume

El-Andaloussi

Leuchs

et al. 2012

J Virol

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The rat parvovirus H-1PV is a promising anticancer agent given its oncosuppressive properties and the absence of known side effects in humans. H-1PV replicates preferentially in transformed cells, but the virus can enter both normal and cancer cells. Uptake by normal cells sequesters a significant portion of the administered viral dose away from the tumor target. Hence, targeting H-1PV entry specifically to tumor cells is important to increase the efficacy of parvovirus-based treatments. In this study, we first found that sialic acid plays a key role in H-1PV entry. We then genetically engineered the H-1PV capsid to improve its affinity for human tumor cells. By analogy with the resolved crystal structure of the closely related parvovirus minute virus of mice, we developed an in silico three-dimensional (3D) model of the H-1PV wild-type capsid. Based on this model, we identified putative amino acids involved in cell membrane recognition and virus entry at the level of the 2-fold axis of symmetry of the capsid, within the so-called dimple region. In situ mutagenesis of these residues significantly reduced the binding and entry of H-1PV into permissive cells. We then engineered an entry-deficient viral capsid and inserted a cyclic RGD-4C peptide at the level of its 3-fold axis spike. This peptide binds ␣ v ␤ 3 and ␣ v ␤ 5 integrins, which are overexpressed in cancer cells and growing blood vessels. The insertion of the peptide rescued viral infectivity toward cells overexpressing ␣ v ␤ 5 integrins, resulting in the efficient killing of these cells by the reengineered virus. This work demonstrates that H-1PV can be genetically retargeted through the modification of its capsid, showing great promise for a more efficient use of this virus in cancer therapy.

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“…Certain autonomous parvoviruses were assumed to induce apoptosis in infected cells (4,11,12,15,18,26). Our previous study (15) demonstrated caspase-3-dependent apoptosis in H-1 virus-infected C6 cells.…”

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confidence: 99%

Propagation of Rat Parvovirus in Thymic Lymphoma Cell Line C58(NT)D and Subsequent Appearance of a Resistant Cell Clone after Lytic Infection

Ueno

Harada

Iseki

et al. 2001

J Virol

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Rat parvovirus (RPV) is nonpathogenic in rats but causes persistent lymphocytotropic infection. We found that RPV was propagated in rat thymic lymphoma cell line C58(NT)D and induced apoptosis. Interestingly, a resistant subclone, C58(NT)D/R, from surviving cells after lytic infection had differentiated phenotypic modifications, such as increased cell adherence, resistance to apoptosis, and suppressed tumorigenicity.Recent molecular studies on parvoviral pathogenicity suggest that the viral nonstructural (NS) protein in which coded genes are highly homologous among parvoviruses correlates with cytotoxicity (2,8,17). The productive and cytotoxic activity of the NS protein is modulated by cellular factors that may vary with the host cell type, particularly in oncogene-transformed cells (2, 13). We have shown that a transcriptional coactivator, CREB binding protein, is required for NS1-mediated viral and cellular transcription in parvovirus-infected cells, resulting in cell proliferation and differentiation to achieve its lytic cycle (16).Newly recognized rodent parvoviruses, also called orphan parvoviruses, are widespread among laboratory mice and rats (21,23,24), and viruses isolated have been classified as mouse parvovirus and rat parvovirus (RPV) (1, 5, 6). Although mouse parvovirus grows well in a murine T-cell clone (L3), causing a cytopathic effect (CPE) (10), effective in vitro RPV propagation has not been established. We found that RPV involved lytic infection in the rat thymic lymphoma cell line C58(NT)D and developed in vitro propagation for RPV. Interestingly, virus-resistant cell clones isolated from subcultures of surviving cells acquired differentiated phenotypes such as reduced tumorigenicity and sensitivity to apoptosis. Viral propagation in cell lines. The RPV strain was isolated from rats infected spontaneously at our facility (23) and passaged three times in specific-pathogen-free newborn SpragueDawley rats. The virus stock was prepared from infected spleens at 7 days postinoculation (p.i.) We initially inoculated a supernatant of 5% infected spleen homogenate into cell lines and primary cell cultures of rats and hamsters to find cells permitting virus propagation. C6 (rat glioma), BRL-3A (Buffalo rat liver), RBL-2H3 (rat basophilic leukemia), BHK-21 (Syrian hamster kidney), and Y3-Ag1.2.3 (rat myeloma) cells were obtained from the Riken Cell Bank, Tsukuba, Japan, and C58(NT)D (rat thymic lymphoma) cells were purchased from the American Type Culture Collection, Manassas, Va. Infected cells were observed for appearance of the viral CPE and examined for presence of the viral antigen by immunofluorescent antibody (IFA) and hemagglutination (HA) ability assays. The isolated virus was propagated only in C58(NT)D cells, not in other cells tested (Table 1). Parvoviral DNA was also detected only in infected C58(NT)D cells (data not shown). In contrast, prototype RV-13 (7) was propagated, with titers in C58(NT)D cells lower than those in other cells. Propagation of the isolate in C58(NT)D cells and a dif...

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Induction of apoptosis in vitro and in vivo by H-1 parvovirus infection.

Cited by 45 publications

References 23 publications

Effective infection, apoptotic cell killing and gene transfer of human hepatoma cells but not primary hepatocytes by parvovirus H1 and derived vectors

Effective infection, apoptotic cell killing and gene transfer of human hepatoma cells but not primary hepatocytes by parvovirus H1 and derived vectors

Retargeting of Rat Parvovirus H-1PV to Cancer Cells through Genetic Engineering of the Viral Capsid

Propagation of Rat Parvovirus in Thymic Lymphoma Cell Line C58(NT)D and Subsequent Appearance of a Resistant Cell Clone after Lytic Infection

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