A Cytoplasmic RNA Vector Derived from Nontransmissible Sendai Virus with Efficient Gene Transfer and Expression

Hai-ou, LI; Zhu, Yong; Asakawa, Makoto; Kuma, Hidekazu; Hirata, Takahiro; Ueda, Yasuji; Lee, Yun-Sik; Fukumura, Masayuki; Iida, Akihiro; Kato, Atsushi; Nagai, Yoshinori; Hasegawa, Mamoru

doi:10.1128/jvi.74.14.6564-6569.2000

Cited by 231 publications

(197 citation statements)

References 34 publications

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“…1 A transmission-incompetent SeV vector has been developed by deleting the F-protein, which is essential for cellular entry of the viral genome (DF/SeV). 4 Moreover, this modification does not reduce transfection efficiency of the virus. 1,4 In general the level of transgene expression achieved from repeat delivery of a viral vector is greatly reduced when compared with that from a single dose due to induction of effective immune responses in the recipient.…”

Section: Introductionmentioning

confidence: 99%

“…4 Moreover, this modification does not reduce transfection efficiency of the virus. 1,4 In general the level of transgene expression achieved from repeat delivery of a viral vector is greatly reduced when compared with that from a single dose due to induction of effective immune responses in the recipient. 5,6 We have previously shown that SeV-mediated gene expression is reduced by 60% on second dose, and that tolerization of mice against immune-dominant SeV epitopes does not improve efficacy.…”

Section: Introductionmentioning

confidence: 99%

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Validation of recombinant Sendai virus in a non-natural host model

et al. 2010

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We have previously shown that recombinant Sendai virus (SeV) vector, derived from murine parainfluenza virus, is one of the most efficient vectors for airway gene transfer. We have also shown that SeV-mediated transfection on second administration, although reduced by 60% when compared with levels achieved after a single dose, is still high because of the efficient transfection achieved by SeV vector in murine airways. Here, we show that these levels further decrease on subsequent doses. In addition, we validated SeV vector repeat administration in a non-natural host model, the sheep. As part of these studies we first assessed viral stability in a Pari LC Plus nebuliser, a polyethylene catheter (PEC) and the Trudell AeroProbe. We also compared the distribution of gene expression after PEC and Trudell AeroProbe administration and quantified virus shedding after sheep transduction. In addition, we show that bronchial brushings and biopsies, collected in anaesthetized sheep, can be used to assess SeV-mediated gene expression over time. Similar to mice, gene expression in sheep was transient and had returned to baseline values by day 14. In conclusion, the SeV vector should be strongly considered for lung-related applications requiring a single administration of the vector even though it might not be suitable for diseases requiring repeat administration.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Validation of recombinant Sendai virus in a non-natural host model

et al. 2010

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“…Site-directed mutagenesis was performed using the Quick-Change Mutagenesis Kit (Stratagene), and mutated F gene was returned to the pSeV18+/DF-GFP backbone. 4 Recovery and amplification of the SeV vector were generated essentially as described before; 4,5 briefly, LLC-MK2 cells were transfected at room temperature with a plasmid mixture containing each plasmid [pSeV+18/F(MMP-C1 MMP-G1/MTS, G2/MTS, G3/MTS, G4/MTS, uPA1, uPA2 and tPA)DM-GFP], pSeV+18/Fct14(MMP-G4/MTS, uPA2) DM-GFP, pGEM-NP, pGEM-P and pGEM-L in 110 ml of Superfect transfection reagent (Qiagen, Tokyo, Japan). The transfected cells were maintained for 3 h in 3 ml of Opti-MEM (Gibco-BRL) plus 3% fetal calf serum, washed three times with modified Eagles' medium (MEM) and incubated for 60 h in MEM containing araC (40 mg ml À1 ).…”

Section: Construction and Recovery Of Improved Oncolytic Sev Vectormentioning

confidence: 99%

“…Virus yield is expressed in cell infection units, as described previously. 4 Fusion assay and cytotoxicity assay Cells were plated in 96-well dishes at a density of 5 Â 10 3 cells per well and infected as described above with each SeV vector at an MOI of 0.3. On day 4, cells were fixed with 4% paraformaldehyde.…”

Section: Construction and Recovery Of Improved Oncolytic Sev Vectormentioning

confidence: 99%

“…This restricted tropism primarily depends on the specific tissue proteases required for cleavage activation of viral fusion (F) glycoprotein, and thus the proteases needed for infectivity of progeny (the capacity to penetrate into and initiate infection in the next cell) are available only on the surface of those limited types of tissue. 3 Using sophisticated manipulating technology of the genome of SeV, namely 'Reverse Genetics, ' we recently generated SeV vectors lacking envelope-related genes, including fusion (F) gene-deleted (SeV/DF), 4 matrix (M) gene-deleted (SeV/DM), 5 hemagglutinin/neuraminidase (HN) gene-deleted (SeV/DHN), both M and F genesdeleted (SeV/DMDF) 6 and all of the envelope-related genes-deleted (SeV/DMDFDHN). 7 Among the proteins contained in the viral lipid bilayer, M protein plays a central role in secondary virus assembly and budding from infected cells.…”

Section: Introductionmentioning

confidence: 99%

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Generation of optimized and urokinase-targeted oncolytic Sendai virus vectors applicable for various human malignancies

et al. 2008

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We previously reported the development of a prototype 'oncolytic Sendai virus (SeV) vector' formed by introducing two major genomic modifications to the original SeV, namely deletion of the matrix (M) gene to avoid budding of secondary viral particles and manipulation of the trypsin-dependent cleavage site of the fusion (F) gene to generate proteasespecific sequences. As a result, the 'oncolytic SeV' that was susceptible to matrix metalloproteinases (MMPs) was shown to selectively kill MMP-expressing tumors through syncytium formation in vitro and in vivo. However, its efficacy has been relatively limited because of the requirement of higher expression of MMPs and smaller populations of MMP-expressing tumors. To overcome these limitations, we have designed an optimized and dramatically powerful oncolytic SeV vector. Truncation of 14-amino acid residues of the cytoplasmic domain of F protein resulted in dramatic enhancement of cell-killing activities of oncolytic SeV, and the combination with replacement of the trypsin cleavage site with the new urokinase type plasminogen activator (uPA)-sensitive sequence (SGRS) led a variety of human tumors, including prostate (PC-3), renal (CAKI-I), pancreatic (BxPC3) and lung (PC14) cancers, to extensive death through massive cell-tocell spreading without significant dissemination to the surrounding noncancerous tissue in vivo. These results indicate a dramatic improvement of antitumor activity; therefore, extensive utility of the newly designed uPA-targeted oncolytic SeV has significant potential for treating patients bearing urokinase-expressing cancers in clinical settings.

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iPSC screening for drug repurposing identifies anti‐RNA virus agents modulating host cell susceptibility

et al. 2021

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Human pathogenic RNA viruses are threats to public health because they are prone to escaping the human immune system through mutations of genomic RNA, thereby causing local outbreaks and global pandemics of emerging or re‐emerging viral diseases. While specific therapeutics and vaccines are being developed, a broad‐spectrum therapeutic agent for RNA viruses would be beneficial for targeting newly emerging and mutated RNA viruses. In this study, we conducted a screen of repurposed drugs using Sendai virus (an RNA virus of the family Paramyxoviridae ), with human‐induced pluripotent stem cells (iPSCs) to explore existing drugs that may present anti‐RNA viral activity. Selected hit compounds were evaluated for their efficacy against two important human pathogens: Ebola virus (EBOV) using Huh7 cells and severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) using Vero E6 cells. Selective estrogen receptor modulators (SERMs), including raloxifene, exhibited antiviral activities against EBOV and SARS‐CoV‐2. Pioglitazone, a PPARγ agonist, also exhibited antiviral activities against SARS‐CoV‐2, and both raloxifene and pioglitazone presented a synergistic antiviral effect. Finally, we demonstrated that SERMs blocked entry steps of SARS‐CoV‐2 into host cells. These findings suggest that the identified FDA‐approved drugs can modulate host cell susceptibility against RNA viruses.

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A Cytoplasmic RNA Vector Derived from Nontransmissible Sendai Virus with Efficient Gene Transfer and Expression

Cited by 231 publications

References 34 publications

Validation of recombinant Sendai virus in a non-natural host model

Validation of recombinant Sendai virus in a non-natural host model

Generation of optimized and urokinase-targeted oncolytic Sendai virus vectors applicable for various human malignancies

iPSC screening for drug repurposing identifies anti‐RNA virus agents modulating host cell susceptibility

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