14084
2011
eng
1-14
36
9
article
1
2016-11-25
--
--
Insertion of the human sodium iodide symporter to facilitate deep tissue imaging does not alter oncolytic or replication capability of a novel vaccinia virus
Introduction:
Oncolytic viruses show promise for treating cancer. However, to assess therapeutic efficacy and potential toxicity, a noninvasive imaging modality is needed. This study aimed to determine if insertion of the human sodium iodide symporter (hNIS) cDNA as a marker for non-invasive imaging of virotherapy alters the replication and oncolytic capability of a novel vaccinia virus, GLV-1h153.
Methods:
GLV-1h153 was modified from parental vaccinia virus GLV-1h68 to carry hNIS via homologous recombination. GLV-1h153 was tested against human pancreatic cancer cell line PANC-1 for replication via viral plaque assays and flow cytometry. Expression and transportation of hNIS in infected cells was evaluated using Westernblot and immunofluorescence. Intracellular uptake of radioiodide was assessed using radiouptake assays. Viral cytotoxicity and tumor regression of treated PANC-1tumor xenografts in nude mice was also determined. Finally, tumor radiouptake in xenografts was assessed via positron emission tomography (PET) utilizing carrier-free (124)I radiotracer.
Results:
GLV-1h153 infected, replicated within, and killed PANC-1 cells as efficiently as GLV-1h68. GLV-1h153 provided dose-dependent levels of hNIS expression in infected cells. Immunofluorescence detected transport of the protein to the cell membrane prior to cell lysis, enhancing hNIS-specific radiouptake (P < 0.001). In vivo, GLV-1h153 was as safe and effective as GLV-1h68 in regressing pancreatic cancer xenografts (P < 0.001). Finally, intratumoral injection of GLV-1h153 facilitated imaging of virus replication in tumors via (124)I-PET.
Conclusion:
Insertion of the hNIS gene does not hinder replication or oncolytic capability of GLV-1h153, rendering this novel virus a promising new candidate for the noninvasive imaging and tracking of oncolytic viral therapy.
Journal of Translational Medicine
10.1186/1479-5876-9-36
urn:nbn:de:bvb:20-opus-140847
Journal of Translational Medicine 2011 9:36., doi:10.1186/1479-5876-9-36
Dana Haddad
Nanhai G. Chen
Qian Zhang
Chun-Hao Chen
Yong A. Yu
Lorena Gonzalez
Susanne G. Carpenter
Joshua Carson
Joyce Au
Arjun Mittra
Mithat Gonen
Pat B. Zanzonico
Yuman Fong
Aladar A. Szalay
eng
uncontrolled
Human Sodium/Iodide symporter
eng
uncontrolled
Reporter gene
eng
uncontrolled
NA+/I-symporter
eng
uncontrolled
Nude-mice
eng
uncontrolled
Cancer
eng
uncontrolled
In-Vivo
eng
uncontrolled
Expression
eng
uncontrolled
Therapy
eng
uncontrolled
Transporter
eng
uncontrolled
GLV-1H68
Biochemie
open_access
Lehrstuhl für Biochemie
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/14084/077_Dana_JOURNAL-OF-TRANSLATIONAL-MEDICINE.pdf
13531
2011
eng
e22069
7
6
article
1
2016-06-22
--
--
Efficient Colonization and Therapy of Human Hepatocellular Carcinoma (HCC) Using the Oncolytic Vaccinia Virus Strain GLV-1h68
Virotherapy using oncolytic vaccinia virus strains is one of the most promising new strategies for cancer therapy. In this study, we analyzed for the first time the therapeutic efficacy of the oncolytic vaccinia virus GLV-1h68 in two human hepatocellular carcinoma cell lines HuH7 and PLC/PRF/5 (PLC) in cell culture and in tumor xenograft models. By viral proliferation assays and cell survival tests, we demonstrated that GLV-1h68 efficiently colonized, replicated in, and did lyse these cancer cells in culture. Experiments with HuH7 and PLC xenografts have revealed that a single intravenous injection (i.v.) of mice with GLV-1h68 resulted in a significant reduction of primary tumor sizes compared to uninjected controls. In addition, replication of GLV-1h68 in tumor cells led to strong inflammatory and oncolytic effects resulting in intense infiltration of MHC class II-positive cells like neutrophils, macrophages, B cells and dendritic cells and in up-regulation of 13 pro-inflammatory cytokines. Furthermore, GLV-1h68 infection of PLC tumors inhibited the formation of hemorrhagic structures which occur naturally in PLC tumors. Interestingly, we found a strongly reduced vascular density in infected PLC tumors only, but not in the non-hemorrhagic HuH7 tumor model. These data demonstrate that the GLV-1h68 vaccinia virus may have an enormous potential for treatment of human hepatocellular carcinoma in man.
PLOS ONE
10.1371/journal.pone.0022069
urn:nbn:de:bvb:20-opus-135319
PLoS ONE 6(7): e22069. doi:10.1371/journal.pone.0022069
false
true
Ivaylo Gentschev
Meike Müller
Marion Adelfinger
Stephanie Weibel
Friedrich Grummt
Martina Zimmermann
Michael Bitzer
Martin Heisig
Qian Zhang
Yong A. Yu
Nanhai G. Chen
Jochen Stritzker
Ulrich M. Lauer
Aladar A. Szalay
eng
uncontrolled
Breast-tumors
eng
uncontrolled
Nude-mice
eng
uncontrolled
In-vivo
eng
uncontrolled
Cancer
eng
uncontrolled
Inhibitor
eng
uncontrolled
Tissue
eng
uncontrolled
Agent
eng
uncontrolled
COX-2
Medizin und Gesundheit
open_access
Institut für Molekulare Infektionsbiologie
Rudolf-Virchow-Zentrum
Lehrstuhl für Biochemie
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/13531/044_Gentschev_PLoSONE.pdf
14226
2011
eng
1-11
164
9
article
1
2016-12-21
--
--
Replication efficiency of oncolytic vaccinia virus in cell cultures prognosticates the virulence and antitumor efficacy in mice
Background:
We have shown that insertion of the three vaccinia virus (VACV) promoter-driven foreign gene expression cassettes encoding Renilla luciferase-Aequorea GFP fusion protein, beta-galactosidase, and beta-glucuronidase into the F14.5L, J2R, and A56R loci of the VACV LIVP genome, respectively, results in a highly attenuated mutant strain GLV 1h68. This strain shows tumor specific replication and is capable of eradicating tumors with little or no virulence in mice. This study aimed to distinguish the contribution of added VACV promoter-driven transcriptional units as inserts from the effects of insertional inactivation of three viral genes, and to determine the correlation between replication efficiency of oncolytic vaccinia virus in cell cultures and the virulence and antitumor efficacy in mice
Methods:
A series of recombinant VACV strains was generated by replacing one, two, or all three of the expression cassettes in GLV 1h68 with short non coding DNA sequences. The replication efficiency and tumor cell killing capacity of these newly generated VACV strains were compared with those of the parent virus GLV-1h68 in cell cultures. The virus replication efficiency in tumors and antitumor efficacy as well as the virulence were evaluated in nu/nu (nude) mice bearing human breast tumor xenografts.
Results:
we found that virus replication efficiency increased with removal of each of the expression cassettes. The increase in virus replication efficiency was proportionate to the strength of removed VACV promoters linked to foreign genes. The replication efficiency of the new VACV strains paralleled their cytotoxicity in cell cultures. The increased replication efficiency in tumor xenografts resulted in enhanced antitumor efficacy in nude mice. Similarly, the enhanced virus replication efficiency was indicative of increased virulence in nude mice.
Conclusions:
These data demonstrated that insertion of VACV promoter-driven transcriptional units into the viral genome for the purpose of insertional mutagenesis did modulate the efficiency of virus replication together with antitumor efficacy as well as virulence. Replication efficiency of oncolytic VACV in cell cultures can predict the virulence and therapeutic efficacy in nude mice. These findings may be essential for rational design of safe and potent VACV strains for vaccination and virotherapy of cancer in humans and animals.
Journal of Translational Medicine
10.1186/1479-5876-9-164
urn:nbn:de:bvb:20-opus-142268
Journal of Translational Medicine 2011 9:164.
Nanhai G. Chen
Yong A. Yu
Qian Zhang
Aladar A. Szalay
eng
uncontrolled
Recombinant vaccinia
eng
uncontrolled
Nude-mice
eng
uncontrolled
Cancer
eng
uncontrolled
GLV-1H68
eng
uncontrolled
Therapy
eng
uncontrolled
Agent
eng
uncontrolled
Regression
eng
uncontrolled
Carcinoma
eng
uncontrolled
Deletion
eng
uncontrolled
Protein
eng
uncontrolled
modulation of virus replication
eng
uncontrolled
GI-101A tumor xenografts
eng
uncontrolled
oncolytic virotherapy
Medizin und Gesundheit
open_access
Institut für Molekulare Infektionsbiologie
Rudolf-Virchow-Zentrum
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/14226/124_Chen_JOURNAL-OF-TRANSLATIONAL-MEDICINE.pdf