Limits...
Oncolytic vaccinia therapy of squamous cell carcinoma.

Yu Z, Li S, Brader P, Chen N, Yu YA, Zhang Q, Szalay AA, Fong Y, Wong RJ - Mol. Cancer (2009)

Bottom Line: All six cell lines supported viral transgene expression (beta-galactosidase, green fluorescent protein, and luciferase) as early as 6 hours after viral exposure.Even at a very low MOI of 0.01, three cell lines still demonstrated >60% cell death over 6 days.A single injection of GLV-1h68 (5 x 10(6) pfu) intratumorally into MSKQLL2 xenografts in mice exhibited localized intratumoral luciferase activity peaking at days 2-4, with gradual resolution over 10 days and no evidence of spread to normal organs.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY, USA. zhenkunyu@yahoo.com.cn

ABSTRACT

Background: Novel therapies are necessary to improve outcomes for patients with squamous cell carcinomas (SCC) of the head and neck. Historically, vaccinia virus was administered widely to humans as a vaccine and led to the eradication of smallpox. We examined the therapeutic effects of an attenuated, replication-competent vaccinia virus (GLV-1h68) as an oncolytic agent against a panel of six human head and neck SCC cell lines.

Results: All six cell lines supported viral transgene expression (beta-galactosidase, green fluorescent protein, and luciferase) as early as 6 hours after viral exposure. Efficient transgene expression and viral replication (>150-fold titer increase over 72 hrs) were observed in four of the cell lines. At a multiplicity of infection (MOI) of 1, GLV-1h68 was highly cytotoxic to the four cell lines, resulting in > or = 90% cytotoxicity over 6 days, and the remaining two cell lines exhibited >45% cytotoxicity. Even at a very low MOI of 0.01, three cell lines still demonstrated >60% cell death over 6 days. A single injection of GLV-1h68 (5 x 10(6) pfu) intratumorally into MSKQLL2 xenografts in mice exhibited localized intratumoral luciferase activity peaking at days 2-4, with gradual resolution over 10 days and no evidence of spread to normal organs. Treated animals exhibited near-complete tumor regression over a 24-day period without any observed toxicity, while control animals demonstrated rapid tumor progression.

Conclusion: These results demonstrate significant oncolytic efficacy by an attenuated vaccinia virus for infecting and lysing head and neck SCC both in vitro and in vivo, and support its continued investigation in future clinical trials.

Show MeSH

Related in: MedlinePlus

In vitro quantification of gene expression and viral replication by GLV-1h68 in human head and neck squamous cell carcinoma. A. Cell lines were infected with GLV-1h68 at an MOI of 5, and six hours later quantitative β-galactosidase assays were performed. B. Cell lines were infected with GLV-1h68 at an MOI of 1, and 12 hours later coelenterazine was added. Bioluminescence was imaged with a cooled CCD camera and quantified with software analysis. C. Cell lines were exposed to GLV-1h68 at an MOI of 0.1 and incubated for 72 hours. Supernatants were collected and viral titers quantified by plaque assays on confluent CV-1 cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2714037&req=5

Figure 2: In vitro quantification of gene expression and viral replication by GLV-1h68 in human head and neck squamous cell carcinoma. A. Cell lines were infected with GLV-1h68 at an MOI of 5, and six hours later quantitative β-galactosidase assays were performed. B. Cell lines were infected with GLV-1h68 at an MOI of 1, and 12 hours later coelenterazine was added. Bioluminescence was imaged with a cooled CCD camera and quantified with software analysis. C. Cell lines were exposed to GLV-1h68 at an MOI of 0.1 and incubated for 72 hours. Supernatants were collected and viral titers quantified by plaque assays on confluent CV-1 cells.

Mentions: Quantitative assays for β-galactosidase expression by GLV-1h68 at an MOI of 5 reflected qualitative visual findings from X-gal staining, with MSKQLL2 showing the highest expression and MSK922 the lowest expression (Figure 2a). All of the cell lines demonstrated some degree of susceptibility to infection by GLV-1h68. Luciferase expression measured by software quantification of acquired photon emission images demonstrated very similar findings as the quantitative β-galactosidase assays, with the same relative order of expression noted (Figure 2b).


Oncolytic vaccinia therapy of squamous cell carcinoma.

Yu Z, Li S, Brader P, Chen N, Yu YA, Zhang Q, Szalay AA, Fong Y, Wong RJ - Mol. Cancer (2009)

In vitro quantification of gene expression and viral replication by GLV-1h68 in human head and neck squamous cell carcinoma. A. Cell lines were infected with GLV-1h68 at an MOI of 5, and six hours later quantitative β-galactosidase assays were performed. B. Cell lines were infected with GLV-1h68 at an MOI of 1, and 12 hours later coelenterazine was added. Bioluminescence was imaged with a cooled CCD camera and quantified with software analysis. C. Cell lines were exposed to GLV-1h68 at an MOI of 0.1 and incubated for 72 hours. Supernatants were collected and viral titers quantified by plaque assays on confluent CV-1 cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2714037&req=5

Figure 2: In vitro quantification of gene expression and viral replication by GLV-1h68 in human head and neck squamous cell carcinoma. A. Cell lines were infected with GLV-1h68 at an MOI of 5, and six hours later quantitative β-galactosidase assays were performed. B. Cell lines were infected with GLV-1h68 at an MOI of 1, and 12 hours later coelenterazine was added. Bioluminescence was imaged with a cooled CCD camera and quantified with software analysis. C. Cell lines were exposed to GLV-1h68 at an MOI of 0.1 and incubated for 72 hours. Supernatants were collected and viral titers quantified by plaque assays on confluent CV-1 cells.
Mentions: Quantitative assays for β-galactosidase expression by GLV-1h68 at an MOI of 5 reflected qualitative visual findings from X-gal staining, with MSKQLL2 showing the highest expression and MSK922 the lowest expression (Figure 2a). All of the cell lines demonstrated some degree of susceptibility to infection by GLV-1h68. Luciferase expression measured by software quantification of acquired photon emission images demonstrated very similar findings as the quantitative β-galactosidase assays, with the same relative order of expression noted (Figure 2b).

Bottom Line: All six cell lines supported viral transgene expression (beta-galactosidase, green fluorescent protein, and luciferase) as early as 6 hours after viral exposure.Even at a very low MOI of 0.01, three cell lines still demonstrated >60% cell death over 6 days.A single injection of GLV-1h68 (5 x 10(6) pfu) intratumorally into MSKQLL2 xenografts in mice exhibited localized intratumoral luciferase activity peaking at days 2-4, with gradual resolution over 10 days and no evidence of spread to normal organs.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY, USA. zhenkunyu@yahoo.com.cn

ABSTRACT

Background: Novel therapies are necessary to improve outcomes for patients with squamous cell carcinomas (SCC) of the head and neck. Historically, vaccinia virus was administered widely to humans as a vaccine and led to the eradication of smallpox. We examined the therapeutic effects of an attenuated, replication-competent vaccinia virus (GLV-1h68) as an oncolytic agent against a panel of six human head and neck SCC cell lines.

Results: All six cell lines supported viral transgene expression (beta-galactosidase, green fluorescent protein, and luciferase) as early as 6 hours after viral exposure. Efficient transgene expression and viral replication (>150-fold titer increase over 72 hrs) were observed in four of the cell lines. At a multiplicity of infection (MOI) of 1, GLV-1h68 was highly cytotoxic to the four cell lines, resulting in > or = 90% cytotoxicity over 6 days, and the remaining two cell lines exhibited >45% cytotoxicity. Even at a very low MOI of 0.01, three cell lines still demonstrated >60% cell death over 6 days. A single injection of GLV-1h68 (5 x 10(6) pfu) intratumorally into MSKQLL2 xenografts in mice exhibited localized intratumoral luciferase activity peaking at days 2-4, with gradual resolution over 10 days and no evidence of spread to normal organs. Treated animals exhibited near-complete tumor regression over a 24-day period without any observed toxicity, while control animals demonstrated rapid tumor progression.

Conclusion: These results demonstrate significant oncolytic efficacy by an attenuated vaccinia virus for infecting and lysing head and neck SCC both in vitro and in vivo, and support its continued investigation in future clinical trials.

Show MeSH
Related in: MedlinePlus