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AAVrh.10-mediated genetic delivery of bevacizumab to the pleura to provide local anti-VEGF to suppress growth of metastatic lung tumors.

Watanabe M, Boyer JL, Crystal RG - Gene Ther. (2010)

Bottom Line: We hypothesized that an intrapleural administration of an adeno-associated virus (AAV) vector expressing an anti-VEGF-A antibody equivalent of bevacizumab would result in sustained anti-VEGF-A localized expression within the lung and suppress metastatic tumor growth.Intrapleural administration of AAVrh.10alphaVEGF directed long-term expression of the anti-human VEGF-A antibody in lung, as shown by sustained, high-level anti-human VEGF titers in lung epithelial lining fluid for 40 weeks, which was the duration of the study.Thus, intrapleural administration of an AAVrh.10 vector, encoding the murine monoclonal antibody equivalent of bevacizumab, effectively suppresses the growth of metastatic lung tumors, suggesting AAV-mediated gene transfer to the pleura to deliver bevacizumab locally to the lung as a novel alternative platform to conventional monoclonal antibody therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Medicine, Weill Cornell Medical College, New York, NY 10065, USA.

ABSTRACT
Vascular endothelial growth factor (VEGF) produced by tumor cells has a central role in stimulating angiogenesis required for tumor growth. Humanized monoclonal anti-VEGF antibody (bevacizumab, Avastin), approved as a treatment for non-squamous, non-small cell lung cancer, requires administration every 3 weeks. We hypothesized that an intrapleural administration of an adeno-associated virus (AAV) vector expressing an anti-VEGF-A antibody equivalent of bevacizumab would result in sustained anti-VEGF-A localized expression within the lung and suppress metastatic tumor growth. The AAV vector AAVrh.10alphaVEGF encodes the light chain and heavy chain complementary DNAs of monoclonal antibody A.4.6.1, a murine antibody that specifically recognizes human VEGF-A with the same antigen-binding site as bevacizumab. A metastatic lung tumor model was established in severe combined immunodeficient mice by intravenous administration of human DU145 prostate carcinoma cells. Intrapleural administration of AAVrh.10alphaVEGF directed long-term expression of the anti-human VEGF-A antibody in lung, as shown by sustained, high-level anti-human VEGF titers in lung epithelial lining fluid for 40 weeks, which was the duration of the study. In the AAVrh.10alphaVEGF-treated animals, tumor growth was significantly suppressed (P<0.05), the numbers of blood vessels and mitotic nuclei in the tumor was decreased (P<0.05) and there was increased survival (P<0.05). Thus, intrapleural administration of an AAVrh.10 vector, encoding the murine monoclonal antibody equivalent of bevacizumab, effectively suppresses the growth of metastatic lung tumors, suggesting AAV-mediated gene transfer to the pleura to deliver bevacizumab locally to the lung as a novel alternative platform to conventional monoclonal antibody therapy.

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Anti-VEGF-A antibody levels after local administration of the AAVrh.10αVEGF vector. A. Comparison of lung ELF anti-VEGF-A antibody levels after local administration of AAVrh.10αVEGF. AAVrh.10αVEGF (1011 gc) was administered to C57BL/6 mice by intrapleural (Ipl) or intratracheal (IT) route. AAVrh.10EGFP and AAVrh.10αPA were administered by the intrapleural route as controls. Eight weeks after vector administration, lung ELF anti-VEGF-A antibody levels were measured by human VEGF-A specific ELISA. Data were obtained from n=5 animals per group. B. Lung ELF and serum anti-VEGF-A antibody levels over 40 wk after intrapleural administration of AAVrh.10αVEGF. AAVrh.10EGFP and AAVrh.10αPA were administered by the intrapleural route as controls. Data were obtained from n=4–5 animals per group.
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Figure 2: Anti-VEGF-A antibody levels after local administration of the AAVrh.10αVEGF vector. A. Comparison of lung ELF anti-VEGF-A antibody levels after local administration of AAVrh.10αVEGF. AAVrh.10αVEGF (1011 gc) was administered to C57BL/6 mice by intrapleural (Ipl) or intratracheal (IT) route. AAVrh.10EGFP and AAVrh.10αPA were administered by the intrapleural route as controls. Eight weeks after vector administration, lung ELF anti-VEGF-A antibody levels were measured by human VEGF-A specific ELISA. Data were obtained from n=5 animals per group. B. Lung ELF and serum anti-VEGF-A antibody levels over 40 wk after intrapleural administration of AAVrh.10αVEGF. AAVrh.10EGFP and AAVrh.10αPA were administered by the intrapleural route as controls. Data were obtained from n=4–5 animals per group.

Mentions: To determine which local administration route of AAVrh.10αVEGF would achieve the highest lung anti-VEGF-A antibody levels, lung ELF antibody levels were assessed 8 wk after intratrapleural or intratracheal administration of the same dose (1011 gc) of the vector. Intrapleural administration of AAVrh.10αVEGF resulted in high anti-VEGF-A antibody expression levels in lung ELF (Figure 2A; p<0.05 compared with all other groups). No anti-VEGF-A antibody was detected in the lung ELF from mice that received intratracheal AAVrh.10αVEGF, intrapleural AAVrh.10αPA or intrapleural AAVrh.10EGFP. Assessment of the expression profile of anti-VEGF-A antibody in lung ELF and serum over 40 wk after intrapleural administration of AAVrh.10αVEGF showed that antibody levels peaked at 4 and 8 wk, respectively, with a mild, then gradual decrease over a period of 24 wk, and then these levels were sustained throughout the 40 wk (Figure 2). Evaluation of BALF total protein levels as a marker of lung inflammation demonstrated no marked elevation after intrapleural administration of AAVrh.10αVEGF compared to naive mice throughout a 40 wk time period [naïve at 40 wk, 477.5 ± 59.6 μg/ml (p>0.4 compared with all time points); maximum level at 16 wk, 531.2 ± 46.7 μg/ml]. The data suggest that no significant lung inflammation was elicited during the experimental period by intrapleural AAVrh.10αVEGF administration.


AAVrh.10-mediated genetic delivery of bevacizumab to the pleura to provide local anti-VEGF to suppress growth of metastatic lung tumors.

Watanabe M, Boyer JL, Crystal RG - Gene Ther. (2010)

Anti-VEGF-A antibody levels after local administration of the AAVrh.10αVEGF vector. A. Comparison of lung ELF anti-VEGF-A antibody levels after local administration of AAVrh.10αVEGF. AAVrh.10αVEGF (1011 gc) was administered to C57BL/6 mice by intrapleural (Ipl) or intratracheal (IT) route. AAVrh.10EGFP and AAVrh.10αPA were administered by the intrapleural route as controls. Eight weeks after vector administration, lung ELF anti-VEGF-A antibody levels were measured by human VEGF-A specific ELISA. Data were obtained from n=5 animals per group. B. Lung ELF and serum anti-VEGF-A antibody levels over 40 wk after intrapleural administration of AAVrh.10αVEGF. AAVrh.10EGFP and AAVrh.10αPA were administered by the intrapleural route as controls. Data were obtained from n=4–5 animals per group.
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Related In: Results  -  Collection

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Show All Figures
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Figure 2: Anti-VEGF-A antibody levels after local administration of the AAVrh.10αVEGF vector. A. Comparison of lung ELF anti-VEGF-A antibody levels after local administration of AAVrh.10αVEGF. AAVrh.10αVEGF (1011 gc) was administered to C57BL/6 mice by intrapleural (Ipl) or intratracheal (IT) route. AAVrh.10EGFP and AAVrh.10αPA were administered by the intrapleural route as controls. Eight weeks after vector administration, lung ELF anti-VEGF-A antibody levels were measured by human VEGF-A specific ELISA. Data were obtained from n=5 animals per group. B. Lung ELF and serum anti-VEGF-A antibody levels over 40 wk after intrapleural administration of AAVrh.10αVEGF. AAVrh.10EGFP and AAVrh.10αPA were administered by the intrapleural route as controls. Data were obtained from n=4–5 animals per group.
Mentions: To determine which local administration route of AAVrh.10αVEGF would achieve the highest lung anti-VEGF-A antibody levels, lung ELF antibody levels were assessed 8 wk after intratrapleural or intratracheal administration of the same dose (1011 gc) of the vector. Intrapleural administration of AAVrh.10αVEGF resulted in high anti-VEGF-A antibody expression levels in lung ELF (Figure 2A; p<0.05 compared with all other groups). No anti-VEGF-A antibody was detected in the lung ELF from mice that received intratracheal AAVrh.10αVEGF, intrapleural AAVrh.10αPA or intrapleural AAVrh.10EGFP. Assessment of the expression profile of anti-VEGF-A antibody in lung ELF and serum over 40 wk after intrapleural administration of AAVrh.10αVEGF showed that antibody levels peaked at 4 and 8 wk, respectively, with a mild, then gradual decrease over a period of 24 wk, and then these levels were sustained throughout the 40 wk (Figure 2). Evaluation of BALF total protein levels as a marker of lung inflammation demonstrated no marked elevation after intrapleural administration of AAVrh.10αVEGF compared to naive mice throughout a 40 wk time period [naïve at 40 wk, 477.5 ± 59.6 μg/ml (p>0.4 compared with all time points); maximum level at 16 wk, 531.2 ± 46.7 μg/ml]. The data suggest that no significant lung inflammation was elicited during the experimental period by intrapleural AAVrh.10αVEGF administration.

Bottom Line: We hypothesized that an intrapleural administration of an adeno-associated virus (AAV) vector expressing an anti-VEGF-A antibody equivalent of bevacizumab would result in sustained anti-VEGF-A localized expression within the lung and suppress metastatic tumor growth.Intrapleural administration of AAVrh.10alphaVEGF directed long-term expression of the anti-human VEGF-A antibody in lung, as shown by sustained, high-level anti-human VEGF titers in lung epithelial lining fluid for 40 weeks, which was the duration of the study.Thus, intrapleural administration of an AAVrh.10 vector, encoding the murine monoclonal antibody equivalent of bevacizumab, effectively suppresses the growth of metastatic lung tumors, suggesting AAV-mediated gene transfer to the pleura to deliver bevacizumab locally to the lung as a novel alternative platform to conventional monoclonal antibody therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Medicine, Weill Cornell Medical College, New York, NY 10065, USA.

ABSTRACT
Vascular endothelial growth factor (VEGF) produced by tumor cells has a central role in stimulating angiogenesis required for tumor growth. Humanized monoclonal anti-VEGF antibody (bevacizumab, Avastin), approved as a treatment for non-squamous, non-small cell lung cancer, requires administration every 3 weeks. We hypothesized that an intrapleural administration of an adeno-associated virus (AAV) vector expressing an anti-VEGF-A antibody equivalent of bevacizumab would result in sustained anti-VEGF-A localized expression within the lung and suppress metastatic tumor growth. The AAV vector AAVrh.10alphaVEGF encodes the light chain and heavy chain complementary DNAs of monoclonal antibody A.4.6.1, a murine antibody that specifically recognizes human VEGF-A with the same antigen-binding site as bevacizumab. A metastatic lung tumor model was established in severe combined immunodeficient mice by intravenous administration of human DU145 prostate carcinoma cells. Intrapleural administration of AAVrh.10alphaVEGF directed long-term expression of the anti-human VEGF-A antibody in lung, as shown by sustained, high-level anti-human VEGF titers in lung epithelial lining fluid for 40 weeks, which was the duration of the study. In the AAVrh.10alphaVEGF-treated animals, tumor growth was significantly suppressed (P<0.05), the numbers of blood vessels and mitotic nuclei in the tumor was decreased (P<0.05) and there was increased survival (P<0.05). Thus, intrapleural administration of an AAVrh.10 vector, encoding the murine monoclonal antibody equivalent of bevacizumab, effectively suppresses the growth of metastatic lung tumors, suggesting AAV-mediated gene transfer to the pleura to deliver bevacizumab locally to the lung as a novel alternative platform to conventional monoclonal antibody therapy.

Show MeSH
Related in: MedlinePlus