Non-invasive gamma camera imaging of gene transfer using an adenoviral vector encoding an epitope-tagged receptor as a reporter

B. E. Rogers; T. R. Chaudhuri; P. N. Reynolds; D. Della Manna; K. R. Zinn

doi:10.1038/sj.gt.3301853

Non-invasive gamma camera imaging of gene transfer using an adenoviral vector encoding an epitope-tagged receptor as a reporter

B. E. Rogers, T. R. Chaudhuri, P. N. Reynolds, D. Della Manna, K. R. Zinn

Research output: Contribution to journal › Article › peer-review

29 Scopus citations

Abstract

A model epitope-tagged receptor was constructed by fusing the hemagglutinin (HA) sequence on the extracellular N-terminus of the human somatostatin receptor subtype 2 (hSSTr2) gene. This construct was placed in an adenoviral (Ad-HAhSSTr2) vector. This study evaluated Ad-HAhSSTr2 in vitro and in vivo using FACS, fluorescent microscopy, radioactive binding assays, and gamma camera imaging techniques. Infection of A-427 non-small cell lung cancer cells with Ad-HAhSSTr2 or Ad-hSSTr2 resulted in similar expression of hSSTr2 by FACS analysis and binding assays using a ^99mTc-labeled somatostatin analogue (^99mTc-P2045 . HAhSSTr2 expression in A-427 cells was specific for infection with Ad-HAhSSTr2. FITC-labeled anti-HA antibody (FITC-HA) confirmed surface expression in live A-427 cells and the absence of internalization. Gamma camera imaging and gamma counter analysis of normal mice showed significantly greater (P<0.05) liver uptake of ^99mTc-labeled anti-HA antibody (^99mTc-anti-HA) in mice injected i.v. 48 h earlier with Ad-HAhSSTr2 (53.6 ± 6.9% ID/g) as compared to mice similarly injected with Ad-hSSTr2 (9.0 ± 1.3% ID/g). in a mouse tumor model, imaging detected increased tumor localization of ^99mTc-anti-HA due to direct intratumor injection Ad-HAhSSTr2. Gamma counter analysis confirmed significantly greater (P<0.05) uptake of ^99mTc-anti-HA in tumors injected with Ad-HAhSSTr2 (12.5 ± 4.1% ID/g) as compared to Ad-hSSTr2-infected tumors (5.1 ± 1.5% ID/g). These studies demonstrate the feasibility of using an epitope-tagged reporter receptor for non-invasively imaging gene transfer.

Original language	English
Pages (from-to)	105-114
Number of pages	10
Journal	Gene therapy
Volume	10
Issue number	2
DOIs	https://doi.org/10.1038/sj.gt.3301853
State	Published - Jan 2003

Keywords

Gene transfer
Hemagglutinin
Imaging
Somatostatin receptor
Tc

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@article{0c94c7b924574ea5be3bb41efa5c15b8,

title = "Non-invasive gamma camera imaging of gene transfer using an adenoviral vector encoding an epitope-tagged receptor as a reporter",

abstract = "A model epitope-tagged receptor was constructed by fusing the hemagglutinin (HA) sequence on the extracellular N-terminus of the human somatostatin receptor subtype 2 (hSSTr2) gene. This construct was placed in an adenoviral (Ad-HAhSSTr2) vector. This study evaluated Ad-HAhSSTr2 in vitro and in vivo using FACS, fluorescent microscopy, radioactive binding assays, and gamma camera imaging techniques. Infection of A-427 non-small cell lung cancer cells with Ad-HAhSSTr2 or Ad-hSSTr2 resulted in similar expression of hSSTr2 by FACS analysis and binding assays using a 99mTc-labeled somatostatin analogue (99mTc-P2045 . HAhSSTr2 expression in A-427 cells was specific for infection with Ad-HAhSSTr2. FITC-labeled anti-HA antibody (FITC-HA) confirmed surface expression in live A-427 cells and the absence of internalization. Gamma camera imaging and gamma counter analysis of normal mice showed significantly greater (P<0.05) liver uptake of 99mTc-labeled anti-HA antibody (99mTc-anti-HA) in mice injected i.v. 48 h earlier with Ad-HAhSSTr2 (53.6 ± 6.9% ID/g) as compared to mice similarly injected with Ad-hSSTr2 (9.0 ± 1.3% ID/g). in a mouse tumor model, imaging detected increased tumor localization of 99mTc-anti-HA due to direct intratumor injection Ad-HAhSSTr2. Gamma counter analysis confirmed significantly greater (P<0.05) uptake of 99mTc-anti-HA in tumors injected with Ad-HAhSSTr2 (12.5 ± 4.1% ID/g) as compared to Ad-hSSTr2-infected tumors (5.1 ± 1.5% ID/g). These studies demonstrate the feasibility of using an epitope-tagged reporter receptor for non-invasively imaging gene transfer.",

keywords = "Gene transfer, Hemagglutinin, Imaging, Somatostatin receptor, Tc",

author = "Rogers, {B. E.} and Chaudhuri, {T. R.} and Reynolds, {P. N.} and {Della Manna}, D. and Zinn, {K. R.}",

note = "Funding Information: This work was supported by the NIH grant CA80104 and grants from the Avon Products Foundation and the American Heart Association. The authors appreciate the generous supply of P2045 from Diatide Research Laboratories (Londonberry, NH, USA). The authors thank Sheila Bright, Ashley Davis, Charlotte Hammond, Synethia Kidd, Richard Kirkman, Gloria Robinson, and Leisa Seay for excellent technical assistance.",

year = "2003",

month = jan,

doi = "10.1038/sj.gt.3301853",

language = "English",

volume = "10",

pages = "105--114",

journal = "Gene therapy",

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AU - Rogers, B. E.

AU - Chaudhuri, T. R.

AU - Reynolds, P. N.

AU - Della Manna, D.

AU - Zinn, K. R.

N1 - Funding Information: This work was supported by the NIH grant CA80104 and grants from the Avon Products Foundation and the American Heart Association. The authors appreciate the generous supply of P2045 from Diatide Research Laboratories (Londonberry, NH, USA). The authors thank Sheila Bright, Ashley Davis, Charlotte Hammond, Synethia Kidd, Richard Kirkman, Gloria Robinson, and Leisa Seay for excellent technical assistance.

PY - 2003/1

Y1 - 2003/1

N2 - A model epitope-tagged receptor was constructed by fusing the hemagglutinin (HA) sequence on the extracellular N-terminus of the human somatostatin receptor subtype 2 (hSSTr2) gene. This construct was placed in an adenoviral (Ad-HAhSSTr2) vector. This study evaluated Ad-HAhSSTr2 in vitro and in vivo using FACS, fluorescent microscopy, radioactive binding assays, and gamma camera imaging techniques. Infection of A-427 non-small cell lung cancer cells with Ad-HAhSSTr2 or Ad-hSSTr2 resulted in similar expression of hSSTr2 by FACS analysis and binding assays using a 99mTc-labeled somatostatin analogue (99mTc-P2045 . HAhSSTr2 expression in A-427 cells was specific for infection with Ad-HAhSSTr2. FITC-labeled anti-HA antibody (FITC-HA) confirmed surface expression in live A-427 cells and the absence of internalization. Gamma camera imaging and gamma counter analysis of normal mice showed significantly greater (P<0.05) liver uptake of 99mTc-labeled anti-HA antibody (99mTc-anti-HA) in mice injected i.v. 48 h earlier with Ad-HAhSSTr2 (53.6 ± 6.9% ID/g) as compared to mice similarly injected with Ad-hSSTr2 (9.0 ± 1.3% ID/g). in a mouse tumor model, imaging detected increased tumor localization of 99mTc-anti-HA due to direct intratumor injection Ad-HAhSSTr2. Gamma counter analysis confirmed significantly greater (P<0.05) uptake of 99mTc-anti-HA in tumors injected with Ad-HAhSSTr2 (12.5 ± 4.1% ID/g) as compared to Ad-hSSTr2-infected tumors (5.1 ± 1.5% ID/g). These studies demonstrate the feasibility of using an epitope-tagged reporter receptor for non-invasively imaging gene transfer.

AB - A model epitope-tagged receptor was constructed by fusing the hemagglutinin (HA) sequence on the extracellular N-terminus of the human somatostatin receptor subtype 2 (hSSTr2) gene. This construct was placed in an adenoviral (Ad-HAhSSTr2) vector. This study evaluated Ad-HAhSSTr2 in vitro and in vivo using FACS, fluorescent microscopy, radioactive binding assays, and gamma camera imaging techniques. Infection of A-427 non-small cell lung cancer cells with Ad-HAhSSTr2 or Ad-hSSTr2 resulted in similar expression of hSSTr2 by FACS analysis and binding assays using a 99mTc-labeled somatostatin analogue (99mTc-P2045 . HAhSSTr2 expression in A-427 cells was specific for infection with Ad-HAhSSTr2. FITC-labeled anti-HA antibody (FITC-HA) confirmed surface expression in live A-427 cells and the absence of internalization. Gamma camera imaging and gamma counter analysis of normal mice showed significantly greater (P<0.05) liver uptake of 99mTc-labeled anti-HA antibody (99mTc-anti-HA) in mice injected i.v. 48 h earlier with Ad-HAhSSTr2 (53.6 ± 6.9% ID/g) as compared to mice similarly injected with Ad-hSSTr2 (9.0 ± 1.3% ID/g). in a mouse tumor model, imaging detected increased tumor localization of 99mTc-anti-HA due to direct intratumor injection Ad-HAhSSTr2. Gamma counter analysis confirmed significantly greater (P<0.05) uptake of 99mTc-anti-HA in tumors injected with Ad-HAhSSTr2 (12.5 ± 4.1% ID/g) as compared to Ad-hSSTr2-infected tumors (5.1 ± 1.5% ID/g). These studies demonstrate the feasibility of using an epitope-tagged reporter receptor for non-invasively imaging gene transfer.

KW - Gene transfer

KW - Hemagglutinin

KW - Imaging

KW - Somatostatin receptor

KW - Tc

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EP - 114

JO - Gene therapy

JF - Gene therapy

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ER -

Non-invasive gamma camera imaging of gene transfer using an adenoviral vector encoding an epitope-tagged receptor as a reporter

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