Genetic incorporation of human metallothionein into the adenovirus protein IX for non-invasive SPECT imaging

James Michael Mathis, Shilpa Bhatia, Alok Khandelwal, Imre Kovesdi, Stephen J. Lokitz, Yoshi Odaka, Amol M. Takalkar, Tracee Terry, David T. Curiel

Research output: Contribution to journalArticleResearchpeer-review

11 Citations (Scopus)

Abstract

As the limits of existing treatments for cancer are recognized, clearly novel therapies must be considered for successful treatment; cancer therapy using adenovirus vectors is a promising strategy. However tracking the biodistribution of adenovirus vectors in vivo is limited to invasive procedures such as biopsies, which are error prone, non-quantitative, and do not give a full representation of the pharmacokinetics involved. Current non-invasive imaging strategies using reporter gene expression have been applied to analyze adenoviral vectors. The major drawback to approaches that tag viruses with reporter genes is that these systems require initial viral infection and subsequent cellular expression of a reporter gene to allow non-invasive imaging. As an alternative to conventional vector detection techniques, we developed a specific genetic labeling system whereby an adenoviral vector incorporates a fusion between capsid protein IX and human metallothionein. Our study herein clearly demonstrates our ability to rescue viable adenoviral particles that display functional metallothionein (MT) as a component of their capsid surface. We demonstrate the feasibility of 99mTc binding in vitro to the pIX-MT fusion on the capsid of adenovirus virions using a simple transchelation reaction. SPECT imaging of a mouse after administration of a 99mTc-radiolabeled virus showed clear localization of radioactivity to the liver. This result strongly supports imaging using pIX-MT, visualizing the normal biodistribution of Ad primarily to the liver upon injection into mice. The ability we have developed to view real-time biodistribution in their physiological milieu represents a significant tool to study adenovirus biology in vivo.

Original languageEnglish
Article numbere16792
JournalPLoS ONE
Volume6
Issue number2
DOIs
StatePublished - 24 Feb 2011

Fingerprint

Metallothionein
metallothionein
Medical Genetics
Adenoviridae
Single-Photon Emission-Computed Tomography
Reporter Genes
reporter genes
image analysis
Imaging techniques
capsid
Capsid
Proteins
Viruses
proteins
Liver
Fusion reactions
viruses
therapeutics
liver
Genes

Cite this

Mathis, James Michael ; Bhatia, Shilpa ; Khandelwal, Alok ; Kovesdi, Imre ; Lokitz, Stephen J. ; Odaka, Yoshi ; Takalkar, Amol M. ; Terry, Tracee ; Curiel, David T. / Genetic incorporation of human metallothionein into the adenovirus protein IX for non-invasive SPECT imaging. In: PLoS ONE. 2011 ; Vol. 6, No. 2.
@article{080f8a0f67e14554bebfc61dc4a4275a,
title = "Genetic incorporation of human metallothionein into the adenovirus protein IX for non-invasive SPECT imaging",
abstract = "As the limits of existing treatments for cancer are recognized, clearly novel therapies must be considered for successful treatment; cancer therapy using adenovirus vectors is a promising strategy. However tracking the biodistribution of adenovirus vectors in vivo is limited to invasive procedures such as biopsies, which are error prone, non-quantitative, and do not give a full representation of the pharmacokinetics involved. Current non-invasive imaging strategies using reporter gene expression have been applied to analyze adenoviral vectors. The major drawback to approaches that tag viruses with reporter genes is that these systems require initial viral infection and subsequent cellular expression of a reporter gene to allow non-invasive imaging. As an alternative to conventional vector detection techniques, we developed a specific genetic labeling system whereby an adenoviral vector incorporates a fusion between capsid protein IX and human metallothionein. Our study herein clearly demonstrates our ability to rescue viable adenoviral particles that display functional metallothionein (MT) as a component of their capsid surface. We demonstrate the feasibility of 99mTc binding in vitro to the pIX-MT fusion on the capsid of adenovirus virions using a simple transchelation reaction. SPECT imaging of a mouse after administration of a 99mTc-radiolabeled virus showed clear localization of radioactivity to the liver. This result strongly supports imaging using pIX-MT, visualizing the normal biodistribution of Ad primarily to the liver upon injection into mice. The ability we have developed to view real-time biodistribution in their physiological milieu represents a significant tool to study adenovirus biology in vivo.",
author = "Mathis, {James Michael} and Shilpa Bhatia and Alok Khandelwal and Imre Kovesdi and Lokitz, {Stephen J.} and Yoshi Odaka and Takalkar, {Amol M.} and Tracee Terry and Curiel, {David T.}",
year = "2011",
month = "2",
day = "24",
doi = "10.1371/journal.pone.0016792",
language = "English",
volume = "6",
journal = "PLoS ONE",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "2",

}

Mathis, JM, Bhatia, S, Khandelwal, A, Kovesdi, I, Lokitz, SJ, Odaka, Y, Takalkar, AM, Terry, T & Curiel, DT 2011, 'Genetic incorporation of human metallothionein into the adenovirus protein IX for non-invasive SPECT imaging', PLoS ONE, vol. 6, no. 2, e16792. https://doi.org/10.1371/journal.pone.0016792

Genetic incorporation of human metallothionein into the adenovirus protein IX for non-invasive SPECT imaging. / Mathis, James Michael; Bhatia, Shilpa; Khandelwal, Alok; Kovesdi, Imre; Lokitz, Stephen J.; Odaka, Yoshi; Takalkar, Amol M.; Terry, Tracee; Curiel, David T.

In: PLoS ONE, Vol. 6, No. 2, e16792, 24.02.2011.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Genetic incorporation of human metallothionein into the adenovirus protein IX for non-invasive SPECT imaging

AU - Mathis, James Michael

AU - Bhatia, Shilpa

AU - Khandelwal, Alok

AU - Kovesdi, Imre

AU - Lokitz, Stephen J.

AU - Odaka, Yoshi

AU - Takalkar, Amol M.

AU - Terry, Tracee

AU - Curiel, David T.

PY - 2011/2/24

Y1 - 2011/2/24

N2 - As the limits of existing treatments for cancer are recognized, clearly novel therapies must be considered for successful treatment; cancer therapy using adenovirus vectors is a promising strategy. However tracking the biodistribution of adenovirus vectors in vivo is limited to invasive procedures such as biopsies, which are error prone, non-quantitative, and do not give a full representation of the pharmacokinetics involved. Current non-invasive imaging strategies using reporter gene expression have been applied to analyze adenoviral vectors. The major drawback to approaches that tag viruses with reporter genes is that these systems require initial viral infection and subsequent cellular expression of a reporter gene to allow non-invasive imaging. As an alternative to conventional vector detection techniques, we developed a specific genetic labeling system whereby an adenoviral vector incorporates a fusion between capsid protein IX and human metallothionein. Our study herein clearly demonstrates our ability to rescue viable adenoviral particles that display functional metallothionein (MT) as a component of their capsid surface. We demonstrate the feasibility of 99mTc binding in vitro to the pIX-MT fusion on the capsid of adenovirus virions using a simple transchelation reaction. SPECT imaging of a mouse after administration of a 99mTc-radiolabeled virus showed clear localization of radioactivity to the liver. This result strongly supports imaging using pIX-MT, visualizing the normal biodistribution of Ad primarily to the liver upon injection into mice. The ability we have developed to view real-time biodistribution in their physiological milieu represents a significant tool to study adenovirus biology in vivo.

AB - As the limits of existing treatments for cancer are recognized, clearly novel therapies must be considered for successful treatment; cancer therapy using adenovirus vectors is a promising strategy. However tracking the biodistribution of adenovirus vectors in vivo is limited to invasive procedures such as biopsies, which are error prone, non-quantitative, and do not give a full representation of the pharmacokinetics involved. Current non-invasive imaging strategies using reporter gene expression have been applied to analyze adenoviral vectors. The major drawback to approaches that tag viruses with reporter genes is that these systems require initial viral infection and subsequent cellular expression of a reporter gene to allow non-invasive imaging. As an alternative to conventional vector detection techniques, we developed a specific genetic labeling system whereby an adenoviral vector incorporates a fusion between capsid protein IX and human metallothionein. Our study herein clearly demonstrates our ability to rescue viable adenoviral particles that display functional metallothionein (MT) as a component of their capsid surface. We demonstrate the feasibility of 99mTc binding in vitro to the pIX-MT fusion on the capsid of adenovirus virions using a simple transchelation reaction. SPECT imaging of a mouse after administration of a 99mTc-radiolabeled virus showed clear localization of radioactivity to the liver. This result strongly supports imaging using pIX-MT, visualizing the normal biodistribution of Ad primarily to the liver upon injection into mice. The ability we have developed to view real-time biodistribution in their physiological milieu represents a significant tool to study adenovirus biology in vivo.

UR - http://www.scopus.com/inward/record.url?scp=79951778035&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0016792

DO - 10.1371/journal.pone.0016792

M3 - Article

VL - 6

JO - PLoS ONE

JF - PLoS ONE

SN - 1932-6203

IS - 2

M1 - e16792

ER -