Construction and radiolabeling of adenovirus variants that incorporate human metallothionein into protein IX for analysis of biodistribution

Lei Liu, Buck E. Rogers, Natalia Aladyshkina, Bing Cheng, Stephen J. Lokitz, David T. Curiel, James Michael Mathis

Research output: Contribution to journalArticleResearchpeer-review

4 Citations (Scopus)

Abstract

Using adenovirus (Ad)-based vectors is a promising strategy for novel cancer treatments; however, current tracking approaches in vivo are limited. The C-terminus of the Ad minor capsid protein IX (pIX) can incorporate heterologous reporters to monitor biodistribution. We incorporated metallothionein (MT), a low-molecular-weight metal-binding protein, as a fusion to pIX. We previously demonstrated 99mTc binding in vitro to a pIX-MT fusion on the Ad capsid. We investigated different fusions of MT within pIX to optimize functional display. We identified a dimeric MT construct fused to pIX that showed significantly increased radiolabeling capacity. After Ad radiolabeling, we characterized metal binding in vitro. We explored biodistribution in vivo in control mice, mice pretreated with warfarin, mice preimmunized with wild-type Ad, and mice that received both warfarin pretreatment and Ad preimmunization. Localization of activity to liver and bladder was seen, with activity detected in spleen, intestine, and kidneys. Afterwards, the mice were euthanized and selected organs were dissected for further analysis. Similar to the imaging results, most of the radioactivity was found in the liver, spleen, kidneys, and bladder, with significant differences between the groups observed in the liver. These results demonstrate this platform application for following Ad dissemination in vivo.

Original languageEnglish
JournalMolecular Imaging
Volume13
Issue number7
DOIs
StatePublished - 1 Sep 2014

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Metallothionein
Adenoviridae
Proteins
Warfarin
Liver
Urinary Bladder
Spleen
Metals
Kidney
Capsid
Capsid Proteins
Radioactivity
Intestines
Carrier Proteins
Molecular Weight
Neoplasms

Cite this

Liu, Lei ; Rogers, Buck E. ; Aladyshkina, Natalia ; Cheng, Bing ; Lokitz, Stephen J. ; Curiel, David T. ; Mathis, James Michael. / Construction and radiolabeling of adenovirus variants that incorporate human metallothionein into protein IX for analysis of biodistribution. In: Molecular Imaging. 2014 ; Vol. 13, No. 7.
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abstract = "Using adenovirus (Ad)-based vectors is a promising strategy for novel cancer treatments; however, current tracking approaches in vivo are limited. The C-terminus of the Ad minor capsid protein IX (pIX) can incorporate heterologous reporters to monitor biodistribution. We incorporated metallothionein (MT), a low-molecular-weight metal-binding protein, as a fusion to pIX. We previously demonstrated 99mTc binding in vitro to a pIX-MT fusion on the Ad capsid. We investigated different fusions of MT within pIX to optimize functional display. We identified a dimeric MT construct fused to pIX that showed significantly increased radiolabeling capacity. After Ad radiolabeling, we characterized metal binding in vitro. We explored biodistribution in vivo in control mice, mice pretreated with warfarin, mice preimmunized with wild-type Ad, and mice that received both warfarin pretreatment and Ad preimmunization. Localization of activity to liver and bladder was seen, with activity detected in spleen, intestine, and kidneys. Afterwards, the mice were euthanized and selected organs were dissected for further analysis. Similar to the imaging results, most of the radioactivity was found in the liver, spleen, kidneys, and bladder, with significant differences between the groups observed in the liver. These results demonstrate this platform application for following Ad dissemination in vivo.",
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Construction and radiolabeling of adenovirus variants that incorporate human metallothionein into protein IX for analysis of biodistribution. / Liu, Lei; Rogers, Buck E.; Aladyshkina, Natalia; Cheng, Bing; Lokitz, Stephen J.; Curiel, David T.; Mathis, James Michael.

In: Molecular Imaging, Vol. 13, No. 7, 01.09.2014.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Mathis, James Michael

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