In vivo magnetic resonance imaging of transgene expression

Weissleder, Ralph; Moore, Anna; Mahmood, Umar; Bhorade, Rajeev; Benveniste, Helene; Chiocca, E. Antonio; Basilion, James P.

doi:10.1038/73219

Cited by 769 publications

(479 citation statements)

References 15 publications

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“…The TfR1 iron response element was removed prior to cloning to ensure efficient translation of the TfR1 protein; this well‐established strategy allows circumvention of the negative feedback loop that prevents the accumulation of TfR1 protein when the concentration of intracellular iron is high 19, 20, 21, 22. Flow cytometry showed that the transduction efficiency with both TfR1:GFP and GFP control lentivirus was almost 100% (Fig.…”

Section: Resultsmentioning

confidence: 99%

Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene

Pereira

Herrmann

Moss

et al. 2016

Contrast Media & Molecular

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Magnetic resonance (MR) reporter genes have the potential for tracking the biodistribution and fate of cells in vivo, thus allowing the safety, efficacy and mechanisms of action of cell‐based therapies to be comprehensively assessed. In this study, we evaluate the effectiveness of the iron importer transferrin receptor‐1 (TfR1) as an MR reporter gene in the model cell line CHO‐K1. Overexpression of the TfR1 transgene led to a reduction in the levels of endogenous TfR1 mRNA, but to a 60‐fold increase in total TfR1 protein levels. Although the mRNA levels of ferritin heavy chain‐1 (Fth1) did not change, Fth1 protein levels increased 13‐fold. The concentration of intracellular iron increased significantly, even when cells were cultured in medium that was not supplemented with iron and the amount of iron in the extracellular environment was thus at physiological levels. However, we found that, by supplementing the cell culture medium with ferric citrate, a comparable degree of iron uptake and MR contrast could be achieved in control cells that did not express the TfR1 transgene. Sufficient MR contrast to enable the cells to be detected in vivo following their administration into the midbrain of chick embryos was obtained irrespective of the reporter gene. We conclude that TfR1 is not an effective reporter and that, to track the biodistribution of cells with MR imaging in the short term, it is sufficient to simply culture cells in the presence of ferric citrate. Copyright © 2016 The Authors Contrast Media & Molecular Imaging Published by John Wiley & Sons Ltd.

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Section: Resultsmentioning

confidence: 99%

Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene

Pereira

Herrmann

Moss

et al. 2016

Contrast Media & Molecular

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“…The only report of a successful in vivo system for imaging reporter gene expression with magnetic resonance 27 requires receptor-driven internalization of the reporting ligand -iron-bound transferrin -to accumulate sufficient reporter to produce a signal. Ectopic overexpression of the transferrin receptor and accumulation of large amounts of superparamagnetic nanoparticles during the imaging process have the same potential consequences that our studies with the mutated D2R are designed to avoid, that ectopic receptor gene expression could change target cell physiology.…”

Section: Discussionmentioning

confidence: 99%

Noninvasive, quantitative imaging in living animals of a mutant dopamine D2 receptor reporter gene in which ligand binding is uncoupled from signal transduction

Liang¹,

Satyamurthy²,

Barrio³

et al. 2001

Gene Ther

168

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“…Multiple marker genes are under investigation for in vivo imaging. Optical candidates include luciferase, 32 GFP, 33 protease activatable fluoroscopic dyes, 34 while MR techniques include engineered internalizing receptors 35 or tyrosinase. 36 PET agents include reporter probes such as FESP (an agonist for dopamine receptors) 37 and 18-FIAU or 18-F gancyclovir/pencyclovir for imaging thymidine kinase.…”

Section: Discussionmentioning

confidence: 99%

Quantitation of HSV mass distribution in a rodent brain tumor model

Schellingerhout¹,

Rainov

Breakefield

et al. 2000

Gene Ther

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A number of different viral vectors have been used for gene therapy of tumors, with many more under construction, ultimately designed to improve tumor targeting and transduction efficiency. It has become apparent that insufficient viral delivery can be a key limitation to treatment efficacy. We have studied the in vivo mass distribution of a herpes simplex virus type 1 (HSV) vector, hrR3, by radiolabeling it with 111 In-oxine. The virus was administered to intracerebral 9L glioma bearing Fisher (F-344) rats by intracarotid and intratumoral injection. The blood half-life of the virus was 1 min (fast component, 10% contribution) and 180 min (slow component, 90% contribution). Approximately 20% of activity had been excreted by 24 h. With intracarotid injection, the total amount of virus that accumulated in tumor was 0.10 ± 0.07% of the injected dose (ID)/g at 1 h and 0.19 ± 0.01% ID/g at 24 h. By comparison, co-injection of RMP-7,

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In vivo magnetic resonance imaging of transgene expression

Cited by 769 publications

References 15 publications

Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene

Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene

Noninvasive, quantitative imaging in living animals of a mutant dopamine D2 receptor reporter gene in which ligand binding is uncoupled from signal transduction

Quantitation of HSV mass distribution in a rodent brain tumor model

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