Overexpression of the MRI Reporter Genes Ferritin and Transferrin Receptor Affect Iron Homeostasis and Produce Limited Contrast in Mesenchymal Stem Cells

Pereira, Sofia M.; Moss, Darren M.; Williams, S. R.; Murray, Patricia; Taylor, Arthur

doi:10.3390/ijms160715481

Cited by 46 publications

(63 citation statements)

References 36 publications

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“…MRI was then performed to see if overexpression of the TfR1 reporter and/or FC supplementation enabled these same cells to be visualized with both the fluorescence and MR imaging modalities. As we have previously shown 7, 17 cells injected into a single site at E3 are often found in the form of several clusters at E5 (Fig. 5), which is a consequence of the quick brain development during this period.…”

Section: Resultssupporting

confidence: 62%

“…Therefore, the observed increase in Fth1 protein in the CHO‐K1 reporter line is most probably due to increased intracellular iron concentrations resulting from the overexpression of TfR1 leading to its storage within ferritin, which is ultimately responsible for the generation of T 2 contrast. However, a recent study from our group showed that overexpression of TfR1 in mMSCs did not lead to an increase in Fth1 protein levels 17. This is probably because TfR1 protein increased by only 15‐fold in the mMSCs, compared with more than 60‐fold in the CHO‐K1 cells.…”

Section: Discussionmentioning

confidence: 68%

“…However, this did not lead to any noticeable changes in MR contrast ( T 2 relaxation), which was probably because the increase was very low and close to background levels. The increase in intracellular iron in TfR1 ‐expressing CHO‐K1 cells is in contrast to mMSCs, where under the same FC supplementation conditions overexpression of TfR1 did not lead to any increase in intracellular iron levels 17. It is important to note that, for both cell types, supplementing the culture medium with FC increased intracellular iron concentrations to a greater extent than did overexpressing TfR1 ; for CHO‐K1 cells supplementation with 0.2 mM and 2 mM FC led to a respective fivefold and 10‐fold increase in intracellular iron, and for mMSCs to a respective 10‐fold and 40‐fold increase 17.…”

Section: Discussionmentioning

confidence: 78%

“…The increase in intracellular iron in TfR1 ‐expressing CHO‐K1 cells is in contrast to mMSCs, where under the same FC supplementation conditions overexpression of TfR1 did not lead to any increase in intracellular iron levels 17. It is important to note that, for both cell types, supplementing the culture medium with FC increased intracellular iron concentrations to a greater extent than did overexpressing TfR1 ; for CHO‐K1 cells supplementation with 0.2 mM and 2 mM FC led to a respective fivefold and 10‐fold increase in intracellular iron, and for mMSCs to a respective 10‐fold and 40‐fold increase 17. This was also the case for mNSCs, where supplementation with 1 mM FC led to an approximate 10‐fold increase in intracellular iron 19.…”

Section: Discussionmentioning

confidence: 78%

“…We have recently investigated if overexpression of ferritin heavy chain‐1 ( Fth1 ) and transferrin receptor‐1 ( TfR 1) in mouse mesenchymal stem cells (mMSCs) affects iron homeostasis and cell phenotype 17. In that study, we reported that, at physiological concentrations of extracellular iron (i.e.…”

Section: Introductionmentioning

confidence: 99%

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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: Resultssupporting

confidence: 62%

Section: Discussionmentioning

confidence: 68%

Section: Discussionmentioning

confidence: 78%

Section: Discussionmentioning

confidence: 78%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

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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Genetically Controlled Lysosomal Entrapment of Superparamagnetic Ferritin for Multimodal and Multiscale Imaging and Actuation with Low Tissue Attenuation

Massner

Sigmund

Pettinger

et al. 2018

Adv Funct Materials

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Nanomaterials are of enormous value for biomedical applications because of their customizable features. However, the material properties of nanomaterials can be altered substantially by interactions with tissue thus making it important to assess them in the specific biological context to understand and tailor their effects. Here, a genetically controlled system is optimized for cellular uptake of superparamagnetic ferritin and subsequent trafficking to lysosomes. High local concentrations of photoabsorbing magnetoferritin give robust contrast in optoacoustic imaging and allow for selective photoablation of cells overexpressing ferritin receptors. Genetically controlled uptake of the biomagnetic nanoparticles also strongly enhances third-harmonic generation due to the change of refractive index caused by the magnetiteprotein interface of ferritins entrapped in lysosomes. Selective uptake of magnetoferritin furthermore enables sensitive detection of receptor-expressing cells by magnetic resonance imaging, as well as efficient magnetic cell sorting and manipulation. Surprisingly, a substantial increase in the blocking temperature of lysosomally entrapped magnetoferritin is observed, which allows for specific ablation of genetically defined cell populations by local magnetic hyperthermia. The subcellular confinement of superparamagnetic ferritins thus enhances their physical properties to empower genetically controlled interrogation of cellular processes with deep tissue penetration.The interest in overexpressed ferritin has recently been revived in attempts to create biomagnetic actuators aimed at evoking controlled cellular responses to magnetic fields. In this regard, ferritin was used to manipulate cellular processes via magnetic hyperthermia or magnetomechanical transduction, [4] although the biophysical mechanisms underlying these effects are so far not understood well on a theoretical level. [5] Fully genetically encoded sensors and actuators, which do not necessitate supplementation of synthetic components, have become the frequently preferred solutions for, e.g., fluorescent Ca 2+ imaging and optogenetics. However, with regard to noninvasive techniques with deep tissue penetration such as optoacoustic (OA) imaging or MRI, synthetic nanostructures often still possess superior material properties as compared to genetically encodable biomaterials.Semigenetic approaches, which consist of a genetic component that interacts with an exogenous compound, can exploit the superior physical properties of synthetic nanostructures for remote cell actuation and deep tissue imaging and at the same time benefit from genetic targetability. [6] A recent study showcased the use of ferritin in a semigenetic approach to obtain cellular MR-contrast. [7] It was hypothesized that the enhanced contrast resulted from ferritin agglomeration inside lysosomes after cellular uptake through murine T cell immunoglobulin domain and mucin domain 2 (Tim-2) receptor. This contrastenhancing effect could be exploited even more effectively if the...

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Biogenic Imaging Contrast Agents

et al. 2023

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Imaging contrast agents are widely investigated in preclinical and clinical studies, among which biogenic imaging contrast agents (BICAs) are developing rapidly and playing an increasingly important role in biomedical research ranging from subcellular level to individual level. The unique properties of BICAs, including expression by cells as reporters and specific genetic modification, facilitate various in vitro and in vivo studies, such as quantification of gene expression, observation of protein interactions, visualization of cellular proliferation, monitoring of metabolism, and detection of dysfunctions. Furthermore, in human body, BICAs are remarkably helpful for disease diagnosis when the dysregulation of these agents occurs and can be detected through imaging techniques. There are various BICAs matched with a set of imaging techniques, including fluorescent proteins for fluorescence imaging, gas vesicles for ultrasound imaging, and ferritin for magnetic resonance imaging. In addition, bimodal and multimodal imaging can be realized through combining the functions of different BICAs, which helps overcome the limitations of monomodal imaging. In this review, the focus is on the properties, mechanisms, applications, and future directions of BICAs.

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Overexpression of the MRI Reporter Genes Ferritin and Transferrin Receptor Affect Iron Homeostasis and Produce Limited Contrast in Mesenchymal Stem Cells

Cited by 46 publications

References 36 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

Genetically Controlled Lysosomal Entrapment of Superparamagnetic Ferritin for Multimodal and Multiscale Imaging and Actuation with Low Tissue Attenuation

Biogenic Imaging Contrast Agents

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