Genetically encoded iron‐associated proteins as <scp>MRI</scp> reporters for molecular and cellular imaging

Naumova, Anna V.; Velde, Greetje Vande

doi:10.1002/wnan.1482

Cited by 10 publications

(10 citation statements)

References 221 publications

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“…Due to their iron oxide storage capacity, ferritins have been studied as intrinsic MRI contrast agents (Naumova and Vande Velde, 2018). Researchers have been also eager to use ferritin for in vitro production of MNPs.…”

Section: Ferritinmentioning

confidence: 99%

Intrinsically Magnetic Cells: A Review on Their Natural Occurrence and Synthetic Generation

Pekarsky

Spadiut

2020

Front. Bioeng. Biotechnol.

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The magnetization of non-magnetic cells has great potential to aid various processes in medicine, but also in bioprocess engineering. Current approaches to magnetize cells with magnetic nanoparticles (MNPs) require cellular uptake or adsorption through in vitro manipulation of cells. A relatively new field of research is "magnetogenetics" which focuses on in vivo production and accumulation of magnetic material. Natural intrinsically magnetic cells (IMCs) produce intracellular, MNPs, and are called magnetotactic bacteria (MTB). In recent years, researchers have unraveled function and structure of numerous proteins from MTB. Furthermore, protein engineering studies on such MTB proteins and other potentially magnetic proteins, like ferritins, highlight that in vivo magnetization of non-magnetic hosts is a thriving field of research. This review summarizes current knowledge on recombinant IMC generation and highlights future steps that can be taken to succeed in transforming non-magnetic cells to IMCs.

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Section: Ferritinmentioning

confidence: 99%

Intrinsically Magnetic Cells: A Review on Their Natural Occurrence and Synthetic Generation

Pekarsky

Spadiut

2020

Front. Bioeng. Biotechnol.

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“…T2*-weighted MRI sequences are less sensitive to edema, and are mostly used for detection of deoxygenated hemoglobin, hemosiderin in lesions and tissues, and intracranial hemorrhage [58]. Signal-void areas on T2*-weighted images are most commonly used for detection of iron accumulation in cells overexpressing ferritin [23].…”

Section: In Vivo Mri Studiesmentioning

confidence: 99%

“…Another way to create MRI contrast is to use MRI-reporters [14]. Different reporter proteins, including the iron storage protein, ferritin, in combination with different viral vectors, were proposed as genetically-based reporters, [23]. In some studies, lentiviral and adenoviral vectors, carrying the ferritin-complementary deoxyribonucleic acid(cDNA), were injected into the SVZ, leading to the ferritin synthesis in the migrating neuroblasts [11,14].…”

Section: Introductionmentioning

confidence: 99%

Tissue-Specific Ferritin- and GFP-Based Genetic Vectors Visualize Neurons by MRI in the Intact and Post-Ischemic Rat Brain

Khodanovich

Akulov

Anan’ina

et al. 2020

IJMS

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(1) Background: Neurogenesis is considered to be a potential brain repair mechanism and is enhanced in stroke. It is difficult to reconstruct the neurogenesis process only from the histological sections taken from different animals at different stages of brain damage and restoration. Study of neurogenesis would greatly benefit from development of tissue-specific visualization probes. (2) Purpose: The study aimed to explore if overexpression of ferritin, a nontoxic iron-binding protein, under a doublecortin promoter can be used for non-invasive visualization of neurogenesis using magnetic resonance imaging (MRI). (3) Methods: Ferritin heavy chain (FerrH) was expressed in the adeno-associated viral backbone (AAV) under the doublecortin promoter (pDCX), specific for young neurons, in the viral construct AAV-pDCX-FerrH. Expression of the enhanced green fluorescent protein (eGFP) was used as an expression control (AAV-pDCX-eGFP). The viral vectors or phosphate-buffered saline (PBS) were injected intracerebrally into 18 adult male Sprague–Dawley rats. Three days before injection, rats underwent transient middle-cerebral-artery occlusion or sham operation. Animals were subjected to In vivo MRI study before surgery and on days 7, 14, 21, and 28 days after injection using a Bruker BioSpec 11.7 T scanner. Brain sections obtained on day 28 after injection were immunostained for ferritin, young (DCX) and mature (NeuN) neurons, and activated microglia/macrophages (CD68). Additionally, RT-PCR was performed to confirm ferritin expression. (4) Results: T2* images in post-ischemic brains of animals injected with AAV-pDCX-FerrH showed two distinct zones of MRI signal hypointensity in the ipsilesioned hemisphere starting from 14 days after viral injection—in the ischemic lesion and near the lateral ventricle and subventricular zone (SVZ). In sham-operated animals, only one zone of hypointensity near the lateral ventricle and SVZ was revealed. Immunochemistry showed that ferritin-expressing cells in ischemic lesions were macrophages (88.1%), while ferritin-expressing cells near the lateral ventricle in animals both after ischemia and sham operation were mostly mature (55.7% and 61.8%, respectively) and young (30.6% and 7.1%, respectively) neurons. RT-PCR confirmed upregulated expression of ferritin in the caudoputamen and corpus callosum. Surprisingly, in animals injected with AAV-pDCX-eGFP we similarly observed two zones of hypointensity on T2* images. Cellular studies also showed the presence of mature (81.5%) and young neurons (6.1%) near the lateral ventricle in both postischemic and sham-operated animals, while macrophages in ischemic lesions were ferritin-positive (98.2%). (5) Conclusion: Ferritin overexpression induced by injection of AAV-pDCX-FerrH was detected by MRI using T2*-weighted images, which was confirmed by immunochemistry showing ferritin in young and mature neurons. Expression of eGFP also caused a comparable reduced MR signal intensity in T2*-weighted images. Additional studies are needed to investigate the potential and tissue-specific features of the use of eGFP and ferritin expression in MRI studies.

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“…MRI has the advantage of deep tissue penetration with relatively high (up to 100 µm) spatial resolution [23]. As for genetically encoded reporters for MRI [24], various metalloproteins, such as methemoglobin [25], transferrin [26], cytochrome P450-BM3 [23], and ferritin [27], are overexpressed for T 2 -weighted contrast generation. Ferritin, which was the first protein to induce MRI contrast in the absence of external reagents, is still the most studied in vivo [28,29].…”

Section: Introductionmentioning

confidence: 99%

Genetically Encoded Self-Assembling Iron Oxide Nanoparticles as a Possible Platform for Cancer-Cell Tracking

et al. 2021

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The study of growth and possible metastasis in animal models of tumors would benefit from reliable cell labels for noninvasive whole-organism imaging techniques such as magnetic resonance imaging. Genetically encoded cell-tracking reporters have the advantage that they are contrast-selective for viable cells with intact protein expression machinery. Besides, these reporters do not suffer from dilution during cell division. Encapsulins, which are bacterial protein nanocompartments, can serve as genetically controlled labels for multimodal detection of cells. Such nanocompartments can host various guest molecules inside their lumen. These include, for example, fluorescent proteins or enzymes with ferroxidase activity leading to biomineralization of iron oxide inside the encapsulin nanoshell. The aim of this work was to implement heterologous expression of encapsulin systems from Quasibacillus thermotolerans using the fluorescent reporter protein mScarlet-I and ferroxidase IMEF in the human hepatocellular carcinoma cell line HepG2. The successful expression of self-assembled encapsulin nanocompartments with functional cargo proteins was confirmed by fluorescence microscopy and transmission electron microscopy. Also, coexpression of encapsulin nanoshells, ferroxidase cargo, and iron transporter led to an increase in T2-weighted contrast in magnetic resonance imaging of HepG2 cells. The results demonstrate that the encapsulin cargo system from Q. thermotolerans may be suitable for multimodal imaging of cancer cells and could contribute to further in vitro and in vivo studies.

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Genetically encoded iron‐associated proteins as MRI reporters for molecular and cellular imaging

Cited by 10 publications

References 221 publications

Intrinsically Magnetic Cells: A Review on Their Natural Occurrence and Synthetic Generation

Intrinsically Magnetic Cells: A Review on Their Natural Occurrence and Synthetic Generation

Tissue-Specific Ferritin- and GFP-Based Genetic Vectors Visualize Neurons by MRI in the Intact and Post-Ischemic Rat Brain

Genetically Encoded Self-Assembling Iron Oxide Nanoparticles as a Possible Platform for Cancer-Cell Tracking

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