The Advantages of Using Fluorescent Proteins for In Vivo Imaging

Hoffman, Robert M.

doi:10.1002/cpet.12

Cited by 3 publications

(5 citation statements)

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“…These exosomes loaded with PAI contrast agent improve tumor diagnosis as exosomes accumulated in melanoma tissues within 2 h of inoculation with increasing PA signals up to 8 h [321]. Likewise, luminescent-and fluorescent-protein-labeled exosomes can visualize biological processes regarding their cellular interactions, uptake, biodistribution and communication [322]. For instance, Takahashi et al used Gaussia luciferase (Gluc) and lactadherin to transduce B16-BL6 murine melanoma cells using a plasmid transduction method, and isolated labeled exosomes from those cells by UC.…”

Section: Mrimentioning

confidence: 99%

Advances in engineered exosomes towards cancer diagnosis and therapeutics

Tanziela

Dong

et al. 2022

Prog. Biomed. Eng.

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Exosomes have been emerged as natural nano-carriers and advantageous in the field of nano-medicines owing to lipid bilayer membrane comprising many proteins, nucleic acids and cell debris. Exosomes be secreted from all types of living cells and play a role in cancer diagnosis and therapy because their biological properties such as intercellular communication, modulation of immune responses, biocompatibility and target specificity. Many studies have shown that exosomes can be engineered or modified with different therapeutic substances including nucleic acids, proteins, drugs and other nanomaterials, to improve their specificity, efficiency and safety in nanomedicines. In this review, we summarize the methodologies for isolation of exosomes, the possible mechanisms of cellular uptake and the role of exosomes in cancer diagnosis, followed by the efficiency of modified exosomes in cancer therapy, and discuss them with appropriate examples. We strongly suggest that a clear articulation of the fundamental principles for making exosome-based theranostic platforms will help reveal the unique powers of exosomes in early cancer diagnosis and therapeutics including chemotherapy, gene therapy, immunotherapy and phototherapy.

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Section: Mrimentioning

confidence: 99%

Advances in engineered exosomes towards cancer diagnosis and therapeutics

Tanziela

Dong

et al. 2022

Prog. Biomed. Eng.

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“…The endogenous labeling technique hijacks cell biosynthesis to favor production of specific endogenous fluorescent markers or fluorescent proteins, which then become part of the exosome membrane. This type of labeling marks the exosome conclusively so that tracking these agents definitely implies tracking of the exosomes (Hoffman, 2017). The exogenous labeling technique uses an external agent that resides within the exosome, such as nanoparticles and fluorescent dyes.…”

Section: Labeling Exosomesmentioning

confidence: 99%

“…Fluorescent proteins (such as GFP and RFP) are mostly introduced to the parent cells-indirect labeling, while lipophilic dyes (such as Cy7, PKH26, luminogens) are incorporated into the exosomes directly. Labeling exosomes with lipophilic dyes provides insights mainly regarding their biodistribution and organotropic uptake while labeling exosomes with fluorescent proteins, can shed light on the biological mechanisms that occur in the cell and involve exosome biogenesis, secretion, and communication (Hoffman, 2017). FLI is considered a straightforward, efficient, and relatively low-cost labeling and imaging technique (Hoffman, 2017;Thomas, 2015).…”

Section: Optical Imagingmentioning

confidence: 99%

“…Labeling exosomes with lipophilic dyes provides insights mainly regarding their biodistribution and organotropic uptake while labeling exosomes with fluorescent proteins, can shed light on the biological mechanisms that occur in the cell and involve exosome biogenesis, secretion, and communication (Hoffman, 2017). FLI is considered a straightforward, efficient, and relatively low-cost labeling and imaging technique (Hoffman, 2017;Thomas, 2015).…”

Section: Optical Imagingmentioning

confidence: 99%

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Advances in imaging strategies for in vivo tracking of exosomes

Betzer

Barnoy

Sadan

et al. 2019

WIREs Nanomed Nanobiotechnol

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Exosomes have many biological functions as short‐ and long distance nanocarriers for cell‐to‐cell communication. They allow the exchange of complex information between cells, and thereby modulate various processes such as homeostasis, immune response and angiogenesis, in both physiological and pathological conditions. In addition, due to their unique abilities of migration, targeting, and selective internalization into specific cells, they are promising delivery vectors. As such, they provide a potentially new field in diagnostics and treatment, and may serve as an alternative to cell‐based therapeutic approaches. However, a major drawback for translating exosome treatment to the clinic is that current understanding of these endogenous vesicles is insufficient, especially in regards to their in vivo behavior. Tracking exosomes in vivo can provide important knowledge regarding their biodistribution, migration abilities, toxicity, biological role, communication capabilities, and mechanism of action. Therefore, the development of efficient, sensitive and biocompatible exosome labeling and imaging techniques is highly desired. Recent studies have developed different methods for exosome labeling and imaging, which have allowed for in vivo investigation of their bio‐distribution, physiological functions, migration, and targeting mechanisms. These improved imaging capabilities are expected to greatly advance exosome‐based nanomedicine applications. This article is categorized under: Therapeutic Approaches and Drug Discovery > Emerging Technologies Diagnostic Tools > In Vivo Nanodiagnostics and Imaging Nanotechnology Approaches to Biology > Nanoscale Systems in Biology

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“…Together with the development of new systems for whole-body imaging, uorescent proteins can be used to visualize many types of cancer processes, including primary tumor growth, tumors cell motility and invasion, metastases and the interaction between tumor and its microenvironment (tumor-host interaction) [3][4][5]. For deep imaging of animal tissues, the optical window favorable for light penetration is in near-infrared wavelengths, which are less absorbed by tissue, requiring proteins with emission spectra in the far-red wavelengths [3,[5][6][7][8][9]. Tumor cell lines stably expressing detectable uorescent proteins enable to evaluate the e cacy of therapeutic anticancer treatments through whole-body imaging without the need for any invasive procedures in tumor-bearing xenograft models.…”

Section: Introductionmentioning

confidence: 99%

Ablation of Red Stable Transfected Prostate Cancer Cell Lines by C-CPE Gold-Nanoparticle Mediated Laser Intervention

Alnajjar

Nolte

Becker

et al. 2021

Preprint

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Background: Claudin (CLDN) proteins have been described to be found and accordingly targeted to evaluate novel therapeutic approaches. C-terminus of Clostridium perfringens enterotoxin (C-CPE) binds efficiently several claudins and thus recombinant C-CPE conjugated to gold nanoparticles (AuNPs) has been used for cancer cell targeting using gold nanoparticle- mediated laser perforation (GNOME-LP). Cancer cells inoculation is routinely used to generate in vivo models to evaluate novel therapeutic approaches in prostate cancer. However, detailed characterization of cancer spreading and early tumor development and therapeutic response is often limited as conventional cell lines do not allow advanced deep tissue imaging.Methods: two canine prostate cancer cell lines were stably transfected with red fluorescent protein (RFP), followed by G418 selection. RFP marker as well as CLDN3, -4 and -7 expression was comparatively confirmed by flow cytometry, qPCR and immunofluorescences. For cancer cells targeting, GNOME-LP at a laser fluence of 72 mJ/cm² and a scanning speed of 0.5 cm/s was used. Statistical analysis was performed using SAS software 7.1, Dunnett´s Multiple Comparison Test and Student´s two-sided t-test. Differences were considered statistically significant for p<0.05.Results: we established two canine prostate carcinoma cell lines, stably expressing RFP allowing perspective deep tissue imaging. Directed C-CPE-AuNP binding to native and RFP transfected cells verified the capability to specifically target CLDN receptors. Cancer cell ablation was demonstrated in vitro setting using a combination of gold nanoparticle mediated laser perforation and C-CPE-AuNPs treatment reducing tumor cell viability to less than 10 % depending on cell line. Conclusion: the results confirm that this therapeutic approach can be used efficiently to target prostate carcinoma cells carrying a marker protein allowing deep tissue imaging. The established cell lines and the verified proof of concept in vitro study provide the basis for perspective Xenograft model in vivo studies. The introduce red fluorescence enables deep tissue imaging in living animals and therefore detailed characterization of tumor growth and subsequently possible tumor ablation through C-CPE-AuNPs treatment.

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The Advantages of Using Fluorescent Proteins for In Vivo Imaging

Cited by 3 publications

References 113 publications

Advances in engineered exosomes towards cancer diagnosis and therapeutics

Advances in engineered exosomes towards cancer diagnosis and therapeutics

Advances in imaging strategies for in vivo tracking of exosomes

Ablation of Red Stable Transfected Prostate Cancer Cell Lines by C-CPE Gold-Nanoparticle Mediated Laser Intervention

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The Advantages of Using Fluorescent Proteins for In Vivo Imaging

Cited by 3 publications

References 113 publications

Advances in engineered exosomes towards cancer diagnosis and therapeutics

Advances in engineered exosomes towards cancer diagnosis and therapeutics

Advances in imaging strategies for in vivo tracking of exosomes

Ablation of Red Stable Transfected Prostate Cancer Cell Lines by C-CPE&nbsp;Gold-Nanoparticle Mediated Laser Intervention

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Product

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Ablation of Red Stable Transfected Prostate Cancer Cell Lines by C-CPE Gold-Nanoparticle Mediated Laser Intervention