Near-Infrared Fluorescence Imaging of Mammalian Cells and Xenograft Tumors with SNAP-Tag

Gong, Haibiao; Kovar, Joy L.; Baker, Brenda; Zhang, Aihua; Cheung, Lael; Draney, Daniel R.; Corrêa, Ivan R.; Xu, Ming‐Qun; Olive, D. Michael

doi:10.1371/journal.pone.0034003

Cited by 31 publications

(61 citation statements)

References 40 publications

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“…192 Currently, antibodies against biomarkers and therapeutic targets of cancer have already been developed. The crosslinking of monoclonal antibodies and NIRF dyes has also been applied for selective cancer theranostics, 193 such as cutaneous tumor, 194,195 breast cancer, 196 ovarian cancer, 193 gastric cancer, 130 and prostate cancer. 197 This controlled drug delivery may potentially address the described limitations of the aforementioned chemotherapy.…”

Section: Drug Deliverymentioning

confidence: 99%

Near-infrared fluorescent probes in cancer imaging and therapy: an emerging field

Yi¹,

Wang²,

Qin³

et al. 2014

IJN

213

149

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Near-infrared fluorescence (NIRF) imaging is an attractive modality for early cancer detection with high sensitivity and multi-detection capability. Due to convenient modification by conjugating with moieties of interests, NIRF probes are ideal candidates for cancer targeted imaging. Additionally, the combinatory application of NIRF imaging and other imaging modalities that can delineate anatomical structures extends fluorometric determination of biomedical information. Moreover, nanoparticles loaded with NIRF dyes and anticancer agents contribute to the synergistic management of cancer, which integrates the advantage of imaging and therapeutic functions to achieve the ultimate goal of simultaneous diagnosis and treatment. Appropriate probe design with targeting moieties can retain the original properties of NIRF and pharmacokinetics. In recent years, great efforts have been made to develop new NIRF probes with better photostability and strong fluorescence emission, leading to the discovery of numerous novel NIRF probes with fine photophysical properties. Some of these probes exhibit tumoricidal activities upon light radiation, which holds great promise in photothermal therapy, photodynamic therapy, and photoimmunotherapy. This review aims to provide a timely and concise update on emerging NIRF dyes and multifunctional agents. Their potential uses as agents for cancer specific imaging, lymph node mapping, and therapeutics are included. Recent advances of NIRF dyes in clinical use are also summarized.

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Section: Drug Deliverymentioning

confidence: 99%

Near-infrared fluorescent probes in cancer imaging and therapy: an emerging field

Yi¹,

Wang²,

Qin³

et al. 2014

IJN

213

149

View full text Add to dashboard Cite

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“…Protein detection is achieved by chemical cross-linking of fluorescent benzylguanine derivatives, permitting direct imaging of protein bands in the polyacrylamide gel (31). Accordingly, the three human ␣ 1 -AR subtypes were subcloned into the pSNAP vector to add N-terminal human O 6 -alkylguanine-DNA alkyltransferase (SNAP) epitope tags.…”

Section: Adra1d N-terminal Domain Is Endogenously Cleaved Inmentioning

confidence: 99%

Endogenous N-terminal Domain Cleavage Modulates α1D-Adrenergic Receptor Pharmacodynamics

Kountz¹,

Lee²,

Aggarwal-Howarth

et al. 2016

Journal of Biological Chemistry

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The ␣ 1D -adrenergic receptor (ADRA1D) is a key regulator of cardiovascular, prostate, and central nervous system functions. This clinically relevant G protein-coupled receptor has proven difficult to study, as it must form an obligate modular homodimer containing the PDZ proteins scribble and syntrophin or become retained in the endoplasmic reticulum as nonfunctional protein. We previously determined that targeted removal of the N-terminal (NT) 79 amino acids facilitates ADRA1D plasma membrane expression and agonist-stimulated functional responses. However, whether such an event occurs in physiological contexts was unknown. Herein, we report the ADRA1D is subjected to innate NT processing in cultured human cells. SNAP near-infrared imaging and tandem-affinity purification revealed the ADRA1D is expressed as both fulllength and NT truncated forms in multiple human cell lines. Serial truncation mapping identified the cleavage site as Leu 90 / Val 91 in the 95-amino acid ADRA1D NT domain, suggesting human cells express a ⌬1-91 ADRA1D species. Tandem-affinity purification MS/MS and co-immunoprecipitation analysis indicate NT processing of ADRA1D is not required to form scribble-syntrophin macromolecular complexes. Yet, label-free dynamic mass redistribution signaling assays demonstrate that ⌬1-91 ADRA1D agonist responses were greater than WT ADRA1D. Mutagenesis of the cleavage site nullified the processing event, resulting in ADRA1D agonist responses less than the WT receptor. Thus, we propose that processing of the ADRA1D NT domain is a physiological mechanism employed by cells to generate a functional ADRA1D isoform with optimal pharmacodynamic properties.␣ 1 -Adrenergic receptors (ARs) 6 belong to the superfamily of class A G protein-coupled receptors (GPCRs). Stimulated by the endogenous catecholamines norepinephrine and epinephrine, ␣ 1 -ARs help coordinate sympathetic nervous function along with the ␣ 2 -and ␤-AR subtypes. This mode of GPCR signaling is particularly relevant during stress, exercise, or lifethreatening situations, as it permits an organism to respond to environmental stimuli supra-maximally and thereby enhance survival probability.Significant gaps remain in our understanding of ␣ 1 -AR biology. Of particular note is the ␣ 1D -AR subtype (ADRA1D). Unlike the closely related ␣ 1A (ADRA1A) and ␣ 1B (ADRA1B) subtypes, which achieve significant plasma membrane expression and robustly respond to agonists in cultured cells, the ADRA1D is cumbersome to study in vitro (1). Following its initial cloning and pharmacological characterization (2-4), numerous studies revealed the ADRA1D is sequestered intracellularly in myriad cell lines (5-11), where it has limited access to agonists and displays minimal functional activity. In a key study, Fan et al. (12) demonstrated ADRA1D functional expression is lost in cultured aortic vascular muscle cells ϳ48 h postdissection. They concluded that factors required for ADRA1D functional expression in vivo are absent in cell culture conditions (12), which may explain ...

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“…Thus, fluorescent agents in the NIR spectrum may find more widespread clinical use in the future. 18,31,34 Current handheld endomicroscopy systems can only excite fluorophores at fixed wavelengths (488 and 780 nm). The number of emission filters is also limited.…”

Section: Limitations Of Fluorophores and Imaging Parametersmentioning

confidence: 99%

Potential application of a handheld confocal endomicroscope imaging system using a variety of fluorophores in experimental gliomas and normal brain

Martirosyan

Georges

Eschbacher

et al. 2014

FOC

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Object The authors sought to assess the feasibility of a handheld visible-wavelength confocal endomicroscope imaging system (Optiscan 5.1, Optiscan Pty., Ltd.) using a variety of rapid-acting fluorophores to provide histological information on gliomas, tumor margins, and normal brain in animal models. Methods Mice (n = 25) implanted with GL261 cells were used to image fluorescein sodium (FNa), 5-aminolevulinic acid (5-ALA), acridine orange (AO), acriflavine (AF), and cresyl violet (CV). A U251 glioma xenograft model in rats (n = 5) was used to image sulforhodamine 101 (SR101). A swine (n = 3) model with AO was used to identify confocal features of normal brain. Images of normal brain, obvious tumor, and peritumoral zones were collected using the handheld confocal endomicroscope. Histological samples were acquired through biopsies from matched imaging areas. Samples were visualized with a benchtop confocal microscope. Histopathological features in corresponding confocal images and photomicrographs of H & E–stained tissues were reviewed. Results Fluorescence induced by FNa, 5-ALA, AO, AF, CV, and SR101 and detected with the confocal endomicroscope allowed interpretation of histological features. Confocal endomicroscopy revealed satellite tumor cells within peritumoral tissue, a definitive tumor border, and striking fluorescent cellular and subcellular structures. Fluorescence in various tumor regions correlated with standard histology and known tissue architecture. Characteristic features of different areas of normal brain were identified as well. Conclusions Confocal endomicroscopy provided rapid histological information precisely related to the site of microscopic imaging with imaging characteristics of cells related to the unique labeling features of the fluorophores. Although experimental with further clinical trial validation required, these data suggest that intraoperative confocal imaging can help to distinguish normal brain from tumor and tumor margin and may have application in improving intraoperative decisions during resection of brain tumors.

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Near-Infrared Fluorescence Imaging of Mammalian Cells and Xenograft Tumors with SNAP-Tag

Cited by 31 publications

References 40 publications

Near-infrared fluorescent probes in cancer imaging and therapy: an emerging field

Near-infrared fluorescent probes in cancer imaging and therapy: an emerging field

Endogenous N-terminal Domain Cleavage Modulates α1D-Adrenergic Receptor Pharmacodynamics

Potential application of a handheld confocal endomicroscope imaging system using a variety of fluorophores in experimental gliomas and normal brain

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