Synthesis of Stable Multifunctional Perfluorocarbon Nanoemulsions for Cancer Therapy and Imaging

Fernandes, Donald A.; Fernandes, Dennis D.; Li, Yuchong; Wang, Yan; Zhang, Zhenfu; Rousseau, Dérick; Gradinaru, Claudiu C.; Kolios, Michael C.

doi:10.1021/acs.langmuir.6b01867

Cited by 78 publications

(62 citation statements)

References 59 publications

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“…More recently, led by multidisciplinary advances in cancer nanomedicine, several nanosized PFC emulsions have been designed to accumulate in solid tumors through the enhanced permeability and retention (EPR) effect 4,20,45-47. When applied to hypoxic tumors, several recent studies have shown improved radiotherapeutic outcomes using PFC agents, even without HBO/carbogen breathing 48-50.…”

Section: Introductionmentioning

confidence: 99%

Perfluorocarbon nanodroplets can reoxygenate hypoxic tumors in vivo without carbogen breathing

Yun¹,

Bernards²,

Hoang³

et al. 2019

Nanotheranostics

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Nanoscale perfluorocarbon (PFC) droplets have enormous potential as clinical theranostic agents. They are biocompatible and are currently used in vivo as contrast agents for a variety of medical imaging modalities, including ultrasound, computed tomography, photoacoustic and 19 F-magnetic resonance imaging. PFC nanodroplets can also carry molecular and nanoparticulate drugs and be activated in situ by ultrasound or light for targeted therapy. Recently, there has been renewed interest in using PFC nanodroplets for hypoxic tumor reoxygenation towards radiosensitization based on the high oxygen solubility of PFCs. Previous studies showed that tumor oxygenation using PFC agents only occurs in combination with enhanced oxygen breathing. However, recent studies suggest that PFC agents that accumulate in solid tumors can contribute to radiosensitization, presumably due to tumor reoxygenation without enhanced oxygen breathing. In this study, we quantify the impact of oxygenation due to PFC nanodroplet accumulation in tumors alone in comparison with other reoxygenation methodologies, in particular, carbogen breathing. Methods: Lipid-stabilized, PFC (i.e., perfluorooctyl bromide, CF 3 (CF 2 ) 7 Br, PFOB) nanoscale droplets were synthesized and evaluated in xenograft prostate (DU145) tumors in male mice. Biodistribution assessment of the nanodroplets was achieved using a fluorescent lipophilic indocarbocyanine dye label (i.e., DiI dye) on the lipid shell in combination with fluorescence imaging in mice (n≥3 per group). Hypoxia reduction in tumors was measured using PET imaging and a known hypoxia radiotracer, [ 18 F]FAZA (n≥ 3 per group). Results: Lipid-stabilized nanoscale PFOB emulsions (mean diameter of ~250 nm), accumulated in the xenograft prostate tumors in mice 24 hours post-injection. In vivo PET imaging with [ 18 F]FAZA showed that the accumulation of the PFOB nanodroplets in the tumor tissues alone significantly reduced tumor hypoxia, without enhanced oxygen (i.e., carbogen) breathing. This reoxygenation effect was found to be comparable with carbogen breathing alone. Conclusion: Accumulation of nanoscale PFOB agents in solid tumors alone successfully reoxygenated hypoxic tumors to levels comparable with carbogen breathing alone, an established tumor oxygenation method. This study confirms that PFC agents can be used to reoxygenate hypoxic tumors in addition to their current applications as multifunctional theranostic agents.

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Section: Introductionmentioning

confidence: 99%

Perfluorocarbon nanodroplets can reoxygenate hypoxic tumors in vivo without carbogen breathing

Yun¹,

Bernards²,

Hoang³

et al. 2019

Nanotheranostics

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“…To characterize the hydrodynamic radius and the photophysical properties of FlAsH-EDT 2 before and after labelling onto the FCM-ybbR peptide, Fluorescence Correlation Spectroscopy (FCS) was performed on a custom built confocal microscope 57 . Experiments were performed using a 488 nm blue laser (TECBL-488nm, WorldStarTech, Canada) with an excitation intensity of ~200 W/cm 2 , in order to avoid laser-induced dark states of the fluorophore.…”

Section: Methodsmentioning

confidence: 99%

“…Two-color imaging of FlAsH-FCM-ybbR immobilized on PEG-biotin-neutravidin-coated glass coverslips and labelled in situ with CoA-A647 was performed on a custom-built TIRF microscope 57 The co-localization between FlAsH and AF647 fluorescent molecules was probed using alternating excitation with a 633 nm laser for AF647 fluorescence and a 473 nm laser (Cobolt Blue, Cobolt AB, Sweden) for FlAsH fluorescence. Modulation of laser excitation was achieved using an acousto-optic tunable filter (TF625–350–2–11-BR1A, Gooch & Housego).…”

Section: Methodsmentioning

confidence: 99%

Characterization of Fluorescein Arsenical Hairpin (FlAsH) as a Probe for Single-Molecule Fluorescence Spectroscopy

Fernandes

Bamrah

Kailasam

et al. 2017

Sci Rep

Self Cite

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In recent years, new labelling strategies have been developed that involve the genetic insertion of small amino-acid sequences for specific attachment of small organic fluorophores. Here, we focus on the tetracysteine FCM motif (FLNCCPGCCMEP), which binds to fluorescein arsenical hairpin (FlAsH), and the ybbR motif (TVLDSLEFIASKLA) which binds fluorophores conjugated to Coenzyme A (CoA) via a phosphoryl transfer reaction. We designed a peptide containing both motifs for orthogonal labelling with FlAsH and Alexa647 (AF647). Molecular dynamics simulations showed that both motifs remain solvent-accessible for labelling reactions. Fluorescence spectra, correlation spectroscopy and anisotropy decay were used to characterize labelling and to obtain photophysical parameters of free and peptide-bound FlAsH. The data demonstrates that FlAsH is a viable probe for single-molecule studies. Single-molecule imaging confirmed dual labeling of the peptide with FlAsH and AF647. Multiparameter single-molecule Förster Resonance Energy Transfer (smFRET) measurements were performed on freely diffusing peptides in solution. The smFRET histogram showed different peaks corresponding to different backbone and dye orientations, in agreement with the molecular dynamics simulations. The tandem of fluorophores and the labelling strategy described here are a promising alternative to bulky fusion fluorescent proteins for smFRET and single-molecule tracking studies of membrane proteins.

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“…The nanodroplets are composed of a biocompatible phospholipid shell encapsulating an inert, non-toxic perfluorocarbon such as perfluoropentane (PFP). Hydrophobic drugs can be incorporated within the lipid shell for transport through circulation [4]. Nanoemulsions accumulate in tumors via the enhanced permeability and retention (EPR) effect, which occurs due to the inherently leaky vascular structure of tumors (Fig 1) [ [5][6][7][8].…”

Section: Introductionmentioning

confidence: 99%

Delivery of thymoquinone to cancer cells with as1411-conjugated nanodroplets

et al. 2020

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Systemic delivery of conventional chemotherapies can cause negative systemic toxicity, including reduced immunity and damage to organs such as the heart and kidneys-limiting the maximum dose that can be administered. Targeted therapies appear to address this problem by having a specific target while mitigating off-target effects. Biocompatible perfluorocarbon-based nanodroplet emulsions encapsulated by a phospholipid shell are in development for delivery of molecular compounds and hold promise as vehicles for targeted delivery of chemotherapeutics to tumors. When ultrasound is applied, perfluorocarbon will undergo a phase change-ultimately inducing transient perforation of the cell membrane when in close proximity, which is more commonly known as "sonoporation." Sonoporation allows enhanced intracellular delivery of molecular compounds and will reseal to encapsulate the molecular compound intracellularly. In this study, we investigated delivery of thymoquinone (TQ), a natural hydrophobic phytochemical compound with bioactivity in cancer cells. In addition, we conjugated a G-quadruplex aptamer, 'AS1411', to TQ-loaded nanodroplets and explored their effects on multiple human cancer cell lines. AS1411 binds nucleolin, which is over-expressed on the surface of cancer cells, and in addition to its tumor-targeting properties AS1411 has also been shown to induce anti-cancer effects. Thymoquinone was loaded onto AS1411-conjugated nanodroplet emulsion to assess activity against cancer cells. Confocal microscopy indicated uptake of AS1411-conjugated nanodroplets by cancer cells. Furthermore, AS1411-conjugated nanoemulsions loaded with TQ significantly enhanced cytotoxicity in cancer cells compared to free compound. These results demonstrate that AS1411 can be conjugated onto nanodroplet emulsions for targeted delivery to human cancer cells. This novel formulation offers significant potential for targeted delivery of hydrophobic chemotherapeutics to tumors for cancer treatment.

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Synthesis of Stable Multifunctional Perfluorocarbon Nanoemulsions for Cancer Therapy and Imaging

Cited by 78 publications

References 59 publications

Perfluorocarbon nanodroplets can reoxygenate hypoxic tumors in vivo without carbogen breathing

Perfluorocarbon nanodroplets can reoxygenate hypoxic tumors in vivo without carbogen breathing

Characterization of Fluorescein Arsenical Hairpin (FlAsH) as a Probe for Single-Molecule Fluorescence Spectroscopy

Delivery of thymoquinone to cancer cells with as1411-conjugated nanodroplets

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