Biodegradable Nanoparticles Combining Cancer Cell Targeting and Anti-Angiogenic Activity for Synergistic Chemotherapy in Epithelial Cancer

Abstract A biodegradable engineered nanoplatform combining anti-angiogenic activity and targeting of cancer cells to improve the anticancer activity of docetaxel (DTX) is here proposed. Indeed, we have developed biodegradable nanoparticles (NPs) of poly(ethylene glycol)-poly(ε-caprolactone), exposing on the surface both folate motifs (Fol) for recognition in cells overexpressing Folate Receptor- a (FRa) and the anti-angiogenic hexapeptide aFLT1. The presence of Fol on NPs did not impair the anti-angiogenic activity of aFLT1, as assessed by in vitro tube formation assay in HUVEC endothelial cells. In both 2D and 3D KB cell cultures in vitro , the cytotoxicity of DTX loaded in NPs was not significantly affected by Fol/aFLT1 double decoration as compared to free DTX. Remarkably, NPs distributed differently in 3D multicellular spheroids of FRa-positive KB cancer cells depending on the type of ligand displayed on the surface. When tested in vivo in zebrafish embryos xenografted with KB cells, NPs displaying Fol/aFLT1 reduced DTX systemic toxicity and inhibited in a synergistic way the growth of the tumor mass and associated vasculature. Overall, nanotechnology offers excellent ground for combining therapeutic concepts in cancer, paving the way to the development of novel multifunctional nanopharmaceuticals where surface decoration with bioactive elements can significantly improve therapeutic outcomes.

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Biodegradable nanoparticles combining cancer cell targeting and anti-angiogenic activity for synergistic chemotherapy in epithelial cancer

Drug Delivery and Translational Research ◽

10.1007/s13346-021-01090-6 ◽

2022 ◽

Author(s):

Francesca Moret ◽

Claudia Conte ◽

Diletta Esposito ◽

Giovanni Dal Poggetto ◽

Concetta Avitabile ◽

...

Keyword(s):

Cancer Cells ◽

Folate Receptor ◽

Biological Fluids ◽

Tube Formation ◽

Multicellular Spheroids ◽

Angiogenic Activity ◽

Biodegradable Nanoparticles ◽

Therapeutic Concepts ◽

Poly Ethylene

AbstractA biodegradable engineered nanoplatform combining anti-angiogenic activity and targeting of cancer cells to improve the anticancer activity of docetaxel (DTX) is here proposed. Indeed, we have developed biodegradable nanoparticles (NPs) of poly(ethylene glycol)-poly(ε-caprolactone), exposing on the surface both folate motifs (Fol) for recognition in cells overexpressing Folate receptor-α (FRα) and the anti-angiogenic hexapeptide aFLT1. NPs showed a size around 100 nm, the exposure of 60% of Fol moieties on the surface, and the ability to entrap DTX and sustain its release with time. NPs were stable in simulated biological fluids and slightly interacted with Fetal Bovine serum, especially in the formulation decorated with Fol and aFLT1. The presence of Fol on NPs did not impair the anti-angiogenic activity of aFLT1, as assessed by in vitro tube formation assay in HUVEC endothelial cells. In both 2D and 3D KB cell cultures in vitro, the cytotoxicity of DTX loaded in NPs was not significantly affected by Fol/aFLT1 double decoration compared to free DTX. Remarkably, NPs distributed differently in 3D multicellular spheroids of FRα-positive KB cancer cells depending on the type of ligand displayed on the surface. In particular, NPs unmodified on the surface were randomly distributed in the spheroid, whereas the presence of Fol promoted the accumulation in the outer rims of the spheroid. Finally, NPs with Fol and aFLT1 gave a uniform distribution throughout the spheroid structure. When tested in zebrafish embryos xenografted with KB cells, NPs displaying Fol/aFLT1 reduced DTX systemic toxicity and inhibited the growth of the tumor mass and associated vasculature synergistically. Overall, nanotechnology offers excellent ground for combining therapeutic concepts in cancer, paving the way to novel multifunctional nanopharmaceuticals decorated with bioactive elements that can significantly improve therapeutic outcomes. Graphical abstract

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The Functional Implications of Endothelial Gap Junctions and Cellular Mechanics in Vascular Angiogenesis

Cancers ◽

10.3390/cancers11020237 ◽

2019 ◽

Vol 11 (2) ◽

pp. 237 ◽

Cited By ~ 15

Author(s):

Takayuki Okamoto ◽

Haruki Usuda ◽

Tetsuya Tanaka ◽

Koichiro Wada ◽

Motomu Shimaoka

Keyword(s):

Endothelial Cells ◽

Cell Migration ◽

Gap Junctions ◽

Gap Junction ◽

Cancer Cells ◽

Tumor Development ◽

Tube Formation ◽

Cellular Mechanics

Angiogenesis—the sprouting and growth of new blood vessels from the existing vasculature—is an important contributor to tumor development, since it facilitates the supply of oxygen and nutrients to cancer cells. Endothelial cells are critically affected during the angiogenic process as their proliferation, motility, and morphology are modulated by pro-angiogenic and environmental factors associated with tumor tissues and cancer cells. Recent in vivo and in vitro studies have revealed that the gap junctions of endothelial cells also participate in the promotion of angiogenesis. Pro-angiogenic factors modulate gap junction function and connexin expression in endothelial cells, whereas endothelial connexins are involved in angiogenic tube formation and in the cell migration of endothelial cells. Several mechanisms, including gap junction function-dependent or -independent pathways, have been proposed. In particular, connexins might have the potential to regulate cell mechanics such as cell morphology, cell migration, and cellular stiffness that are dynamically changed during the angiogenic processes. Here, we review the implication for endothelial gap junctions and cellular mechanics in vascular angiogenesis.

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Evaluation of Angiogenesis Assays

Biomedicines ◽

10.3390/biomedicines7020037 ◽

2019 ◽

Vol 7 (2) ◽

pp. 37 ◽

Cited By ~ 14

Author(s):

Zachary I. Stryker ◽

Mehdi Rajabi ◽

Paul J. Davis ◽

Shaker A. Mousa

Keyword(s):

Endothelial Cells ◽

Tube Formation ◽

Angiogenic Activity ◽

In Vitro Methods ◽

Current State ◽

Angiogenesis Process ◽

Pertinent Information ◽

Exogenous Factors

Angiogenesis assays allow for the evaluation of pro- or anti-angiogenic activity of endogenous or exogenous factors (stimulus or inhibitors) through investigation of their pro-or anti- proliferative, migratory, and tube formation effects on endothelial cells. To model the process of angiogenesis and the effects of biomolecules on that process, both in vitro and in vivo methods are currently used. In general, in vitro methods monitor specific stages in the angiogenesis process and are used for early evaluations, while in vivo methods more accurately simulate the living microenvironment to provide more pertinent information. We review here the current state of angiogenesis assays as well as their mechanisms, advantages, and limitations.

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Redox-Sensitive and Folate-Receptor-Mediated Targeting of Cervical Cancer Cells for Photodynamic Therapy Using Nanophotosensitizers Composed of Chlorin e6-Conjugated β-Cyclodextrin via Diselenide Linkage

Cells ◽

10.3390/cells10092190 ◽

2021 ◽

Vol 10 (9) ◽

pp. 2190

Author(s):

Howard Kim ◽

Mi Woon Kim ◽

Young-IL Jeong ◽

Hoe Saeng Yang

Keyword(s):

Cervical Cancer ◽

Photodynamic Therapy ◽

Hela Cells ◽

Folate Receptor ◽

Chlorin E6 ◽

Ros Generation ◽

Cervical Cancer Cells ◽

Poly Ethylene

The aim of this study was to fabricate a reactive oxygen species (ROS)-sensitive and folate-receptor-targeted nanophotosensitizer for the efficient photodynamic therapy (PDT) of cervical carcinoma cells. Chlorin e6 (Ce6) as a model photosensitizer was conjugated with succinyl β-cyclodextrin via selenocystamine linkages. Folic acid (FA)-poly(ethylene glycol) (PEG) (FA-PEG) conjugates were attached to these conjugates and then FA-PEG-succinyl β-cyclodextrin-selenocystamine-Ce6 (FAPEGbCDseseCe6) conjugates were synthesized. Nanophotosensitizers of FaPEGbCDseseCe6 conjugates were fabricated using dialysis membrane. Nanophotosensitizers showed spherical shapes with small particle sizes. They were disintegrated in the presence of hydrogen peroxide (H2O2) and particle size distribution changed from monomodal distribution pattern to multimodal pattern. The fluorescence intensity and Ce6 release rate also increased due to the increase in H2O2 concentration, indicating that the nanophotosensitizers displayed ROS sensitivity. The Ce6 uptake ratio, ROS generation and cell cytotoxicity of the nanophotosensitizers were significantly higher than those of the Ce6 itself against HeLa cells in vitro. Furthermore, the nanophotosensitizers showed folate-receptor-specific delivery capacity and phototoxicity. The intracellular delivery of nanophotosensitizers was inhibited by folate receptor blocking, indicating that they have folate-receptor specificity in vitro and in vivo. Nanophotosensitizers showed higher efficiency in inhibition of tumor growth of HeLa cells in vivo compared to Ce6 alone. These results show that nanophotosensitizers of FaPEGbCDseseCe6 conjugates are promising candidates as PDT of cervical cancer.

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Anti-Angiogenic Activity of Rotenoids from the Stems of Derris trifoliata

Planta Medica ◽

10.1055/s-0044-100797 ◽

2018 ◽

Vol 84 (11) ◽

pp. 779-785 ◽

Cited By ~ 3

Author(s):

Yanisa Mittraphab ◽

Nattaya Ngamrojanavanich ◽

Kuniyoshi Shimizu ◽

Kiminori Matsubara ◽

Khanitha Pudhom

Keyword(s):

Endothelial Cells ◽

Cancer Cells ◽

Ex Vivo ◽

Umbilical Vein ◽

Tube Formation ◽

Ex Vivo Model ◽

Cell Growth And Migration ◽

Vitro Assay ◽

Angiogenic Activity

The plants in the genus Derris have proven to be a rich source of rotenoids, of which cytotoxic effect against cancer cells seem to be pronounced. However, their effect on angiogenesis playing a crucial role in both cancer growth and metastasis has been seldom investigated. This study aimed at investigating the effect of the eight rotenoids (1–8) isolated from Derris trifoliata stems on three cancer cells and angiogenesis. Among them, 12a-hydroxyrotenone (2) exhibited potent inhibition on both cell growth and migration of HCT116 colon cancer cells. Further, anti-angiogenic assay in an ex vivo model was carried out to determine the effect of the isolated rotenoids on angiogenesis. Results revealed that 12a-hydroxyrotenone (2) displayed the most potent suppression of microvessel sprouting. The in vitro assay on human umbilical vein endothelial cells was performed to determine whether compound 2 elicits anti-angiogenic effect and its effect was found to occur via suppression of endothelial cells proliferation and tube formation, but not endothelial cells migration. This study provides the first evidence that compound 2 could potently inhibit HCT116 cancer migration and anti-angiogenic activity, demonstrating that 2 might be a potential agent or a lead compound for cancer therapy.

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Nanophotosensitizers for Folate Receptor-Targeted and Redox-Sensitive Delivery of Chlorin E6 against Cancer Cells

Materials ◽

10.3390/ma13122810 ◽

2020 ◽

Vol 13 (12) ◽

pp. 2810

Author(s):

Min-Suk Kook ◽

Chang-Min Lee ◽

Young-Il Jeong ◽

Byung-Hoon Kim

Keyword(s):

Folate Receptor ◽

Chlorin E6 ◽

Dependent Manner ◽

Oxygen Species ◽

Poly Ethylene Glycol ◽

Specific Manner ◽

Poly Ethylene ◽

End Group

In this study, FA–PEG3500-ss-Ce6tri copolymer was synthesized to deliver photosensitizers via redox-sensitive and folate receptor-specific manner. Folic acid (FA) was attached to amine end of poly (ethylene glycol) (PEG3500) (FA–PEG3500 conjugates) and cystamine-conjugated chlorin e6 (Ce6) (Ce6-cystamine conjugates). FA–PEG3500 was further conjugated with Ce6-cystamine to produce FA–PEG3500-ss-Ce6 conjugates. To the remaining amine end group of Ce6-cystamine conjugates, Ce6 was attached to produce FA–PEG3500-ss-Ce6tri. Nanophotosensitizers of FA–PEG3500-ss-Ce6tri copolymer were smaller than 200 nm. Their shapes were disintegrated by treatment with GSH and then Ce6 released by GSH-dependent manner. Compared to Ce6 alone, FA–PEG3500-ss-Ce6tri copolymer nanophotosensitizers recorded higher Ce6 uptake ratio, reactive oxygen species (ROS) production and cellular cytotoxicity against KB and YD-38 cells. The in vitro and in vivo study approved that delivery of nanophotosensitizers is achieved by folate receptor-sensitive manner. These results indicated that FA–PEG3500-ss-Ce6tri copolymer nanophotosensitizers are superior candidate for treatment of oral cancer.

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Delivery of disulfiram into breast cancer cells using folate-receptor-targeted PLGA-PEG nanoparticles: in vitro and in vivo investigations

Journal of Nanobiotechnology ◽

10.1186/s12951-016-0183-z ◽

2016 ◽

Vol 14 (1) ◽

Cited By ~ 47

Author(s):

Hamidreza Fasehee ◽

Rassoul Dinarvand ◽

Ardeshir Ghavamzadeh ◽

Mehdi Esfandyari-Manesh ◽

Hanieh Moradian ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Folate Receptor

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Folate Receptor Targeted Imaging Using Poly (ethylene glycol)-folate: In Vitro and In Vivo Studies

Journal of Korean Medical Science ◽

10.3346/jkms.2007.22.3.405 ◽

2007 ◽

Vol 22 (3) ◽

pp. 405 ◽

Cited By ~ 26

Author(s):

Se-Lim Kim ◽

Hwan-Jeong Jeong ◽

Eun-Mi Kim ◽

Chang-Moon Lee ◽

Tae-Hyoung Kwon ◽

...

Keyword(s):

Ethylene Glycol ◽

Folate Receptor ◽

In Vivo Studies ◽

Targeted Imaging ◽

Poly Ethylene Glycol ◽

Poly Ethylene

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The Anti-Angiogenic Activity of a Cystatin F Homologue from the Buccal Glands of Lampetra morii

Marine Drugs ◽

10.3390/md16120477 ◽

2018 ◽

Vol 16 (12) ◽

pp. 477 ◽

Cited By ~ 1

Author(s):

Mingru Zhu ◽

Bowen Li ◽

Jihong Wang ◽

Rong Xiao

Keyword(s):

Cdna Sequence ◽

Umbilical Vein ◽

Tube Formation ◽

Mass Spectrometry Analysis ◽

Cysteine Protease Inhibitors ◽

Angiogenic Activity ◽

Spectrometry Analysis ◽

Umbilical Vein Endothelial Cells

Cystatins are a family of cysteine protease inhibitors which are associated with a variety of physiological and pathological processes in vivo. In the present study, the cDNA sequence of a cystatin F homologue called Lm-cystatin F was cloned from the buccal glands of Lampetra morii. Although Lm-cystatin F shares a lower homology with cystatin superfamily members, it is also composed of a signal peptide and three highly conserved motifs, including the G in the N-terminal, QXVXG, as well as the PW in the C-terminal of the sequence. After sequence optimization and recombination, the recombinant protein was expressed as a soluble protein in Escherichia coli with a molecular weight of 19.85 kDa. Through affinity chromatography and mass spectrometry analysis, the purified protein was identified as a recombinant Lm-cystatin F (rLm-cystatin F). Additionally, rLm-cystatin F could inhibit the activity of papain. Based on MTT assay, rLm-cystatin F inhibited the proliferation of human umbilical vein endothelial cells (HUVECs) dose dependently with an IC50 of 5 μM. In vitro studies show that rLm-cystatin F suppressed the adhesion, migration, invasion, and tube formation of HUVECs, suggesting that rLm-cystatin F possesses anti-angiogenic activity, which provides information on the feeding mechanisms of Lampetra morii and insights into the application of rLm-cystatin F as a potential drug in the future.

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