Unraveling the hypoxia modulating potential of VEGF family genes in pan-cancer.

Tumor hypoxia, oxygen deprivation state, occurs in most cancers and promotes angiogenesis, enhancing the potential for metastasis. The vascular endothelial growth factor (VEGF) family genes play crucial roles in tumorigenesis by promoting angiogenesis. To investigate the malignant processes triggered by hypoxia-induced angiogenesis across pan-cancers, we comprehensively analyzed the relationships between the expression of VEGF family genes and hypoxic microenvironment based on integrated bioinformatics methods. Our results suggest that the expression of VEGF family genes differs significantly among various cancers, highlighting their heterogeneity effect on human cancers. Across the 33 cancers, VEGFB and VEGFD showed the highest and lowest expression levels, respectively. The survival analysis showed that VEGFA and placental growth factor (PGF) were correlated with poor prognosis in many cancers, including kidney renal cell and liver hepatocellular carcinoma. VEGFC expression was positively correlated with glioma and stomach cancer. VEGFA and PGF showed distinct positive correlations with hypoxia scores in most cancers, indicating a potential correlation with tumor aggressiveness. The expression of miRNAs targeting VEGF family genes, including hsa-miR-130b-5p and hsa-miR-940, was positively correlated with hypoxia. In immune subtypes analysis, VEGFC was highly expressed in C3 (inflammatory) and C6 (transforming growth factor ß dominant) across various cancers, indicating its potential role as a tumor promotor. VEGFC expression exhibited positive correlations with immune infiltration scores, suggesting low tumor purity. High expression of VEGFA and VEGFC showed favorable responses to various drugs, including BLU-667, which abrogates RET signaling, an oncogenic driver in liver and thyroid cancers. Our findings suggest potential roles of VEGF family genes in malignant processes related with hypoxia-induced angiogenesis.

Prostate cancer risk prediction based on clinical factors and prostate-specific antigen.

INTRODUCTION: The incidence rate of prostate cancer (PCa) has continued to rise in Korea. This study aimed to construct and evaluate a 5-year PCa risk prediction model using a cohort with PSA < 10 ng/mL by incorporating PSA levels and individual factors. METHODS: The PCa risk prediction model including PSA levels and individual risk factors was constructed using a cohort of 69,319 participants from the Kangbuk Samsung Health Study. 201 registered PCa incidences were observed. A Cox proportional hazards regression model was used to generate the 5-year risk of PCa. The performance of the model was assessed using standards of discrimination and calibration. RESULTS: The risk prediction model included age, smoking status, alcohol consumption, family history of PCa, past medical history of dyslipidemia, cholesterol levels, and PSA level. Especially, an elevated PSA level was a significant risk factor of PCa (hazard ratio [HR]: 1.77, 95% confidence interval [CI]: [1.67-1.88]). This model performed well with sufficient discrimination ability and satisfactory calibration (C-statistic: 0.911, 0.874; Nam-D'Agostino test statistic:19.76, 4.21 in the development and validation cohort, respectively). CONCLUSIONS: Our risk prediction model was effective in predicting PCa in a population according to PSA levels. When PSA levels are inconclusive, an assessment of both PSA and specific individual risk factors (e.g., age, total cholesterol, and family history of PCa) could provide further information in predicting PCa.

Probabilistic evaluation of surface-enhanced localized surface plasmon resonance biosensing.

In this paper, we investigate detection characteristics of localized surface plasmon resonance biosensing based on a probabilistic Poisson distribution of target molecules. The model uses random nanoislands for localization of near-fields in three detection scenarios of non-specific, non-colocalized, and colocalized detection. Optical signatures were found to increase monotonically with target concentration and size regardless of the detection scenarios. The signatures were largest in colocalized detection of target interactions to localized fields, followed by non-colocalized and non-specific detection. The confidence interval was the narrowest in the colocalized detection due to the increased spatial certainty by localization. Based on the relative confidence interval, it was found that limit of detection can be enhanced by more than four orders of magnitude through colocalization.

Multifunctional magnetic nanoparticles modified with polyethylenimine and folic acid for biomedical theranostics.

This paper describes the preparation of magnetic nanoparticles modified with polyethylenimine (PEI)-folic acid (PF) conjugate and their potential biomedical applications. Magnetic nanoparticles modified with (3-(2-aminoethylamino)propyltrimethoxysilane) (AEAPS) were first prepared using a ligand exchange method to provide biocompatibility and hydrophilicity, and further conjugated with PF to carry gene and enhance specific uptake into cancer cells. We demonstrated the feasibility of the multifunctional magnetic nanoparticles as contrast agents in magnetic resonance imaging (MRI) and as gene carriers for gene delivery. In vitro results revealed that the cytotoxicity of the multifunctional magnetic nanoparticles was lower compared to that of pristine magnetic nanoparticles. Furthermore, we demonstrated the specific uptake of the magnetic nanoparticles modified with PF to KB cells using WI-38 cells as comparison by confocal microscopy. The PF-modified magnetic nanoparticles can potentially be employed as theranostic nanoplatforms for targeted gene delivery to cancer cells and simultaneous magnetic resonance imaging.

Synthesis and characterization of multifunctional Fe3O4/poly(fluorescein O-methacrylate) core/shell nanoparticles.

Multifunctional fluorescent and superparamagnetic Fe(3)O(4)/poly(fluorescein O-methacrylate) [Fe(3)O(4)/poly(FMA)] nanoparticles with core/shell structure were synthesized via surface-initiated polymerization. First, polymerizable double bonds were introduced onto the surface of Fe(3)O(4) nanoparticles via ligand exchange and a condensation reaction. A fluorescent monomer, FMA, was then polymerized to the double bonds at the surface via free-radical polymerization, leading to form a fluorescent polymer shell around the superparamagnetic Fe(3)O(4) core. The resultant Fe(3)O(4)/poly(FMA) nanoparticles were characterized by Fourier transform infrared, nuclear magnetic resonance, and X-ray diffraction spectroscopy to confirm the reactions. Transmission electron microscopy images showed that the Fe(3)O(4)/poly(FMA) nanoparticles have a spherical and monodisperse core/shell morphology. Photoluminescence spectroscopy and superconducting quantum interference device magnetometer analyses confirmed that the Fe(3)O(4)/poly(FMA) nanoparticles exhibited fluorescent and superparamagnetic properties, respectively. In addition, we demonstrated the potential bioimaging application of the Fe(3)O(4)/poly(FMA) nanoparticles by visualizing the cellular uptake of the nanoparticles into A549 lung cancer cells.

Fabrication of cross-linked alginate beads using electrospraying for adenovirus delivery.

Cross-linked alginate beads containing adenovirus (Ad) were successfully fabricated using an electrospraying method to achieve the protection and release of Ad in a controlled manner. An aqueous alginate solution containing Ad was electrosprayed into an aqueous phase containing a cross-linking agent (calcium chloride) at different process variables (voltages, alginate concentrations, and flow rates). Alginate beads containing Ad were used for transduction of U343 glioma cells and the transduction efficiency of the alginate beads was measured by quantification of gene expression using a fluorescence-activated cell sorter at different time points. In vitro results of gene expression revealed that the Ad encapsulated in the alginate beads with 0.5 wt% of alginate concentration exhibited a high activity for a long period (over 7 days) and was released in a sustained manner from the alginate beads. The Ad-encapsulating alginate beads could be promising materials for local delivery of Ad at a high concentration into target sites.

Ionically crosslinked Ad/chitosan nanocomplexes processed by electrospinning for targeted cancer gene therapy.

For effective cancer gene therapy, systemic administration of tumor-targeting adenoviral (Ad) complexes is critical for delivery to both primary and metastatic lesions. Electrospinning was used to generate nanocomplexes of Ad, chitosan, poly(ethylene glycol) (PEG), and folic acid (FA) for effective FA receptor-expressing tumor-specific transduction. The chemical structure of the Ad/chitosan-PEG-FA nanocomplexes was characterized by NMR and FT-IR, and the diameter and surface charge were analyzed by dynamic light scattering and zeta potentiometry, respectively. The average size of Ad/chitosan-PEG-FA nanocomplexes was approximately 140 nm, and the surface charge was 2.1 mV compared to -4.9 mV for naked Ad. Electron microscopy showed well-dispersed, individual Ad nanocomplexes without aggregation or degradation. Ad/chitosan nanocomplexes retained biological activity without impairment of the transduction efficiency of naked Ad. The transduction efficiency of Ad/chitosan-PEG-FA was increased as a function of FA ratio in FA receptor-expressing KB cells, but not in FA receptor-negative U343 cells, demonstrating FA receptor-targeted viral transduction. In addition, the transduction efficiency of Ad/chitosan-PEG-FA was 57.2% higher than chitosan-encapsulated Ad (Ad/chitosan), showing the superiority of FA receptor-mediated endocytosis for viral transduction. The production of inflammatory cytokine, IL-6 from macrophages was significantly reduced by Ad/chitosan-PEG-FA nanocomplexes, implying the potential for use in systemic administration. These results clearly demonstrate that cancer cell-targeted viral transduction by Ad/chitosan-PEG-FA nanocomplexes can be used effectively for metastatic tumor treatment with reduced immune reaction against Ad.

A chitosan hydrogel-based cancer drug delivery system exhibits synergistic antitumor effects by combining with a vaccinia viral vaccine.

Cancer treatment combining chemotherapy and immunotherapy has been vigorously exploited to further improve cancer therapeutic efficacy. This study investigated a new chemoimmunotherapy approach utilizing hydrogel as a local anti-cancer drug delivery system. Chitosan hydrogel containing doxorubicin (CH-DOX) and vaccinia virus vaccine expressing Sig/E7/LAMP-1 (Vac-Sig/E7/LAMP-1) were used as chemoimmunotherapeutic agents. It was found that intratumoral injection of CH-DOX effectively inhibited tumor growth itself and, in addition, exhibited a synergistic antitumor effect in combination with a vaccinia virus-based vaccine. This combination did not decrease but rather increased the number of tumor-specific CD8(+) T cells primed by vaccinia virus-mediated vaccination; the resulting antitumor effects were further improved up to 60 days as compared with monotherapy after tumor challenge, and the survival of tumor-bearing mice was dramatically prolonged. This study is a pioneer report that demonstrates the use of a biodegradable hydrogel system as an anti-cancer drug delivery system for successful chemoimmunotherapy. It is hoped that, this study can provide a foundation for a rational approach to improve antitumor efficacy of chemoimmunotherapy.

Enhanced circulation time and antitumor activity of doxorubicin by comblike polymer-incorporated liposomes.

Polymer incorporation on liposomal membranes has been extensively studied as a method of enhancing the circulation time of liposomes in the bloodstream. In this study, we investigated the in vitro and in vivo characteristics of liposomes whose surface was modified using a comblike polymer comprised of a poly(methyl methacrylate) (PMMA) backbone and short poly(ethylene oxide) (PEO) side chains. Doxorubicin (DOX)-loaded liposomes incorporating with the comblike polymer were prepared and their circulation time, biodistribution and antitumor activity were evaluated in B16F10 melanoma tumor-bearing mice. The circulation half-life time in the bloodstream of the comblike polymer-incorporated liposomes (CPILs) was approximately 14- or 2-fold higher than those of the conventional or polyethyleneglycol-fixed liposomes (PEG-liposomes), respectively. Additionally, in the biodistribution assay, the accumulation of the CPILs in the tumor was higher than those of the other liposomes. Based on this result, the antitumor activities of the CPILs were higher than those of conventional liposome formulation of DOX or free DOX due to the higher passive targeting efficiency of the long-circulating CPILs to tumor. This study suggests that the incorporation of the comblike polymer on the liposomal membrane is a promising tool to further improve circulation time of liposomes in tumor-bearing mice.

Anticancer Drug-Phospholipid Conjugate for Enhancement of Intracellular Drug Delivery.

Tumor specific delivery of anti-cancer drugs is one of the major challenges faced by drug development processes. In this study, we prepared a doxorubicin (DOX)-conjugated liposome (DCL) by incorporating the newly synthesized DSPE-PEG2000-DOX (DPD) into liposomes as a lipid component and tested its anti-tumor activity in vivo. DPD was synthesized by coupling DOX to DSPE-PEG2000-COOH via amide linkage and the chemical structure of resulting DPD was confirmed by (1)H-NMR analysis. DCL having liposome size of 130 nm was prepared through thin film cast-hydration method. DCL was found to have significantly higher cellular uptake than conventional liposomes as confirmed by flow cytometry analysis. Anti-tumor activity of DCL against murine B16F10 melanoma tumor-bearing mice revealed that DCL inhibits tumor growth more efficiently than the conventional liposomes, presumably attributed to DOX mediated endocytosis process.

Hyperthermia-induced antitumor activity of thermosensitive polymer modified temperature-sensitive liposomes.

Temperature-sensitive liposomes (TS-liposomes) have been studied for chemotherapeutic purposes to enhance the release of anticancer drugs at tumor sites. In this study, we prepared poly(N-isopropylacrylamide-co-acrylamide) (PNIPAM-AAM) and polyethylene glycol (PEG)-modified TS-liposomes (PETS-liposomes). PETS-liposomes significantly increased in vitro drug release in serum compared with PEG-fixed or PNIPAM-AAM-modified liposomes. Furthermore, incorporation of both PNIPAM-AAM and PEG into PETS-liposomes enhanced the stabilities of liposomes in serum by inhibiting protein adsorption. In addition, to investigate the therapeutic efficacy of doxorubicin (DOX)-loaded PETS-liposomes, the in vivo antitumor activity of liposomes in combination with hyperthermia was evaluated in a B16F10 melanoma tumor-bearing mouse model. PETS-liposomes showed much higher levels of tumor growth inhibition than PEG-fixed or PNIPAM-AAM-modified TS-liposomes. Moreover, the antitumor activity of PETS-liposomes was enhanced significantly when they were administered in combination with hyperthermia. PETS-liposomes were found to be highly efficacious carriers for the in vivo delivery of anticancer drugs, and to have potential anticancer applications in combination with hyperthermia.

In vivo distribution and antitumor activity of heparin-stabilized doxorubicin-loaded liposomes.

The purpose of this study was to investigate the effect of heparin conjugation to the surface of doxorubicin (DOX)-loaded liposomes on the circulation time, biodistribution and antitumor activity after intravenous injection in murine B16F10 melanoma tumor-bearing mice. The heparin-conjugated liposomes (heparin-liposomes) were prepared by fixation of the negatively charged heparin to the positively charged liposomes. The existence of heparin on the liposomal surface was confirmed by measuring the changes in the particle size, zeta potential and heparin amount of the liposomes. The stability of the heparin-liposomes in serum was higher than that of the control liposomes, due to the heparin-liposomes being better protected from the adsorption of serum proteins. The DOX-loaded heparin-liposomes showed high drug levels for up to 64 h after the intravenous injection and the half-life of DOX was approximately 8.4- or 1.5-fold higher than that of the control liposomes or polyethyleneglycol-fixed liposomes (PEG-liposomes), respectively. The heparin-liposomes accumulated to a greater extent in the tumor than the control or PEG-liposomes as a result of their lower uptake by the reticuloendothelial system cells in the liver and spleen. In addition, the DOX-loaded heparin-liposomes retarded the growth of the tumor effectively compared with the control or PEG-liposomes. These results indicate the promising potential of heparin-liposomes as a new sterically stabilized liposomal delivery system for the enhancement of the therapeutic efficacy of chemotherapeutic agents.