Poly (ADP-ribose) polymerase inhibitors combined with other small-molecular compounds for the treatment of ovarian cancer.

Ovarian cancer is a heterogeneous disease with complex molecular and genetic hallmarks. Benefitting from profound understanding of molecular mechanisms in ovarian cancer pathogenesis, novel targeted drugs have been actively explored in preclinical studies and clinical trials. Considered as one of the most potent and effective targeted therapies for the treatment of ovarian cancer, poly (ADP-ribose) polymerase (PARP) inhibitors (PARPis) take advantages of synthetic lethality mechanisms to prevent DNA damage repair in cancer cells and cause their death, especially in cancers with BRCA mutations. Mounting evidence has indicated that the combination of PARPis with cytotoxic drugs or other targeted drugs has shown favorable synergistic effects. Excitingly, the antitumor activity of combination therapy of PARPis has been actively tested in multiple clinical trials and in-vitro or in-vivo experiments. In this review, we will briefly discuss the molecular mechanisms of PARPis combined with other therapeutic small-molecular compounds for the treatment of ovarian cancer.

Differential Modulating Effect of MoS2 on Amyloid Peptide Assemblies.

The abnormal fibrillogenesis of amyloid peptides such as amyloid fibril and senior amyloid plaques, is associated with the pathogenesis of many amyloid diseases. Hence, modulation of amyloid assemblies is related to the possible pathogenesis of some diseases. Some two-dimensional nanomaterials, that is, graphene oxide, tungsten disulfide, exhibit strong modulation effects on the amyloid fibrillogenesis. Herein, the modulation effect of molybdenum disulfide on two amyloid peptide assemblies based on the label-free techniques is presented, including quartz crystal microbalance (QCM), AFM, and CD spectroscopy. MoS2 presents different modulating effects on the assembly of amyloid-ß peptide (33-42) [Aß (33-42)] and amylin (20-29), mainly owing to the distinct affinity between amyloid peptides and MoS2 . This is to our knowledge the first report of MoS2 as a modulator for amyloid aggregation. It enriches the variety of 2D nanomodulators of amyloid fibrillogenesis and explains the mechanism for the self-assembly of amyloid peptides, and expands the applications of MoS2 in biology.

Correlation between Membrane Partitioning and Functional Activity in a Single Lipid Vesicle Assay Establishes Design Guidelines for Antiviral Peptides.

The nanometer-scale discrimination of virus-rupturing peptides is tested using lipid membrane platforms. In combination with single-vesicle analysis of peptide-induced vesicle rupture, a correlation between membrane partitioning and biologically relevant activities is established. Taken together, the findings support that the degree of rupture activity should be balanced by membrane curvature-selectivity for optimal therapeutic properties of antiviral peptides.

Elucidating the nanostructure of small interfering RNA-loaded lipidoid-polymer hybrid nanoparticles.

We analyzed the structural and material properties of small interfering RNA (siRNA)-loaded lipid-polymer hybrid nanoparticles (LPNs) containing ionizable lipidoid and poly(dl-lactic-co-glycolic acid) (PLGA) using small-angle X-ray scattering, cryogenic transmission electron microscopy, polarized light microscopy, the Langmuir monolayer methodology, differential scanning calorimetry, and attenuated total reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy. Scattering analyses showed that bulk lipidoid self-assemble into lamellar structures with a d-spacing of 38 Å, whereas lipidoid-siRNA lipoplexes display an in-plane lateral organization of siRNA in between lipidoid bilayers with a repeat distance of approximately 55 Å. The siRNA-loaded LPNs adopted a core-shell structure with an interaxial alignment of siRNA between lipidoid shell bilayers. Langmuir monolayer experiments showed a distinct interaction between the lipidoid headgroups and siRNA, which was dependent on buffer subphase pH. Thermal analyses suggested that PLGA and lipidoid interact, which was evident from a shift in the phase transition temperature of lipidoid, and the thermotropic phase behavior of lipidoid was affected by inclusion of siRNA. ATR-FTIR data confirmed the shift or disappearance of characteristic absorption bands of siRNA after lipidoid binding. In conclusion, siRNA-loaded LPNs display a core-shell structure, wherein the polymeric core functions as a colloid matrix support for siRNA-loaded lipidoid shell layers.

Inhaled RNA Therapeutics for Obstructive Airway Diseases: Recent Advances and Future Prospects.

Obstructive airway diseases, e.g., chronic obstructive pulmonary disease (COPD) and asthma, represent leading causes of morbidity and mortality worldwide. However, the efficacy of currently available inhaled therapeutics is not sufficient for arresting disease progression and decreasing mortality, hence providing an urgent need for development of novel therapeutics. Local delivery to the airways via inhalation is promising for novel drugs, because it allows for delivery directly to the target site of action and minimizes systemic drug exposure. In addition, novel drug modalities like RNA therapeutics provide entirely new opportunities for highly specific treatment of airway diseases. Here, we review state of the art of conventional inhaled drugs used for the treatment of COPD and asthma with focus on quality attributes of inhaled medicines, and we outline the therapeutic potential and safety of novel drugs. Subsequently, we present recent advances in manufacturing of thermostable solid dosage forms for pulmonary administration, important quality attributes of inhalable dry powder formulations, and obstacles for the translation of inhalable solid dosage forms to the clinic. Delivery challenges for inhaled RNA therapeutics and delivery technologies used to overcome them are also discussed. Finally, we present future prospects of novel inhaled RNA-based therapeutics for treatment of obstructive airways diseases, and highlight major knowledge gaps, which require further investigation to advance RNA-based medicine towards the bedside.

PEGylated phospholipid membrane on polymer cushion and its interaction with cholesterol.

By employing poly(ethylene glycol) (PEG) shielding and a polymer cushion to achieve air stability of the lipid membrane, we have analyzed PEG influence on dried membranes and the interaction with cholesterol. Small unilamellar vesicles (SUVs) formed by the mixture of 1,2-dimyristoylphosphatidylcholine (DMPC) with different molar fraction of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(poly(ethylene glycol))-2000] (DSPE-PEG(2000)) adsorb and fuse into membranes on different polymer-modified silicon dioxide surfaces, including chitosan, poly(L-lysine) (PLL), and hyaluronic acid. Dried membranes are further examined by ellipsometer and atomic force microscopy (AFM). Only chitosan can support a visible and uniform lipid array. The thickness of dry PEGylated lipid membrane is reduced gradually as the molar fraction of PEG increases. AFM scanning confirms the lipid membrane stacking for vesicles containing low PEG, and only a proper amount of PEG can maintain a single lipid bilayer; however, the air stability of the membrane will be destroyed if overloading PEG. Cholesterol incorporation can greatly improve the structural stability of lipid membrane, especially for those containing high molar fraction of PEG. Different amounts of cholesterol influence the thickness and surface morphology of dried membrane.

The Long Road Toward COVID-19 Herd Immunity: Vaccine Platform Technologies and Mass Immunization Strategies.

There is an urgent need for effective countermeasures against the current emergence and accelerating expansion of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Induction of herd immunity by mass vaccination has been a very successful strategy for preventing the spread of many infectious diseases, hence protecting the most vulnerable population groups unable to develop immunity, for example individuals with immunodeficiencies or a weakened immune system due to underlying medical or debilitating conditions. Therefore, vaccination represents one of the most promising counter-pandemic measures to COVID-19. However, to date, no licensed vaccine exists, neither for SARS-CoV-2 nor for the closely related SARS-CoV or Middle East respiratory syndrome-CoV. In addition, a few vaccine candidates have only recently entered human clinical trials, which hampers the progress in tackling COVID-19 infection. Here, we discuss potential prophylactic interventions for SARS-CoV-2 with a focus on the challenges existing for vaccine development, and we review pre-clinical progress and ongoing human clinical trials of COVID-19 vaccine candidates. Although COVID-19 vaccine development is currently accelerated via so-called fast-track programs, vaccines may not be timely available to have an impact on the first wave of the ongoing COVID-19 pandemic. Nevertheless, COVID-19 vaccines will be essential in the future for reducing morbidity and mortality and inducing herd immunity, if SARS-CoV-2 becomes established in the population like for example influenza virus.

One-pot synthesis and characterization of ovalbumin-conjugated gold nanoparticles: A comparative study of adjuvanticity against the physical mixture of ovalbumin and gold nanoparticles.

Only few adjuvants are licensed for use in humans and there is a need to develop safe and improved vaccine adjuvants. In this study, we report the one-pot synthesis of antigen ovalbumin (OVA)-conjugated gold nanoparticles (OVA@GNPs). A systematical study was performed by comparing OVA@GNPs with the simple mixture of OVA and gold nanoparticles (OVA+GNPs), including their physiochemical properties through spectrometric and electrophoretic analysis, in vitro stability, cytotoxicity and cellular uptake, and in vivo humoral immune responses following subcutaneous and transcutaneous immunization in mice. The results demonstrate a much stronger interaction between protein and GNPs in OVA@GNPs than OVA+GNPs, which makes OVA@GNPs more stable under in vitro conditions than OVA+GNPs with the ability to induce 4 times higher OVA-specific serum IgG titers following subcutaneous immunization. We also show the dose sparing of OVA@GNPs, as the dosage for aluminum adjuvant required to reach to an equivalent OVA-specific antibody titer was almost five times higher than OVA@GNPs. However, we found that the co-administration of small-sized GNPs had a limited ability for the transcutaneous delivery of OVA. These results demonstrate the potential application of one-pot synthesis approach for producing antigen protein-conjugated gold nanoparticles for vaccine delivery.

NOD-like receptor signaling in inflammation-associated cancers: From functions to targeted therapies.

BACKGROUND: Recently, many studies have reported that some botanicals and natural products were able to regulate NOD-like receptor signaling. NOD-like receptors (NLRs) have been established as crucial regulators in inflammation-associated tumorigenesis, angiogenesis, cancer cell stemness and chemoresistance. NLRs specifically sense pathogen-associated molecular patterns and respond by activating other signaling regulators, including Rip2 kinase, NF-κB, MAPK and ASC/caspase-1, leading to the secretion of various cytokines. PURPOSE: The aim of this article is to review the molecular mechanisms of NOD-like receptor signaling in inflammation-associated cancers and the NLRs-targeted botanicals and synthetic small molecules in cancer intervention. RESULTS: Aberrant activation of NLRs occurs in various cancers, orchestrating the tissue microenvironment and potentiating neoplastic risk. Blocking NLR inflammasome activation by botanicals or synthetic small molecules may be a valuable way to prevent cancer progression. Moreover, due to the roles of NLRs in regulating cytokine production, NLR signaling may be correlated with senescence-associated secretory phenotype. CONCLUSION: In this review, we discuss how NLR signaling is involved in inflammation-associated cancers, and highlight the NLR-targeted botanicals and synthetic small molecules in cancer intervention.

Targeting Membrane Receptors of Ovarian Cancer Cells for Therapy.

Ovarian cancer is a leading cause of death worldwide from gynecological malignancies, mainly because there are few early symptoms and the disease is generally diagnosed at an advanced stage. In addition, despite the effectiveness of cytoreductive surgery for ovarian cancer and the high response rates to chemotherapy, survival has improved little over the last 20 years. The management of patients with ovarian cancer also remains similar despite studies showing striking differences and heterogeneity among different subtypes. It is therefore clear that novel targeted therapeutics are urgently needed to improve clinical outcomes for ovarian cancer. To that end, several membrane receptors associated with pivotal cellular processes and often aberrantly overexpressed in ovarian cancer cells have emerged as potential targets for receptor-mediated therapeutic strategies including specific agents and multifunctional delivery systems based on ligand-receptor binding. This review focuses on the profiles and potentials of such strategies proposed for ovarian cancer treatment and imaging.

Formulation and in vitro stability evaluation of ethosomal carbomer hydrogel for transdermal vaccine delivery.

The primary objective of this study was to develop, evaluate and compare the effectiveness and stability of ethosomal carbomer gels in different solvents. The optimal ethosomal formulation was isolated to create ethosomal gels using carbomer in either pure water (water gel) or PBS containing 30% ethanol (PBS gel). In vitro release of the ethosomal gels were tested using Franz apparatus on hydrophilic and hydrophobic artificial membranes. In vitro stability of two ethosomal gels was systematically evaluated. Transdermal antigen delivery of ethosomal gel was finally performed on the skin of hair removal mice. Both solvent and concentration effects on the in vitro release performance of ethosomal gel of carbomer have been confirmed. Penetration depth has been found to be dependent on the nature of the membranes such that penetration rate is higher in the hydrophobic membrane than the hydrophilic ones. Furthermore, in vitro stability test indicated that ethosomal PBS gel was more stable than ethosomal water gel. In vivo immunoassay confirmed that the ethosomal PBS gel could deliver the antigenic molecules into the skin of mice and stimulate specific IgG secretion. Using the same solvent for lipid vesicular formulation when making polymeric hydrogel may help to provide a more conducive environment for lipid vesicles and hence enhance their roles in transdermal antigen delivery.

In Vivo SELEX of an Inhibitory NSCLC-Specific RNA Aptamer from PEGylated RNA Library.

Aptamers are widely used in numerous biochemical, bioanalytical, and biological studies. Most aptamers are developed through an in vitro selection process called SELEX against either purified targets or living cells expressing targets of interest. We report here an in vivo SELEX in mice using a PEGylated RNA library for the identification of a 2'-F RNA aptamer (RA16) that specifically binds to NCI-H460 non-small-cell lung cancer cells with an affinity (KD) of 9 ± 2 nM. Interestingly, RA16 potently inhibited cancer cell proliferation in a dose-dependent manner with an IC50 of 116.7 nM. When tested in vivo in xenografted mice, RA16 showed gradual migration toward tumor and accumulation at tumor site over time. An in vivo anti-cancer study showed that the average inhibition rate for mouse tumors in the RA16-treated group was 54.26% ± 5.87% on day 16 versus the control group. The aptamer RA16 adducted with epirubicin (RA16-epirubicin) showed significantly higher toxicity against targeted NCI-H460 cells and low toxicity against non-targeted tumor cells. Furthermore, RA16-epirubicin adduct exhibited in vivo anti-cancer efficacy, with an inhibition rate of 64.38% ± 7.92% when administrated in H460 xenograft mouse model. In summary, a specific bi-functional RNA aptamer RA16 was selected targeting and inhibiting toward NCI-H460 in vitro and in vivo.

Lipid vesicular nanocarrier: Quick encapsulation efficiency determination and transcutaneous application.

Nanoscale delivery systems have been widely investigated to overcome the penetration barrier of stratum corneum for effective transcutaneous application. The aim of this study is the development of effective vesicular formulations of ovalbumin and saponin which are able to promote penetration through the skin layers. Three kinds of vesicular formulations have been investigated as carriers, including liposomes, transfersomes and ethosomes, in which cholesterol and/or cationic lipid stearylamine are incorporated. The impact of membrane composition variations on the protein entrapment has been evaluated for each vesicle type. Formulations were characterized for particle size, polydispersity and encapsulation efficiency. The best formulations for each type of vesicle were subjected to in vivo transdermal immunization in mice. Among the three kinds of vesicular carrier, ethosomal nano carrier not only showed the best stability over a two months' storage, but also enabled the highest increase in the titer of serum antibody. In this regard, cationic nano-ethosomes can be considered as a promising vesicular carrier for transdermal vaccines. Meanwhile, we have developed a simple method to determine encapsulation efficiency of vesicular systems, which has potential application as a high throughput screening for vesicular formulations.

[Expression and characterization of human sTNFR II-IgG Fc fusion protein in Pichia pastoris].

AIM: To find if human soluble tumor necrosis factor receptor II (p75) fused IgG Fc protein (sTNFR II-IgG Fc) could be expressed in Pichia pastoris with an active dimmer form and characterize its N-linked oligosaccharides. METHODS: Two gene fragment, human sTNFR II and IgGFc, were got by RT-PCR from leucocytes stimulated with LPS. And the chimeric gene sTNFR II-IgG Fc achieved through gene splicing by over lap extension (SOE) method was cloned into pPIC9 and transformed into methanotropic yeast Pichia pastoris. The fusion protein purified by Protein A affinity column was analyzed with SDS-PAGE electrophoresis under reducing or non-reducing conditions and immunological methods. The anti-TNF-alpha biological activity assay of fusion protein was performed with L929 cells and detected with MTT colorimetry. The N-linked oligosaccharides hydrolyzed from fusion protein were labeled with 8-amino-1, 3, 6-naphthalene trisulfonic acid (ANTS) were analyzed with fluorophore-assisted carbohydrate eletrophoresis (FACE) as well. RESULTS: The recombinant P. pastoris strain that expressed human sTNFR II-IgG Fc fusion protein was constructed. The expression level of fusion protein in 2 L flask reached 2 mg/L. SDS-PAGE and Western blot showed the expressed fusion protein purified by protein was a dimer linked with inter-molecular disulfide linkage. The fusion protein neutralized cytotoxic activity of TNF-alpha to L929 cells, and the EC(50) of the fusion protein to inhibit 5 x 10(4) U/L of TNF-alpha was 170 microg/L. The FACE analysis showed there are 11 to 13 hexoses on each N-linked oligosaccharide. CONCLUSION: The human sTNFR II-IgG Fc fusion protein is expressed successfully in P. pastoris and it could be a reference for the future expression of other Fc fusion proteins or immunoglobulins in Pichia pastoris.