Direct interaction of platelet with tumor cell aggravates hepatocellular carcinoma metastasis by activating TLR4/ADAM10/CX3CL1 axis.

Metastasis is the main culprit of cancer-related death and account for the poor prognosis of hepatocellular carcinoma. Although platelets have been shown to accelerate tumor cell metastasis, the exact mechanism remained to be fully understood. Here, we found that high blood platelet counts and increased tumor tissue ADAM10 expression indicated the poor prognosis of HCC patients. Meanwhile, blood platelet count has positive correlation with tumor tissue ADAM10 expression. In vitro, we revealed that platelet increased ADAM10 expression in tumor cell through TLR4/NF-κB signaling pathway. ADAM10 catalyzed the shedding of CX3CL1 which bound to CX3CR1 receptor, followed by inducing epithelial to mesenchymal transition and activating RhoA signaling in cancer cells. Moreover, knockdown HCC cell TLR4 (Tlr4) or inhibition of ADAM10 prevented platelet-increased tumor cell migration, invasion and endothelial permeability. In vivo, we further verified in mice lung metastatic model that platelet accelerated tumor metastasis via cancer cell TLR4/ADAM10/CX3CL1 axis. Overall, our study provides new insights into the underlying mechanism of platelet-induced HCC metastasis. Therefore, targeting the TLR4/ADAM10/CX3CL1 axis in cancer cells hold promise for the inhibition of platelet-promoted lung metastasis of HCC.

Investigation on Filtration Control of Zwitterionic Polymer AADN in High Temperature High Pressure Water-Based Drilling Fluids.

With the exploration and development of high-temperature and high-salt deep oil and gas, more rigorous requirements are warranted for the performance of water-based drilling fluids (WBDFs). In this study, acrylamide, 2-acrylamide-2-methylpropanesulfonic acid, diallyl dimethyl ammonium chloride, and N-vinylpyrrolidone were synthesized by free radical copolymerization in an aqueous solution to form a temperature and salt-resistant zwitterionic polymer gel filtration loss reducer (AADN). The zwitterionic polymer had excellent adsorption and hydration groups, which could effectively combine with bentonite through hydrogen bonds and electrostatic attraction, strengthening the hydration film thickness on the surface of bentonite, and promoting the stable dispersion of drilling fluid. In addition, the reverse polyelectrolyte effect of zwitterionic polymers strengthened the drilling fluid's ability to resist high-temperature and high-salt. The AADN-based drilling fluid showed excellent rheological and filtration control properties (FLAPI < 8 mL, FLHTHP < 29.6 mL) even after aging at high-temperature (200 °C) and high-salt (20 wt% NaCl) conditions. This study provides a new strategy for simultaneously improving the high-temperature and high-salt tolerance of WBDFs, presenting the potential for application in drilling in high-temperature and high-salt deep formations.

New Zwitterionic Polymer as a Highly Effective Salt- and Calcium-Resistant Fluid Loss Reducer in Water-Based Drilling Fluids.

To control the filtration loss of drilling fluids in salt-gypsum formations, a novel type of zwitterionic polymer gel (DNDAP) was synthesized by free radical polymerization, which was used as a salt- and calcium-resistant fluid loss reducer for water-based drilling fluids (WBDF). DNDAP was prepared with N, N-dimethylacrylamide (DMAA), N-vinylpyrrolidone (NVP), Diallyl dimethyl ammonium chloride (DMDAAC), 2-acrylamide-2-methylpropaneonic acid (AMPS), and isopentenol polyether (TPEG) as raw materials. Fourier transform infrared spectroscopy (FT-IR) and proton nuclear magnetic resonance (1H-NMR) were used to characterize the composition and structure of the DNDAP copolymer. The thermal stability of DNDAP was evaluated by the use of thermogravimetric analysis (TGA). WBDF with DNDAP was analyzed for zeta potential and particle size and the corresponding filter cake underwent energy dispersive spectrum (EDS) analysis and scanning electron microscope (SEM) analysis. The results showed that the thermal decomposition of DNDAP mainly occurred above 303 °C. DNDAP exhibits excellent rheological and filtration properties in water-based drilling fluids, even under high-temperature aging (up to 200 °C) and high salinity (20 wt% NaCl or 5 wt% CaCl2) environments. The strong adsorption effect of DNDAP makes the particle size of bentonite reasonably distributed to form a dense mud cake that reduces filtration losses.

Platelet-Derived TGF (Transforming Growth Factor)-ß1 Enhances the Aerobic Glycolysis of Pulmonary Arterial Smooth Muscle Cells by PKM2 (Pyruvate Kinase Muscle Isoform 2) Upregulation.

BACKGROUND: Metabolic reprogramming is a hallmark of pulmonary arterial hypertension. Platelet activation has been implicated in pulmonary arterial hypertension (PAH), whereas the role of platelet in the pathogenesis of PAH remains unclear. METHODS: First, we explored the platelet function of semaxanib' a inhibitor of VEGF receptor (SU5416)/hypoxia mice and monocrotaline-injected rats PAH model. Then we investigated pulmonary arterial smooth muscle cell aerobic glycolysis after being treated with platelet supernatant. TGF (transforming growth factor)-ßRI, pyruvate kinase muscle 2, and other antagonists were applied to identify the underlying mechanism. In addition, platelet-specific deletion TGF-ß1 mice were exposed to chronic hypoxia and SU5416. Cardiopulmonary hemodynamics, vascular remodeling, and aerobic glycolysis of pulmonary arterial smooth muscle cell were determined. RESULTS: Here, we demonstrate that platelet-released TGF-ß1 enhances the aerobic glycolysis of pulmonary arterial smooth muscle cells after platelet activation via increasing pyruvate kinase muscle 2 expression. Mechanistically, platelet-derived TGF-ß1 regulate spyruvate kinase muscle 2 expression through mTOR (mammalian target of rapamycin)/c-Myc/PTBP-1(polypyrimidine tract binding protein 1)/hnRNPA-1(heterogeneous nuclear ribonucleoprotein A1) pathway. Platelet TGF-ß1 deficiency mice are significantly protected from SU5416 plus chronic hypoxia-induced PAH, including attenuated increases in right ventricular systolic pressure and less pulmonary vascular remodeling. Also, in Pf4cre+ Tgfb1fl/fl mice, pulmonary arterial smooth muscle cells showed lower glycolysis capacity and their pyruvate kinase muscle 2 expression decreased. CONCLUSIONS: Our data demonstrate that TGF-ß1 released by platelet contributes to the pathogenesis of PAH and further highlights the role of platelet in PAH.

Nanocarriers for pancreatic cancer imaging, treatments, and immunotherapies.

Pancreatic tumors are highly desmoplastic and immunosuppressive. Delivery and distribution of drugs within pancreatic tumors are compromised due to intrinsic physical and biochemical stresses that lead to increased interstitial fluid pressure, vascular compression, and hypoxia. Immunotherapy-based approaches, including therapeutic vaccines, immune checkpoint inhibition, CAR-T cell therapy, and adoptive T cell therapies, are challenged by an immunosuppressive tumor microenvironment. Together, extensive fibrosis and immunosuppression present major challenges to developing treatments for pancreatic cancer. In this context, nanoparticles have been extensively studied as delivery platforms and adjuvants for cancer and other disease therapies. Recent advances in nanotechnology have led to the development of multiple nanocarrier-based formulations that not only improve drug delivery but also enhance immunotherapy-based approaches for pancreatic cancer. This review discusses and critically analyzes the novel nanoscale strategies that have been used for drug delivery and immunomodulation to improve treatment efficacy, including newly emerging immunotherapy-based approaches. This review also presents important perspectives on future research directions that will guide the rational design of novel and robust nanoscale platforms to treat pancreatic tumors, particularly with respect to targeted therapies and immunotherapies. These insights will inform the next generation of clinical treatments to help patients manage this debilitating disease and enhance survival rates.

Biomaterial nanocarrier-driven mechanisms to modulate anti-tumor immunity.

Cancer immunotherapy approaches that utilize or enhance patients' inherent immunity have received extensive attention in the past decade. Biomaterial-based nanocarriers with tunable physicochemical properties offer significant promise in cancer immunotherapies. They can lower payload toxicity, provide sustained release of diverse payloads, and target specific disease site(s). Furthermore, nanocarrier-mediated immunotherapies can induce antigen-specific T lymphocytes, tissue-directed immune activation, and apoptosis of cancer cells all of which may comprise a new paradigm in cancer immunotherapy. This review describes key steps in biomaterial-mediated immune activation ranging from biomaterial surface protein adsorption, antigen presenting cell processing, and T cell activation. Nanocarrier-based immunomodulatory mechanisms including inherent adjuvanticity, enhanced cellular internalization, lymph node delivery, cross-presentation, and immunogenic cell death are discussed. In addition, studies that synergistically influence outcomes of nanocarrier-based combination immunotherapies are presented.

Polymeric Nanoparticle-Based Vaccine Adjuvants and Delivery Vehicles.

As vaccine formulations have progressed from including live or attenuated strains of pathogenic components for enhanced safety, developing new adjuvants to more effectively generate adaptive immune responses has become necessary. In this context, polymeric nanoparticles have emerged as a promising platform with multiple advantages, including the dual capability of adjuvant and delivery vehicle, administration via multiple routes, induction of rapid and long-lived immunity, greater shelf-life at elevated temperatures, and enhanced patient compliance. This comprehensive review describes advances in nanoparticle-based vaccines (i.e., nanovaccines) with a particular focus on polymeric particles as adjuvants and delivery vehicles. Examples of the nanovaccine approach in respiratory infections, biodefense, and cancer are discussed.

Polyanhydride nanoparticles stabilize pancreatic cancer antigen MUC4ß.

Pancreatic cancer (PC) is one of the most lethal malignancies and represents an increasing and challenging threat, especially with an aging population. The identification of immunogenic PC-specific upregulated antigens and an enhanced understanding of the immunosuppressive tumor microenvironment have provided opportunities to enable the immune system to recognize cancer cells. Due to its differential upregulation and functional role in PC, the transmembrane mucin MUC4 is an attractive target for immunotherapy. In the current study we characterized the antigen stability, antigenicity and release kinetics of a MUC4ß-nanovaccine to guide further optimization and, in vivo evaluation. Amphiphilic polyanhydride copolymers based on 20 mol % 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane and 80 mol % 1,6-bis(p-carboxyphenoxy)hexane were used to synthesize nanoparticles. Structurally stable MUC4ß protein was released from the particles in a sustained manner and characterized by gel electrophoresis and fluorescence spectroscopy. Modest levels of protein degradation were observed upon release. The released protein was also analyzed by MUC4ß-specific monoclonal antibodies using ELISA and showed no significant loss of epitope availability. Further, mice immunized with multiple formulations of combination vaccines containing MUC4ß-loaded nanoparticles generated MUC4ß-specific antibody responses. These results indicate that polyanhydride nanoparticles are viable MUC4ß vaccine carriers, laying the foundation for evaluation of this platform for PC immunotherapy.

Polyanhydride Nanoparticles Induce Low Inflammatory Dendritic Cell Activation Resulting in CD8+ T Cell Memory and Delayed Tumor Progression.

INTRODUCTION: Adjuvants and immunotherapies designed to activate adaptive immunity to eliminate infectious disease and tumors have become an area of interest aimed at providing a safe and effective strategy to prevent or eliminate disease. Existing approaches would benefit from the development of immunization regimens capable of inducing efficacious cell-mediated immunity directed toward CD8+ T cell-specific antigens. This goal is critically dependent upon appropriate activation of antigen-presenting cells (APCs) most notably dendritic cells (DCs). In this regard, polyanhydride particles have been shown to be effectively internalized by APCs and induce activation. METHODS: Here, a prophylactic vaccine regimen designed as a single-dose polyanhydride nanovaccine encapsulating antigen is evaluated for the induction of CD8+ T cell memory in a model system where antigen-specific protection is restricted to CD8+ T cells. Bone marrow-derived dendritic cells (BMDCs) are used as an in vitro model system to evaluate the magnitude and phenotype of APC activation. Primary DCs, particularly those with described ability to activate CD8+ T cells, are also evaluated for their in vitro responses to polyanhydride nanoparticles. RESULTS: Herein, polyanhydride nanoparticles are shown to induce potent in vitro upregulation of costimulatory molecules on the cell surface of BMDCs. In contrast to the classically used TLR agonists, nanoparticles did not induce large amounts of pro-inflammatory cytokines, did not induce characteristic metabolic response of DCs, nor produce innate antimicrobial effector molecules, such as nitric oxide (NO). The polyanhydride nanovaccine results in protective CD8+ T cell responses as measured by inhibition of tumor progression and survival. DISCUSSION: Together, these results suggest that the use of a polyanhydride-based nanovaccine can be an effective approach to inducing antigen-specific CD8+ T cell memory by providing antigen delivery and DC activation while avoiding overt inflammatory responses typically associated with traditional adjuvants.

Cyclic Stiffness Modulation of Cell-Laden Protein-Polymer Hydrogels in Response to User-Specified Stimuli including Light.

Although mechanical signals presented by the extracellular matrix are known to regulate many essential cell functions, the specific effects of these interactions, particularly in response to dynamic and heterogeneous cues, remain largely unknown. Here, we introduce a modular semisynthetic approach to create protein-polymer hydrogel biomaterials that undergo reversible stiffening in response to user-specified inputs. Employing a novel dual-chemoenzymatic modification strategy, we create fusion protein-based gel crosslinkers that exhibit stimuli-dependent intramolecular association. Linkers based on calmodulin yield calcium-sensitive materials, while those containing the photosensitive LOV2 (light, oxygen, and voltage sensing domain 2) protein give phototunable constructs whose moduli can be cycled on demand with spatiotemporal control about living cells. We exploit these unique materials to demonstrate the significant role that cyclic mechanical loading plays on fibroblast-to-myofibroblast transdifferentiation in three-dimensional (3D) space. Our moduli-switchable materials should prove useful for studies in mechanobiology, providing new avenues to probe and direct matrix-driven changes in 4D cell physiology.

Investigation of Mesoporous Graphitic Carbon Nitride as the Adsorbent to Remove Ni (II) Ions.

The mesoporous graphitic carbon nitride (mpg-C3N4/r, r was defined as the initial silica/dicyandiamide mass ratio) was successfully synthesized by heating the mixture of silica and dicyandiamide in a nitrogen atmosphere. The morphology and structure of mpg-C3N4/r were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), Brunauer-Emmett-Teller surface area measurement (BET), X-ray powder diffraction (XRD), and Fourier Transform Infrared spectroscopy (FT-IR). The adsorption performances of Ni (II) ions by mpg-C3N4/r were investigated. With increasing of r value, the BET specific surface area of the synthesized mpg-C3N4/r increased; the highest specific surface area of mpg-C3N4/1.5 increased up to 169.3 m2/g. This work shows that mpg-C3N4/1.5 is a promising, high-efficiency adsorbent that can be used to purify the water of a low Ni (II) ions concentration. The maximum adsorption capacity of Ni(II) ions by mpg-C3N4/1.5 was 15.26 mg/g. The adsorption properties of Ni (II) ions by mpg-C3N4/r complied well with pseudo-second-order kinetics and Langmuir isotherm model.