Development of resistance to chemo- and immunotherapies often occurs following treatment of melanoma brain metastasis (MBM). The brain microenvironment (BME), particularly astrocytes, cooperate ...toward MBM progression by upregulating secreted factors, among which we found that monocyte chemoattractant protein-1 (MCP-1) and its receptors, CCR2 and CCR4, were overexpressed in MBM compared with primary lesions. Among other sources of MCP-1 in the brain, we show that melanoma cells altered astrocyte secretome and evoked MCP-1 expression and secretion, which in turn induced CCR2 expression in melanoma cells, enhancing in vitro tumorigenic properties, such as proliferation, migration, and invasion of melanoma cells. In vivo pharmacological blockade of MCP-1 or molecular knockout of CCR2/CCR4 increased the infiltration of cytotoxic CD8+ T cells and attenuated the immunosuppressive phenotype of the BME as shown by decreased infiltration of Tregs and tumor-associated macrophages/microglia in several models of intracranially injected MBM. These in vivo strategies led to decreased MBM outgrowth and prolonged the overall survival of the mice. Our findings highlight the therapeutic potential of inhibiting interactions between BME and melanoma cells for the treatment of this disease.
Abstract Effective antibody responses are essential to generate protective humoral immunity. Different inflammatory signals polarize T cells towards appropriate effector phenotypes during an ...infection or immunization. Th1 and Th2 cells have been associated with the polarization of humoral responses. However, T follicular helper cells (Tfh) have a unique ability to access the B cell follicle and support the germinal center (GC) responses by providing B cell help. We investigated the specialization of Tfh cells induced under type-1 and type-2 conditions. We first studied homogenous Tfh cell populations generated by adoptively transferred TCR-transgenic T cells in mice immunized with type-1 and type-2 adjuvants. Using a machine learning approach, we established a gene expression signature that discriminates Tfh cells polarized towards type-1 and type-2 response, defined as Tfh1 and Tfh2 cells. The distinct signatures of Tfh1 and Tfh2 cells were validated against datasets of Tfh cells induced following lymphocytic choriomeningitis virus (LCMV) or helminth infection. We generated single-cell and spatial transcriptomics datasets to dissect the heterogeneity of Tfh cells and their localization under the two immunizing conditions. Besides a distinct specialization of GC Tfh cells under the two immunizations and in different regions of the lymph nodes, we found a population of Gzmk + Tfh cells specific for type-1 conditions. In human individuals, we could equally identify CMV-specific Tfh cells that expressed Gzmk. Our results show that Tfh cells acquire a specialized function under distinct types of immune responses and with particular properties within the B cell follicle and the GC.
Cancer is a heterogeneous disease that results from a multi-step process, being characterized by uncontrolled proliferation, invasion and metastasis. The understanding that tumor cells can be ...recognized by host immune cells has highlighted the potential advantages of using vaccination purposes to eliminate cancer cells, while avoiding severe side effects associated to conventional cancer treatments. Interesting outcomes have been obtained with the new identified tumor associated antigens (TAAs), including recombinant proteins and peptides. However, these molecules are weakly immunogenic, demanding the concomitant use of adjuvants to boost and achieve a strong tumor-specific immune response. Different classes of nanosystems have been used to protect and deliver several vaccine components. In vitro and preclinical studies have emphasized their promising role to attain a prolonged eradication of cancer cells, including metastasis. However, some studies support the co-entrapment of multiple adjuvants and TAAs within a single particulate carrier, while others indicate that stronger immune responses were obtained using a mixture of nanocarriers entrapping different combinations of TAAs and adjuvants. These apparently contradictory results may be related to nanocarrier physicochemical properties, which have a profound impact on their interaction with targeted cells and consequent biological effects. This review discusses the application of nanoscale systems as cancer vaccines, highlighting the particular characteristics of tumor biology and immunology that have been used to guide the design of these nanodelivery tools. We also aim to explore the major weaknesses that have prevented their wide application in the clinic to overcome the delivery, efficacy and safety issues associated to biological entities.
Produce biodegradable nanoparticles to target antigen-presenting cells (APCs) and evaluate their potential to be used as a vaccine delivery system.
Untargeted PEGylated ...poly(d,l-lactic-co-glycolide)-based nanoparticles and mannose-grafted nanoparticles were formulated and physicochemically characterized. Immortalized and primary APCs were used to study nanoparticle internalization patterns. The endocytic pathways and intracellular trafficking followed by nanoparticles were also investigated.
Nanoparticles displayed mannose residues available for binding at the nanoparticle surface. Different nanoparticle internalization patterns by immortalized and primary APCs were verified. Macropinocytosis, clathrin-mediated endocytosis, caveolin- and lipid raft-dependent endocytosis are involved in nanoparticles internalization. Nanoparticles demonstrate both endolysosomal and cytosolic localizations and a tendency to accumulate nearby the endoplasmic reticulum.
The developed nanoparticles might drive antigens to be presented through MHC class I and II molecules to both CD8(+) and CD4(+) T cells, favoring a complete and coordinated immune response.
Regulatory aspects on nanomedicines Sainz, Vanessa; Conniot, João; Matos, Ana I. ...
Biochemical and biophysical research communications,
12/2015, Letnik:
468, Številka:
3
Journal Article
Recenzirano
Nanomedicines have been in the forefront of pharmaceutical research in the last decades, creating new challenges for research community, industry, and regulators. There is a strong demand for the ...fast development of scientific and technological tools to address unmet medical needs, thus improving human health care and life quality. Tremendous advances in the biomaterials and nanotechnology fields have prompted their use as promising tools to overcome important drawbacks, mostly associated to the non-specific effects of conventional therapeutic approaches. However, the wide range of application of nanomedicines demands a profound knowledge and characterization of these complex products. Their properties need to be extensively understood to avoid unpredicted effects on patients, such as potential immune reactivity.
Research policy and alliances have been bringing together scientists, regulators, industry, and, more frequently in recent years, patient representatives and patient advocacy institutions. In order to successfully enhance the development of new technologies, improved strategies for research-based corporate organizations, more integrated research tools dealing with appropriate translational requirements aiming at clinical development, and proactive regulatory policies are essential in the near future.
This review focuses on the most important aspects currently recognized as key factors for the regulation of nanomedicines, discussing the efforts under development by industry and regulatory agencies to promote their translation into the market. Regulatory Science aspects driving a faster and safer development of nanomedicines will be a central issue for the next years.
•Nanotechnology has emerged as a “new technological revolution”.•Development of engineered nanosystems is challenging industry and regulatory agencies.•Global regulatory trends for nanopharmaceuticals are yet to be defined.•New quality control assays and robust techniques for nanosystems׳ characterization are needed.•Relation between physicochemical properties and biological effect to foster marketed nano-based products.
Dendrimers are hyperbranched polymers with a multifunctional architecture that can be tailored for the use in various biomedical applications. Peptide dendrimers are particularly relevant for drug ...delivery applications due to their versatility and safety profile. The overall lack of knowledge of their three-dimensional structure, conformational behavior and structure-activity relationship has slowed down their development. Fluorophores are often conjugated to dendrimers to study their interaction with biomolecules and provide information about their mechanism of action at the molecular level. However, these probes can change dendrimer surface properties and have a direct impact on their interactions with biomolecules and with lipid membranes. In this study, we have used computer-aided molecular design and molecular dynamics simulations to identify optimal topology of a poly(l-glutamic acid) (PG) backbone dendrimer that allows incorporation of fluorophores in the core with minimal availability for undesired interactions. Extensive all-atom molecular dynamic simulations with the CHARMM force field were carried out for different generations of PG dendrimers with the core modified with a fluorophore (nitrobenzoxadiazole and Oregon Green 488) and various surface groups (glutamic acid, lysine and tryptophan). Analysis of structural and topological features of all designed dendrimers provided information about their size, shape, internal distribution and dynamic behavior. We have found that four generations of a PG dendrimer are needed to ensure minimal exposure of a core-conjugated fluorophore to external environment and absence of undesired interactions regardless of the surface terminal groups. Our findings suggest that NBD-PG-G4 can provide a suitable scaffold to be used for biophysical studies of surface-modified dendrimers to provide a deeper understanding of their intermolecular interactions, mechanisms of action and trafficking in a biological system.
Fiumicino town in the Tiber River delta, near Rome International Airport (Italy), is historically affected by large amounts of carbon dioxide (CO2) in the ground and gas eruptions triggered by ...shallow drilling. While it is known that CO2 originates from carbonate thermometamorphism and/or mantle degassing, the origin of methane (CH4) associated with CO2 is uncertain and the outgassing spatial distribution is unknown. Combining isotope gas geochemistry, soil gas, and structural‐stratigraphic analyses, we provide evidence for a hybrid fluid source system, classifiable as Sediment‐Hosted Geothermal System (SHGS), where biotic CH4 from sedimentary rocks is carried by deep geothermic CO2 through active segments of a half‐graben. Molecular and isotopic composition of CH4 and concentration of heavier alkanes (ethane and propane), obtained from gas vents and soil gas throughout the delta area, reveal that thermogenic CH4 (up to 3.7 vol% in soil gas; δ13CCH4: −37 to −40‰ VPDB‐Vienna Peedee Belemnite, and δ2HCH4: −162 to −203‰ VSMOW ‐ Vienna Standard Mean Ocean Water in gas vents) prevails over possible microbial and abiotic components. The hydrocarbons likely result from known Meso‐Cenozoic petroleum systems of the Latium Tyrrhenian coast. Overmaturation of source rocks or molecular fractionation induced by gas migration are likely responsible for increased C1/C2+ ratios. CO2 and CH4 soil gas anomalies are scattered along NW‐SE and W‐E alignments, which, based on borehole, geomorphologic, and structural‐stratigraphic analyses, coincide with active faults of a half‐graben that seems to have controlled the recent evolution of the Tiber delta. This SHGS can be a source of considerable greenhouse gas emissions to the atmosphere and hazards for humans and buildings.
Key Points
Hybrid inorganic CO2 + biotic CH4 degassing system in the Tiber delta
Thermogenic CH4 (and CO2+ alkanes) transported by geothermal CO2 crossing a petroleum system
Degassing occurs along neotectonic fault systems forming the Fiumicino half‐graben
We hypothesized that the co-entrapment of melanoma-associated antigens and the Toll-like receptor (TLR) ligands Poly(I:C) and CpG, known to be Th1-immunopotentiators, in mannose-functionalized ...aliphatic polyester-based nanoparticles (NPs) could be targeted to mannose receptors on antigen-presenting cells and induce anti-tumor immune responses. High entrapment efficiencies of antigens and immunopotentiators in 150nm NPs were obtained. The co-entrapment of the model antigen ovalbumin and the TLR ligands was crucial to induce high IgG2c/IgG1 ratios and high levels of IFN-γ and IL-2. Mannose-functionalization of NPs potentiated the Th1 immune response. The nanoparticulate vaccines decreased the growth rate of murine B16F10 melanoma tumors in therapeutic and prophylatic settings. The combination of mannose-functionalized NPs containing MHC class I- or class II-restricted melanoma antigens and the TLR ligands induced the highest tumor growth delay. Overall, we demonstrate that the multifunctional properties of NPs in terms of targeting and antigen/adjuvant delivery have high cancer immunotherapeutic potential.
Display omitted
We use 13 new 40Ar/39Ar and 4 new 14C datings of volcanic deposits and organic material found within near-coastal aggradational successions deposited by the Tiber River near Rome, Italy, to integrate ...a larger dataset previously achieved in order to offer independent age constraints to the sea-level fluctuations associated with Late Quaternary glacial cycles during the last 450 ka. Results are compared with the chronologically independently constrained Red Sea relative sea-level curve, and with the astronomically tuned deep-sea benthic δ18O record. We find good agreements for the timings of change, and in several cases for both the amplitudes and timings of change during glacial terminations T-1, T-2, T-3, and T-5. There is one striking exception, namely for glacial termination T-4 that led into interglacial Marine Isotope Stage (MIS) 9. T-4 in our results is dated a full 18 ka earlier than in the Red Sea and deep-sea benthic δ18O records (which are in good agreement with each other in spite of their independent chronological constraints). The observed discrepancy is beyond the scale of the combined age uncertainties. One possible explanation is that the documented aggradation represents an early phase, triggered by a smaller event in the sea-level record, but the thickness of the aggradational sediment sequence then suggests that the amplitude of this earlier sea-level rise is underestimated in the Red Sea and benthic δ18O records. Also, this would imply that the aggradational succession of the main T-4 deglaciation has not yet been located in the study region, which is hard to reconcile with our extensive fieldwork and borehole coverage, unless unlikely non-deposition or complete erosion. Resolving this discrepancy will improve understanding of the timing of deglaciations relative to the orbitally modulated insolation forcing of climate and will require further focused research, both into the nature and chronology of the Tiber sequences of this period, and into the chronologies of the Red Sea and deep-sea benthic δ18O records.
•We provide age constraints to the aggradational successions of the Tiber River.•We demonstrate the glacio-eustatic forcing on sediment aggradation in the last 450 ka.•We use a conceptual sedimentary model as a proxy for glacial terminations.•We provide radioisotopic independent ages for the last five glacial terminations.•We discuss a possible misfit with astrochronological calibration of T-4 at 330 ka.
PDF
