The neurological devastation of neurodegenerative and cerebrovascular diseases reinforces our perseverance to find advanced treatments to deal with these fatal pathologies.High-performance preclinical results have fai...The neurological devastation of neurodegenerative and cerebrovascular diseases reinforces our perseverance to find advanced treatments to deal with these fatal pathologies.High-performance preclinical results have failed at clinical level,as it has been the case for a wide variety of neuroprotective agents and cell-based therapies employed to treat high prevalent brain pathologies such as stroke,Alzheimer’s and Parkinson’s diseases.An unquestionable reality is the current absence of effective therapies to neuroprotect the brain,to arrest neurodegeneration and rewire the impaired brain circuits.Part of the problem might arise from the lack of adequate in vitro and in vivo models and that most of the underlying pathophysiological mechanisms are not yet clarified.Another contributing factor is the lack of efficient systems to sustain drug release at therapeutic concentrations and enhance the survival and function of grafted cells in transplantation procedures.For medical applications the use of biomaterials of different compositions and formats has experienced a boom in the last decades.Although the greater complexity of central nervous system has probably conditioned their extensive use with respect to other organs,the number of biomaterials-based applications to treat the injured brain or in the process of being damaged has grown exponentially.Hydrogel-based biomaterials have constituted a turning point in the treatment of cerebral disorders using a new form of advanced therapy.Hydrogels show mechanical properties in the range of cerebral tissue resulting very suitable for local implantation of drugs and cells.It is also possible to fabricate three-dimensional hydrogel constructs with adaptable mesh size to facilitate axonal guidance and elongation.Along this article,we review the current trends in this area highlighting the positive impact of hydrogel-based biomaterials over the exhaustive control of drug delivery,cell engraftment and axonal reinnervation in brain pathologies.展开更多
Human neural stem cells（h NSCs） derived from the ventral mesencephalon are powerful research tools and candidates for cell therapies in Parkinson＇s disease. However, their clinical translation has not been fully re...Human neural stem cells（h NSCs） derived from the ventral mesencephalon are powerful research tools and candidates for cell therapies in Parkinson＇s disease. However, their clinical translation has not been fully realized due, in part, to the limited ability to track stem cell regional localization and survival over long periods of time after in vivo transplantation. Magnetic resonance imaging provides an excellent non-invasive method to study the fate of transplanted cells in vivo. For magnetic resonance imaging cell tracking, cells need to be labeled with a contrast agent, such as magnetic nanoparticles, at a concentration high enough to be easily detected by magnetic resonance imaging. Grafting of human neural stem cells labeled with magnetic nanoparticles allows cell tracking by magnetic resonance imaging without impairment of cell survival, proliferation, self-renewal, and multipotency. However, the results reviewed here suggest that in long term grafting, activated microglia and macrophages could contribute to magnetic resonance imaging signal by engulfing dead labeled cells or iron nanoparticles dispersed freely in the brain parenchyma over time.展开更多
Venous thromboembolism(VTE) is an illness that has a potentially life-threatening condition that affects a large percentage of the global population. VTE with pulmonary embolism(PE) is the third leading cause of death...Venous thromboembolism(VTE) is an illness that has a potentially life-threatening condition that affects a large percentage of the global population. VTE with pulmonary embolism(PE) is the third leading cause of death after myocardial infarction and stroke. In the first three months after an acute PE, there is an estimated 15% mortality among submassive PE, and 68% mortality in massive PE. Current guidelines suggest fibrinolytic therapy regarding the clinical severity, however some studies suggest a more aggressive treatment approach. This review will summarize the available endovascular treatments and the different techniques with its indications and outcomes.展开更多
AIM To assess in rodent and human adipocytes the antilipolytic capacity of hexaquis(benzylammonium)decavanadate(B6V10),previously shown to exert antidiabetic effects in rodent models,such as lowering free fatty acids(...AIM To assess in rodent and human adipocytes the antilipolytic capacity of hexaquis(benzylammonium)decavanadate(B6V10),previously shown to exert antidiabetic effects in rodent models,such as lowering free fatty acids(FFA)and glucose circulating levels.METHODS Adipose tissue(AT)samples were obtained after informed consent from overweight women undergoing plastic surgery.Comparison of the effects of B6V10 and reference antilipolytic agents(insulin,benzylamine,vanadate)on the lipolytic activity was performed on adipocytes freshly isolated from rat,mouse and human AT.Glycerol release was measured using colorimetric assay as an index of lipolytic activity.The influence of B6V10 and reference agents on glucose transport into human fat cells was determined using the radiolabelled 2-deoxyglucose uptake assay.RESULTS In all the species studied,B6V10 exhibited a dosedependent inhibition of adipocyte lipolysis when triglyceride breakdown was moderately enhanced byβ-adrenergic receptor stimulation.B6V10 exerted on human adipocyte a maximal lipolysis inhibition of glycerol release that was stronger than that elicited by insulin.However,B6V10 did not inhibit basal and maximally stimulated lipolysis.When incubated at dose≥10μmol/L,B6V10 stimulated by twofold the glucose uptake in human fat cells,but-similarly to benzylamine-without reaching the maximal effect of insulin,while it reproduced one-half of the insulin-stimulation of lipogenesis in mouse fat cells.CONCLUSION B6V10 exerts insulin-like actions in adipocytes,including lipolysis inhibition and glucose transport activation.B6V10 may be useful in limiting lipotoxicity related to obesity and insulin resistance.展开更多
Diabetes mellitus is a leading cause of acquired vision loss and one of the world＇s fastest growing chronic diseases. Diabetic retinopathy （DR）, a specific complication of chronic hyperglycemia, is the leading caus...Diabetes mellitus is a leading cause of acquired vision loss and one of the world＇s fastest growing chronic diseases. Diabetic retinopathy （DR）, a specific complication of chronic hyperglycemia, is the leading cause of acquired vision loss worldwide in middle-aged and there- fore economically active people that also increases the medical and economic burden on the society （Klein, 2007）.展开更多
Balancing the risks and benefits of organophosphate pesticides(OPs)on human and environmental health relies partly on their accurate measurement.A highly sensitive fluorescence anti-quenching multi-residue bio-barcode...Balancing the risks and benefits of organophosphate pesticides(OPs)on human and environmental health relies partly on their accurate measurement.A highly sensitive fluorescence anti-quenching multi-residue bio-barcode immunoassay was developed to detect OPs(triazophos,parathion,and chlorpyrifos)in apples,turnips,cabbages,and rice.Gold nanoparticles were functionalized with monoclonal antibodies against the tested OPs.DNA oligonucleotides were complementarily hybridized with an RNA fluorescent label for signal amplification.The detection signals were generated by DNA-RNA hybridization and ribonuclease H dissociation of the fluorophore.The resulting fluorescence signal enables multiplexed quantification of triazophos,parathion,and chlorpyrifos residues over the concentration range of 0.01-25,0.01-50,and 0.1-50 ng/mL with limits of detection of 0.014,0.011,and 0.126 ng/mL,respectively.The mean recovery ranged between 80.3% and 110.8% with relative standard deviations of 7.3%-17.6%,which correlate well with results obtained by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The proposed bio-barcode immunoassay is stable,reproducible and reliable,and is able to detect low residual levels of multi-residue OPs in agricultural products.展开更多
The brain is a high-energy demanding organ,consuming around 20%of the metabolic energy generated.To fulfill this demand,cerebral blood flow(CBF)supplies oxygen and glucose continuously through the intricate network of...The brain is a high-energy demanding organ,consuming around 20%of the metabolic energy generated.To fulfill this demand,cerebral blood flow(CBF)supplies oxygen and glucose continuously through the intricate network of cerebral blood vessels.Although for many years brain activity and blood flow were conceived as independent processes,MRI-based functional brain imaging demonstrated that there is a coupling between them.展开更多
Among all contaminants of emerging interest,drugs are the ones that give rise to the greatest concern.Any of the multiple stages of the drug's life cycle(production,consumption and waste management)is a possible e...Among all contaminants of emerging interest,drugs are the ones that give rise to the greatest concern.Any of the multiple stages of the drug's life cycle(production,consumption and waste management)is a possible entry point to the different environmental matrices.Psychiatric drugs have received special attention because of two reasons.First,their use is increasing.Second,many of them act on phylogenetically highly conserved neuroendocrine systems,so they have the potential to affect many non-target organisms.Currently,wastewater is considered the most important source of drugs to the environment.Furthermore,the currently available wastewater treatment plants are not specifically prepared to remove drugs,so they reach practically all environmental matrices,even tap water.As drugs are designed to produce pharmacological effects at low concentrations,they are capable of producing ecotoxicological effects on microorganisms,flora and fauna,even on human health.It has also been observed that certain antidepressants and antipsychotics can bioaccumulate along the food chain.Drug pollution is a complicated and diffuse problem characterized by scientific uncertainties,a large number of stakeholders with different values and interests,and enormous complexity.Possible solutions consist on acting at source,using medicines more rationally,eco-prescribing or prescribing greener drugs,designing pharmaceuticals that are more readily biodegraded,educating both health professionals and citizens,and improving coordination and collaboration between environmental and healthcare sciences.Besides,end of pipe measures like improving or developing new purification systems(biological,physical,chemical,combination)that eliminate these residues efficiently and at a sustainable cost should be a priority.Here,we describe and discuss the main aspects of drug pollution,highlighting the specific issues of psychiatric drugs.展开更多
The characterization and isolation of various stem cell populations, from embryonic to tissue-derived stem cells and induced pluripotent stem cells (iPSCs), have led to a rapid growth in the field of stem cell researc...The characterization and isolation of various stem cell populations, from embryonic to tissue-derived stem cells and induced pluripotent stem cells (iPSCs), have led to a rapid growth in the field of stem cell research and its potentially clinical application in the field of regenerative medicine and tissue repair. Stem cell therapy has recently progressed from the preclinical to the early clinical trial arena for a variety of diseases states, although further knowledge on action mechanisms, long-term safety issues, and standardization and characterization of the therapeutic cell products remains to be thoroughly elucidated. In this paper we summarize the current state of the art of basic and clinical research that were highlighted at the 2012 meeting of the Spanish Cell Therapy Network. This includes the current research involving in genomic and transcriptomic characterization of selected stem cell populations, studies of the role of resident and transplanted stem cells during tissue regeneration and their mechanism of action, improved new strategies of tissue engineering, transplantation of mesenchymal stem cells (MSCs) in different animal models of disease, disease correction by iPSCs, and preliminary results of cell therapy in human clinical trials.展开更多
3D-printing has emerged as a leading technology for fabricating personalized scaffolds for bone regeneration.Among the 3D-printing technologies,vat photopolymerization(VP)stands out for its high precision and versatil...3D-printing has emerged as a leading technology for fabricating personalized scaffolds for bone regeneration.Among the 3D-printing technologies,vat photopolymerization(VP)stands out for its high precision and versatility.It enables the creation of complex,patient-specific scaffolds with advanced pore architectures that enhance mechanical stability and promote cell growth,key factors for effective bone regeneration.This review provides an overview of the advances made in vat photopolymerization printing of calcium phosphates,covering both the fabrication of full ceramic bodies and polymer-calcium phosphate composites.The review examines key aspects of the fabrication process,including slurry composition,architectural design,and printing accuracy,highlighting their impact on the mechanical and biological performance of 3D-printed scaffolds.The need to tailor porosity,pore size,and geometric design to achieve both mechanical integrity and biological functionality is emphasized by a review of data published in the recent literature.This review demonstrates that advanced geometries like Triply Periodic Minimal Surfaces and nature-inspired designs,achievable with exceptional precision by this technology,enhance mechanical and osteogenic performance.In summary,VP’s versatility,driven by the diversity of material options,consolidation methods,and precision opens new horizons for scaffold-based bone regeneration.展开更多
Although breast cancer(BC)predominantly affects women over 50,about 5%of cases occur in very young women,aged 35 years or younger,presenting significant clinical challenges and poor outcomes.To explore ageassociated m...Although breast cancer(BC)predominantly affects women over 50,about 5%of cases occur in very young women,aged 35 years or younger,presenting significant clinical challenges and poor outcomes.To explore ageassociated molecular differences in BC,we evaluated the transcriptomic profiles of tumors from very young and older patients,focusing on the tumor microenvironment(TME).Methods are described in the Supplementary file.This study enrolled 66 patients from Hospital Clínico Universitario of Valencia,stratified into very young(≤35 years,premenopausal)and older(>50 years,postmenopausal)BC patients.RNA-sequencing was performed on untreated primary tumors,and data were collected from 22 very young(median:32.5 years;range:22-35 years)and 27 older(median:66 years;range:50-94 years)BC patients who met quality criteria(clinicopathological characteristics in Supplementary Table S1).Principal component analysis illustrated increased heterogeneity among very young HR^(+)/HER2^(−)BC samples(Figure 1A).Differential expression analysis revealed 82 downregulated and 12 upregulated genes in very young BC samples(log2 fold change≥|2|;adjusted P<0.05)(Figure 1B-C,Supplementary Table S2).展开更多
Over the past years, significant efforts have been devoted to explore novel drug delivery and detection strategies for simultaneous therapy and diagnostics. The development of biotinylated polyurethane-urea nanopartic...Over the past years, significant efforts have been devoted to explore novel drug delivery and detection strategies for simultaneous therapy and diagnostics. The development of biotinylated polyurethane-urea nanoparticles as theranostic nanocarriers for targeted drug and plasmid delivery, for fluorescence detection of human hepatocellular carcinoma cells, is described herein. These targeted nanoparticles are specifically designed to incorporate biotin into the polymeric matrix, since many tumor types overexpress receptors for biotin as a mechanism to boost uncontrolled cell growth. The obtained nanoparticles were spherical, exhibited an average diameter ranging 110-145 nm, and showed no cytotoxicity in healthy endothelial cells. Biotinylated nanoparticles are selectively incorporated into the perinuclear and nuclear area of the human hepatocellular carcinoma cell line, HepG2, in division, but not into growing, healthy, human endothelial cells. Indeed, the simultaneous incorporation of the anticancer drugs, phenoxodiol or sunitinib, together with plasmid DNA encoding green fluorescent protein, into these nanoparticles allows a targeted pharmacological antitumor effect and furthermore, selective transfection of a reporter gene, to detect these cancer cells. The combined targeted therapy and detection strategy described here could be exploited for liver cancer therapy and diagnostics, with a moderate safety profile, and may also be a potential tool for other types of cancer.展开更多
The generation of functional retinal pigment epithelium （RPE） is of great therapeutic interest to the field of regenerative medicine and may provide possible cures for retinal degenerative diseases, including age-re...The generation of functional retinal pigment epithelium （RPE） is of great therapeutic interest to the field of regenerative medicine and may provide possible cures for retinal degenerative diseases, including age-related macular degeneration （AMD）. Although RPE cells can be produced from either embryonic stem cells or induced pluripotent stem cells, direct cell reprogramming driven by lineage-determining transcription factors provides an immediate route to their generation. By monitoring a human RPE specific Bestl：：GFP reporter, we report the conversion of human fibroblasts into RPE lineage using defined sets of transcription factors. We found that Bestl：：GFP positive cells formed colonies and exhibited morphological and molecular features of early stage RPE cells. Moreover, they were able to obtain pigmen- tation upon activation of Retinoic acid （RA） and Sonic Hedgehog （SHH） signaling pathways. Our study not only established an ideal platform to investigate the tran- scriptional network regulating the RPE cell fate deter- mination, but also provided an alternative strategy to generate functional RPE cells that complement the useof pluripotent stem cells for disease modeling, drug screening, and cell therapy of retinal degeneration.展开更多
The ultrastructure of the bone provides a unique mechanical strength against compressive, torsional andtensional stresses. An elastin-like recombinamer (ELR) with a nucleation sequence for hydroxyapatitewas incorporat...The ultrastructure of the bone provides a unique mechanical strength against compressive, torsional andtensional stresses. An elastin-like recombinamer (ELR) with a nucleation sequence for hydroxyapatitewas incorporated into films prepared from a collagen-silk fibroin blend carrying microchannel patternsto stimulate anisotropic osteogenesis. SEM and fluorescence microscopy showed the alignment ofadipose-derived stem cells (ADSCs) and the human osteoblasts (HOBs) on the ridges and in the groovesof microchannel patterned collagen-fibroin-ELR blend films. The Young's modulus and the ultimatetensile strength (UTS) of untreated films were 0.58 ± 0.13 MPa and 0.18 ± 0.05 MPa, respectively. After 28days of cell culture, ADSC seeded film had a Young's modulus of 1.21 ± 0.42 MPa and UTS of0.32 ± 0.15 MPa which were about 3 fold higher than HOB seeded films. The difference in Young'smodulus was statistically significant (p: 0.02). ADSCs attached, proliferated and mineralized better thanthe HOBs. In the light of these results, ADSCs served as a better cell source than HOBs for bone tissueengineering of collagen-fibroin-ELR based constructs used in this study. We have thus shown theenhancement in the tensile mechanical properties of the bone tissue engineered scaffolds by usingADSCs.展开更多
The integration of implants or medical devices into the body tissues requires of good cell–material interactions.However,most polymeric materials used for these applications lack on biological cues,which enhanced mid...The integration of implants or medical devices into the body tissues requires of good cell–material interactions.However,most polymeric materials used for these applications lack on biological cues,which enhanced mid-and long-term implant failure due to weak integration with the surrounding tissue.Commonly used strategies for tissue–material integration focus on functionalization of the material surface by means of natural proteins or short peptides.However,the use of these biomolecules involves major drawbacks such as immunogenic problems and oversimplification of the constructs.Here,designed elastin-like recombinamers(ELRs)are used to enhance poly(methyl methacrylate)surface properties and compared against the use of short peptides.In this study,cell response has been analysed for different functionalization conditions in the presence and absence of a competing protein,which interferes on surface–cell interaction by unspecific adsorption on the interface.The study has shown that ELRs can induce higher rates of cell attachment and stronger cell anchorages than short peptides,being a better choice for surface functionalization.展开更多
The aim of the present study was the in vitro and in vivo analysis of a bi-layered 3D-printed scaffold combining a PLA layer and a biphasic PLA/bioglass G5 layer for regeneration of osteochondral defects in vivo Focus...The aim of the present study was the in vitro and in vivo analysis of a bi-layered 3D-printed scaffold combining a PLA layer and a biphasic PLA/bioglass G5 layer for regeneration of osteochondral defects in vivo Focus of the in vitro analysis was on the(molecular)weight loss and the morphological and mechanical variations after immersion in SBF.The in vivo study focused on analysis of the tissue reactions and differences in the implant bed vascularization using an established subcutaneous implantation model in CD-1 mice and established histological and histomorphometrical methods.Both scaffold parts kept their structural integrity,while changes in morphology were observed,especially for the PLA/G5 scaffold.Mechanical properties decreased with progressive degradation,while the PLA/G5 scaffolds presented higher compressive modulus than PLA scaffolds.The tissue reaction to PLA included low numbers of BMGCs and minimal vascularization of its implant beds,while the addition of G5 lead to higher numbers of BMGCs and a higher implant bed vascularization.Analysis revealed that the use of a bi-layered scaffold shows the ability to observe distinct in vivo response despite the physical proximity of PLA and PLA/G5 layers.Altogether,the results showed that the addition of G5 enables to reduce scaffold weight loss and to increase mechanical strength.Furthermore,the addition of G5 lead to a higher vascularization of the implant bed required as basis for bone tissue regeneration mediated by higher numbers of BMGCs,while within the PLA parts a significantly lower vascularization was found optimally for chondral regeneration.Thus,this data show that the analyzed bi-layered scaffold may serve as an ideal basis for the regeneration of osteochondral tissue defects.Additionally,the results show that it might be able to reduce the number of experimental animals required as it may be possible to analyze the tissue response to more than one implant in one experimental animal.展开更多
The current study investigated a triad,which comprises of adipose tissue derived stem cells isolated from infrapatellar fat pad and gelatin/polyvinyl alcohol(PVA)-based matrix with exclusive ascorbic acid signalling.T...The current study investigated a triad,which comprises of adipose tissue derived stem cells isolated from infrapatellar fat pad and gelatin/polyvinyl alcohol(PVA)-based matrix with exclusive ascorbic acid signalling.Though,the bio-mechanical properties of the gelatin–PVA blended scaffolds in wet condition are equivalent to the ECM of soft tissues in general,in this study,the triad was tested as a model for neural tissue engineering.Apart from being cytocompatible and biocompatible,the porosity of the scaffold has been designed in such a manner that it facilitates the cell signalling and enables the exchange of nutrients and gases.The highly proliferative stem cells from Passage 2 were characterized using both,mesenchymal and embryonic stem cell markers.As an initial exploration the mesenchymal stem cells at Passage 4 were exposed to ascorbic acid and basic fibroblast growth factor signalling for neuronal differentiation in 2D environment independently.The MSCs successfully differentiated and acquired neuron specific markers related to cytoskeleton and synapses.Subsequently,three phases of experiments have been conducted on the 3D gelatin/PVA matrix to prove their efficacy,the growth of stem cells,growth of differentiated neurons and the in situ growth and differentiation of MSCs.The scaffold was conducive and directed MSCs to neuronal lineage under specific signalling.Overall,this organotypic model triad could open a new avenue in the field of soft tissue engineering as a simple and effective tissue construct.展开更多
Loco-regional recurrences and distant metastases represent the main cause of head and neck squamous cell carcinoma(HNSCC) mortality. The overexpression of chemokine receptor 4(CXCR4) in HNSCC primary tumors associates...Loco-regional recurrences and distant metastases represent the main cause of head and neck squamous cell carcinoma(HNSCC) mortality. The overexpression of chemokine receptor 4(CXCR4) in HNSCC primary tumors associates with higher risk of developing loco-regional recurrences and distant metastases, thus making CXCR4 an ideal entry pathway for targeted drug delivery. In this context, our group has generated the self-assembling protein nanocarrier T22-GFP-H6, displaying multiple T22 peptidic ligands that specifically target CXCR4. This study aimed to validate T22-GFP-H6 as a suitable nanocarrier to selectively deliver cytotoxic agents to CXCR4^(+)tumors in a HNSCC model. Here we demonstrate that T22-GFP-H6 selectively internalizes in CXCR4^(+)HNSCC cells, achieving a high accumulation in CXCR4^(+)tumors in vivo, while showing negligible nanocarrier distribution in non-tumor bearing organs. Moreover, this T22-empowered nanocarrier can incorporate bacterial toxin domains to generate therapeutic nanotoxins that induce cell death in CXCR4-overexpressing tumors in the absence of histological alterations in normal organs. Altogether, these results show the potential use of this T22-empowered nanocarrier platform to incorporate polypeptidic domains of choice to selectively eliminate CXCR4^(+)cells in HNSCC. Remarkably, to our knowledge, this is the first study testing targeted proteinonly nanoparticles in this cancer type, which may represent a novel treatment approach for HNSCC patients.展开更多
Poly-histidine peptides such as H6(HHHHHH)are used in protein biotechnologies as purification tags,protein-assembling agents and endosomal-escape entities.The pleiotropic properties of such peptides make them appealin...Poly-histidine peptides such as H6(HHHHHH)are used in protein biotechnologies as purification tags,protein-assembling agents and endosomal-escape entities.The pleiotropic properties of such peptides make them appealing to design protein-based smart materials or nanoparticles for imaging or drug delivery to be produced in form of recombinant proteins.However,the clinical applicability of H6-tagged proteins is restricted by the potential immunogenicity of these segments.In this study,we have explored several humanized histidine-rich peptides in tumor-targeted modular proteins,which can specifically bind and be internalized by the target cells through the tumoral marker CXCR4.We were particularly interested in exploring how protein purification,self-assembling and endosomal escape perform in proteins containing the variant histidine-rich tags.Among the tested candidates,the peptide H5 E(HEHEHEHEH)is promising as a good promoter of endosomal escape of the associated fulllength protein upon endosomal internalization.The numerical modelling of cell penetration and endosomal escape of the tested proteins has revealed a negative relationship between the amount of protein internalized into target cells and the efficiency of cytoplasmic release.This fact demonstrates that the His-mediated,proton sponge-based endosomal escape saturates at moderate amounts of internalized protein,a fact that might be critical for the design of protein materials for cytosolic molecular delivery.展开更多
Individuals infected with human immunodeficiency virus type-1(HIV-1)usually show a general dysregulation and hyper-activation of the immune system.A direct influence of HIV-1 particles on B-cell phenotypes and functio...Individuals infected with human immunodeficiency virus type-1(HIV-1)usually show a general dysregulation and hyper-activation of the immune system.A direct influence of HIV-1 particles on B-cell phenotypes and functions has been previously described.However,the consequences of B-cell dysregulation are still poorly understood.We evaluated the phenotypic changes in primary B cells after direct contact with HIV-1 particles in comparison with different types of stimuli.The functionality of treated B cells was challenged in co-culture experiments with autologous CD4+and CD8+T cells.We demonstrated that HIV-1 induces a phenotypic change in B cells towards a regulatory B-cell phenotype,showing a higher level of IL-10,TGF-β1,EBI3 or IL-12(p35)mRNA expression and acquiring an immunosuppressive profile.The acquisition of a Breg phenotype was confirmed by co-culture experiments where HIVtreated B cells reduced the proliferation and the TNFαproduction of CD4+or CD8+T cells.This suppressive ability of HIV-treated B cells was dependent on cell-to-cell contact between these B cells and effector cells.To our knowledge,these data provide the first evidence that HIV-1 can directly induce a regulatory B cell-like immunosuppressive phenotype,which could have the ability to impair specific immune responses.This dysregulation could constitute one of the mechanisms underlying unsuccessful efforts to develop an efficient vaccine against HIV-1.展开更多
基金supported by the Ministerio de Economia y Competitividad(MAT2016-79832-R,to DGN)the Community of Madrid(Neurocentro-B2017/BMD-3760,to DGN)
文摘The neurological devastation of neurodegenerative and cerebrovascular diseases reinforces our perseverance to find advanced treatments to deal with these fatal pathologies.High-performance preclinical results have failed at clinical level,as it has been the case for a wide variety of neuroprotective agents and cell-based therapies employed to treat high prevalent brain pathologies such as stroke,Alzheimer’s and Parkinson’s diseases.An unquestionable reality is the current absence of effective therapies to neuroprotect the brain,to arrest neurodegeneration and rewire the impaired brain circuits.Part of the problem might arise from the lack of adequate in vitro and in vivo models and that most of the underlying pathophysiological mechanisms are not yet clarified.Another contributing factor is the lack of efficient systems to sustain drug release at therapeutic concentrations and enhance the survival and function of grafted cells in transplantation procedures.For medical applications the use of biomaterials of different compositions and formats has experienced a boom in the last decades.Although the greater complexity of central nervous system has probably conditioned their extensive use with respect to other organs,the number of biomaterials-based applications to treat the injured brain or in the process of being damaged has grown exponentially.Hydrogel-based biomaterials have constituted a turning point in the treatment of cerebral disorders using a new form of advanced therapy.Hydrogels show mechanical properties in the range of cerebral tissue resulting very suitable for local implantation of drugs and cells.It is also possible to fabricate three-dimensional hydrogel constructs with adaptable mesh size to facilitate axonal guidance and elongation.Along this article,we review the current trends in this area highlighting the positive impact of hydrogel-based biomaterials over the exhaustive control of drug delivery,cell engraftment and axonal reinnervation in brain pathologies.
基金To AMS:Instituto de Salud Carlos-III(RETICS Ter Cel RD12/0019/0013)Comunidad Autónoma de Madrid(S2010-BMD-2336)+3 种基金MINECO(SAF2010-17167)the institutional grant of the Fundación Ramón Areces to the CBMSOTo MRG:Reina Sofia FoundationComunidad Autónoma Madrid(S2010-BMD-2460)
文摘Human neural stem cells（h NSCs） derived from the ventral mesencephalon are powerful research tools and candidates for cell therapies in Parkinson＇s disease. However, their clinical translation has not been fully realized due, in part, to the limited ability to track stem cell regional localization and survival over long periods of time after in vivo transplantation. Magnetic resonance imaging provides an excellent non-invasive method to study the fate of transplanted cells in vivo. For magnetic resonance imaging cell tracking, cells need to be labeled with a contrast agent, such as magnetic nanoparticles, at a concentration high enough to be easily detected by magnetic resonance imaging. Grafting of human neural stem cells labeled with magnetic nanoparticles allows cell tracking by magnetic resonance imaging without impairment of cell survival, proliferation, self-renewal, and multipotency. However, the results reviewed here suggest that in long term grafting, activated microglia and macrophages could contribute to magnetic resonance imaging signal by engulfing dead labeled cells or iron nanoparticles dispersed freely in the brain parenchyma over time.
文摘Venous thromboembolism(VTE) is an illness that has a potentially life-threatening condition that affects a large percentage of the global population. VTE with pulmonary embolism(PE) is the third leading cause of death after myocardial infarction and stroke. In the first three months after an acute PE, there is an estimated 15% mortality among submassive PE, and 68% mortality in massive PE. Current guidelines suggest fibrinolytic therapy regarding the clinical severity, however some studies suggest a more aggressive treatment approach. This review will summarize the available endovascular treatments and the different techniques with its indications and outcomes.
基金Supported by Institut National de la Santéet de la Recherche Médicale to the INSERM U1048
文摘AIM To assess in rodent and human adipocytes the antilipolytic capacity of hexaquis(benzylammonium)decavanadate(B6V10),previously shown to exert antidiabetic effects in rodent models,such as lowering free fatty acids(FFA)and glucose circulating levels.METHODS Adipose tissue(AT)samples were obtained after informed consent from overweight women undergoing plastic surgery.Comparison of the effects of B6V10 and reference antilipolytic agents(insulin,benzylamine,vanadate)on the lipolytic activity was performed on adipocytes freshly isolated from rat,mouse and human AT.Glycerol release was measured using colorimetric assay as an index of lipolytic activity.The influence of B6V10 and reference agents on glucose transport into human fat cells was determined using the radiolabelled 2-deoxyglucose uptake assay.RESULTS In all the species studied,B6V10 exhibited a dosedependent inhibition of adipocyte lipolysis when triglyceride breakdown was moderately enhanced byβ-adrenergic receptor stimulation.B6V10 exerted on human adipocyte a maximal lipolysis inhibition of glycerol release that was stronger than that elicited by insulin.However,B6V10 did not inhibit basal and maximally stimulated lipolysis.When incubated at dose≥10μmol/L,B6V10 stimulated by twofold the glucose uptake in human fat cells,but-similarly to benzylamine-without reaching the maximal effect of insulin,while it reproduced one-half of the insulin-stimulation of lipogenesis in mouse fat cells.CONCLUSION B6V10 exerts insulin-like actions in adipocytes,including lipolysis inhibition and glucose transport activation.B6V10 may be useful in limiting lipotoxicity related to obesity and insulin resistance.
基金supported by EU Program FP7-PEOPLE-2013-IAPP(612218/3D-NET)FEDER-CICYT MAT2013-47501-CO2-1-R(Ministry of Economy and Competitiveness,Spain)Grants+1 种基金supported by RETICS(RD12/0034/0001)Instituto de Salud CarlosⅢ,Spain
文摘Diabetes mellitus is a leading cause of acquired vision loss and one of the world＇s fastest growing chronic diseases. Diabetic retinopathy （DR）, a specific complication of chronic hyperglycemia, is the leading cause of acquired vision loss worldwide in middle-aged and there- fore economically active people that also increases the medical and economic burden on the society （Klein, 2007）.
基金supported by the Central Public Interest Scientific Institution Basal Research Fund for the Chinese Academy of Agricultural Sciences(Grant No.:Y2021PT05)National Institute of Environmental Health Science Superfund Research Program(Grant No.:P42 ES004699)+1 种基金National Academy of Sciences(Subaward No.:2000009144)Ningbo Innovation Project for Agro-Products Quality and Safety(Grant No.:2019CXGC007).
文摘Balancing the risks and benefits of organophosphate pesticides(OPs)on human and environmental health relies partly on their accurate measurement.A highly sensitive fluorescence anti-quenching multi-residue bio-barcode immunoassay was developed to detect OPs(triazophos,parathion,and chlorpyrifos)in apples,turnips,cabbages,and rice.Gold nanoparticles were functionalized with monoclonal antibodies against the tested OPs.DNA oligonucleotides were complementarily hybridized with an RNA fluorescent label for signal amplification.The detection signals were generated by DNA-RNA hybridization and ribonuclease H dissociation of the fluorophore.The resulting fluorescence signal enables multiplexed quantification of triazophos,parathion,and chlorpyrifos residues over the concentration range of 0.01-25,0.01-50,and 0.1-50 ng/mL with limits of detection of 0.014,0.011,and 0.126 ng/mL,respectively.The mean recovery ranged between 80.3% and 110.8% with relative standard deviations of 7.3%-17.6%,which correlate well with results obtained by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The proposed bio-barcode immunoassay is stable,reproducible and reliable,and is able to detect low residual levels of multi-residue OPs in agricultural products.
基金funded by the Ministry of Science,Innovation and Universities(MICIU)through the project NEUR-ON-A-CHIP(RTI2018-097038-B-C21 and RTI2018-097038-B-C22)(to MM,AL)the project UNIBBB(PDC2022-133918-C21)(to MM,AL)+4 种基金supported by Networking Biomedical Research Center(CIBER),Spain(to MM,AL)CIBER is an initiative funded by the VI National R&D&i Plan 2008–2011,Iniciativa Ingenio 2010,Consolider Program,CIBER Actions,and the Instituto de Salud Carlos III,with the support of the European Regional Development Fundfunded by the CERCA Programby the Commission for Universities and Research of the Department of Innovation,Universities,and Enterprise of the Generalitat de Catalunya(2017 SGR 1079)(to MM,AL)support from the program for predoctoral contracts for the training of doctors of the State Training Subprogram for the Promotion of Talent and its Employability in R+D+I(PRE2019-088286)by the Ministry of Science,Innovation and Universities(MICIU)。
文摘The brain is a high-energy demanding organ,consuming around 20%of the metabolic energy generated.To fulfill this demand,cerebral blood flow(CBF)supplies oxygen and glucose continuously through the intricate network of cerebral blood vessels.Although for many years brain activity and blood flow were conceived as independent processes,MRI-based functional brain imaging demonstrated that there is a coupling between them.
文摘Among all contaminants of emerging interest,drugs are the ones that give rise to the greatest concern.Any of the multiple stages of the drug's life cycle(production,consumption and waste management)is a possible entry point to the different environmental matrices.Psychiatric drugs have received special attention because of two reasons.First,their use is increasing.Second,many of them act on phylogenetically highly conserved neuroendocrine systems,so they have the potential to affect many non-target organisms.Currently,wastewater is considered the most important source of drugs to the environment.Furthermore,the currently available wastewater treatment plants are not specifically prepared to remove drugs,so they reach practically all environmental matrices,even tap water.As drugs are designed to produce pharmacological effects at low concentrations,they are capable of producing ecotoxicological effects on microorganisms,flora and fauna,even on human health.It has also been observed that certain antidepressants and antipsychotics can bioaccumulate along the food chain.Drug pollution is a complicated and diffuse problem characterized by scientific uncertainties,a large number of stakeholders with different values and interests,and enormous complexity.Possible solutions consist on acting at source,using medicines more rationally,eco-prescribing or prescribing greener drugs,designing pharmaceuticals that are more readily biodegraded,educating both health professionals and citizens,and improving coordination and collaboration between environmental and healthcare sciences.Besides,end of pipe measures like improving or developing new purification systems(biological,physical,chemical,combination)that eliminate these residues efficiently and at a sustainable cost should be a priority.Here,we describe and discuss the main aspects of drug pollution,highlighting the specific issues of psychiatric drugs.
基金supported by grants from the Ministry of Economy and Competitiveness(FIS PI10/02529,FIS EC07/90762,FIS PI12/00760,FIS PI13/00666)the Ministry of Science and Technology(BIO2009-13903-C02-02)+4 种基金the Andalusian Government(P07-CVI-2781,PAIDI BIO-217,PI-0729-2010)Spanish Cell Therapy Network(TerCel)and CIBER-BBN are an initiative funded by the VI National R&D&I Plan 2008-2011(RD06/0010/0023,RD12/0019/0001)Advanced Therapies and Transplant General Direction(Health Ministry,Spain)(TRA-137),Iniciativa Ingenio 2010,Consolider Program,CIBER Actions,and financed by the Instituto de Salud Carlos Ⅲ(ISC-Ⅲ)with assistance from the European Regional Development FundWork in Munoz-Chapuli’s laboratory is supported by grants BFU2011-25304,BFU2012-35799,P11-CTS-7564,and PITN-GA-2011-289600in Raya’s laboratory by grants SAF2012-33526,ACI2010-1117,and ISC-Ⅲ(TerCel,RD12/0019/0019).
文摘The characterization and isolation of various stem cell populations, from embryonic to tissue-derived stem cells and induced pluripotent stem cells (iPSCs), have led to a rapid growth in the field of stem cell research and its potentially clinical application in the field of regenerative medicine and tissue repair. Stem cell therapy has recently progressed from the preclinical to the early clinical trial arena for a variety of diseases states, although further knowledge on action mechanisms, long-term safety issues, and standardization and characterization of the therapeutic cell products remains to be thoroughly elucidated. In this paper we summarize the current state of the art of basic and clinical research that were highlighted at the 2012 meeting of the Spanish Cell Therapy Network. This includes the current research involving in genomic and transcriptomic characterization of selected stem cell populations, studies of the role of resident and transplanted stem cells during tissue regeneration and their mechanism of action, improved new strategies of tissue engineering, transplantation of mesenchymal stem cells (MSCs) in different animal models of disease, disease correction by iPSCs, and preliminary results of cell therapy in human clinical trials.
基金funded by the European Union-ERC grant(BAMBBI,101055053,doi:10.3030/101055053)The research group is part of Maria de Maeztu Units of Excellence Programme CEX2023-001300-M/funded by MCIN/AEI/10.13039/501100011033,Spanish Ministry of Science and Innovation.
文摘3D-printing has emerged as a leading technology for fabricating personalized scaffolds for bone regeneration.Among the 3D-printing technologies,vat photopolymerization(VP)stands out for its high precision and versatility.It enables the creation of complex,patient-specific scaffolds with advanced pore architectures that enhance mechanical stability and promote cell growth,key factors for effective bone regeneration.This review provides an overview of the advances made in vat photopolymerization printing of calcium phosphates,covering both the fabrication of full ceramic bodies and polymer-calcium phosphate composites.The review examines key aspects of the fabrication process,including slurry composition,architectural design,and printing accuracy,highlighting their impact on the mechanical and biological performance of 3D-printed scaffolds.The need to tailor porosity,pore size,and geometric design to achieve both mechanical integrity and biological functionality is emphasized by a review of data published in the recent literature.This review demonstrates that advanced geometries like Triply Periodic Minimal Surfaces and nature-inspired designs,achievable with exceptional precision by this technology,enhance mechanical and osteogenic performance.In summary,VP’s versatility,driven by the diversity of material options,consolidation methods,and precision opens new horizons for scaffold-based bone regeneration.
基金supported by grants from Fundación FERO,Asociación Española Contra el Cáncer(PRYCO211372RODR)the Biomedical Research Networking Centre for Oncology(CIBERONC)(CB16/12/00481).I.Garrido-Cano.was funded by the Margarita Salas postdoctoral grant(European Union-Next generation EU),M.Tapia was funded by SEOM-Río Hortega 2023,and A.L.and A.A.were funded by predoctoral fellowships from Asociación Española Contra el Cancer.J.M.Cejalvo was funded by Joan Rodes contract.
文摘Although breast cancer(BC)predominantly affects women over 50,about 5%of cases occur in very young women,aged 35 years or younger,presenting significant clinical challenges and poor outcomes.To explore ageassociated molecular differences in BC,we evaluated the transcriptomic profiles of tumors from very young and older patients,focusing on the tumor microenvironment(TME).Methods are described in the Supplementary file.This study enrolled 66 patients from Hospital Clínico Universitario of Valencia,stratified into very young(≤35 years,premenopausal)and older(>50 years,postmenopausal)BC patients.RNA-sequencing was performed on untreated primary tumors,and data were collected from 22 very young(median:32.5 years;range:22-35 years)and 27 older(median:66 years;range:50-94 years)BC patients who met quality criteria(clinicopathological characteristics in Supplementary Table S1).Principal component analysis illustrated increased heterogeneity among very young HR^(+)/HER2^(−)BC samples(Figure 1A).Differential expression analysis revealed 82 downregulated and 12 upregulated genes in very young BC samples(log2 fold change≥|2|;adjusted P<0.05)(Figure 1B-C,Supplementary Table S2).
文摘Over the past years, significant efforts have been devoted to explore novel drug delivery and detection strategies for simultaneous therapy and diagnostics. The development of biotinylated polyurethane-urea nanoparticles as theranostic nanocarriers for targeted drug and plasmid delivery, for fluorescence detection of human hepatocellular carcinoma cells, is described herein. These targeted nanoparticles are specifically designed to incorporate biotin into the polymeric matrix, since many tumor types overexpress receptors for biotin as a mechanism to boost uncontrolled cell growth. The obtained nanoparticles were spherical, exhibited an average diameter ranging 110-145 nm, and showed no cytotoxicity in healthy endothelial cells. Biotinylated nanoparticles are selectively incorporated into the perinuclear and nuclear area of the human hepatocellular carcinoma cell line, HepG2, in division, but not into growing, healthy, human endothelial cells. Indeed, the simultaneous incorporation of the anticancer drugs, phenoxodiol or sunitinib, together with plasmid DNA encoding green fluorescent protein, into these nanoparticles allows a targeted pharmacological antitumor effect and furthermore, selective transfection of a reporter gene, to detect these cancer cells. The combined targeted therapy and detection strategy described here could be exploited for liver cancer therapy and diagnostics, with a moderate safety profile, and may also be a potential tool for other types of cancer.
文摘The generation of functional retinal pigment epithelium （RPE） is of great therapeutic interest to the field of regenerative medicine and may provide possible cures for retinal degenerative diseases, including age-related macular degeneration （AMD）. Although RPE cells can be produced from either embryonic stem cells or induced pluripotent stem cells, direct cell reprogramming driven by lineage-determining transcription factors provides an immediate route to their generation. By monitoring a human RPE specific Bestl：：GFP reporter, we report the conversion of human fibroblasts into RPE lineage using defined sets of transcription factors. We found that Bestl：：GFP positive cells formed colonies and exhibited morphological and molecular features of early stage RPE cells. Moreover, they were able to obtain pigmen- tation upon activation of Retinoic acid （RA） and Sonic Hedgehog （SHH） signaling pathways. Our study not only established an ideal platform to investigate the tran- scriptional network regulating the RPE cell fate deter- mination, but also provided an alternative strategy to generate functional RPE cells that complement the useof pluripotent stem cells for disease modeling, drug screening, and cell therapy of retinal degeneration.
基金The authors would like to thank METU(BAP-07.02.2013.101)for the financial support of the study by E.S.the Scientific and Technological Research Council of Turkey(TUBITAK)for the scholarship to E.S.through BIDEB 2211C+1 种基金We are grateful to Ministry of Development of Turkey for funding BIOMATEN through Grant DPT2011K120350J.C.R.C.acknowledges the funding from the EC(HEALTH-F4-2011-278557,PITN-GA-2012-317306,MSCA-ITN-2014-642687 and NMP-2014-646075),MINECO(MAT2013-42473-R and MAT2015-68901R)and JCyL(VA244U13,VA313U14 and VA015U16).
文摘The ultrastructure of the bone provides a unique mechanical strength against compressive, torsional andtensional stresses. An elastin-like recombinamer (ELR) with a nucleation sequence for hydroxyapatitewas incorporated into films prepared from a collagen-silk fibroin blend carrying microchannel patternsto stimulate anisotropic osteogenesis. SEM and fluorescence microscopy showed the alignment ofadipose-derived stem cells (ADSCs) and the human osteoblasts (HOBs) on the ridges and in the groovesof microchannel patterned collagen-fibroin-ELR blend films. The Young's modulus and the ultimatetensile strength (UTS) of untreated films were 0.58 ± 0.13 MPa and 0.18 ± 0.05 MPa, respectively. After 28days of cell culture, ADSC seeded film had a Young's modulus of 1.21 ± 0.42 MPa and UTS of0.32 ± 0.15 MPa which were about 3 fold higher than HOB seeded films. The difference in Young'smodulus was statistically significant (p: 0.02). ADSCs attached, proliferated and mineralized better thanthe HOBs. In the light of these results, ADSCs served as a better cell source than HOBs for bone tissueengineering of collagen-fibroin-ELR based constructs used in this study. We have thus shown theenhancement in the tensile mechanical properties of the bone tissue engineered scaffolds by usingADSCs.
基金This work was supported by the Spain’s Ministerio de Economı´a y Competitividad[projects MAT2008-06887-C03-01,MAT2010-15310,MAT2013-41723-R,MAT2013-42473-R,VA313U14 and VA244U13].X.P.acknowledges grant BES-2009-027524 from the Spain’s Ministerio de Economı´a y Competitividad.
文摘The integration of implants or medical devices into the body tissues requires of good cell–material interactions.However,most polymeric materials used for these applications lack on biological cues,which enhanced mid-and long-term implant failure due to weak integration with the surrounding tissue.Commonly used strategies for tissue–material integration focus on functionalization of the material surface by means of natural proteins or short peptides.However,the use of these biomolecules involves major drawbacks such as immunogenic problems and oversimplification of the constructs.Here,designed elastin-like recombinamers(ELRs)are used to enhance poly(methyl methacrylate)surface properties and compared against the use of short peptides.In this study,cell response has been analysed for different functionalization conditions in the presence and absence of a competing protein,which interferes on surface–cell interaction by unspecific adsorption on the interface.The study has shown that ELRs can induce higher rates of cell attachment and stronger cell anchorages than short peptides,being a better choice for surface functionalization.
基金We thank the Spanish MINECO for supporting the project MAT2012-38793 and for funding MN through the Ramon y Cajal program and TS through the“Personal Tecnico de Apoyo”subprogram.
文摘The aim of the present study was the in vitro and in vivo analysis of a bi-layered 3D-printed scaffold combining a PLA layer and a biphasic PLA/bioglass G5 layer for regeneration of osteochondral defects in vivo Focus of the in vitro analysis was on the(molecular)weight loss and the morphological and mechanical variations after immersion in SBF.The in vivo study focused on analysis of the tissue reactions and differences in the implant bed vascularization using an established subcutaneous implantation model in CD-1 mice and established histological and histomorphometrical methods.Both scaffold parts kept their structural integrity,while changes in morphology were observed,especially for the PLA/G5 scaffold.Mechanical properties decreased with progressive degradation,while the PLA/G5 scaffolds presented higher compressive modulus than PLA scaffolds.The tissue reaction to PLA included low numbers of BMGCs and minimal vascularization of its implant beds,while the addition of G5 lead to higher numbers of BMGCs and a higher implant bed vascularization.Analysis revealed that the use of a bi-layered scaffold shows the ability to observe distinct in vivo response despite the physical proximity of PLA and PLA/G5 layers.Altogether,the results showed that the addition of G5 enables to reduce scaffold weight loss and to increase mechanical strength.Furthermore,the addition of G5 lead to a higher vascularization of the implant bed required as basis for bone tissue regeneration mediated by higher numbers of BMGCs,while within the PLA parts a significantly lower vascularization was found optimally for chondral regeneration.Thus,this data show that the analyzed bi-layered scaffold may serve as an ideal basis for the regeneration of osteochondral tissue defects.Additionally,the results show that it might be able to reduce the number of experimental animals required as it may be possible to analyze the tissue response to more than one implant in one experimental animal.
基金National Foundation for Liver Research(NFLR)for financially supporting the cell-based work in this project.CIBER-BBN is an initiative funded by the VI National R&D&I Plan 2008–2011,Iniciativa Ingenio 2010,Consolider Program.CIBER actions are financed by the Instituto de Salud CarlosⅢwith assistance from the European Regional Development Fundthe University Grants Commission,India for the award of the Basic Scientific Research(UGC-BSR)Faculty Fellowship[No.F.4-5(11)2019(BSR)].
文摘The current study investigated a triad,which comprises of adipose tissue derived stem cells isolated from infrapatellar fat pad and gelatin/polyvinyl alcohol(PVA)-based matrix with exclusive ascorbic acid signalling.Though,the bio-mechanical properties of the gelatin–PVA blended scaffolds in wet condition are equivalent to the ECM of soft tissues in general,in this study,the triad was tested as a model for neural tissue engineering.Apart from being cytocompatible and biocompatible,the porosity of the scaffold has been designed in such a manner that it facilitates the cell signalling and enables the exchange of nutrients and gases.The highly proliferative stem cells from Passage 2 were characterized using both,mesenchymal and embryonic stem cell markers.As an initial exploration the mesenchymal stem cells at Passage 4 were exposed to ascorbic acid and basic fibroblast growth factor signalling for neuronal differentiation in 2D environment independently.The MSCs successfully differentiated and acquired neuron specific markers related to cytoskeleton and synapses.Subsequently,three phases of experiments have been conducted on the 3D gelatin/PVA matrix to prove their efficacy,the growth of stem cells,growth of differentiated neurons and the in situ growth and differentiation of MSCs.The scaffold was conducive and directed MSCs to neuronal lineage under specific signalling.Overall,this organotypic model triad could open a new avenue in the field of soft tissue engineering as a simple and effective tissue construct.
基金supported by Instituto de Salud CarlosⅢ(ISCIII,SpainCo-funding from FEDER,European Union)[PI18/00650,PIE15/00028,PI15/00378 and EU COST Action CA 17140 to Ramon Mangues,PI19/01661 to Xavier León,and PI17/00584 to Miquel Quer]+7 种基金Agencia Estatal de Investigación(AEI,Spain)and Fondo Europeo de Desarrollo Regional(FEDER,European Union)[grant BIO2016-76063-R,AEI/FEDER,UE to Antonio Villaverde and grant PID2019-105416RB-I00/AEI/10.13039/501100011033 to Esther Vazquez]CIBER-BBN(Spain)[CB06/01/1031 and 4NanoMets to Ramon Mangues,VENOM4CANCER to Antonio Villaverde,NANOREMOTE to Esther Vazquez,and NANOSCAPE to Ugutz Unzueta]AGAUR(Spain)2017-SGR865 to Ramon Mangues,and 2017SGR-229 to Antonio VillaverdeJosep Carreras Leukemia Research Institute(Spain)[P/AG to Ramon Mangues]supported by a predoctoral fellowship from AGAUR(Spain)(2020FI_B200168 and 2018FI_B2_00051)co-funded by European Social Fund(ESF investing in your future,European Union)supported by a postdoctoral fellowship from AECC(Spanish Association of Cancer Research,Spain)Antonio Villaverde received an Icrea Academia Award(Spain)supported by Grant PERIS SLT006/17/00093 from la Generalitat de Catalunya(Spain)and Miguel Servet fellowship(CP19/00028)from Instituto de Salud CarlosⅢ(Spain)co-funded by European Social Fund(ESF investing in your future,European Union)。
文摘Loco-regional recurrences and distant metastases represent the main cause of head and neck squamous cell carcinoma(HNSCC) mortality. The overexpression of chemokine receptor 4(CXCR4) in HNSCC primary tumors associates with higher risk of developing loco-regional recurrences and distant metastases, thus making CXCR4 an ideal entry pathway for targeted drug delivery. In this context, our group has generated the self-assembling protein nanocarrier T22-GFP-H6, displaying multiple T22 peptidic ligands that specifically target CXCR4. This study aimed to validate T22-GFP-H6 as a suitable nanocarrier to selectively deliver cytotoxic agents to CXCR4^(+)tumors in a HNSCC model. Here we demonstrate that T22-GFP-H6 selectively internalizes in CXCR4^(+)HNSCC cells, achieving a high accumulation in CXCR4^(+)tumors in vivo, while showing negligible nanocarrier distribution in non-tumor bearing organs. Moreover, this T22-empowered nanocarrier can incorporate bacterial toxin domains to generate therapeutic nanotoxins that induce cell death in CXCR4-overexpressing tumors in the absence of histological alterations in normal organs. Altogether, these results show the potential use of this T22-empowered nanocarrier platform to incorporate polypeptidic domains of choice to selectively eliminate CXCR4^(+)cells in HNSCC. Remarkably, to our knowledge, this is the first study testing targeted proteinonly nanoparticles in this cancer type, which may represent a novel treatment approach for HNSCC patients.
基金Agencia Estatal de Investigación(AEI)and to Fondo Europeo de Desarrollo Regional(FEDER)(BIO2016-76063-R,AEI/FEDER,UE)to Villaverde A,AGAUR(2017SGR-229)to Villaverde A and 2017SGR-865 GRCISCⅢ(PI15/00272 co-founding FEDER)to Vázquez E and ISCⅢ(Co-founding FEDER)PIE15//00028 and PI18/00650 to Mangues R,and to EU COST Action CA 17140+3 种基金funded by theⅥNational R&D&I Plan 2008–2011,Iniciativa Ingenio 2010,Consolider Program,CIBER Actionsfinanced by the Instituto de Salud CarlosⅢ,with assistance from the European Regional Development Fundsupported by a predoctoral fellowship from AGAUR(2019 FI_B 00352)PERIS program from the Health Department of the Generalitat de Catalunya
文摘Poly-histidine peptides such as H6(HHHHHH)are used in protein biotechnologies as purification tags,protein-assembling agents and endosomal-escape entities.The pleiotropic properties of such peptides make them appealing to design protein-based smart materials or nanoparticles for imaging or drug delivery to be produced in form of recombinant proteins.However,the clinical applicability of H6-tagged proteins is restricted by the potential immunogenicity of these segments.In this study,we have explored several humanized histidine-rich peptides in tumor-targeted modular proteins,which can specifically bind and be internalized by the target cells through the tumoral marker CXCR4.We were particularly interested in exploring how protein purification,self-assembling and endosomal escape perform in proteins containing the variant histidine-rich tags.Among the tested candidates,the peptide H5 E(HEHEHEHEH)is promising as a good promoter of endosomal escape of the associated fulllength protein upon endosomal internalization.The numerical modelling of cell penetration and endosomal escape of the tested proteins has revealed a negative relationship between the amount of protein internalized into target cells and the efficiency of cytoplasmic release.This fact demonstrates that the His-mediated,proton sponge-based endosomal escape saturates at moderate amounts of internalized protein,a fact that might be critical for the design of protein materials for cytosolic molecular delivery.
基金This work was partially supported by Ministry of Economy and Competitiveness ISCIII-FIS grants PI12/01763,PI12/00934 and PI15/00923co-financed by ERDF(FEDER)Funds from the European Commission,‘A way of making Europe’+3 种基金JL-A was supported by a Grant from IiSGMAP-S was supported by the Youth Employment Program co-financed by the Madrid community and FEDER FoundsRC-R was supported by the‘Miguel Servet’program(CPII13/00033)MP by the Spanish MICINN through the Ramón y Cajal(RYC-2009-05486).
文摘Individuals infected with human immunodeficiency virus type-1(HIV-1)usually show a general dysregulation and hyper-activation of the immune system.A direct influence of HIV-1 particles on B-cell phenotypes and functions has been previously described.However,the consequences of B-cell dysregulation are still poorly understood.We evaluated the phenotypic changes in primary B cells after direct contact with HIV-1 particles in comparison with different types of stimuli.The functionality of treated B cells was challenged in co-culture experiments with autologous CD4+and CD8+T cells.We demonstrated that HIV-1 induces a phenotypic change in B cells towards a regulatory B-cell phenotype,showing a higher level of IL-10,TGF-β1,EBI3 or IL-12(p35)mRNA expression and acquiring an immunosuppressive profile.The acquisition of a Breg phenotype was confirmed by co-culture experiments where HIVtreated B cells reduced the proliferation and the TNFαproduction of CD4+or CD8+T cells.This suppressive ability of HIV-treated B cells was dependent on cell-to-cell contact between these B cells and effector cells.To our knowledge,these data provide the first evidence that HIV-1 can directly induce a regulatory B cell-like immunosuppressive phenotype,which could have the ability to impair specific immune responses.This dysregulation could constitute one of the mechanisms underlying unsuccessful efforts to develop an efficient vaccine against HIV-1.
