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Search terms: exosomes OR "extracellular vesicles" OR microvesicles OR microparticles. Direct link to the PubMed search here.

On an effective approach to improve the properties and the drug release of chitosan-based microparticles.

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On an effective approach to improve the properties and the drug release of chitosan-based microparticles.

Int J Biol Macromol. 2020 Jul 06;:

Authors: Arnaldi P, Pastorino L, Monticelli O

Abstract
The work investigated the possibility to develop an easy scalable treatment capable of modifying only the surface of chitosan-based materials, limiting the degradation of the bulk and the burst release of a drug, without compromising the properties of the polymeric matrix. To this aim, microparticles of CHI were superficially coated with poly-(styrene-co-maleic anhydride) (PSMA), taking advantage of the potential reactivity of chitosan amino groups and maleic functionalities of PSMA. The specific reactions/interactions occurring between the two polymers were studied by IR measurements, while FE-SEM analysis evidenced the modification of the morphology of the particles contacted with PSMA. Contact angle measurements demonstrated the change of wettability in the modified systems and TGA analysis allowed to estimate the amount of the deposited PSMA. The above treatment turned out to improve the particle stability both in an acidic environment and in an enzymatic system. The release properties of the treated and of the untreated particles, over a period of 10 h, were tested using, as model drug, the protein Bovine Serum Albumin (BSA). Finally, the cytocompatibility of the developed composite microparticles was assessed on MCF-7 human breast cancer cells, which measurements demonstrated the non-toxicity of the treatment.

PMID: 32645491 [PubMed - as supplied by publisher]

Stem Cell Derived Extracellular Vesicles for Vascular Elastic Matrix Regenerative Repair.

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Stem Cell Derived Extracellular Vesicles for Vascular Elastic Matrix Regenerative Repair.

Acta Biomater. 2020 Jul 06;:

Authors: Sajeesh S, Broekelman T, Mecham RP, Ramamurthi A

Abstract
Abdominal aortic aneurysms (AAA) are localized expansions of the abdominal aorta that develop due to chronic proteolytic disruption of the structural extracellular matrix (ECM) components (elastin and collagen) within the aorta wall. Major limitations in arresting or reversing AAAs lie in naturally poor and aberrant regeneration and repair of elastic matrix structures in the aorta wall. Bone marrow derived mesenchymal stem cells (BM-MSCs) have emerged as a promising regenerative tool and their therapeutic effects are also known to be effected through their paracrine secretions. Extracellular vesicles (EVs) present in these secretions have emerged as critical cellular component in facilitating many therapeutic benefits of MSCs. EV treatment is thus potentially appealing as a stem cell-inspired cell-free approach to avoid possible phenotypic plasticity of MSCs in vivo. In this study, we investigated the thus far unknown effects of BM-MSC derived EVs on vascular elastic matrix repair in the context of AAA treatment. EVs isolated from BM-MSC source were characterized and their pro-regenerative and their anti-proteolytic effects were evaluated on our established in vitro experimental conditions derived from AAA rat model. Our studies revealed the efficacy of BM-MSC derived EVs in attenuating the proteolytic activity and also in imparting elastic matrix regenerative benefits under aneurysmal environment. Interestingly, compared to cell culture conditioned media (CCM), EVs demonstrated superior regenerative and anti-proteolytic benefits in a proteolytic injury culture model of AAA. From these studies, it appears that EVs derived from BM-MSCs could be beneficial in undertaking a reparative effort in AAA induced degeneration of vascular tissue. Statement of Significance Abdominal aortic aneurysms (AAAs) are localized, rupture-prone expansions of the aorta which result from loss of wall flexibility due to enzymatic breakdown of elastic fibers. There are no established alternatives to surgery, which possess high risk for the mostly elderly patients. Our previous studies have established the elastic regenerative and reparative effect of cell culture secretions derived from adult stem cell source. In this study, we propose to isolate extracellular vesicles (exosomes) from these secretions and evaluate their regenerative benefits in AAA smooth muscle cell culture model. This simple and innovative treatment approach has the potential to arrest or reverse AAA growth to rupture, not possible so far.

PMID: 32645438 [PubMed - as supplied by publisher]

Vaterite Submicron Particles Design for Photodynamic Therapy in Cells.

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Vaterite Submicron Particles Design for Photodynamic Therapy in Cells.

Photodiagnosis Photodyn Ther. 2020 Jul 06;:101913

Authors: Souza EF, Ambrósio JAR, Pinto BCS, Beltrame M, Sakane KK, Pinto JG, Ferreira-Strixino J, Gonçalves EP, Simioni AR

Abstract
BACKGROUND: Calcium carbonate (CaCO3) is one of the most abundant materials in the world. It has several different crystalline phases as present in the minerals: calcite, aragonite and vaterite, which are anhydrous crystalline polymorphs. Regarding the preparation of these microparticles, the most important aspect is the control of the polymorphism, particle size and material morphology. This study aimed to develop porous microparticles of calcium carbonate in the vaterite phase for the encapsulation of chloro-aluminum phthalocyanine (ClAlPc) as a photosensitizer (PS) for application in Photodynamic Therapy (TFD).
METHODS: In this study, spherical vaterite composed of microparticles are synthesized by precipitation route assisted by polycarboxylate superplasticizer (PSS). The calcium carbonate was prepared by reacting a mixed solution of Na2CO3 with a CaCl2 solution at an ambient temperature, 25 °C, in the presence of polycarboxylate superplasticizer as a stabilizer. The photosensitizer was incorporated by adsorption technique in the CaCO3 microparticles. The CaCO3 microparticles were studied by scanning electron microscopy, steady-state, and their biological activity was evaluated using in vitro cancer cell lines by trypan blue exclusion method. The intracellular localization of ClAlPc was examined by confocal microscopy.
RESULTS: The CaCO3 microparticles obtained are uniform and homogeneously sized, non-aggregated, and highly porous microparticles. The calcium carbonate microparticles show an average size of 3 µm average pore size of about 30-40 nm. The phthalocyanine derivative loaded-microparticles maintained their photophysical behavior after encapsulation. The captured carriers have provided dye localization inside cells. The in vitro experiments with ClAlPc-loaded CaCO3 microparticles showed that the system is not cytotoxic in darkness, but exhibits a substantial phototoxicity at 3 µmol.L-1 of photosensitizer concentration and 10 J.cm-2 of light. These conditions are sufficient to kill about 80% of the cells.
CONCLUSIONS: All the performed physical-chemical, photophysical, and photobiological measurements indicated that the phthalocyanine-loaded CaCO3 microparticles are a promising drug delivery system for photodynamic therapy and photoprocesses.

PMID: 32645435 [PubMed - as supplied by publisher]

Development of nimesulide amorphous solid dispersions via supercritical anti-solvent process for dissolution enhancement.

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Development of nimesulide amorphous solid dispersions via supercritical anti-solvent process for dissolution enhancement.

Eur J Pharm Sci. 2020 Jul 06;:105457

Authors: Liu G, Gong L, Zhang J, Wu Z, Deng H, Deng S

Abstract
Formulating amorphous solid dispersions (ASDs) is one of the most promising strategies to overcome solubility limitations in drug development. In this work, development of nimesulide (NIM) ASDs via supercritical anti-solvent (SAS) process was proposed, where the mixtures of dichloromethane (DCM) and methanol (MeOH) were selected as the liquid solvent, and the mixtures of hydroxypropyl methylcellulose (HPMC) and polyvinylpyrrolidone (PVP) were the dispersing materials. The effects of NIM/HPMC/PVP (w/w/w) ratio and DCM/MeOH (v/v) ratio on particle solid-state properties were investigated to identify successful operating conditions. NIM-ASDs powders were formed by well separated spherical microparticles, where NIM crystals had transformed into amorphous state completely; the production yield was 93.6±1.14%; and the reproducibility was very high. For NIM-ASDs, intermolecular interactions between NIM and dispersing materials were formed; the residual solvent was far below the ICH limit; and the chemical structure of NIM did not be degraded or disrupted. Moreover, NIM-ASDs increased the NIM solubility in PBS (pH=6.8) more than 5-folds; the dissolution of NIM from NIM-ASDs granules was faster and more complete than that from commercial Aulin® granules in PBS (pH=6.8). Also, NIM-ASDs well hindered the aging in the recrystallization of amorphous NIM during 12-month sealed storage. Overall, development of NIM-ASDs via SAS process presents an opportunity that as a modified product to increase the efficacy of NIM.

PMID: 32645426 [PubMed - as supplied by publisher]

Continuous in-line homogenization process for scale-up production of naltrexone-loaded PLGA microparticles.

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Continuous in-line homogenization process for scale-up production of naltrexone-loaded PLGA microparticles.

J Control Release. 2020 Jul 06;:

Authors: Sharifi F, Otte A, Yoon G, Park K

Abstract
Injectable, long-acting drug delivery systems provide effective drug concentrations in the blood for up to 6 months. Naltrexone-loaded poly(lactide-co-glycolide) (PLGA) microparticles were prepared using an in-line homogenization method. It allows the transition from a laboratory scale to scale-up production. This research was designed to understand how the processing parameters affect the properties of the microparticles, such as microparticle size distributions, surface and internal morphologies, drug loadings, and drug release kinetics, and thus, to control them. The in-line homogenization system was used at high flow rates for the oil- and water-phases, e.g., 100 mL/min and 400 mL/min, respectively, to continuously generate microparticles. A high molecular weight (148 kDa) PLGA at various concentrations was used to generate oil phases with a range of viscosities and also to compare with a 64 and 79 kDa at a single, high concentration. The uniformity of the microparticles was found to be related to the viscosity of the oil phase. As the viscosity of the oil phase increased from 52.6 mPa∙s to 4046 mPa∙s, the span value (a measure of uniformity) increased from 1.24 to 3.1 for the microparticles generated at the homogenization speed of 2000 RPM. Increasing the PLGA concentration from 5.58% to 16.85% showed a corresponding rise in the encapsulation efficiency (EE) from 74.0% to 85.8% and drug loading (DL) from 27.4% to 31.7% for the microparticles made with the homogenization speed of 2000 RPM. These increases may be due to a faster shell formulation, enabling PLGA microparticles to entrap more naltrexone into the structure. A higher DL, however, shortened the drug release duration from 56 to 42 days. The changes in morphology of the microparticles during different phases of the in vitro release study were also studied for three types of microparticles made with different PLGA concentrations and MWs. As PLGA microparticles went through structural changes, the surface showed raisin-like wrinkled morphologies within the first 10 days. Then, the microparticles swelled to form smooth surfaces. The in-line approach produced PLGA microparticles with a highly reproducible size distribution, DL, and naltrexone release rate.

PMID: 32645336 [PubMed - as supplied by publisher]

Ionic Strength Influences on Biofunctional Au-Decorated Microparticles for Enhanced Performance in Multiplexed Colorimetric Sensors.

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Ionic Strength Influences on Biofunctional Au-Decorated Microparticles for Enhanced Performance in Multiplexed Colorimetric Sensors.

ACS Appl Mater Interfaces. 2020 Jul 09;:

Authors: Díaz-Amaya S, Zhao M, Allebach JP, Chiu GT, Stanciu LA

Abstract
The rising development of biosensors offers a great potential for health, food, and environmental monitoring. However, in many colorimetric platforms, there is a performance limitation stemming from the tendency of traditional Au nanoparticles toward nonspecific aggregation in response to changing ionic strength (salt concentration). This work puts forward a new type of colorimetric aptamer-functionalized labeling of microparticles, which allows to leverage an increase in ionic strength as a positive driver of enhanced detection performance of analytical targets. The resulting device is a cost-effective, instrument-free, portable, and reliable aptasensor that serves as basis for the fabrication of universal paper-based colorimetric platforms with the capability of multiplex, multireplicates and provides quantitative colorimetric detection. A controlled fabrication process was demonstrated by keeping 90% of the signal obtained from the as-fabricated devices (n = 40) within ± 1 standard deviation (SD) (relative SD = 5.69%) and following a mesokurtic normal-like distribution (p = 0.385). We propose for the first time a salt-induced aggregation mechanism for highly stable multilayered label particles (ssDNA-PEI-Au-PS) as the basis of the detection scheme. The use of DNA aptamers as capture biomolecules and PEI as an encapsulating agent allows for a sensitive and highly specific colorimetric response. As a proof of concept, multiplexed detection of mercury (Hg2+) and arsenic (As3+) was demonstrated. In addition, we introduced a robust image analysis algorithm for testing zone segmentation and color signal quantification that allowed for analytical detection, reaching a limit of detection of 1 ppm for both targeted analytes, with enough evidence (p > 0.05) to prove the high specificity of the fabricated device versus a pool of possible interferent ions.

PMID: 32645268 [PubMed - as supplied by publisher]

Exosomes derived from microglia exposed to elevated pressure amplify the neuroinflammatory response in retinal cells.

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Exosomes derived from microglia exposed to elevated pressure amplify the neuroinflammatory response in retinal cells.

Glia. 2020 Jul 09;:

Authors: Aires ID, Ribeiro-Rodrigues T, Boia R, Catarino S, Girão H, Ambrósio AF, Santiago AR

Abstract
Glaucoma is a degenerative disease that causes irreversible loss of vision and is characterized by retinal ganglion cell (RGC) loss. Others and we have demonstrated that chronic neuroinflammation mediated by reactive microglial cells plays a role in glaucomatous pathology. Exosomes are extracellular vesicles released by most cells, including microglia, that mediate intercellular communication. The role of microglial exosomes in glaucomatous degeneration remains unknown. Taking the prominent role of microglial exosomes in brain neurodegenerative diseases, we studied the contribution of microglial-derived exosomes to the inflammatory response in experimental glaucoma. Microglial cells were exposed to elevated hydrostatic pressure (EHP), to mimic elevated intraocular pressure, the main risk factor for glaucoma. Naïve microglia (BV-2 cells or retinal microglia) were exposed to exosomes derived from BV-2 cells under EHP conditions (BV-Exo-EHP) or cultured in control pressure (BV-Exo-Control). We found that BV-Exo-EHP increased the production of pro-inflammatory cytokines, promoted retinal microglia motility, phagocytic efficiency, and proliferation. Furthermore, the incubation of primary retinal neural cell cultures with BV-Exo-EHP increased cell death and the production of reactive oxygen species. Exosomes derived from retinal microglia (MG-Exo-Control or MG-Exo-EHP) were injected in the vitreous of C57BL/6J mice. MG-Exo-EHP sustained activation of retinal microglia, mediated cell death, and impacted RGC number. Herein, we show that exosomes derived from retinal microglia have an autocrine function and propagate the inflammatory signal in conditions of elevated pressure, contributing to retinal degeneration in glaucomatous conditions.

PMID: 32645245 [PubMed - as supplied by publisher]

COVID-19 comorbidities, associated pro-coagulant extracellular vesicles and venous thromboembolisms: a possible link with ethnicity?

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COVID-19 comorbidities, associated pro-coagulant extracellular vesicles and venous thromboembolisms: a possible link with ethnicity?

Br J Haematol. 2020 Jul 09;:

Authors: Inal J

Abstract
Several comorbidities predict mortality in patients with COVID-19, some of which are more prevalent in Black Asian and Minority Ethnic (BAME) groups. Extracellular vesicles (EVs) released from cells are nano-sized membrane-bound vesicles that carry nucleic acids and proteins and mediate intercellular communication. In COVID-19 associated comorbidities including diabetes, CVD and risk factors such as hypertension, elevated angiotensin II and obesity, levels of circulating EVs are raised and their cargo modified. Importantly, EVs released from damaged endothelial cells are known to be procoagulant. EVs in Black and White populations differentially express phosphorylated insulin signalling proteins, associated with impaired insulin signalling, insulin resistance and an increased risk of venous thromboembolism; these EV proteins are also associated with clinical mortality risk markers. If plasma EV levels are raised, and cargo modified in comorbidities associated with COVID-19 such as diabetes, and account for excess deaths in BAME patients, this raises the possibility for important roles of EVs, also relating to ethnicity. This places EVs as hitherto understudied key factors, filling the current knowledge gap in COVID-19, amongst BAME patients.

PMID: 32645205 [PubMed - as supplied by publisher]

Sickle particulars of microparticles.

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Sickle particulars of microparticles.

Blood. 2020 Jul 09;136(2):154-155

Authors: Kato GJ

PMID: 32645168 [PubMed - as supplied by publisher]

Exosomal Circ-HIPK3 Facilitates Tumor Progression and Temozolomide Resistance by Regulating miR-421/ZIC5 Axis in Glioma.

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Exosomal Circ-HIPK3 Facilitates Tumor Progression and Temozolomide Resistance by Regulating miR-421/ZIC5 Axis in Glioma.

Cancer Biother Radiopharm. 2020 Jul 09;:

Authors: Han C, Wang S, Wang H, Zhang J

Abstract
Background: The resistance of glioma patients to temozolomide (TMZ) treatment is a limiting factor in clinical treatment. Circular RNA HIPK3 (circ-HIPK3) was found to be highly expressed in glioma, however, the role and potential mechanism of exosomal circ-HIPK3 from TMZ-resistant cells remain poorly unclear. Methods: Exosomes were characterized by transmission electron microscopy. The levels of all protein were detected by western blot. Expression levels of circ-HIPK3, microRNA-421 (miR-421), and zinc finger protein of the cerebellum 5 (ZIC5) were measured by quantitative real-time polymerase chain reaction. The cell's 50% inhibitory concentration (IC50) of TMZ, apoptosis, and invasion were determined by methyl thiazolyl tetrazolium (MTT), flow cytometry, and Transwell assays, respectively. The correlation between miR-421 and circ-HIPK3 or ZIC5 was identified by dual-reporter luciferase and RNA immunoprecipitation (RIP) assays. The xenograft model was established to explore the effect of circ-HIPK3 in vivo. Results: Circ-HIPK3 was obviously increased in TMZ-resistant glioma cells and their exosomes, miR-421, was downregulated in TMZ-resistant glioma. Circ-HIPK3 directly targeted miR-421 and their expressions were negatively correlated in glioma tissues. Besides, circ-HIPK3 knockdown hampered the IC50 of TMZ, cell invasion, TMZ resistance, and triggered cell apoptosis, whereas these effects were reversed by transfection of anti-miR-421. ZIC5 was the target of miR-421 and ZIC5 overexpression weakened the inhibition effects of miR-421 on cell progression and TMZ resistance. More importantly, circ-HIPK3 depletion inhibited tumor growth by decreasing ZIC5 through sponging miR-421 in vivo. Conclusion: Exosomal circ-HIPK3 could promote cell progression and TMZ resistance by regulating miR-421/ZIC5 axis in TMZ-resistant glioma.

PMID: 32644821 [PubMed - as supplied by publisher]

Inhalable nano-composite microparticles with enhanced dissolution and superior aerosol performance.

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Inhalable nano-composite microparticles with enhanced dissolution and superior aerosol performance.

Mol Pharm. 2020 Jul 09;:

Authors: Zhu C, Chen J, Yu S, Que C, Taylor LS, Tan W, Wu C, Zhou QT

Abstract
Previous studies have shown that combining colistin (Col), a cationic polypeptide antibiotic, with ivacaftor (Iva), a cystic fibrosis (CF) drug, could achieve synergistic antibacterial effects against Pseudomonas aeruginosa. The purpose of this study was to develop dry powder inhaler (DPI) formulations for co-delivery of Col and Iva, aiming to treat CF and lung infection simultaneously. In order to improve solubility and dissolution for the water insoluble Iva, Iva was encapsulated into bovine serum albumin (BSA) nanoparticles (Iva-BSA-NPs). Inhalable composite microparticles of Iva-BSA-NPs were produced by spray-freeze-drying using water-soluble Col as the matrix material and L-leucine as an aerosol enhancer. The optimal formulation showed irregular-shaped morphology with fine particle fraction (FPF) values of 73.8 ± 5.2% for Col and 80.9 ± 4.1% for Iva. Correlations between "D×√(ρ_tapped )" and FPF were established for both Iva and Col. The amorphous solubility of Iva is 66 times higher than the crystalline solubility in the buffer. Iva-BSA-NPs were amorphous and remained in the amorphous state after spray-freeze-drying as examined by powder X-ray diffraction. In-vitro dissolution profiles of the selected DPI formulation indicated that Col and Iva were almost completely released within 3 hours, which was substantially faster regarding Iva release than the jet milled physical mixture of the two drugs. In summary, this study developed a novel inhalable nano-composite microparticle using a synergistic water-soluble drug as the matrix material, which achieved reduced use of excipients for high-dose medications, improved dissolution rate for the water-insoluble drug and superior aerosol performance.

PMID: 32643939 [PubMed - as supplied by publisher]

Alveolar macrophage-derived extracellular vesicles inhibit endosomal fusion of influenza virus.

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Alveolar macrophage-derived extracellular vesicles inhibit endosomal fusion of influenza virus.

EMBO J. 2020 Jul 09;:e105057

Authors: Schneider DJ, Smith KA, Latuszek CE, Wilke CA, Lyons DM, Penke LR, Speth JM, Marthi M, Swanson JA, Moore BB, Lauring AS, Peters-Golden M

Abstract
Alveolar macrophages (AMs) and epithelial cells (ECs) are the lone resident lung cells positioned to respond to pathogens at early stages of infection. Extracellular vesicles (EVs) are important vectors of paracrine signaling implicated in a range of (patho)physiologic contexts. Here we demonstrate that AMs, but not ECs, constitutively secrete paracrine activity localized to EVs which inhibits influenza infection of ECs in vitro and in vivo. AMs exposed to cigarette smoke extract lost the inhibitory activity of their secreted EVs. Influenza strains varied in their susceptibility to inhibition by AM-EVs. Only those exhibiting early endosomal escape and high pH of fusion were inhibited via a reduction in endosomal pH. By contrast, strains exhibiting later endosomal escape and lower fusion pH proved resistant to inhibition. These results extend our understanding of how resident AMs participate in host defense and have broader implications in the defense and treatment of pathogens internalized within endosomes.

PMID: 32643835 [PubMed - as supplied by publisher]

On the potential role of exosomes in the COVID-19 reinfection/reactivation opportunity.

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On the potential role of exosomes in the COVID-19 reinfection/reactivation opportunity.

J Biomol Struct Dyn. 2020 Jul 09;:1-12

Authors: Elrashdy F, Aljaddawi AA, Redwan EM, Uversky VN

Abstract
We propose here that one of the potential mechanisms for the relapse of the COVID-19 infection could be a cellular transport pathway associated with the release of the SARS-CoV-2-loaded exosomes and other extracellular vesicles. It is possible that this "Trojan horse" strategy represents possible explanation for the re-appearance of the viral RNA in the recovered COVID-19 patients 7-14 day post discharge, suggesting that viral material was hidden within such exosomes or extracellular vesicles during this "silence" time period and then started to re-spread again. Communicated by Ramaswamy H. Sarma.

PMID: 32643586 [PubMed - as supplied by publisher]

Exosomes as mediators and biomarkers in fibrosis.

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Exosomes as mediators and biomarkers in fibrosis.

Biomark Med. 2020 Jul 09;:

Authors: Qin XJ, Zhang JX, Wang RL

Abstract
Fibrosis is characterized by aberrant myofibroblast accumulation and excessive extracellular matrix deposition, which leads to organ failure and significantly contributes to mortality worldwide. Exosomes, which are extracellular nanovesicles with a diameter of 30-100 nm that are secreted into the extracellular space by various types of cells, facilitate intercellular communication by delivering different cargos such as proteins, mRNAs and microRNAs. Growing evidence indicates that exosomes play an important role in various fibrotic diseases. A deeper understanding of the effects of exosomes in fibrosis may help in exploring new diagnostic and therapeutic targets. In this review, we summarize recent findings on exosomes in fibrotic diseases, with a special focus on exosomal cargo dysregulation and their potential diagnostic and therapeutic value in fibrosis.

PMID: 32643390 [PubMed - as supplied by publisher]

Insulin-like growth factor 2 (IGF2) protects against Huntington's disease through the extracellular disposal of protein aggregates.

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Insulin-like growth factor 2 (IGF2) protects against Huntington's disease through the extracellular disposal of protein aggregates.

Acta Neuropathol. 2020 Jul 08;:

Authors: García-Huerta P, Troncoso-Escudero P, Wu D, Thiruvalluvan A, Cisternas-Olmedo M, Henríquez DR, Plate L, Chana-Cuevas P, Saquel C, Thielen P, Longo KA, Geddes BJ, Lederkremer GZ, Sharma N, Shenkman M, Naphade S, Sardi SP, Spichiger C, Richter HG, Court FA, Tshilenge KT, Ellerby LM, Wiseman RL, Gonzalez-Billault C, Bergink S, Vidal RL, Hetz C

Abstract
Impaired neuronal proteostasis is a salient feature of many neurodegenerative diseases, highlighting alterations in the function of the endoplasmic reticulum (ER). We previously reported that targeting the transcription factor XBP1, a key mediator of the ER stress response, delays disease progression and reduces protein aggregation in various models of neurodegeneration. To identify disease modifier genes that may explain the neuroprotective effects of XBP1 deficiency, we performed gene expression profiling of brain cortex and striatum of these animals and uncovered insulin-like growth factor 2 (Igf2) as the major upregulated gene. Here, we studied the impact of IGF2 signaling on protein aggregation in models of Huntington's disease (HD) as proof of concept. Cell culture studies revealed that IGF2 treatment decreases the load of intracellular aggregates of mutant huntingtin and a polyglutamine peptide. These results were validated using induced pluripotent stem cells (iPSC)-derived medium spiny neurons from HD patients and spinocerebellar ataxia cases. The reduction in the levels of mutant huntingtin was associated with a decrease in the half-life of the intracellular protein. The decrease in the levels of abnormal protein aggregation triggered by IGF2 was independent of the activity of autophagy and the proteasome pathways, the two main routes for mutant huntingtin clearance. Conversely, IGF2 signaling enhanced the secretion of soluble mutant huntingtin species through exosomes and microvesicles involving changes in actin dynamics. Administration of IGF2 into the brain of HD mice using gene therapy led to a significant decrease in the levels of mutant huntingtin in three different animal models. Moreover, analysis of human postmortem brain tissue and blood samples from HD patients showed a reduction in IGF2 level. This study identifies IGF2 as a relevant factor deregulated in HD, operating as a disease modifier that buffers the accumulation of abnormal protein species.

PMID: 32642868 [PubMed - as supplied by publisher]

Molecular profiles and immunomodulatory activities of glioblastoma-derived exosomes.

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Molecular profiles and immunomodulatory activities of glioblastoma-derived exosomes.

Neurooncol Adv. 2020 Jan-Dec;2(1):vdaa056

Authors: Azambuja JH, Ludwig N, Yerneni S, Rao A, Braganhol E, Whiteside TL

Abstract
Background: Glioblastoma is one of the most immunosuppressive human tumors. Emerging data suggest that glioblastoma-derived exosomes (GBex) reprogram the tumor microenvironment into a tumor-promoting milieu by mechanisms that not yet understood.
Methods: Exosomes were isolated from supernatants of glioblastoma cell lines by size exclusion chromatography. The GBex endosomal origin, size, protein cargos, and ex vivo effects on immune cell functions were determined. GBex were injected intravenously into mice to evaluate their ability to in vivo modulate normal immune cell subsets.
Results: GBex carried immunosuppressive proteins, including FasL, TRAIL, CTLA-4, CD39, and CD73, but contained few immunostimulatory proteins. GBex co-incubated with primary human immune cells induced simultaneous activation of multiple molecular pathways. In CD8+ T cells, GBex suppressed TNF-α and INF-γ release and mediated apoptosis. GBex suppressed natural killer (NK) and CD4+ T-cell activation. GBex activated the NF-κB pathway in macrophages and promoted their differentiation into M2 cells. Inhibition of the NF-κB pathway in macrophages reversed the GBex-mediated effects. GBex-driven reprogramming of macrophages involved the release of soluble factors that promoted tumor proliferation in vitro. In mice injected with GBex, the frequency of splenic CD8+ T cells, NK cells, and M1-like macrophages was reduced, while that of naïve and M2-like macrophages increased (P < .05).
Conclusions: GBex reprogrammed functions of all types of immune cells in vitro and altered their frequency in vivo. By creating and sustaining a highly immunosuppressive environment, GBex play a key role in promoting tumor progression.

PMID: 32642708 [PubMed]

RNU6-1 in circulating exosomes differentiates GBM from non-neoplastic brain lesions and PCNSL but not from brain metastases.

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RNU6-1 in circulating exosomes differentiates GBM from non-neoplastic brain lesions and PCNSL but not from brain metastases.

Neurooncol Adv. 2020 Jan-Dec;2(1):vdaa010

Authors: Puigdelloses M, González-Huárriz M, García-Moure M, Martínez-Vélez N, Esparragosa Vázquez I, Bruna J, Zandio B, Agirre A, Marigil M, Petrirena G, Nuñez-Córdoba JM, Tejada-Solís S, Díez-Valle R, Gállego-Culleré J, Martínez-Vila E, Patiño-García A, Alonso MM, Gállego Pérez-Larraya J

Abstract
Background: Glioblastoma (GBM) is the most common malignant primary brain tumor in adults. Circulating biomarkers may assist in the processes of differential diagnosis and response assessment. GBM cells release extracellular vesicles containing a subset of proteins and nucleic acids. We previously demonstrated that exosomes isolated from the serum of GBM patients had an increased expression of RNU6-1 compared to healthy subjects. In this exploratory study, we investigated the role of this small noncoding RNA as a diagnostic biomarker for GBM versus other brain lesions with some potential radiological similarities.
Methods: We analyzed the expression of RNU6-1 in circulating exosomes of GBM patients (n = 18), healthy controls (n = 30), and patients with subacute stroke (n = 30), acute/subacute hemorrhage (n = 30), acute demyelinating lesions (n = 18), brain metastases (n = 21), and primary central nervous system lymphoma (PCNSL; n = 12) using digital droplet PCR.
Results: Expression of RNU6-1 was significantly higher in GBM patients than in healthy controls (P = .002). RNU6-1 levels were also significantly higher in exosomes from GBM patients than from patients with non-neoplastic lesions (stroke [P = .05], hemorrhage [P = .01], demyelinating lesions [P = .019]) and PCNSL (P = .004). In contrast, no significant differences were found between patients with GBM and brain metastases (P = .573). Receiver operator characteristic curve analyses supported the role of this biomarker in differentiating GBM from subacute stroke, acute/subacute hemorrhage, acute demyelinating lesions, and PCNSL (P < .05), but again not from brain metastases (P = .575).
Conclusions: Our data suggest that the expression of RNU6-1 in circulating exosomes could be useful for the differentiation of GBM from non-neoplastic brain lesions and PCNSL, but not from brain metastases.

PMID: 32642678 [PubMed]

Biomarkers and smart intracranial devices for the diagnosis, treatment, and monitoring of high-grade gliomas: a review of the literature and future prospects.

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Biomarkers and smart intracranial devices for the diagnosis, treatment, and monitoring of high-grade gliomas: a review of the literature and future prospects.

Neurooncol Adv. 2019 May-Dec;1(1):vdz013

Authors: Hafeez U, Cher LM

Abstract
Glioblastoma (GBM) is the most common primary brain neoplasm with median overall survival (OS) around 15 months. There is a dearth of effective monitoring strategies for patients with high-grade gliomas. Relying on magnetic resonance images of brain has its challenges, and repeated brain biopsies add significant morbidity. Hence, it is imperative to establish a less invasive way to diagnose, monitor, and guide management of patients with high-grade gliomas. Currently, multiple biomarkers are in various phases of development and include tissue, serum, cerebrospinal fluid (CSF), and imaging biomarkers. Here we review and summarize the potential biomarkers found in blood and CSF, including extracellular macromolecules, extracellular vesicles, circulating tumor cells, immune cells, endothelial cells, and endothelial progenitor cells. The ability to detect tumor-specific biomarkers in blood and CSF will potentially not only reduce the need for repeated brain biopsies but also provide valuable information about the heterogeneity of tumor, response to current treatment, and identify disease resistance. This review also details the status and potential scope of brain tumor-related cranial devices and implants including Ommaya reservoir, microelectromechanical systems-based depot device, Alzet mini-osmotic pump, Metronomic Biofeedback Pump (MBP), ipsum G1 implant, ultra-thin needle implant, and putative devices. An ideal smart cranial implant will overcome the blood-brain barrier, deliver various drugs, provide access to brain tissue, and potentially measure and monitor levels of various biomarkers.

PMID: 32642651 [PubMed]

CD44v8-10 mRNA contained in serum exosomes as a diagnostic marker for docetaxel resistance in prostate cancer patients.

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CD44v8-10 mRNA contained in serum exosomes as a diagnostic marker for docetaxel resistance in prostate cancer patients.

Heliyon. 2020 Jul;6(7):e04138

Authors: Kato T, Mizutani K, Kawakami K, Fujita Y, Ehara H, Ito M

Abstract
Background: Docetaxel is first-line chemotherapy for castration-resistant prostate cancer (CRPC), but most patients acquire docetaxel resistance. CD44 has been shown to be involved in drug resistance of cancers including prostate cancer. We hypothesized that CD44 in serum exosomes could be a diagnostic marker for docetaxel resistance in CRPC patients. In this study, we examined CD44 protein and mRNA expression in cell lysates and exosomes isolated from prostate cancer cells, evaluated the effect of CD44v8-10 knockdown on docetaxel sensitivity and measured CD44 mRNA copy numbers contained in serum exosomes in prostate cancer patients.
Materials and methods: Docetaxel-sensitive PC-3 prostate cancer cells and docetaxel-resistant PC-3R cells established previously from parental PC-3 cells were used. CD44v8-10 knockdown was performed by siRNA transfection. Blood was collected from 50 docetaxel-naïve and 10 docetaxel-resistant patients and 15 control males. CD44 protein expression was evaluated by Western blotting. CD44 mRNA expression was measured by RT-digital PCR.
Results: The levels of CD44v8-10 protein and mRNA in cell lysates and exosomes were higher in PC-3R cells than in PC-3 cells. CD44v8-10 knockdown significantly increased docetaxel sensitivity of PC-3R cells. The CD44v8-10 mRNA copy numbers in serum exosomes were higher in docetaxel-resistant patients than in docetaxel-naïve patients and control males (median 46, 12 and 17 copies/mL serum, respectively, P = 0.032). In contrast, the serum exosomal mRNA copy numbers of CD44 standard isoform (CD44s) were not different among 3 groups (median 25, 14 and 13 copies/mL serum, respectively, P = 0.150).
Conclusions: CD44v8-10 may be involved in docetaxel resistance in prostate cancer and serum exosomal CD44v8-10 mRNA could be a diagnostic marker for docetaxel-resistant CRPC.

PMID: 32642575 [PubMed]

Delivery strategies for macromolecular drugs in cancer therapy.

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Delivery strategies for macromolecular drugs in cancer therapy.

Acta Pharm Sin B. 2020 Jun;10(6):979-986

Authors: Guo Q, Jiang C

Abstract
With the development of biotherapy, biomacromolecular drugs have gained tremendous attention recently, especially in drug development field due to the sophisticated functions in vivo. Over the past few years, a motley variety of drug delivery strategies have been developed for biomacromolecular drugs to overcome the difficulties in the druggability, e.g., the instability and easily restricted by physiologic barriers. The application of novel delivery systems to deliver biomacromolecular drugs can usually prolong the half-life, increase the bioavailability, or improve patient compliance, which greatly improves the efficacy and potentiality for clinical use of biomacromolecular drugs. In this review, recent studies regarding the drug delivery strategies for macromolecular drugs in cancer therapy are summarized, mainly drawing on the development over the last five years.

PMID: 32642406 [PubMed]

Focused Ultrasound Hyperthermia Augments Release of Glioma-derived Extracellular Vesicles with Differential Immunomodulatory Capacity.

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Focused Ultrasound Hyperthermia Augments Release of Glioma-derived Extracellular Vesicles with Differential Immunomodulatory Capacity.

Theranostics. 2020;10(16):7436-7447

Authors: Sheybani ND, Batts AJ, Mathew AS, Thim EA, Price RJ

Abstract
Background: Increasing evidence points to the critical role of extracellular vesicles (EVs) as molecular parcels that carry a diverse array of bioactive payloads for coordination of complex intracellular signaling. Focused ultrasound (FUS) hyperthermia is a technique for non-invasive, non-ionizing sublethal heating of cells in a near-instantaneous manner; while it has been shown to improve drug delivery and immunological recognition of tumors, its impact on EVs has not been explored to date. The goal of this study was to determine whether FUS impacts the release, proteomic profile, and immune-activating properties of tumor-derived EVs. Methods: Monolayered murine glioma cells were seeded within acoustically transparent cell culture chambers, and FUS hyperthermia was applied to achieve complete coverage of the chamber. Glioma-derived EVs (GEVs) were isolated for characterization by Nanoparticle Tracking Analysis, cryo-electron microscopy and mass spectrometry. An in vitro experimental setup was designed to further dissect the impact of GEVs on innate inflammation; immortalized murine dendritic cells (DCs) were pulsed with GEVs (either naïve or FUS hyperthermia-exposed) and assayed for production of IL-12p70, an important regulator of DC maturation and T helper cell polarization toward the interferon-γ-producing type 1 phenotype. Results: We confirmed that FUS hyperthermia significantly augments GEV release (by ~46%) as well as shifts the proteomic profile of these GEVs. Such shifts included enrichment of common EV-associated markers, downregulation of markers associated with cancer progression and resistance and modulation of inflammation-associated markers. When DCs were pulsed with GEVs, we noted that naïve GEVs suppressed IL-12p70 production by DCs in a GEV dose-dependent manner. In contrast, GEVs from cells exposed to FUS hyperthermia promoted a significant upregulation in IL-12p70 production by DCs, consistent with a pro-inflammatory stimulus. Conclusion: FUS hyperthermia triggers release of proteomically distinct GEVs that are capable of facilitating an important component of innate immune activation, lending both to a potential mechanism by which FUS interfaces with the tumor-immune landscape and to a role for GEV-associated biomarkers in monitoring response to FUS.

PMID: 32642004 [PubMed - in process]

Current understanding of the role of Adipose-derived Extracellular Vesicles in Metabolic Homeostasis and Diseases: Communication from the distance between cells/tissues.

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Current understanding of the role of Adipose-derived Extracellular Vesicles in Metabolic Homeostasis and Diseases: Communication from the distance between cells/tissues.

Theranostics. 2020;10(16):7422-7435

Authors: Li CJ, Fang QH, Liu ML, Lin JN

Abstract
Extracellular vesicles (EVs) including exosomes, microvesicles (MVs), and apoptotic bodies, are small membrane vesicular structures that are released during cell activation, senescence, or programmed cell death, including apoptosis, necroptosis, and pyroptosis. EVs serve as novel mediators for long-distance cell-to-cell communications and can transfer various bioactive molecules, such as encapsulated cytokines and genetic information from their parental cells to distant target cells. In the context of obesity, adipocyte-derived EVs are implicated in metabolic homeostasis serving as novel adipokines. In particular, EVs released from brown adipose tissue or adipose-derived stem cells may help control the remolding of white adipose tissue towards browning and maintaining metabolic homeostasis. Interestingly, EVs may even serve as mediators for the transmission of metabolic dysfunction across generations. Also, EVs have been recognized as novel modulators in various metabolic disorders, including insulin resistance, diabetes mellitus, and non-alcoholic fatty liver disease. In this review, we summarize the latest progress from basic and translational studies regarding the novel effects of EVs on metabolic diseases. We also discuss EV-mediated cross-talk between adipose tissue and other organs/tissues that are relevant to obesity and metabolic diseases, as well as the relevant mechanisms, providing insight into the development of new therapeutic strategies in obesity and metabolic diseases.

PMID: 32642003 [PubMed - in process]

Bioenergetic Crosstalk between Mesenchymal Stem Cells and various Ocular Cells through the intercellular trafficking of Mitochondria.

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Bioenergetic Crosstalk between Mesenchymal Stem Cells and various Ocular Cells through the intercellular trafficking of Mitochondria.

Theranostics. 2020;10(16):7260-7272

Authors: Jiang D, Chen FX, Zhou H, Lu YY, Tan H, Yu SJ, Yuan J, Liu H, Meng W, Jin ZB

Abstract
Rationale: Mitochondrial disorders preferentially affect tissues with high energy requirements, such as the retina and corneal endothelium, in human eyes. Mesenchymal stem cell (MSC)-based treatment has been demonstrated to be beneficial for ocular degeneration. However, aside from neuroprotective paracrine actions, the mechanisms underlying the beneficial effect of MSCs on retinal and corneal tissues are largely unknown. In this study, we investigated the fate and associated characteristics of mitochondria subjected to intercellular transfer from MSCs to ocular cells. Methods: MSCs were cocultured with corneal endothelial cells (CECs), 661W cells (a photoreceptor cell line) and ARPE-19 cells (a retinal pigment epithelium cell line). Immunofluorescence, fluorescence activated cell sorting and confocal microscopy imaging were employed to investigate the traits of intercellular mitochondrial transfer and the fate of transferred mitochondria. The oxygen consumption rate of recipient cells was measured to investigate the effect of intercellular mitochondrial transfer. Transcriptome analysis was performed to investigate the expression of metabolic genes in recipient cells with donated mitochondria. Results: Mitochondrial transport is a ubiquitous intercellular mechanism between MSCs and various ocular cells, including the corneal endothelium, retinal pigmented epithelium, and photoreceptors. Additionally, our results indicate that the donation process depends on F-actin-based tunneling nanotubes. Rotenone-pretreated cells that received mitochondria from MSCs displayed increased aerobic capacity and upregulation of mitochondrial genes. Furthermore, living imaging determined the ultimate fate of transferred mitochondria through either degradation by lysosomes or exocytosis as extracellular vesicles. Conclusions: For the first time, we determined the characteristics and fate of mitochondria undergoing intercellular transfer from MSCs to various ocular cells through F-actin-based tunneling nanotubes, helping to characterize MSC-based treatment for ocular tissue regeneration.

PMID: 32641991 [PubMed - in process]

Bacterial extracellular vesicle-coated multi-antigenic nanovaccines protect against drug-resistant Staphylococcus aureus infection by modulating antigen processing and presentation pathways.

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Bacterial extracellular vesicle-coated multi-antigenic nanovaccines protect against drug-resistant Staphylococcus aureus infection by modulating antigen processing and presentation pathways.

Theranostics. 2020;10(16):7131-7149

Authors: Chen G, Bai Y, Li Z, Wang F, Fan X, Zhou X

Abstract
Background: Vaccination provides an alternative to antibiotics in addressing drug-resistant Staphylococcus aureus (S. aureus) infection. However, vaccine potency is often limited by a lack of antigenic breadth and a demand on the generation of antibody responses alone. Methods: In this study, bacterial extracellular vesicles (EVs) coating indocyanine green (ICG)-loaded magnetic mesoporous silica nanoparticles (MSN) were constructed as multi-antigenic vaccines (EV/ICG/MSN) with the ability to modulate antigen presentation pathways in dendritic cells (DCs) to induce cellular immune responses. Results: Exposing the EV/ICG/MSNs to a laser could promote DC maturation and enhance the proteasome-dependent antigen presentation pathway by facilitating endolysosomal escape, improving proteasome activity, and elevating MHC-I expression. Immunization by EV/ICG/MSNs with laser irradiation in vivo triggered improved CD8+ T cell responses while maintaining CD4+ T cell responses and humoral immunity. In addition, in vivo tracking data revealed that the vaccine could be efficiently transported from the injection site into lymph nodes. Skin infection experiments showed that the vaccine not only prevented and treated superficial infection but also decreased bacterial invasiveness, thus strongly suggesting that EV/ICG/MSNs were effective in preventing complications resulting from the introduction of S. aureus infections. Conclusion: This multi-antigenic nanovaccine-based modulation of antigen presentation pathways provides an effective strategy against drug-resistant S. aureus infection.

PMID: 32641983 [PubMed - in process]

Rab27a dependent exosome releasing participated in albumin handling as a coordinated approach to lysosome in kidney disease.

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Rab27a dependent exosome releasing participated in albumin handling as a coordinated approach to lysosome in kidney disease.

Cell Death Dis. 2020 Jul 08;11(7):513

Authors: Feng Y, Zhong X, Tang TT, Wang C, Wang LT, Li ZL, Ni HF, Wang B, Wu M, Liu D, Liu H, Tang RN, Liu BC, Lv LL

Abstract
Exosomes are increasingly recognized as vehicles of intercellular communication. However, the role of exosome in maintaining cellular homeostasis under stress conditions remained unclear. Here we show that Rab27a expression was upregulated exclusively in tubular epithelial cells (TECs) during proteinuria nephropathy established by adriamycin (ADR) injection and 5/6 nephrectomy as well as in chronic kidney disease patients, leading to the increased secretion of exosomes carrying albumin. The active exosome production promoted tubule injury and inflammation in neighboring and the producing cells. Interferon regulatory factor 1 (IRF-1) was found as the transcription factor contributed to the upregulation of Rab27a. Albumin could be detected in exosome fraction and co-localized with exosome marker CD63 indicating the secretion of albumin into extracellular space by exosomes. Interestingly, inhibition of exosome release accelerated albumin degradation which reversed tubule injury with albumin overload, while lysosome suppression augmented exosome secretion and tubule inflammation. Our findings revealed that IRF-1/Rab27a mediated exosome secretion constituted a coordinated approach to lysosome degradation for albumin handling, which lead to the augment of albumin toxicity as a maladaptive response to maintain cell homeostasis. The findings may suggest a novel therapeutic strategy for proteinuric kidney disease by targeting exosome secretion.

PMID: 32641688 [PubMed - in process]

In vivo two-photon microscopy reveals the contribution of Sox9+ cell to kidney regeneration in a mouse model with extracellular vesicle treatment.

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In vivo two-photon microscopy reveals the contribution of Sox9+ cell to kidney regeneration in a mouse model with extracellular vesicle treatment.

J Biol Chem. 2020 Jul 08;:

Authors: Zhang K, Chen S, Sun H, Wang L, Li H, Zhao J, Zhang C, Li N, Guo Z, Han Z, Han ZC, Zheng G, Chen X, Li Z

Abstract
Mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) have been shown to stimulate regeneration in the treatment of kidney injury. Renal regeneration is also thought to be stimulated by the activation of Sox9+ cells. However, whether and how the activation mechanisms underlying EV treatment and Sox9+ cell-dependent regeneration intersect is unclear. We reasoned that a high-resolution imaging platform in living animals could help to untangle this system. To test this idea, we first applied EVs derived from human placenta-derived MSCs (hP-MSCs) to a Sox9-CreERT2; R26mTmG transgenic mouse model of acute kidney injury (AKI). Then, we developed an abdominal imaging window (AIW) in the mouse and tracked the Sox9+ cells in the inducible Sox9 Cre transgenic mice via in vivo lineage tracing with two-photon intravital microscopy. Our results demonstrated that EVs can travel to the injured kidneys post intravenous injection as visualized by Gaussia luciferase imaging and markedly increased the activation of Sox9+ cells. Moreover, the two-photon living imaging of lineage-labeled Sox9+ cells showed that the EVs promoted the expansion of Sox9+ cells in kidneys post AKI. Histological staining results confirmed that the descendants of Sox9+ cells contributed to nephric tubule regeneration which significantly ameliorated the renal function after AKI. In summary, intravital lineage tracing with two-photon microscopy through an embedded AIW provides a practical strategy to investigate the beneficial functions and to clarify the mechanisms of regenerative therapies in AKI.

PMID: 32641493 [PubMed - as supplied by publisher]

Plasma gelsolin inhibits CD8+ T cell function and regulates glutathione production to confer chemoresistance in ovarian cancer.

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Plasma gelsolin inhibits CD8+ T cell function and regulates glutathione production to confer chemoresistance in ovarian cancer.

Cancer Res. 2020 Jul 08;:

Authors: Asare-Werehene M, Communal L, Carmona E, Han Y, Song YS, Burger D, Mes-Masson AM, Tsang BK

Abstract
Although initial treatment of ovarian cancer (OVCA) is successful, tumors typically relapse and become resistant to treatment. Due to poor infiltration of effector T cells, patients are mostly unresponsive to immunotherapy. Plasma gelsolin (pGSN) is transported by exosomes (sEV) and plays a key role in OVCA chemoresistance, yet little is known about its role in immunosurveillance. Here we report the immunomodulatory roles of sEV-pGSN in OVCA chemoresistance. In chemosensitive conditions, secretion of sEV-pGSN was low, allowing for optimal CD8+ T cell function. This resulted in increased T cell secretion of IFNγ, which reduced intracellular glutathione (GSH) production and sensitized chemosensitive cells to cisplatin (CDDP)-induced apoptosis. In chemoresistant conditions, increased secretion of sEV-pGSN by OVCA cells induced apoptosis in CD8+ T cells. IFNγ secretion was therefore reduced, resulting in high GSH production and resistance to CDDP-induced death in OVCA cells. These findings support our hypothesis that sEV-pGSN attenuates immunosurveillance and regulates GSH biosynthesis, a phenomenon that contributes to chemoresistance in OVCA.

PMID: 32641415 [PubMed - as supplied by publisher]

Exosomal noncoding RNAs and tumor drug resistance.

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Exosomal noncoding RNAs and tumor drug resistance.

Cancer Res. 2020 Jul 08;:

Authors: Guo C, Liu J, Zhou Q, Song J, Zhang Z, Li Z, Wang G, Sun Z, Yuan W

Abstract
Tumor drug resistance is a major challenge in the treatment of cancer. Non-coding RNAs (ncRNA) play a role in the progression of drug resistance. Recent studies have indicated that exosomes, with their in vitro and in vivo compatibility, are the best natural carrier of ncRNA, and their transport of ncRNA into cells could regulate drug resistance. Exosomal ncRNA impact drug resistance through participation in drug efflux, regulation of signaling pathways, and modification of the tumor microenvironment. In this review, we evaluate the mechanism of exosomal ncRNA related to tumor drug resistance, their role in different tumors, and potential clinical applications.

PMID: 32641408 [PubMed - as supplied by publisher]

Erythrocytic α-synuclein contained in microvesicles regulates astrocytic glutamate homeostasis: a new perspective on Parkinson's disease pathogenesis.

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Erythrocytic α-synuclein contained in microvesicles regulates astrocytic glutamate homeostasis: a new perspective on Parkinson's disease pathogenesis.

Acta Neuropathol Commun. 2020 Jul 08;8(1):102

Authors: Sheng L, Stewart T, Yang D, Thorland E, Soltys D, Aro P, Khrisat T, Xie Z, Li N, Liu Z, Tian C, Bercow M, Matsumoto J, Zabetian CP, Peskind E, Quinn JF, Shi M, Zhang J

Abstract
Parkinson's disease is a neurodegenerative disorder characterized by the transmission and accumulation of toxic species of α-synuclein (α-syn). Extracellular vesicles (EVs) are believed to play a vital role in the spread of toxic α-syn species. Recently, peripheral α-syn pathology has been investigated, but little attention has been devoted to erythrocytes, which contain abundant α-syn. In this study, we first demonstrated that erythrocyte-derived EVs isolated from Parkinson's disease patients carried elevated levels of oligomeric α-syn, compared to those from healthy controls. Moreover, human erythrocyte-derived EVs, when injected into peripheral blood in a mouse model of Parkinson's disease, were found to readily cross the blood-brain barrier (BBB). These EVs accumulated in astrocyte endfeet, a component of the BBB, where they impaired glutamate uptake, likely via interaction between excitatory amino acid transporter 2 (EAAT2) and oligomeric α-syn. These data suggest that erythrocyte-derived EVs and the oligomeric α-syn carried in them may play critical roles in the progression or even initiation of Parkinson's disease. Additionally, the mechanisms involved are attributable at least in part to dysfunction of astrocytes induced by these EVs. These observations provide new insight into the understanding of the mechanisms involved in Parkinson's disease.

PMID: 32641150 [PubMed - in process]

Optimized method for extraction of exosomes from human primary muscle cells.

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Optimized method for extraction of exosomes from human primary muscle cells.

Skelet Muscle. 2020 Jul 08;10(1):20

Authors: Le Gall L, Ouandaogo ZG, Anakor E, Connolly O, Butler Browne G, Laine J, Duddy W, Duguez S

Abstract
Skeletal muscle is increasingly considered an endocrine organ secreting myokines and extracellular vesicles (exosomes and microvesicles), which can affect physiological changes with an impact on different pathological conditions, including regenerative processes, aging, and myopathies. Primary human myoblasts are an essential tool to study the muscle vesicle secretome. Since their differentiation in conditioned media does not induce any signs of cell death or cell stress, artefactual effects from those processes are unlikely. However, adult human primary myoblasts senesce in long-term tissue culture, so a major technical challenge is posed by the need to avoid artefactual effects resulting from pre-senescent changes. Since these cells should be studied within a strictly controlled pre-senescent division count (<21 divisions), and yields of myoblasts per muscle biopsy are low, it is difficult or impossible to amplify sufficiently large cell numbers (some 250 × 106 myoblasts) to obtain sufficient conditioned medium for the standard ultracentrifugation approach to exosome isolation.Thus, an optimized strategy to extract and study secretory muscle vesicles is needed. In this study, conditions are optimized for the in vitro cultivation of human myoblasts, and the quality and yield of exosomes extracted using an ultracentrifugation protocol are compared with a modified polymer-based precipitation strategy combined with extra washing steps. Both vesicle extraction methods successfully enriched exosomes, as vesicles were positive for CD63, CD82, CD81, floated at identical density (1.15-1.27 g.ml-1), and exhibited similar size and cup-shape using electron microscopy and NanoSight tracking. However, the modified polymer-based precipitation was a more efficient strategy to extract exosomes, allowing their extraction in sufficient quantities to explore their content or to isolate a specific subpopulation, while requiring >30 times fewer differentiated myoblasts than what is required for the ultracentrifugation method. In addition, exosomes could still be integrated into recipient cells such as human myotubes or iPSC-derived motor neurons.Modified polymer-based precipitation combined with extra washing steps optimizes exosome yield from a lower number of differentiated myoblasts and less conditioned medium, avoiding senescence and allowing the execution of multiple experiments without exhausting the proliferative capacity of the myoblasts.

PMID: 32641118 [PubMed - in process]

Exosomes derived from human umbilical cord MSCs rejuvenate aged MSCs and enhance their functions for myocardial repair.

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Exosomes derived from human umbilical cord MSCs rejuvenate aged MSCs and enhance their functions for myocardial repair.

Stem Cell Res Ther. 2020 Jul 08;11(1):273

Authors: Zhang N, Zhu J, Ma Q, Zhao Y, Wang Y, Hu X, Chen J, Zhu W, Han Z, Yu H

Abstract
BACKGROUND: Age and other cardiovascular risk factors have been reported to impair the activities of mesenchymal stem cells (MSCs), which will affect the efficacy of stem cell transplantation. The objective of the study is to investigate whether exosomes derived from human umbilical cord MSCs (UMSCs) could enhance the activities of bone marrow MSCs from old person (OMSCs), and improve their capacity for cardiac repair.
METHODS: Exosomes extracted from conditioned medium of UMSCs were used to treat OMSCs to generate OMSCsExo. The key molecule in the exosomes that have potential to rejuvenate aged MSCs were screened, and the role of OMSC was tested in the mouse model of mycardial infarction (MI).
RESULTS: We found the activity of senescence-associated β-galactosidase and the expression of aging-related factors such as p53, p21, and p16 were significantly higher in OMSCs than those in UMSCs. After treatment with UMSC exosomes, these senescence phenotypes of OMSCs were remarkably reduced. The proliferation, migration, differentiation, and anti-apoptotic and paracrine effect were increased in OMSCsExo. In vivo study, mice with cardiac infarction had significantly better cardiac function, less fibrosis, and more angiogenesis after they were injected with OMSCsExo as compared with those with OMSC. There was more miR-136 expression in UMSCs and OMSCsExo than in OMSCs. Upregulation of miR-136 by transfection of miR-136 mimic into OMSCs significantly attenuated the apoptosis and senescence of OMSCs. Apoptotic peptidase activating factor (Apaf1) was found to be the downstream gene that is negatively regulated by miR-136 via directly targeting at its 3'UTR.
CONCLUSION: Our data suggest that exosomes from young MSCs can improve activities of aged MSCs and enhance their function for myocardial repair by transferring exosomal miR-136 and downregulating Apaf1.

PMID: 32641103 [PubMed - in process]

Protective effect of miRNA-containing extracellular vesicles derived from mesenchymal stromal cells of old rats on renal function in chronic kidney disease.

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Protective effect of miRNA-containing extracellular vesicles derived from mesenchymal stromal cells of old rats on renal function in chronic kidney disease.

Stem Cell Res Ther. 2020 Jul 08;11(1):274

Authors: Wang Y, Guo YF, Fu GP, Guan C, Zhang X, Yang DG, Shi YC

Abstract
INTRODUCTION: Mesenchymal stromal cells (MSCs) play an important role in the prevention of cell and tissue fibrosis. Senescence may decrease the function of MSCs during recovery from tissue and organ damage. Extracellular vesicles (EVs) released from MSCs contribute to the repair of kidney injury. We explored the influence of senescence on EVs derived from MSCs (MSC-EVs) and detected the protective effects of MSC-EVs expressing low levels of miR-294/miR-133 derived from old rats against chronic kidney disease (CKD).
METHODS: The effects of MSC-EVs derived from 3-month-old and 18-month-old male Fisher 344 rats on renal fibrosis were explored in a unilateral ureteral obstruction (UUO) model. pLV-miR-294/pLV-miR-133 mimic/inhibitor were injected into young and old rats before UUO to detect the effects of miR-294/miR-133, which were decreased in MSC-EVs and sera from old rats, on renal function in CKD. Transforming growth factor-β1 (TGF-β1)-induced human renal proximal tubular epithelial (HK2) cells were used to imitate the pathological process of renal fibrosis in vitro. Western blotting was used to assess the expression of epithelial/mesenchymal markers and phosphorylation of proteins in HK2 cells.
RESULTS: The inhibition of UUO-induced CKD by MSC-EVs was weaker in old rats than in young rats. Downregulation of miRNAs (miR-294 and miR-133) in both MSC-EVs and sera from old rats obviously attenuated UUO-induced renal injury in old rats. miR-294 and miR-133 overexpression mitigated TGF-β1-mediated epithelial-mesenchymal transition (EMT) in HK2 cells, and the obvious increase in the phosphorylation of both SMAD2/3 and ERK1/2 induced by TGF-β1 was prevented in miR-294- and miR-133-overexpressing HK2 cells.
CONCLUSIONS: The ability of MSC-EVs to inhibit renal fibrosis decreased with age. miR-294/miR-133 in MSC-EVs and sera had an important effect on renal fibrosis in old rats and on EMT in HK2 cells. Furthermore, miR-294/miR-133 overexpression prevented SMAD2/3 and ERK1/2 phosphorylation in HK2 cells during TGF-β1-mediated EMT. These findings show that miR-294/miR-133 may be therapeutic in renal fibrosis and related renal dysfunction in elderly individuals.

PMID: 32641100 [PubMed - in process]

Renal cancer-derived exosomes induce tumor immune tolerance by MDSCs-mediated antigen-specific immunosuppression.

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Renal cancer-derived exosomes induce tumor immune tolerance by MDSCs-mediated antigen-specific immunosuppression.

Cell Commun Signal. 2020 Jul 08;18(1):106

Authors: Gao Y, Xu H, Li N, Wang H, Ma L, Chen S, Liu J, Zheng Y, Zhang Y

Abstract
BACKGOUND: Although Myeloid-derived suppressor cells (MDSCs) have a prominent ability to suppress the immune responses of T lymphocytes and propel tumor immune escape, a lack of profound systemic immunesuppression in tumor-bearing mice and tumor patients. The underlying mechanism of these remains unclear.
METHODS: For this purpose, renal cancer-derived exosomes (RDEs) were first labeled with PKH67 and been observed the internalization by MDSCs. Flow cytometry analysis showed the proportion and activity change of MDSCs in spleen and bone marrow induced by RDEs. Further, western blot experiments were used to verify triggered mechanism of MDSCs by RDEs. Finally, proliferation and cytotoxicity of cytotoxic T lymphocytes (CTLs) co-cultured with MDSCs in vitro and a series of experiments in vivo were performed to demonstrate the specific inhibitory effect of RDEs-induced MDSCs.
RESULTS: This study suggested that RDEs crucially contributed to presenting antigenic information, activating and driving specific immunosuppressive effect to MDSCs. HSP70, which is highly expressed in RDEs, initiate this process in a toll like receptor 2 (TLR2)-dependent manner. Importantly, RDEs-induced MDSCs could exert an antigen-specific immunosuppression effect on CTL and specific promote renal tumors-growth and immune escape in consequence.
CONCLUSION: The immunosuppression mediated by MDSCs which is induced by RDEs is antigen-specific. HSP70, which is highly expressed in RDEs, plays a pivotal role in this process. Targeted abrogating the function of MDSCs, or eliminating the expression of HSP70 in exosomes, or blocking the crosstalk between them provides a new direction and theoretical support for future immunotherapy. Video abstract.

PMID: 32641056 [PubMed - in process]

Extracellular vesicles: novel communicators in lung diseases.

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Extracellular vesicles: novel communicators in lung diseases.

Respir Res. 2020 Jul 08;21(1):175

Authors: Mohan A, Agarwal S, Clauss M, Britt NS, Dhillon NK

Abstract
The lung is the organ with the highest vascular density in the human body. It is therefore perceivable that the endothelium of the lung contributes significantly to the circulation of extracellular vesicles (EVs), which include exosomes, microvesicles, and apoptotic bodies. In addition to the endothelium, EVs may arise from alveolar macrophages, fibroblasts and epithelial cells. Because EVs harbor cargo molecules, such as miRNA, mRNA, and proteins, these intercellular communicators provide important insight into the health and disease condition of donor cells and may serve as useful biomarkers of lung disease processes. This comprehensive review focuses on what is currently known about the role of EVs as markers and mediators of lung pathologies including COPD, pulmonary hypertension, asthma, lung cancer and ALI/ARDS. We also explore the role EVs can potentially serve as therapeutics for these lung diseases when released from healthy progenitor cells, such as mesenchymal stem cells.

PMID: 32641036 [PubMed - in process]

Cross talk between exosomes and pancreatic β-cells in diabetes.

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Cross talk between exosomes and pancreatic β-cells in diabetes.

Arch Physiol Biochem. 2020 Jul 08;:1-10

Authors: Wu Y, Huang Q, Bu S

Abstract
Exosomes are a class of extracellular vesicles with a diameter of 50-100 nm secreted by various cells. They are generated through complex intracellular production mechanisms before being secreted to the extracellular environment. Due to their inclusion of proteins, lipids, and nucleic acids, exosomes play an important role in intercellular communication. Pancreatic β-cells play an irreplaceable role in the body's glucose metabolism. Their dysfunction is one of the causes of diabetes. Exosomes of various cells regulate the function of β-cells by regulating autoimmunity, delivering non-coding RNAs, or directly regulating intracellular signal pathways. This communication between β-cells and other cells plays an important role in the pathogenesis and development of diabetes, and has potential for clinical application. This paper reviews the biological sources and functions of exosomes, as well as intercellular crosstalk between β-cells and other cells that is involved in β-cell failure and regeneration.

PMID: 32640845 [PubMed - as supplied by publisher]

Development of Artificial Plasma Membranes Derived Nanovesicles Suitable for Drugs Encapsulation.

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Development of Artificial Plasma Membranes Derived Nanovesicles Suitable for Drugs Encapsulation.

Cells. 2020 Jul 06;9(7):

Authors: Martinelli C, Gabriele F, Dini E, Carriero F, Bresciani G, Slivinschi B, Dei Giudici M, Zanoletti L, Manai F, Paolillo M, Schinelli S, Azzalin A, Comincini S

Abstract
Extracellular vesicles (EVs) are considered as promising nanoparticle theranostic tools in many pathological contexts. The increasing clinical employment of therapeutic nanoparticles is contributing to the development of a new research area related to the design of artificial EVs. To this aim, different approaches have been described to develop mimetic biologically functional nanovescicles. In this paper, we suggest a simplified procedure to generate plasma membrane-derived nanovesicles with the possibility to efficiently encapsulate different drugs during their spontaneously assembly. After physical and molecular characterization by Tunable Resistive Pulse Sensing (TRPS) technology, transmission electron microscopy, and flow cytometry, as a proof of principle, we have loaded into mimetic EVs the isoquinoline alkaloid Berberine chloride and the chemotherapy compounds Temozolomide or Givinostat. We demonstrated the fully functionality of these nanoparticles in drug encapsulation and cell delivery, showing, in particular, a similar cytotoxic effect of direct cell culture administration of the anticancer drugs. In conclusion, we have documented the possibility to easily generate scalable nanovesicles with specific therapeutic cargo modifications useful in different drug delivery contexts.

PMID: 32640653 [PubMed - in process]

The circulating exosomal microRNAs related to albuminuria in patients with diabetic nephropathy.

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The circulating exosomal microRNAs related to albuminuria in patients with diabetic nephropathy.

J Transl Med. 2019 07 22;17(1):236

Authors: Kim H, Bae YU, Jeon JS, Noh H, Park HK, Byun DW, Han DC, Ryu S, Kwon SH

Abstract
BACKGROUND: Diabetic nephropathy (DN) is associated with high risk of cardiovascular disease and mortality. Exosomal microRNAs (miRNAs) regulate gene expression in a variety of tissues and play important roles in the pathology of various diseases. We hypothesized that the exosomal miRNA profile would differ between DN patients and patients without nephropathy.
METHODS: We prospectively enrolled 74 participants, including healthy volunteers (HVs), diabetic patients without nephropathy, and those with DN. The serum exosomal miRNA profiles of participants were examined using RNA sequencing.
RESULTS: The expression levels of 107 miRNAs differed between HVs and patients without DN, whereas the expression levels of 95 miRNAs differed between HVs and patients with DN. Among these miRNAs, we found 7 miRNAs (miR-1246, miR-642a-3p, let-7c-5p, miR-1255b-5p, let-7i-3p, miR-5010-5p, miR-150-3p) that were uniquely up-regulated in DN patients compared to HVs, and miR-4449 that was highly expressed in DN patients compared to patients without DN. A pathway analysis revealed that these eight miRNAs are likely involved in MAPK signaling, integrin function in angiogenesis, and regulation of the AP-1 transcription factor. Moreover, they were all significantly correlated with the degree of albuminuria.
CONCLUSIONS: Patients with DN have a different serum exosomal miRNA profile compared to HVs. These miRNAs may be promising candidates for the diagnosis and treatment of DN and cardiovascular disease.

PMID: 31331349 [PubMed - indexed for MEDLINE]

Cholesterol efflux alterations in adolescent obesity: role of adipose-derived extracellular vesical microRNAs.

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Cholesterol efflux alterations in adolescent obesity: role of adipose-derived extracellular vesical microRNAs.

J Transl Med. 2019 07 22;17(1):232

Authors: Barberio MD, Kasselman LJ, Playford MP, Epstein SB, Renna HA, Goldberg M, DeLeon J, Voloshyna I, Barlev A, Salama M, Ferrante SC, Nadler EP, Mehta N, Reiss AB, Freishtat RJ

Abstract
BACKGROUND: Macrophage cholesterol efflux capacity has been identified as a predictor for cardiovascular disease. We assessed the relationship between adipocyte-derived extracellular vesicle microRNAs and macrophage cholesterol efflux capacity.
METHODS: We assessed an adolescent cohort (n = 93, Age, median (IQR) = 17 (3) year, Female = 71, Male = 22) throughout the BMI continuum (BMI = 45.2 (13.2) kg/m2) for: (1) cholesterol efflux capacity and lipoprotein profiles; (2) adipocyte-derived extracellular vesicle microRNAs in serum; (3) the role of visceral adipose tissue extracellular vesicle in regulation of cholesterol efflux and cholesterol efflux gene expression in THP-1 macrophages in vitro.
RESULTS: Efflux capacity was significantly associated with HDL (r = 0.30, p = 0.01) and LDL (r = 0.33, p = 0.005) particle size. Multivariate-analysis identified six microRNAs associated (p < 0.05) with cholesterol efflux capacity: miR-3129-5p (Beta = 0.695), miR-20b (0.430), miR9-5p (0.111), miR-320d (- 0.190), miR301a-5p (0.042), miR-155-5p (0.004). In response to increasing concentrations (1 μg/mL vs. 3 μg/mL) of VAT extracellular vesicle, cholesterol efflux (66% ± 10% vs. 49% ± 2%; p < 0.01) and expression of ABCA1 (FC = 1.9 ± 0.8 vs 0.5 ± 0.2; p < 0.001), CD36 (0.7 ± 0.4 vs. 2.1 ± 0.8, p = 0.02), CYP27A1 (1.4 ± 0.4 vs. 0.9 ± 0.5; p < 0.05), and LXRA (1.8 ± 1.1 vs. 0.5 ± 0.2; p < 0.05) was altered in THP-1 cells in vitro.
CONCLUSION: Adipocyte-derived extracellular vesicle microRNAs may, in part, be involved macrophage cholesterol efflux regulation.

PMID: 31331347 [PubMed - indexed for MEDLINE]

Perspectives in melanoma: meeting report from the Melanoma Bridge (November 29th-1 December 1st, 2018, Naples, Italy).

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Perspectives in melanoma: meeting report from the Melanoma Bridge (November 29th-1 December 1st, 2018, Naples, Italy).

J Transl Med. 2019 07 22;17(1):234

Authors: Ascierto PA, Agarwala SS, Botti G, Budillon A, Davies MA, Dummer R, Ernstoff M, Ferrone S, Formenti S, Gajewski TF, Garbe C, Hamid O, Lo RS, Luke JJ, Michielin O, Palmieri G, Zitvogel L, Marincola FM, Masucci G, Caracò C, Thurin M, Puzanov I

Abstract
Diagnosis of melanocytic lesions, correct prognostication of patients, selection of appropriate adjuvant and systemic therapies, and prediction of response to a given therapy remain very real challenges in melanoma. Recent studies have shown that immune checkpoint blockade that represents a forefront in cancer therapy, provide responses but they are not universal. Improved understanding of the tumor microenvironment, tumor immunity and response to therapy has prompted extensive translational and clinical research in melanoma. Development of novel biomarker platforms may help to improve diagnostics and predictive accuracy for selection of patients for specific treatment. There is a growing evidence that genomic and immune features of pre-treatment tumor biopsies may correlate with response in patients with melanoma and other cancers they have yet to be fully characterized and implemented clinically. For example, advancements in sequencing and the understanding of the tumor microenvironment in melanoma have led to the use of genome sequencing and gene expression for development of multi-marker assays that show association with inflammatory state of the tumor and potential to predict response to immunotherapy. As such, melanoma serves as a model system for understanding cancer immunity and patient response to immunotherapy, either alone or in combination with other treatment modalities. Overall, the aim for the translational and clinical studies is to achieve incremental improvements through the development and identification of optimal treatment regimens, which increasingly involve doublet as well as triplet combinations, as well as through development of biomarkers to improve immune response. These and other topics in the management of melanoma were the focus of discussions at the fourth Melanoma Bridge meeting (November 29th-December 1st, 2018, Naples, Italy), which is summarised in this report.

PMID: 31331337 [PubMed - indexed for MEDLINE]

 

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