Tumor Microenvironment Modulates Invadopodia Activity of Non-Selected and Acid-Selected Pancreatic Cancer Cells and Its Sensitivity to Gemcitabine and C18-Gemcitabine.

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is a deadly disease with high mortality due to early metastatic dissemination and high chemoresistance. All these factors are favored by its extracellular matrix (ECM)-rich microenvironment, which is also highly hypoxic and acidic. Gemcitabine (GEM) is still the first-line therapy in PDAC. However, it is quickly deaminated to its inactive metabolite. Several GEM prodrugs have emerged to improve its cytotoxicity. Here, we analyzed how the acidic/hypoxic tumor microenvironment (TME) affects the response of PDAC cell death and invadopodia-mediated ECM proteolysis to both GEM and its C18 prodrug. METHODS: For this, two PDAC cell lines, PANC-1 and Mia PaCa-2 were adapted to pHe 6.6 or not for 1 month, grown as 3D organotypic cultures and exposed to either GEM or C18 in the presence and absence of acidosis and the hypoxia inducer, deferoxamine. RESULTS: We found that C18 has higher cytotoxic and anti-invadopodia activity than GEM in all culture conditions and especially in acid and hypoxic environments. CONCLUSIONS: We propose C18 as a more effective approach to conventional GEM in developing new therapeutic strategies overcoming PDAC chemoresistance.

Diagnostic Imaging of Agminated Blue Lesions and Blue Lesions with Satellitosis: Case Series with a Concise Review of the Current Literature.

Background: Agmination and/or satellitosis in pigmented blue lesions is a phenomenon rarely mentioned in the literature and not well known. This phenomenon can be expressed by several benign and malignant pigmented blue lesions, such as blue nevi, Spitz nevi, melanocytoma and melanoma. On this spectrum, dermoscopy, reflectance confocal microscopy (RCM) and dynamic Optical coherence tomography (D-OCT) represent non-invasive imaging technologies, which may help clinicians in the diagnosis of melanoma and non-melanoma skin cancers in daily clinical practice. Methods: Currently, in the literature there is a lack of new data about agminated blue lesions and blues lesions with satellitosis, as well as the lack of a recent and updated review of the literature about this topic. Therefore, considering that clinicians must be confident with the diagnosis of these rare skin lesions, we decided to carry out this work. Results: In this paper, four new cases of agminated pigmented cutaneous lesions were described. Moreover, a review of the current literature on this topic was performed. Conclusions: A clinical-pathological correlation is often needed to reach a correct diagnosis; currently, dermoscopy and non-invasive diagnostic techniques, such as reflectance confocal microscopy and optical coherence tomography, due to the depth of these skin lesions in the dermis, can only make a partial and limited contribution.

Normative values and diagnostic optimisation of three social cognition measures for autism and schizophrenia diagnosis in a healthy adolescent and adult sample.

Awareness of the importance of assessing social cognition skills under conditions showing atypical social behaviours has increased over the years. However, the evaluation of the psychometric properties of the measures and the availability of normative values for the clinical context are still limited. This study aims to revise, provide normative values, and evaluate the clinical validity of the Italian version of three social cognition measures: Advanced Theory of Mind (A-ToM) task, the Emotion Attribution Task (EAT), and the Social Situation Task (SST). Measures were administered to 580 adolescents and adult healthy controls (age range 14-50). We performed differential item functioning and Rasch analysis to revise each task, so normative data of the revised measures were calculated. Moreover, the revised measures were administered to 38 individuals with autism spectrum disorder (ASD) and 35 individuals with schizophrenia spectrum disorders (SSD): ASD and SSD were matched by age, gender, and IQ with a control sample to evaluate clinical validity. ROC analysis showed that the SST is the best measure differentiating between healthy and clinical groups, compared to the A-ToM (AUCASD = 0.70; AUCSSD = 0.65) and EAT (AUCASD = 0.67; AUCSSD = 0.50), which showed poorer performance. For SSD diagnosis, two SST subscales (Violation and Gravity score) indicated the best accuracy (AUCs of 0.88 and 0.84, respectively); for the ASD diagnosis, we propose a combined score between the SST subscale and A-ToM (AUC = 0.86). The results suggest that the proposed measures can be used to support the diagnostic process and clinical practice.

Quantitative Diffusion-Weighted MR Imaging: Is There a Prognostic Role in Noninvasively Predicting the Histopathologic Type of Uveal Melanomas?

Histopathologically, uveal melanomas (UMs) can be classified as spindle cell, mixed cell and epithelioid cell type, with the latter having a more severe prognosis. The aim of our study was to assess the correlation between the apparent diffusion coefficient (ADC) and the histologic type of UMs in order to verify the role of diffusion-weighted magnetic resonance imaging (DWI) as a noninvasive prognostic marker. A total of 26 patients with UMs who had undergone MRI and subsequent primary enucleation were retrospectively selected. The ADC of the tumor was compared with the histologic type. The data were compared using both one-way analysis of variance (ANOVA) (assessing the three histologic types separately) and the independent t-test (dichotomizing histologic subtypes as epithelioid versus non-epithelioid). Histologic type was present as follows: the epithelioid cell was n = 4, and the spindle cell was n = 11, the mixed cell type was n = 11. The mean ADC was 1.06 ± 0.24 × 10-3 mm2/s in the epithelioid cells, 0.98 ± 0.19 × 10-3 mm2/s in the spindle cells and 0.96 ± 0.26 × 10-3 mm2/s in the mixed cell type. No significant difference in the mean ADC value of the histopathologic subtypes was found, either when assessing the three histologic types separately (p = 0.76) or after dichotomizing the histologic subtypes as epithelioid and non-epithelioid (p = 0.82). DWI-ADC is not accurate enough to distinguish histologic types of UMs.

Radiotherapy of Orbital and Ocular Adnexa Lymphoma: Literature Review and University of Catania Experience.

Orbital and ocular adnexa lymphomas are rare neoplasms confined to the orbital region. The prognosis is generally favorable, with a high proportion of localized disease, indolent clinical course, prolonged disease-free intervals, and low lymphoma-related mortality rate. We report our experience on eleven patients with confirmed histological diagnosis of lymphoma stage IE-IIE, treated between 2010 and 2021 with radiotherapy alone or in association with chemotherapy or immunotherapy. Eight patients were treated with primary radiotherapy only, while three received previous systemic treatments. Six patients were treated with Proton beam therapy (PBT), and five with external beam radiotherapy (EBRT). The five-year local control rate was 98%; only one patient developed an out-of-field recurrence. We also conducted a comprehensive literature review using electronic databases (PubMed, EMBASE, and Cochrane Library). Articles were selected based on their pertinence to treatment of the ocular and adnexal lymphoma focusing on radiotherapy techniques (electron beam radiotherapy, photon beam radiotherapy, or proton beam radiotherapy), treatment total dose, fractionation schedule, early and late radio-induced toxicities, and patient's clinical outcome. Radiotherapy is an effective treatment option for orbital lymphoma, especially as standard treatment in the early stage of orbital lymphoma, with excellent local control rate and low rates of toxicity.

The Role of Reirradiation in Childhood Progressive Diffuse Intrinsic Pontine Glioma (DIPG): An Ongoing Challenge beyond Radiobiology.

To investigate the clinical impact of multiple courses of irradiation on pediatric patients with progressive diffuse intrinsic pontine glioma (DIPG), we conducted a retrospective case series on three children treated at our institution from 2018 to 2022. All children were candidates to receive systemic therapy with vinorelbine and nimotuzumab. Radiotherapy was administered to a total dose of 54 Gy. At any disease progression, our local tumor board evaluated the possibility of offering a new course of radiotherapy. To determine feasibility and assess toxicity rates, all children underwent clinical and hematological evaluation both during and after the treatment. To assess efficacy, all children performed contrast-enhanced MRI almost quarterly after the end of the treatment. In all children, following any treatment course, neurological improvement (>80%) was associated with a radiological response (41.7-46%). The longest overall survival (24 months) was observed in the child who underwent three courses of radiotherapy, without experiencing significant side effects. Even though it goes beyond the understanding of conventional radiobiology, first and second reirradiation in pediatric patients with progressive DIPG may represent a feasible and safe approach, capable of increasing overall survival and disease-free survival in selected patients and improving their quality of life.

Defining the identity and the niches of epithelial stem cells with highly pleiotropic multilineage potency in the human thymus.

Thymus is necessary for lifelong immunological tolerance and immunity. It displays a distinctive epithelial complexity and undergoes age-dependent atrophy. Nonetheless, it also retains regenerative capacity, which, if harnessed appropriately, might permit rejuvenation of adaptive immunity. By characterizing cortical and medullary compartments in the human thymus at single-cell resolution, in this study we have defined specific epithelial populations, including those that share properties with bona fide stem cells (SCs) of lifelong regenerating epidermis. Thymic epithelial SCs display a distinctive transcriptional profile and phenotypic traits, including pleiotropic multilineage potency, to give rise to several cell types that were not previously considered to have shared origin. Using here identified SC markers, we have defined their cortical and medullary niches and shown that, in vitro, the cells display long-term clonal expansion and self-organizing capacity. These data substantively broaden our knowledge of SC biology and set a stage for tackling thymic atrophy and related disorders.

Extracellular Matrix Collagen I Differentially Regulates the Metabolic Plasticity of Pancreatic Ductal Adenocarcinoma Parenchymal Cell and Cancer Stem Cell.

Pancreatic ductal adenocarcinoma (PDAC) has a 5-year survival rate of less than 10 percent largely due to the intense fibrotic desmoplastic reaction, characterized by high levels of extracellular matrix (ECM) collagen I that constitutes a niche for a subset of cancer cells, the cancer stem cells (CSCs). Cancer cells undergo a complex metabolic adaptation characterized by changes in metabolic pathways and biosynthetic processes. The use of the 3D organotypic model in this study allowed us to manipulate the ECM constituents and mimic the progression of PDAC from an early tumor to an ever more advanced tumor stage. To understand the role of desmoplasia on the metabolism of PDAC parenchymal (CPC) and CSC populations, we studied their basic metabolic parameters in organotypic cultures of increasing collagen content to mimic in vivo conditions. We further measured the ability of the bioenergetic modulators (BMs), 2-deoxyglucose, dichloroacetate and phenformin, to modify their metabolic dependence and the therapeutic activity of paclitaxel albumin nanoparticles (NAB-PTX). While all the BMs decreased cell viability and increased cell death in all ECM types, a distinct, collagen I-dependent profile was observed in CSCs. As ECM collagen I content increased (e.g., more aggressive conditions), the CSCs switched from glucose to mostly glutamine metabolism. All three BMs synergistically potentiated the cytotoxicity of NAB-PTX in both cell lines, which, in CSCs, was collagen I-dependent and the strongest when treated with phenformin + NAB-PTX. Metabolic disruption in PDAC can be useful both as monotherapy or combined with conventional drugs to more efficiently block tumor growth.

ECM Composition Differentially Regulates Intracellular and Extracellular pH in Normal and Cancer Pancreatic Duct Epithelial Cells.

Intracellular pH (pHi) regulation is a challenge for the exocrine pancreas, where the luminal secretion of bicarbonate-rich fluid is accompanied by interstitial flows of acid. This acid-base transport requires a plethora of ion transporters, including bicarbonate transporters and the Na+/H+ exchanger isoform 1 (NHE1), which are dysregulated in Pancreatic Ductal Adenocarcinoma (PDAC). PDAC progression is favored by a Collagen-I rich extracellular matrix (ECM) which exacerbates the physiological interstitial acidosis. In organotypic cultures of normal human pancreatic cells (HPDE), parenchymal cancer cells (CPCs) and cancer stem cells (CSCs) growing on matrices reproducing ECM changes during progression, we studied resting pHi, the pHi response to fluxes of NaHCO3 and acidosis and the role of NHE1 in pHi regulation. Our findings show that: (i) on the physiological ECM, HPDE cells have the most alkaline pHi, followed by CSCs and CPCs, while a Collagen I-rich ECM reverses the acid-base balance in cancer cells compared to normal cells; (ii) both resting pHi and pHi recovery from an acid load are reduced by extracellular NaHCO3, especially in HPDE cells on a normal ECM; (iii) cancer cell NHE1 activity is less affected by NaHCO3. We conclude that ECM composition and the fluctuations of pHe cooperate to predispose pHi homeostasis towards the presence of NaHCO3 gradients similar to that expected in the tumor.

Hypofractionated Radiotherapy in Localized, Low-Intermediate-Risk Prostate Cancer: Current and Future Prospectives.

At the time of diagnosis, the vast majority of prostate carcinoma patients have a clinically localized form of the disease, with most of them presenting with low- or intermediate-risk prostate cancer. In this setting, various curative-intent alternatives are available, including surgery, external beam radiotherapy and brachytherapy. Randomized clinical trials have demonstrated that moderate hypofractionated radiotherapy can be considered as a valid alternative strategy for localized prostate cancer. High-dose-rate brachytherapy can be administered according to different schedules. Proton beam radiotherapy represents a promising strategy, but further studies are needed to make it more affordable and accessible. At the moment, new technologies such as MRI-guided radiotherapy remain in early stages, but their potential abilities are very promising.

Registro argentino de COVID-19 en VIH (Estudio COVIDARE): características clínicas, epidemiológicas y factores asociados a hospitalización

Introducción: Existe controversia con respecto a los factores que determinan un mayor riesgo de gravedad y complicaciones por COVID-19 en personas que viven con VIH (PVVIH). Asimismo, hay datos limitados sobre el impacto de la vacunación contra SARS-CoV-2 en la hospitalización en esta población. Objetivos: Describir las características clínicas y evolutivas de COVID-19 en PVVIH; Evaluar factores de riesgo para hospitalización; Evaluar el impacto de la vacunación en la hospitalización. Pacientes y Métodos: Estudio observacional, prospectivo, multicéntrico (septiembre de 2020 a junio de 2022). Se registraron variables clínicas, inmunovirológicas, tratamiento antirretroviral (TARV), vacunación contra SARS-CoV-2 y hospitalización en PVVIH con COVID-19. Se realizaron análisis uni y multivariados examinando factores asociados a hospitalización utilizando dos modelos: primer modelo (sin vacunación) y segundo modelo (vacunación, mínimo una dosis). Resultados: Se incluyeron 1.201 PVVIH. La mediana de edad fue 45 años. El 65,3% fueron hombres; el 38,7% presentó comorbilidades. Recibía TARV el 92,8% y presentó carga viral (CV) indetectable el 83,1%. La mediana de linfocitos T CD4+ fue de 600 céls/mm3. El 95,7% presentó síntomas. Las tasas de hospitalización, ingreso a UCI, requerimiento de oxígeno y muerte fueron 17,8%, 2,8%, 10,7% y 1,39%, respectivamente. De acuerdo con el análisis multivariado para el primer modelo, la edad > 60 años y las comorbilidades se asociaron a mayor riesgo de hospitalización, mientras que el sexo femenino y un recuento de linfocitos T CD4+ > 500 céls/mm3 tuvieron un efecto protector. En el segundo modelo sólo las comorbilidades se relacionaron con un mayor riesgo de hospitalización mientras que la vacunación y células CD4+ > 500 céls/mm3 la redujeron. Conclusiones: En PVVIH las comorbilidades se asociaron con mayor tasa de hospitalización, mientras que tener linfocitos T CD4+ elevados y estar vacunado tuvieron un efecto protector. El TARV y la CV no tuvieron impacto en modelo alguno mientras que la edad y el sexo solo influyeron cuando no se consideró la vacunación.

Background: There is controversy regarding the factors that determine a greater risk of severity and complications from COVID-19 in people living with HIV (PLHIV). Likewise, there are limited data on the impact of SARS-CoV-2 vaccination on hospitalization in this population. Aims: To describe clinical characteristics and outcome of COVID-19 in PLHIV; To assess risk factors for hospitalization; To evaluate the impact of vaccination on hospitalization. Methods: Multicenter, prospective, observational study (September 2020 to June 2022). Clinical and immunovirological variables, antiretroviral treatment (ART), SARS-CoV-2 vaccination, and hospitalization in PLHIV with COVID-19 were recorded. Univariate and multivariate analyzes were performed examining factors associated with hospitalization using two models: first model (without vaccination) and second model (vaccination, minimum one dose). Results: 1,201 PLHIV were included. The median age was 45 years. 65.3% were men; 38.7% presented comorbidities. 92.8% received ART and 83.1% presented undetectable viral load (VL). The median CD4+ T-cell count was 600/mm3. 95.7% presented symptoms. The rates of hospitalization, ICU admission, oxygen requirement, and death were 17.8 %, 2.8%, 10.7% and 1.39%, respectively. According to the multivariate analysis for the first model, age > 60 years and comorbidities were associated with a higher risk of hospitalization, while female sex and CD4+ > 500/mm3 had a protective effect. In the second model, only the comorbidities were associated with a higher risk of hospitalization, while vaccination and CD4+ > 500/mm3 reduced it. Conclusions: in PLHIV, comorbidities were associated with a higher hospitalization rate, while having elevated CD4+ T-cell counts and being vaccinated had a protective effect. ART and VL had no impact in any model, while age and sex only had an influence when vaccination was not considered.

Tenofovir-Containing Antiretroviral Therapy and Clinical Outcomes of SARS-CoV-2 Infection in People Living with HIV.

Tenofovir has been hypothesized to be effective against COVID-19 and is available as two prodrugs, tenofovir disoproxil fumarate (TDF) and tenofovir alafenamide (TAF), both part of antiretroviral therapy (ART) regimens. People living with human immunodeficiency virus (PLWH) might be at higher risk for COVID-19 progression; however, information about the impact of tenofovir on COVID-19 clinical outcomes remains controversial. The COVIDARE is a prospective observational multicentric study in Argentina. PLWH with COVID-19 were enrolled from September 2020 to mid-June 2022. Patients were stratified according to baseline ART into those with tenofovir (TDF or TAF) and those without. Univariate and multivariate analyses were performed to evaluate the impact of tenofovir vs. non-tenofovir-containing regimens on major clinical outcomes. Of the 1155 subjects evaluated, 927 (80%) received tenofovir-based ART (79% TDF, 21% TAF) whilst the remaining population was under non-tenofovir regimens. The non-tenofovir group had older age and a higher prevalence of heart and kidney disease. Regarding the prevalence of symptomatic COVID-19, tomographic findings, hospitalization, and mortality, no differences were observed. The oxygen therapy requirement was higher in the non-tenofovir group. In the multivariate analyses, a first model with adjustment for viral load, CD4 T-cell count, and overall comorbidities showed that oxygen requirement was associated with non-tenofovir ART. In a second model with adjustment by chronic kidney disease, tenofovir exposure was not statistically significant.

Chemometrics in Protein Formulation: Stability Governed by Repulsion and Protein Unfolding.

Therapeutic proteins can be challenging to develop due to their complexity and the requirement of an acceptable formulation to ensure patient safety and efficacy. To date, there is no universal formulation development strategy that can identify optimal formulation conditions for all types of proteins in a fast and reliable manner. In this work, high-throughput characterization, employing a toolbox of five techniques, was performed on 14 structurally different proteins formulated in 6 different buffer conditions and in the presence of 4 different excipients. Multivariate data analysis and chemometrics were used to analyze the data in an unbiased way. First, observed changes in stability were primarily determined by the individual protein. Second, pH and ionic strength are the two most important factors determining the physical stability of proteins, where there exists a significant statistical interaction between protein and pH/ionic strength. Additionally, we developed prediction methods by partial least-squares regression. Colloidal stability indicators are important for prediction of real-time stability, while conformational stability indicators are important for prediction of stability under accelerated stress conditions at 40 °C. In order to predict real-time storage stability, protein-protein repulsion and the initial monomer fraction are the most important properties to monitor.

Integrated Intensified Chemoradiation in the Setting of Total Neoadjuvant Therapy (TNT) in Patients with Locally Advanced Rectal Cancer: A Retrospective Single-Arm Study on Feasibility and Efficacy.

While surgery is considered the main treatment for early-stage rectal cancer, locally advanced rectal cancer needs to be handled with a multidisciplinary approach. Based on literature data suggesting promising advantages of total neoadjuvant therapy (TNT), we performed a retrospective, single-arm, single-center study on 45 patients affected by histologically and radiologically proven locally advanced rectal cancer, with the aim of analyzing the feasibility and short-term efficacy of an integrated intensified treatment in the setting of TNT. Each analyzed patient performed three cycles of FOLFOX4 or De Gramont induction chemotherapy (iCT), followed by concurrent chemoradiotherapy (CRT) with long course radiotherapy (LCRT) plus concomitant boost and continuous 5-FU infusion, followed by three cycles of FOLFOX4 or De Gramont consolidation chemotherapy (conCT) and then surgery with total mesorectal excision. At a median follow-up of 30 months, this strategy has shown to be feasible and effective in terms of pathological complete response (pCR) and short-term disease-free survival (DFS).

A rubric as a tool for summative and formative evaluation in Laboratory Animals Technologist degree at the University of Buenos Aires (Argentina).

Laboratory Animals Technologist degree (Técnico Universitario en Gestión Integral de Bioterios) was created in 1989. It is offered jointly by the Faculty of Veterinary Sciences and the Faculty of Pharmacy and Biochemistry at the University of Buenos Aires. The complete programme is organized into 15 courses, including basic and applied sciences, along with specific courses (Técnicas de Bioterio I to V) with a solid practice content that students must achieve.An additional challenge occurs when practices consist of animal handling and procedures, which can be stressful for students and animals. Since traditional evaluation methods may not be the best way to determine the acquisition of knowledge and practical performance, an evaluation rubric is proposed to assess the practices carried out by students.The aspects and criteria to be examined were previously known of by students and teachers. The evaluation levels include: 'Daily report and record-keeping', 'Food and water supply', 'Cleaning and disinfection of areas, Standardized Operational Procedures and routines fulfilment', 'Personnel and supplies circulation', 'Use of personal protection equipment and physical care', 'Fulfilment of tasks in established times', 'Animals procedures and organization of needed materials', 'Animal care' and 'Teamwork camaraderie and organization'.The proposed tool was implemented with good results to achieve the academic objectives while augmenting the confidence for practical expertise, minimizing doubts and uncertainties related to traditional evaluation methods, and providing students with an environment that favours knowledge acquisition and practical skills, establishing standard criteria among the teaching staff, and reducing and refining the use of animals for educational purposes.

A Differential Inertial Wearable Device for Breathing Parameter Detection: Hardware and Firmware Development, Experimental Characterization.

Breathing monitoring is crucial for evaluating a patient's health status. The technologies commonly used to monitor respiration are costly, bulky, obtrusive, and inaccurate, mainly when the user moves. Consequently, efforts have been devoted to providing new solutions and methodologies to overcome these limitations. These methods have several uses, including healthcare monitoring, measuring athletic performance, and aiding patients with respiratory diseases, such as COPD (chronic obtrusive pulmonary disease), sleep apnea, etc. Breathing-induced chest movements can be measured noninvasively and discreetly using inertial sensors. This research work presents the development and testing of an inertia-based chest band for breathing monitoring through a differential approach. The device comprises two IMUs (inertial measurement units) placed on the patient's chest and back to determine the differential inertial signal, carrying out information detection about the breathing activity. The chest band includes a low-power microcontroller section to acquire inertial data from the two IMUs and process them to extract the breathing parameters (i.e., RR-respiration rate; TI/TE-inhalation/exhalation time; IER-inhalation-to-exhalation time; V-flow rate), using the back IMU as a reference. A BLE transceiver wirelessly transmits the acquired breathing parameters to a mobile application. Finally, the test results demonstrate the effectiveness of the used dual-inertia solution; correlation and Bland-Altman analyses were performed on the RR measurements from the chest band and the reference, demonstrating a high correlation (r¯ = 0.92) and low mean difference (MD¯ = -0.27 BrPM (breaths per minute)), limits of agreement (LoA¯ = +1.16/-1.75 BrPM), and mean absolute error (MAE¯ = 1.15%). Additionally, the experimental results demonstrated that the developed device correctly measured the other breathing parameters (TI, TE, IER, and V), keeping an MAE of ≤5%. The obtained results indicated that the developed chest band is a viable solution for long-term breathing monitoring, both in stationary and moving users.

PCSK9 Confers Inflammatory Properties to Extracellular Vesicles Released by Vascular Smooth Muscle Cells.

Vascular smooth muscle cells (VSMCs) are key participants in both early- and late-stage atherosclerosis and influence neighbouring cells possibly by means of bioactive molecules, some of which are packed into extracellular vesicles (EVs). Proprotein convertase subtilisin/kexin type 9 (PCSK9) is expressed and secreted by VSMCs. This study aimed to unravel the role of PCSK9 on VSMCs-derived EVs in terms of content and functionality. EVs were isolated from human VSMCs overexpressing human PCSK9 (VSMCPCSK9-EVs) and tested on endothelial cells, monocytes, macrophages and in a model of zebrafish embryos. Compared to EVs released from wild-type VSMCs, VSMCPCSK9-EVs caused a rise in the expression of adhesion molecules in endothelial cells and of pro-inflammatory cytokines in monocytes. These acquired an increased migratory capacity, a reduced oxidative phosphorylation and secreted proteins involved in immune response and immune effector processes. Concerning macrophages, VSMCPCSK9-EVs enhanced inflammatory milieu and uptake of oxidized low-density lipoproteins, whereas the migratory capacity was reduced. When injected into zebrafish embryos, VSMCPCSK9-EVs favoured the recruitment of macrophages toward the site of injection. The results of the present study provide evidence that PCSK9 plays an inflammatory role by means of EVs, at least by those derived from smooth muscle cells of vascular origin.

Upregulation of YKL-40 Promotes Metastatic Phenotype and Correlates with Poor Prognosis and Therapy Response in Patients with Colorectal Cancer.

YKL-40 is a heparin- and chitin-binding glycoprotein that belongs to the family of glycosyl hydrolases but lacks enzymatic properties. It affects different (patho)physiological processes, including cancer. In different tumors, YKL-40 gene overexpression has been linked to higher cell proliferation, angiogenesis, and vasculogenic mimicry, migration, and invasion. Because, in colorectal cancer (CRC), the serological YKL-40 level may serve as a risk predictor and prognostic biomarker, we investigated the underlying mechanisms by which it may contribute to tumor progression and the clinical significance of its tissue expression in metastatic CRC. We demonstrated that high-YKL-40-expressing HCT116 and Caco2 cells showed increased motility, invasion, and proliferation. YKL-40 upregulation was associated with EMT signaling activation. In the AOM/DSS mouse model, as well as in tumors and sera from CRC patients, elevated YKL-40 levels correlated with high-grade tumors. In retrospective analyses of six independent cohorts of CRC patients, elevated YKL-40 expression correlated with shorter survival in patients with advanced CRC. Strikingly, high YKL-40 tissue levels showed a predictive value for a better response to cetuximab, even in patients with stage IV CRC and mutant KRAS, and worse sensitivity to oxaliplatin. Taken together, our findings establish that tissue YKL-40 overexpression enhances CRC metastatic potential, highlighting this gene as a novel prognostic candidate, a predictive biomarker for therapy response, and an attractive target for future therapy in CRC.