Crosstalk between cells with different EMT profiles and endothelial cells in breast cancer progression

Epithelial to mesenchymal transition (EMT) is a developmental trait that is hijacked in some disease conditions such as fibrosis and cancer. In cancer, EMT has been linked to increased invasion and metastasis, and is generally associated with a poor prognosis. In this study, I have compared phenotypic and functional differences between two isogenic cell lines exhibiting an EMT profile, D492M and D492HER2, but to a distinct extent. These cell lines are derived from D492, a breast epithelial cell line with stem cell properties that can generate both luminal and myoepithelial cells, and in 3D culture, it forms branching terminal ductal lobular units (TDLU)-like structures. When co-cultured with breast endothelial cells, D492 undergoes EMT, which is the origin of the D492M subline. Another subline of D492 was generated by overexpressing the HER2 oncogene. D492M has a more fixed EMT phenotype while D492HER2 expresses a more intermediate EMT phenotype. Moreover, D492M and D492HER2 differ in their tumorigenicity, that is, they are non-tumorigenic and tumorigenic, respectively. This study aimed to analyze the expression profile of these cell lines and study how they interact with endothelial cells. This study has prompted the identification of potential oncogenes, YKL-40 and ECM1, and the evaluation of their effects on the cellular phenotype and angiogenic potential in endothelial cells. YKL-40 and ECM1 may provide D492HER2 with increased aggressiveness that supports cancer progression.Bandvefsumbreyting þekjufrumna (e. epithelial to mesenchymal transition (EMT)) er þroskunarfræðilegur ferill sem kemur fyrir í sjúkdómum eins og trefjun (e. fibrosis) og krabbameini. Í krabbameini hefur EMT verið tengt við ífarandi æxlisvöxt og meinvarpamyndun og er ferillinn almennt tengdur slæmum batahorfum. Í þessari rannsókn bar ég saman mun á svipgerð (e. phenotypic) og virkni (e. functional) tveggja frumulína með sama genetíska bakgrunn (e. isogenic) sem sýna mismunandi EMT prófíl, D492M og D492HER2. Þessar frumulínur eru komnar frá D492, brjóstþekjufrumulínu með stofnfrumueiginleika sem getur myndað bæði kirtilþekju- og vöðvaþekjufrumur, og í þrívíðri rækt myndar hún greinótta formgerð. Í samrækt með æðaþeli, undirgekkst D492 frumulínan EMT og við það myndaðist D492M. D492HER2 var hins vegar mynduð með yfirtjáningu á HER2 krabbameinsgeninu. Svipgerð D492M frumulínunnar með tilliti til EMT er lengra komin (e. fixed EMT phenotype) miðað við D492HER2 (e. intermediate EMT phenotype). Ennfremur eru D492M og D492HER2 mismunandi hvað varðar eiginleika til að mynda æxli í músum, það er að segja D492M myndar ekki æxli á meðan D492HER2 myndar æxli. Markmið þessarar rannsóknar var að greina mun í tjáningu gena á milli þessara frumulína og skoða mismun í samskiptum þeirra við æðaþel og stoðvef. Þessi rannsókn hefur leitt af sér greiningu mögulegra krabbameinsgena (e. potential oncogenes) og mat á áhrifum þeirra á frumusvipgerð og æðamyndun í æðaþelsfrumum. YKL-40 og ECM1 geta veitt D492HER2 aukna árásargirni sem styður framrás krabbameins

Playing, we participate! An inclusive look at pedagogical practices

El presente proyecto de carácter investigativo ¡Jugando, participamos! Una mirada inclusiva a las prácticas pedagógicas, articulado a la línea de investigación Discapacidad y Educación Inclusiva, tiene como propósito analizar las prácticas pedagógicas de las docentes de los jardines infantiles Betania y La Fortaleza relacionadas con la implementación de experiencias de juego potenciando practicas inclusivas. Esta propuesta está enmarcada en el paradigma cualitativo y hace parte del enfoque socio crítico, encontrando su accionar en el método de investigación acción tal como lo propone (Martí, 2017) en sus cuatro fases, de las cuales se desarrollaron dos. El objeto de estudio como actores principales fueron las infancias con parálisis cerebral habitantes de la localidad de Usme, agentes pedagógicos, familias y/ o cuidadores. Los instrumentos de recolección de información fueron: diario de campo, entrevistas estructuradas y valoraciones pedagógicas, como resultado se genera un análisis de las practicas pedagógicas existentes, para potenciar practicas inclusivas. De esta manera se diseña la propuesta ¡Jugando, participamos! de tipo planeación pedagógica, que contempla aspectos fundamentales de la educación inclusiva tales como el Diseño Universal para el Aprendizaje, ajustes razonables y sistemas de apoyo en aras de ampliar las posibilidades de la participación efectiva de todos y todas las niñas en los escenarios de la primera infancia.Secretaria de Integración SocialMagíster en Educación Inclusiva e InterculturalMaestríaThe present investigative project, "Playing, We Participate! An Inclusive Look at Pedagogical Practices," is connected to the research line of Disability and Inclusive Education. Its purpose is to analyze the pedagogical practices of teachers at the Betania and La Fortaleza kindergartens concerning the implementation of inclusive play experiences. This proposal is framed within the qualitative paradigm and follows a socio-critical approach, applying the action research method as proposed by (Martí, 2017) in its four phases, of which two were developed. The main subjects of study were children with cerebral palsy residing in the Usme locality, as well as pedagogical agents, families, and/or caregivers. The data collection instruments used were field diaries, structured interviews, and pedagogical assessments. The results generated an analysis of existing pedagogical practices to enhance inclusive practices. As a result, the "Playing, We Participate!" proposal was designed as a pedagogical planning approach, which includes key aspects of inclusive education such as Universal Design for Learning, reasonable adjustments, and support systems, aiming to broaden opportunities for effective participation of all children in early childhood settings

YKL-40/CHI3L1 facilitates migration and invasion in HER2 overexpressing breast epithelial progenitor cells and generates a niche for capillary-like network formation

Publisher's version (útgefin grein).Epithelial to mesenchymal transition (EMT) is a developmental event that is hijacked in some diseases such as fibrosis and cancer. In cancer, EMT has been linked to increased invasion and metastasis and is generally associated with a poor prognosis. In this study, we have compared phenotypic and functional differences between two isogenic cell lines with an EMT profile: D492M and D492HER2 that are both derived from D492, a breast epithelial cell line with stem cell properties. D492M is non-tumorigenic while D492HER2 is tumorigenic. Thus, the aim of this study was to analyze the expression profile of these cell lines, identify potential oncogenes, and evaluate their effects on cellular phenotype. We performed transcriptome and secretome analyses of D492M and D492HER2 and verified expression of selected genes at the RNA and protein level. One candidate, YKL-40 (also known as CHI3L1), was selected for further studies due to its differential expression between D492M and D492HER2, being considerably higher in D492HER2. YKL-40 has been linked to chronic inflammation diseases and cancer, yet its function is not fully understood. Knock-down experiments of YKL-40 in D492HER2 resulted in reduced migration and invasion as well as reduced ability to induce angiogenesis in an in vitro assay, plus changes in the EMT-phenotype. In summary, our data suggest that YKL-40 may provide D492HER2 with increased aggressiveness, supporting cancer progression and facilitating angiogenesis.This work was supported by Grants from Landspitali University Hospital Science Fund, University of Iceland Research Fund, and Icelandic Science and Technology Policy—Grant of Excellence: 152144051. ‘Göngum saman,’ a supporting group for breast cancer research in Iceland ( www.gongumsaman.is ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer Reviewe

YKL-40/CHI3L1 facilitates migration and invasion in HER2 overexpressing breast epithelial progenitor cells and generates a niche for capillary-like network formation

Publisher's version (útgefin grein).Epithelial to mesenchymal transition (EMT) is a developmental event that is hijacked in some diseases such as fibrosis and cancer. In cancer, EMT has been linked to increased invasion and metastasis and is generally associated with a poor prognosis. In this study, we have compared phenotypic and functional differences between two isogenic cell lines with an EMT profile: D492M and D492HER2 that are both derived from D492, a breast epithelial cell line with stem cell properties. D492M is non-tumorigenic while D492HER2 is tumorigenic. Thus, the aim of this study was to analyze the expression profile of these cell lines, identify potential oncogenes, and evaluate their effects on cellular phenotype. We performed transcriptome and secretome analyses of D492M and D492HER2 and verified expression of selected genes at the RNA and protein level. One candidate, YKL-40 (also known as CHI3L1), was selected for further studies due to its differential expression between D492M and D492HER2, being considerably higher in D492HER2. YKL-40 has been linked to chronic inflammation diseases and cancer, yet its function is not fully understood. Knock-down experiments of YKL-40 in D492HER2 resulted in reduced migration and invasion as well as reduced ability to induce angiogenesis in an in vitro assay, plus changes in the EMT-phenotype. In summary, our data suggest that YKL-40 may provide D492HER2 with increased aggressiveness, supporting cancer progression and facilitating angiogenesis.This work was supported by Grants from Landspitali University Hospital Science Fund, University of Iceland Research Fund, and Icelandic Science and Technology Policy—Grant of Excellence: 152144051. ‘Göngum saman,’ a supporting group for breast cancer research in Iceland ( www.gongumsaman.is ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer Reviewe

Os principais tipos e manifestações da Cirrose Hepática: uma atualização clínica

Introdução: A cirrose hepática é um processo patológica crônico, considerado a hepatopatia mais comum, definido como a conversão difusa morfoestrutural por nódulos de arquitetura anômalo envoltos por fibrose. Objetivou-se descrever os tipos mais relevantes de cirrose e suas devidas manifestações. Metodologia: Trata-se de uma revisão bibliográfica, fundamentada nas plataformas do SciELO, PubMed, Scopus, utilizando os termos “hepatical cirrhosis”, “liver disease” e “hepatocellular insufficiency” a qual através da revisão narrativa, abordou amplamente a respeito da contextualização da cirrose e as principais etiologias. Resultados e Discussão: Foi analisado que tal condição afeta qualquer faixa etária, sexo, etnia e independe da classe socioeconômica, mas as diversas etiologias impõem um perfil epidemiológico específico conforme a aparição. As principais origens abordam o tipo alcoólico, hepatite, aplicação crônica de alguns fármacos e esteatose gordurosa ou não. Ademais, estima-se que estas afetam a anatomofuncionalidade do órgão responsável por grande parte da homeostase, culminando em diversas manifestações clínicas.  Conclusão: A cirrose é uma consequência grave de fatores de base em estágio avançado, a qual devido ao seu curso geralmente silencioso culmina no desenvolvimento e progressão clínica. Neste contexto, a atenção aos fatores predisponentes como alimentação rica em lipídios, estilismo, negligência a exames de rotina, sedentarismo e obesidade contribuem constituem medidas eficazes de prevenção primária.&nbsp

ECM1 secreted by HER2-overexpressing breast cancer cells promotes formation of a vascular niche accelerating cancer cell migration and invasion.

To access publisher's full text version of this article click on the hyperlink belowThe tumor microenvironment is increasingly recognized as key player in cancer progression. Investigating heterotypic interactions between cancer cells and their microenvironment is important for understanding how specific cell types support cancer. Forming the vasculature, endothelial cells (ECs) are a prominent cell type in the microenvironment of both normal and neoplastic breast gland. Here, we sought out to analyze epithelial-endothelial cross talk in the breast using isogenic non-tumorigenic vs. tumorigenic breast epithelial cell lines and primary ECs. The cellular model used here consists of D492, a breast epithelial cell line with stem cell properties, and two isogenic D492-derived EMT cell lines, D492M and D492HER2. D492M was generated by endothelial-induced EMT and is non-tumorigenic while D492HER2 is tumorigenic, expressing the ErbB2/HER2 oncogene. To investigate cellular cross talk, we used both conditioned medium (CM) and 2D/3D co-culture systems. Secretome analysis of D492 cell lines was performed using mass spectrometry and candidate knockdown (KD), and overexpression (OE) was done using siRNA and CRISPRi/CRISPRa technology. D492HER2 directly enhances endothelial network formation and activates a molecular axis in ECs promoting D492HER2 migration and invasion, suggesting an endothelial feedback response. Secretome analysis identified extracellular matrix protein 1 (ECM1) as potential angiogenic inducer in D492HER2. Confirming its involvement, KD of ECM1 reduced the ability of D492HER2-CM to increase endothelial network formation and induce the endothelial feedback, while recombinant ECM1 (rECM1) increased both. Interestingly, NOTCH1 and NOTCH3 expression was upregulated in ECs upon treatment with D492HER2-CM or rECM1 but not by CM from D492HER2 with ECM1 KD. Blocking endothelial NOTCH signaling inhibited the increase in network formation and the ability of ECs to promote D492HER2 migration and invasion. In summary, our data demonstrate that cancer-secreted ECM1 induces a NOTCH-mediated endothelial feedback promoting cancer progression by enhancing migration and invasion. Targeting this interaction may provide a novel possibility to improve cancer treatment.Landspitali University Hospital Science Fund University of Iceland Research Fund Icelandic Science and Technology Polic

Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensis

Angiostrongylus costaricensis is a nematode that causes abdominal angiostrongyliasis, a widespread human parasitism in Latin America. This study aimed to characterize the protease profiles of different developmental stages of this helminth. First-stage larvae (L1) were obtained from the faeces of infected Sigmodon hispidus rodents and third-stage larvae (L3) were collected from mollusks Biomphalaria glabrata previously infected with L1. Adult worms were recovered from rodent mesenteric arteries. Protein extraction was performed after repeated freeze-thaw cycles followed by maceration of the nematodes in 40 mM Tris base. Proteolysis of gelatin was observed by zymography and found only in the larval stages. In L3, the gelatinolytic activity was effectively inhibited by orthophenanthroline, indicating the involvement of metalloproteases. The mechanistic class of the gelatinases from L1 could not be precisely determined using traditional class-specific inhibitors. Adult worm extracts were able to hydrolyze haemoglobin in solution, although no activity was observed by zymography. This haemoglobinolytic activity was ascribed to aspartic proteases following its effective inhibition by pepstatin, which also inhibited the haemoglobinolytic activity of L1 and L3 extracts. The characterization of protease expression throughout the A. costaricensis life cycle may reveal key factors influencing the process of parasitic infection and thus foster our understanding of the disease pathogenesis