Cancer associated fibroblasts predict for poor outcome and promote periostin-dependent invasion in oesophageal adenocarcinoma

Interactions between cancer cells and cancer-associated fibroblasts (CAF) play an important role in tumour development and progression. In this study we investigated the functional role of CAF in oesophageal adenocarcinoma (EAC). We used immunochemistry to analyse a cohort of EAC patients (183 patients) for CAF markers related to disease mortality. We characterized CAF and normal oesophageal fibroblasts (NOF) using western blotting, immunofluorescence and gel contraction. Transwell assays, 3-D organotypic culture and xenograft models were used to examine effects on EAC cell function, and dissect molecular mechanisms regulating invasion. Most EAC (93%) contained CAF with a myofibroblastic (?-SMA-positive) phenotype, which correlated significantly with poor survival (p?=?0.016; HR 7. 1 (1.7-29.4). Primary CAF, isolated from EAC, have a contractile, myofibroblastic phenotype, and promote EAC cell invasion in vitro (Transwell assays, p?=?<0.05; organotypic culture, p?<?0.001) and in vivo (p?=?<0.05). In vitro, this pro-invasive effect is modulated through the matricellular protein periostin. Periostin is secreted by CAF, and acts as a ligand for EAC cell integrins ?v?3 and ?v?5, promoting activation of the PI3kinase/Akt pathway. In patient samples, periostin expression at the tumour cell/stromal interface correlates with poor overall and disease-free survival. Our study highlights the importance of the tumour stroma in EAC progression. Paracrine interaction between CAF-secreted periostin and EAC-expressed integrins results in PI3 kinase/Akt activation and increased tumour cell invasion. Most EAC contain a myofibroblastic CAF-rich stroma; this may explain the aggressive, highly infiltrative nature of the disease, and suggests that stromal targeting may produce therapeutic benefit in EAC patient

HPV, tumour metabolism and novel target identification in head and neck squamous cell carcinoma

Background Metabolic changes in tumour cells are used in clinical imaging and may provide potential therapeutic targets. Human papillomavirus (HPV) status is important in classifying head and neck cancers (HNSCC), identifying a distinct clinical phenotype; metabolic differences between these HNSCC subtypes remain poorly understood. Methods We used RNA sequencing to classify the metabolic expression profiles of HPV+ve and HPV−ve HNSCC, performed a meta-analysis on FDG-PET imaging characteristics and correlated results with in vitro extracellular flux analysis of HPV−ve and HPV+ve HNSCC cell lines. The monocarboxylic acid transporter-1 (MCT1) was identified as a potential metabolic target and tested in functional assays. Results Specific metabolic profiles were associated with HPV status, not limited to carbohydrate metabolism. There was dominance of all energy pathways in HPV-negative disease, with elevated expression of genes associated with glycolysis and oxidative phosphorylation. In vitro analysis confirmed comparative increased rates of oxidative phosphorylation and glycolysis in HPV-negative cell lines. PET SUV(max) scores however were unable to reliably differentiate between HPV-positive and HPV-negative tumours. MCT1 expression was significantly increased in HPV-negative tumours, and inhibition suppressed tumour cell invasion, colony formation and promoted radiosensitivity. Conclusion HPV-positive and negative HNSCC have different metabolic profiles which may have potential therapeutic applications

A subset of myofibroblastic cancer-associated fibroblasts regulate collagen fiber elongation, which is prognostic in multiple cancers

Collagen structure has been shown to influence tumor cell invasion, metastasis and clinical outcome in breast cancer. However, it remains unclear how it affects other solid cancers. Here we utilized multi-photon laser scanning microscopy and Second Harmonic Generation to identify alterations to collagen fiber structure within the tumor stroma of head & neck, esophageal and colorectal cancers. Image segmentation algorithms were then applied to quantitatively characterize these morphological changes, showing that elongated collagen fibers significantly correlated with poor clinical outcome (Log Rank p < 0.05). We used TGF-? treatment to model fibroblast conversion to smooth muscle actin SMA-positive cancer associated fibroblasts (CAFs) and found that these cells induce the formation of elongated collagen fibers in vivo. However, proteomic/transcriptomic analysis of SMA-positive CAFs cultured ex-vivo showed significant heterogeneity in the expression of genes with collagen fibril organizing gene ontology. Notably, stratifying patients according to stromal SMA-positivity and collagen fiber elongation was found to provide a highly significant correlation with poor survival in all 3 cancer types (Log Rank p ? 0.003). In summary, we show that increased collagen fiber length correlates with poor patient survival in multiple tumor types and that only a sub-set of SMA-positive CAFs can mediate the formation of this collagen structure

HABITAT: An IoT Solution for Independent Elderly

In this work, a flexible and extensive digital platform for Smart Homes is presented, exploiting the most advanced technologies of the Internet of Things, such as Radio Frequency Identification, wearable electronics, Wireless Sensor Networks, and Artificial Intelligence. Thus, the main novelty of the paper is the system-level description of the platform flexibility allowing the interoperability of different smart devices. This research was developed within the framework of the operative project HABITAT (Home Assistance Based on the Internet of Things for the Autonomy of Everybody), aiming at developing smart devices to support elderly people both in their own houses and in retirement homes, and embedding them in everyday life objects, thus reducing the expenses for healthcare due to the lower need for personal assistance, and providing a better life quality to the elderly users

Induction of fibroblast senescence generates a non-fibrogenic myofibroblast phenotype that differentially impacts on cancer prognosis

Cancer-associated fibroblasts (CAF) remain a poorly characterized, heterogeneous cell population. Here we characterized two previously described tumor-promoting CAF sub-types, smooth muscle actin (SMA)-positive myofibroblasts and senescent fibroblasts, identifying a novel link between the two. Analysis of CAF cultured ex vivo, showed that senescent CAF are predominantly SMA-positive; this was confirmed by immunochemistry in head & neck (HNSCC) and esophageal (EAC) cancers. In vitro, we found that fibroblasts induced to senesce develop molecular, ultrastructural and contractile features typical of myofibroblasts and this is dependent on canonical TGF-? signaling. Similar to TGF-?1-generated myofibroblasts, these cells secrete soluble factors that promote tumor cell motility. However, RNA-sequencing revealed significant transcriptomic differences between the two SMA-positive CAF groups, particularly in genes associated with extracellular matrix (ECM) deposition and organization, which differentially promote tumor cell invasion. Notably, second harmonic generation imaging and bioinformatic analysis of SMA-positive human HNSCC and EAC showed that collagen fiber organization correlates with poor prognosis, indicating that heterogeneity within the SMA-positive CAF population differentially impacts on survival. These results show that non-fibrogenic, SMA-positive myofibroblasts can be directly generated through induction of fibroblast senescence and suggest that senescence and myofibroblast differentiation are closely linked processes

NOX4 inhibition potentiates immunotherapy by overcoming cancer-associated fibroblast-mediated CD8 T-cell exclusion from tumours

Determining mechanisms of resistance to PD-1/PD-L1 immune checkpoint immunotherapy is key to developing new treatment strategies. Cancer-associated fibroblasts (CAF) have many tumor-promoting functions and promote immune evasion through multiple mechanisms, but as yet, there are no CAF-specific inhibitors clinically available. Here we generated CAF-rich murine tumor models (TC1, MC38, 4T1) to investigate how CAF influence the immune microenvironment and affect response to different immunotherapy modalities (anti-cancer vaccination; TC1, [HPV E7 DNA vaccine];PD-1, MC38) and found that CAFs broadly suppressed response by specifically excluding CD8+ T-cells from tumors (not CD4+ T-cells or macrophages); CD8+ T-cell exclusion was similarly present in CAF-rich human tumors. RNA sequencing of CD8+ T-cells from CAF-rich murine tumors and immunochemistry analysis of human tumors identified significant upregulation of CTLA-4 in the absence of other exhaustion markers; inhibiting CTLA-4 with a non-depleting antibody overcame the CD8+ T-cell exclusion effect without affecting T-regs. We then examined the potential for CAF targeting, focusing on the ROS-producing enzyme NOX4, which is upregulated by CAF in many human cancers, and compared this to TGF-062;1 inhibition, a key regulator of the CAF phenotype. siRNA knockdown or pharmacological inhibition (GKT137831 [Setanaxib]) of NOX4 'normalized' CAF to a quiescent phenotype and promoted intratumoral CD8+T-cell infiltration, overcoming the exclusion effect; TGF-062;1 inhibition could prevent, but not reverse, CAF differentiation. Finally, NOX4 inhibition restored immunotherapy response in CAF-rich tumors. These findings demonstrate that CAF-mediated immunotherapy resistance can be effectively overcome through NOX4 inhibition, and could improve outcome in a broad range of cancers

ATM Regulates Differentiation of Myofibroblastic Cancer-Associated Fibroblasts and Can Be Targeted to Overcome Immunotherapy Resistance

Myofibroblastic cancer-associated fibroblast (myoCAF)-rich tumors generally contain few T cells and respond poorly to immune-checkpoint blockade. Although myoCAFs are associated with poor outcome in most solid tumors, the molecular mechanisms regulating myoCAF accumulation remain unclear, limiting the potential for therapeutic intervention. Here, we identify ataxia-telangiectasia mutated (ATM) as a central regulator of the myoCAF phenotype. Differentiating myofibroblasts in vitro and myoCAFs cultured ex vivo display activated ATM signaling, and targeting ATM genetically or pharmacologically could suppress and reverse differentiation. ATM activation was regulated by the reactive oxygen species-producing enzyme NOX4, both through DNA damage and increased oxidative stress. Targeting fibroblast ATM in vivo suppressed myoCAF-rich tumor growth, promoted intratumoral CD8 T-cell infiltration, and potentiated the response to anti-PD-1 blockade and antitumor vaccination. This work identifies a novel pathway regulating myoCAF differentiation and provides a rationale for using ATM inhibitors to overcome CAF-mediated immunotherapy resistance.SignificanceATM signaling supports the differentiation of myoCAFs to suppress T-cell infiltration and antitumor immunity, supporting the potential clinical use of ATM inhibitors in combination with checkpoint inhibition in myoCAF-rich, immune-cold tumors

A cidadania italiana e os descendentes de cidadãos italianos emigrados na Colômbia. A eliminação de uma injustiça social

El Derecho a menudo es fuente de discriminaciones sociales y a la vez puede ser la herramienta para solucionarlas. Los autores tratan de dar un ejemplo de este fenómeno, al analizar el impacto de las normas en materia de ciudadanía italiana sobre los descendientes de ciudadanos italianos emigrados al extranjero y más precisamente a América del Sur.Así, en aplicación de la vieja normativa italiana, solo el padre transmitía iure sanguinis la ciudadanía a sus descendientes: además, las mujeres perdían automáticamente la ciudadanía italiana si hubiesen adquirido una ciudadanía extranjera por matrimonio con cónyuge extranjero. Estas normas discriminaban a la mujer italiana emigrada al extranjero y especialmente a sus descendientes, quienes no podían adquirir dicha ciudadanía. Las normas discriminatorias fueron por fin abrogadas por parte de la Corte Constitucional en los años setenta y ochenta: sin embargo, los efectos de estas seguían persistiendo, considerando que las sentencias de la Corte Constitucional no cobijaban hechos ocurridos anteriormente a la entrada en vigor de la Constitución (01.01.1948) y así no daban solución a las discriminaciones acontecidas antes de esa fecha. En el 2009, la Corte Suprema Italiana, al extender los efectos ratione temporis de las decisiones de la Corte Constitucional, “abría las puertas” de la ciudadanía italiana a un número considerable de ciudadanos italianos nacidos de mujeres italianas antesdel 1º de enero de 1948. Así, los autores focalizan su atención sobre el impacto social de la decisión para todos los potenciales ciudadanos que viven en América del Sur y buscan evaluar sus efectos jurídicos sobre el ordenamiento italiano.O direito com freqüência é fonte de discriminações sociais e à vez pode ser a ferramentapara solucioná-las. Os autores tratam de dar um exemplo deste fenômeno, ao analisaro impacto das normas em matéria de cidadania italiana sobre os descendentes decidadãos italianos emigrados ao estrangeiro e mais precisamente na América do sul.Assim, em aplicação da velha normativa italiana, só o pai transmitia iure sanguinis,a cidadania a seus descendentes; além disso, as mulheres perdiam automaticamente acidadania italiana se houvessem adquirido uma cidadania estrangeira por matrimoniocom cônjuge estrangeiro.Estas normas discriminavam à mulher italiana emigrada ao estrangeiro e especialmentea seus descendentes, os quais não podiam adquirir a cidadania italiana.As normas discriminatórias foram por fim ab-rogadas por parte da Corte Constitucionalnos anos Setenta e Oitenta; no entanto, os efeitos das mesmas seguiam persistindo,considerando que as sentenças da Corte Constitucional não abrigavam fatos ocorridosanteriormente à entrada em vigor da Constituição (01.01.1948) e assim não davamsolução às discriminações acontecidas antes dessa data.Em 2009, a Corte Suprema Italiana, ao estender os efeitos ratione temporis das decisões daCorte Constitucional, “abria as portas” da cidadania italiana a um número considerávelde cidadãos italianos nascidos de mulheres italianas antes do 1° de janeiro de 1948.Assim, os autores focalizam sua atenção sobre o impacto social da decisão paratodos os potenciais cidadãos que moram na América do Sul e procuram avaliarseus efeitos jurídicos sobre o ordenamento italiano.Law is often the source of social discriminations, but, at the same time, it can be thekey to delete these social discriminations. The authors try to give an example of thisphenomenon, by analyzing the impact of the Italian citizenship’s rules over the descendantsof the Italian citizens emigrated abroad and, especially, in South America.Indeed, according to the former Italian law, only fathers could transmit iure sanguinisthe citizenship to their children: moreover, women automatically lost theItalian citizenship if they get a foreign citizenship by concluding a marriage witha foreign husband.These rules hardly discriminate the Italian women emigrated abroad and, especially,their descendants who were prevented to get the Italian’s citizenship.These discriminatory rules were finally deleted by the Italian Constitutional Courtin the Seventies and in the Eighties: however, the effects of those rules still persisted,since the decision of the Constitutional Court could not overcome the temporal limit of the entry into force of the Constitution (01.01.1948) and, therefore, could not“cover” the discriminatory facts occurred before that date.Finally in 2009, the Italian Supreme Court, by extending the effects ratione temporisof the decisions of the Constitutional Court, “reopened the doors” of the Italiancitizenship to a huge number of Italian citizenship born from Italian women beforethe 01.01.1948.Therefore, the authors focus on the social impact of this decision for all the potentialItalian citizens living in South America and try to assess its juridical effects overthe Italian law

La ciudadanía italiana y los descendientes de ciudadanos italianos emigrados en Colombia. La eliminación de una injusticia social

Law is often the source of social discriminations, but, at the same time, it can be thekey to delete these social discriminations. The authors try to give an example of thisphenomenon, by analyzing the impact of the Italian citizenship’s rules over the descendantsof the Italian citizens emigrated abroad and, especially, in South America.Indeed, according to the former Italian law, only fathers could transmit iure sanguinisthe citizenship to their children: moreover, women automatically lost theItalian citizenship if they get a foreign citizenship by concluding a marriage witha foreign husband.These rules hardly discriminate the Italian women emigrated abroad and, especially,their descendants who were prevented to get the Italian’s citizenship.These discriminatory rules were finally deleted by the Italian Constitutional Courtin the Seventies and in the Eighties: however, the effects of those rules still persisted,since the decision of the Constitutional Court could not overcome the temporal limit of the entry into force of the Constitution (01.01.1948) and, therefore, could not“cover” the discriminatory facts occurred before that date.Finally in 2009, the Italian Supreme Court, by extending the effects ratione temporisof the decisions of the Constitutional Court, “reopened the doors” of the Italiancitizenship to a huge number of Italian citizenship born from Italian women beforethe 01.01.1948.Therefore, the authors focus on the social impact of this decision for all the potentialItalian citizens living in South America and try to assess its juridical effects overthe Italian law.O direito com freqüência é fonte de discriminações sociais e à vez pode ser a ferramentapara solucioná-las. Os autores tratam de dar um exemplo deste fenômeno, ao analisaro impacto das normas em matéria de cidadania italiana sobre os descendentes decidadãos italianos emigrados ao estrangeiro e mais precisamente na América do sul.Assim, em aplicação da velha normativa italiana, só o pai transmitia iure sanguinis,a cidadania a seus descendentes; além disso, as mulheres perdiam automaticamente acidadania italiana se houvessem adquirido uma cidadania estrangeira por matrimoniocom cônjuge estrangeiro.Estas normas discriminavam à mulher italiana emigrada ao estrangeiro e especialmentea seus descendentes, os quais não podiam adquirir a cidadania italiana.As normas discriminatórias foram por fim ab-rogadas por parte da Corte Constitucionalnos anos Setenta e Oitenta; no entanto, os efeitos das mesmas seguiam persistindo,considerando que as sentenças da Corte Constitucional não abrigavam fatos ocorridosanteriormente à entrada em vigor da Constituição (01.01.1948) e assim não davamsolução às discriminações acontecidas antes dessa data.Em 2009, a Corte Suprema Italiana, ao estender os efeitos ratione temporis das decisões daCorte Constitucional, “abria as portas” da cidadania italiana a um número considerávelde cidadãos italianos nascidos de mulheres italianas antes do 1° de janeiro de 1948.Assim, os autores focalizam sua atenção sobre o impacto social da decisão paratodos os potenciais cidadãos que moram na América do Sul e procuram avaliarseus efeitos jurídicos sobre o ordenamento italiano.El Derecho a menudo es fuente de discriminaciones sociales y a la vez puede ser la herramienta para solucionarlas. Los autores tratan de dar un ejemplo de este fenómeno, al analizar el impacto de las normas en materia de ciudadanía italiana sobre los descendientes de ciudadanos italianos emigrados al extranjero y más precisamente a América del Sur.Así, en aplicación de la vieja normativa italiana, solo el padre transmitía iure sanguinis la ciudadanía a sus descendientes: además, las mujeres perdían automáticamente la ciudadanía italiana si hubiesen adquirido una ciudadanía extranjera por matrimonio con cónyuge extranjero. Estas normas discriminaban a la mujer italiana emigrada al extranjero y especialmente a sus descendientes, quienes no podían adquirir dicha ciudadanía. Las normas discriminatorias fueron por fin abrogadas por parte de la Corte Constitucional en los años setenta y ochenta: sin embargo, los efectos de estas seguían persistiendo,c onsiderando que las sentencias de la Corte Constitucional no cobijaban hechos ocurridos anteriormente a la entrada en vigor de la Constitución (01.01.1948) y así no daban solución a las discriminaciones acontecidas antes de esa fecha. En el 2009, la Corte Suprema Italiana, al extender los efectos ratione temporis de las decisiones de la Corte Constitucional, “abría las puertas” de la ciudadanía italiana a un número considerable de ciudadanos italianos nacidos de mujeres italianas antesdel 1º de enero de 1948. Así, los autores focalizan su atención sobre el impacto social de la decisión para todos los potenciales ciudadanos que viven en América del Sur y buscan evaluar sus efectos jurídicos sobre el ordenamiento italiano

A cidadania italiana e os descendentes de cidadãos italianos emigrados na Colômbia. A eliminação de uma injustiça social

El Derecho a menudo es fuente de discriminaciones sociales y a la vez puede ser la herramienta para solucionarlas. Los autores tratan de dar un ejemplo de este fenómeno, al analizar el impacto de las normas en materia de ciudadanía italiana sobre los descendientes de ciudadanos italianos emigrados al extranjero y más precisamente a América del Sur.Así, en aplicación de la vieja normativa italiana, solo el padre transmitía iure sanguinis la ciudadanía a sus descendientes: además, las mujeres perdían automáticamente la ciudadanía italiana si hubiesen adquirido una ciudadanía extranjera por matrimonio con cónyuge extranjero. Estas normas discriminaban a la mujer italiana emigrada al extranjero y especialmente a sus descendientes, quienes no podían adquirir dicha ciudadanía. Las normas discriminatorias fueron por fin abrogadas por parte de la Corte Constitucional en los años setenta y ochenta: sin embargo, los efectos de estas seguían persistiendo, considerando que las sentencias de la Corte Constitucional no cobijaban hechos ocurridos anteriormente a la entrada en vigor de la Constitución (01.01.1948) y así no daban solución a las discriminaciones acontecidas antes de esa fecha. En el 2009, la Corte Suprema Italiana, al extender los efectos ratione temporis de las decisiones de la Corte Constitucional, “abría las puertas” de la ciudadanía italiana a un número considerable de ciudadanos italianos nacidos de mujeres italianas antesdel 1º de enero de 1948. Así, los autores focalizan su atención sobre el impacto social de la decisión para todos los potenciales ciudadanos que viven en América del Sur y buscan evaluar sus efectos jurídicos sobre el ordenamiento italiano.O direito com freqüência é fonte de discriminações sociais e à vez pode ser a ferramentapara solucioná-las. Os autores tratam de dar um exemplo deste fenômeno, ao analisaro impacto das normas em matéria de cidadania italiana sobre os descendentes decidadãos italianos emigrados ao estrangeiro e mais precisamente na América do sul.Assim, em aplicação da velha normativa italiana, só o pai transmitia iure sanguinis,a cidadania a seus descendentes; além disso, as mulheres perdiam automaticamente acidadania italiana se houvessem adquirido uma cidadania estrangeira por matrimoniocom cônjuge estrangeiro.Estas normas discriminavam à mulher italiana emigrada ao estrangeiro e especialmentea seus descendentes, os quais não podiam adquirir a cidadania italiana.As normas discriminatórias foram por fim ab-rogadas por parte da Corte Constitucionalnos anos Setenta e Oitenta; no entanto, os efeitos das mesmas seguiam persistindo,considerando que as sentenças da Corte Constitucional não abrigavam fatos ocorridosanteriormente à entrada em vigor da Constituição (01.01.1948) e assim não davamsolução às discriminações acontecidas antes dessa data.Em 2009, a Corte Suprema Italiana, ao estender os efeitos ratione temporis das decisões daCorte Constitucional, “abria as portas” da cidadania italiana a um número considerávelde cidadãos italianos nascidos de mulheres italianas antes do 1° de janeiro de 1948.Assim, os autores focalizam sua atenção sobre o impacto social da decisão paratodos os potenciais cidadãos que moram na América do Sul e procuram avaliarseus efeitos jurídicos sobre o ordenamento italiano.Law is often the source of social discriminations, but, at the same time, it can be thekey to delete these social discriminations. The authors try to give an example of thisphenomenon, by analyzing the impact of the Italian citizenship’s rules over the descendantsof the Italian citizens emigrated abroad and, especially, in South America.Indeed, according to the former Italian law, only fathers could transmit iure sanguinisthe citizenship to their children: moreover, women automatically lost theItalian citizenship if they get a foreign citizenship by concluding a marriage witha foreign husband.These rules hardly discriminate the Italian women emigrated abroad and, especially,their descendants who were prevented to get the Italian’s citizenship.These discriminatory rules were finally deleted by the Italian Constitutional Courtin the Seventies and in the Eighties: however, the effects of those rules still persisted,since the decision of the Constitutional Court could not overcome the temporal limit of the entry into force of the Constitution (01.01.1948) and, therefore, could not“cover” the discriminatory facts occurred before that date.Finally in 2009, the Italian Supreme Court, by extending the effects ratione temporisof the decisions of the Constitutional Court, “reopened the doors” of the Italiancitizenship to a huge number of Italian citizenship born from Italian women beforethe 01.01.1948.Therefore, the authors focus on the social impact of this decision for all the potentialItalian citizens living in South America and try to assess its juridical effects overthe Italian law