MODEL SYSTEMS OF INTESTINAL INFLAMMATION: A STEP TOWARDS PERSONALIZED MEDICINE

Inflammatory bowel disease (IBD) is a lifelong chronic inflammatory condition of the gastrointestinal tract (GIT), with incidence and prevalence increasing worldwide. It is considered a complex, multifactorial disease with no cure. Even though large progress has been made in recent years, current therapies are far from satisfactory, and show extreme variability of outcomes due to patient heterogeneity. The traditional therapy consists of anti-inflammatories, corticosteroids, antibiotics, and immunomodulatory drugs. This non-specific immunosuppression guarantees disease-control in some patients although the long-term use of these drugs is correlated with a significant number of therapy-associated complications and side-effects. A dramatic improvement in disease management was achieved by the introduction of biological agents targeting pro-inflammatory cytokines such as anti-TNF-α. Despite the revolutionary impact of these agents in IBD disease management, treatments such as anti-TNF-α do show several drawbacks – for example, up to 50% of patients do not respond at all or eventually lose response. This variability in clinical outcome is reflecting the variability of individuals due to different genetics, life style and inflammatory state. Therefore, there is a need to define the specific inflammatory state of a given patient, considering individual complications and develop new in-vitro systems and biomarkers that predict drug responsiveness and allow developing patient-specific treatment In this thesis, different in-vitro models were developed addressing different aspects and compartments of IBD pathology including the enteric nervous system, the ECM component fibrillin-1, as well as patient-derived, three dimensional short-term and long-term cultures that will bring us a step closer towards personalized medicine

Anti-inflammatory activity of Wnt signaling in enteric nervous system: in vitro preliminary evidences in rat primary cultures

Background: In the last years, Wnt signaling was demonstrated to regulate inflammatory processes. In particular, an increased expression of Wnts and Frizzled receptors was reported in inflammatory bowel disease (IBD) and ulcerative colitis to exert both anti- and pro-inflammatory functions regulating the intestinal activated nuclear factor \u3baB (NF-\u3baB), TNFa release, and IL10 expression. Methods: To investigate the role of Wnt pathway in the response of the enteric nervous system (ENS) to inflammation, neurons and glial cells from rat myenteric plexus were treated with exogenous Wnt3a and/or LPS with or without supporting neurotrophic factors such as basic fibroblast growth factor (bFGF), epithelial growth factor (EGF), and glial cell-derived neurotrophic factor (GDNF). The immunophenotypical characterization by flow cytometry and the protein and gene expression analysis by qPCR and Western blotting were carried out. Results: Flow cytometry and immunofluorescence staining evidenced that enteric neurons coexpressed Frizzled 9 and toll-like receptor 4 (TLR4) while glial cells were immunoreactive to TLR4 and Wnt3a suggesting that canonical Wnt signaling is active in ENS. Conclusions: The results of this study suggested the existence of neuronal surveillance through FZD9 and Wnt3a in enteric myenteric plexus. Moreover, experimental evidences were provided to clarify the correlation among soluble trophic factors, Wnt signaling, and anti-inflammatory protection of ENS

Nanopatterned acellular valve conduits drive the commitment of blood-derived multipotent cells

Considerable progress has been made in recent years toward elucidating the correlation among nanoscale topography, mechanical properties, and biological behavior of cardiac valve substitutes. Porcine TriCol scaffolds are promising valve tissue engineering matrices with demonstrated self-repopulation potentiality. In order to define an in vitro model for investigating the influence of extracellular matrix signaling on the growth pattern of colonizing blood-derived cells, we cultured circulating multipotent cells (CMC) on acellular aortic (AVL) and pulmonary (PVL) valve conduits prepared with TriCol method and under no-flow condition. Isolated by our group from Vietnamese pigs before heart valve prosthetic implantation, porcine CMC revealed high proliferative abilities, three-lineage differentiative potential, and distinct hematopoietic/endothelial and mesenchymal properties. Their interaction with valve extracellular matrix nanostructures boosted differential messenger RNA expression pattern and morphologic features on AVL compared to PVL, while promoting on both matrices the commitment to valvular and endothelial cell-like phenotypes. Based on their origin from peripheral blood, porcine CMC are hypothesized in vivo to exert a pivotal role to homeostatically replenish valve cells and contribute to hetero- or allograft colonization. Furthermore, due to their high responsivity to extracellular matrix nanostructure signaling, porcine CMC could be useful for a preliminary evaluation of heart valve prosthetic functionality

Informationsbeschaffungs- und Publikationsverhalten von Wissenschaftlerinnen und Wissenschaftlern der natur- und ingenieurwissenschaftlichen Fächern : Auswertung einer Umfrage mit Schwerpunkt auf nicht-textuellen Materialien

[no abstract available

MODEL SYSTEMS OF INTESTINAL INFLAMMATION: A STEP TOWARDS PERSONALIZED MEDICINE

Inflammatory bowel disease (IBD) is a lifelong chronic inflammatory condition of the gastrointestinal tract (GIT), with incidence and prevalence increasing worldwide. It is considered a complex, multifactorial disease with no cure. Even though large progress has been made in recent years, current therapies are far from satisfactory, and show extreme variability of outcomes due to patient heterogeneity. The traditional therapy consists of anti-inflammatories, corticosteroids, antibiotics, and immunomodulatory drugs. This non-specific immunosuppression guarantees disease-control in some patients although the long-term use of these drugs is correlated with a significant number of therapy-associated complications and side-effects. A dramatic improvement in disease management was achieved by the introduction of biological agents targeting pro-inflammatory cytokines such as anti-TNF-α. Despite the revolutionary impact of these agents in IBD disease management, treatments such as anti-TNF-α do show several drawbacks – for example, up to 50% of patients do not respond at all or eventually lose response. This variability in clinical outcome is reflecting the variability of individuals due to different genetics, life style and inflammatory state. Therefore, there is a need to define the specific inflammatory state of a given patient, considering individual complications and develop new in-vitro systems and biomarkers that predict drug responsiveness and allow developing patient-specific treatment In this thesis, different in-vitro models were developed addressing different aspects and compartments of IBD pathology including the enteric nervous system, the ECM component fibrillin-1, as well as patient-derived, three dimensional short-term and long-term cultures that will bring us a step closer towards personalized medicine.Le malattie infiammatorie croniche intestinali (MICI) sono un gruppo di patologie complesse ad eziologia multifattoriale, caratterizzate da uno stato infiammatorio cronico del tratto gastrointestinale, la cui incidenza a livello mondiale è in continuo aumento. Nonostante nel corso degli ultimi anni siano stati fatti numerosi progressi nel controllo della malattia, l’attuale approccio terapeutico rimane ancora lontano dall’essere soddisfacente e l’esito clinico che ne deriva è estremamente variabile a causa della vasta eterogeneità tra i pazienti. La terapia tradizionale, che consiste nella somministrazione di farmaci antinfiammatori, corticosteroidi, antibiotici o farmaci immunomodulatori, garantisce il controllo della malattia in alcuni pazienti, ma, a causa della non-specificità e del lungo periodo di utilizzo, è correlata all’insorgenza di numerose complicanze ed effetti collaterali. Un significativo miglioramento nella gestione della malattia è stato raggiunto attraverso l’introduzione di farmaci biologici, il cui target è rappresentato principalmente dalle citochine pro-infiammatorie implicate nella patologia, come ad esempio il TNF-α. Nonostante il forte impatto clinico, l’utilizzo di farmaci biologici, come l’anti-TNF-α ha mostrato diversi svantaggi, tra cui un’alta percentuale di non-responsività al trattamento oppure la perdita di risposta nel corso del tempo. La grande variabilità che si riscontra nella risposta clinica riflette di fatto la variabilità che sussiste tra i diversi individui, ed è dovuta principalmente a differenze a livello genetico, nello stile di vita e nello stato infiammatorio. Di conseguenza, cresce sempre più la necessità di definire nello specifico lo stato infiammatorio e le complicanze caratteristiche di ciascun paziente e di sviluppare nuovi sistemi di screening in-vitro che possano predire la risposta al trattamento e quindi consentire un approccio terapeutico specifico per ciascun individuo. Nell’ottica di una medicina predittiva e della terapia personalizzata, in questa tesi sono stati sviluppati differenti modelli in-vitro, che prendono in considerazione diversi aspetti e compartimenti implicati nelle malattie infiammatorie croniche intestinali, quali il sistema nervoso enterico, la fibrillina-1, componente della matrice extracellulare, così come colture tridimensionali a breve e lungo termine derivate da campioni bioptici umani. Questi modelli sperimentali di infiammazione cronica intestinale potranno costituire uno strumento clinico utile per applicazioni di medicina personalizzata

Structural and functional failure of fibrillin-1 in human diseases (Review)

Fibrillins (FBNs) are key relay molecules that form the backbone of microfibrils in elastic and non-elastic tissues. Interacting with other components of the extracellular matrix (ECM), these ubiquitous glycoproteins exert pivotal roles in tissue development, homeostasis and repair. In addition to mechanical support, FBN networks also exhibit regulatory activities on growth factor signalling, ECM formation, cell behaviour and the immune response. Consequently, mutations affecting the structure, assembly and stability of FBN microfibrils have been associated with impaired biomechanical tissue properties, altered cell-matrix interactions, uncontrolled growth factor or cytokine activation, and the development of fibrillinopathies and associated severe complications in multiple organs. Beyond a panoramic overview of structural cues of the FBN network, the present review will also describe the pathological implications of FBN disorders in the development of inflammatory and fibrotic conditions

Die TIB: Mehr als eine Bibliothek

Die TIB gestaltet aktiv den digitalen Wandel von Bibliotheken mit – in Form von Open Science, Open Access, Forschungsdaten und digitaler Langzeitarchivierung sowie bei der Entwicklung von Diensten an der Schnittstelle von analogen und digitalen Formaten. Sie baut ihre Rolle als deutsches Informationszentrum für die Digitalisierung von Wissenschaft und Technik in Zukunft weiter aus und setzt auf die Entwicklung neuer digitaler Angebote, die klassische bibliothekarische Dienste und innovative Forschungsergebnisse miteinander verzahnen. Als wissenschaftliche Infrastruktureinrichtung steht sie vor zwei großen Herausforderungen: Als Deutsche Zentrale Fachbibliothek für Technik und Naturwissenschaften bewegt sie sich als Akteurin im Zentrum des digitalen Wandels des wissenschaftlichen Arbeitens, des digitalen Publikationswesens und der Globalisierung von Information und Kommunikation. Gleichzeitig erbringt sie als von Bund und Ländern finanzierte Einrichtung verstärkt eigene Forschung.TIB plays an active role in achieving the digital transformation of libraries – not only in the form of open science, open access, research data and digital preservation, but also in the development of services at the interface between analogue and digital formats. The library will continue to expand its role as the German information centre for the digitalisation of science and technology in the future, focusing on the development of new digital offerings that link traditional library services with innovative research results. As a scientific infrastructure facility, TIB faces two major challenges: being the German national library of science and technology, it operates as a stakeholder at the heart of the transition to the digital production of scientific work, digital publishing, and the globalisation of information and communication. At the same time, TIB increasingly conducts its own research as an institute financed by the German federation and the federal states.Peer Reviewe

STW 5 Herbal Preparation Modulates Wnt3a and Claudin 1 Gene Expression in Zebrafish IBS-like Model

Aim: Irritable bowel syndrome (IBS) is a functional bowel disorder characterized by chronic abdominal pain and stool irregularities. STW 5 has proven clinical efficacy in functional gastrointestinal disorders, including IBS, targeting pathways that suppress inflammation and protect the mucosa. Wnt signaling is known to modulate NF-kβ-dependent inflammatory cytokine production. This sparked the idea of evaluating the impact of STW 5 on the expression of inflammatory-response and Wnt/β catenin-target genes in an IBS-like model. Main methods: We used zebrafish and dextran sodium sulfate (DSS) treatment to model IBS-like conditions in vivo and in vitro and examined the effects of subsequent STW 5 treatment on the intestines of DSS-treated fish and primary cultured intestinal and neuronal cells. Gross gut anatomy, histology, and the expression of Wnt-signaling and cytokine genes were analyzed in treated animals and/or cells, and in controls. Key findings: DSS treatment up-regulated the expression of interleukin-8, tumor necrosis factor-α, wnt3a, and claudin-1 in explanted zebrafish gut. Subsequent STW 5 treatment abolished both the macroscopic signs of gut inflammation, DSS-induced mucosecretory phenotype, and normalized the DSS-induced upregulated expression of il10 and Wnt signaling genes, such as wnt3a and cldn1 in explanted zebrafish gut. Under inflammatory conditions, STW 5 downregulated the expression of the pro-inflammatory cytokine genes il1β, il6, il8, and tnfα while it upregulated the expression of the anti-inflammatory genes il10 and wnt3a in enteric neuronal cells in vitro. Significance: Wnt signaling could be a novel target for the anti-inflammatory and intestinal permeability-restoring effects of STW 5, possibly explaining its clinical efficacy in IBS

Histone deacetylase inhibitors restore IL-10 expression in lipopolysaccharide-induced cell inflammation and reduce IL-1\u3b2 and IL-6 production in breast silicone implant in C57BL/6J wild-type murine model

Among epigenetic enzymes, histone deacetylases (HDACs) are responsible for regulating the expression of an extensive array of genes by reversible deacetylation of nuclear histones as well as a large number of non-histone proteins. Initially proposed for cancer therapy, recently the interest for HDAC inhibitors (HDACi) as orally active, safe, and anti-inflammatory agents is rising due to their ability in reducing the severity of inflammatory and autoimmune diseases. In particular, selective HDAC3, HDAC6, and HDAC8 inhibitors have been described to downregulate the expression of pro-inflammatory cytokines (TNF-\u3b1, TGF-\u3b2, IL-1\u3b2, and IL-6). Herein, using KB31, C2C12, and 3T3-J2 cell lines, we demonstrated that, under lipopolysaccharide-induced in vitro inflammation, HDAC3/6/8 inhibitor MC2625 and HDAC6-selective inhibitor MC2780 were effective at a concentration of 30\u2009ng/mL to downregulate mRNA expression of pro-inflammatory cytokines (IL-1\u3b2 and IL-6) and to promote the transcription of IL-10 gene, without affecting the cell viability. Afterwards, we investigated by immunohistochemistry the activity of MC2625 and MC2780 at a concentration of 60\u2009ng/kg animal weight to regulate silicone-triggered immune response in C57BL/6J female mice. Our findings evidenced the ability of such inhibitors to reduce host inflammation in silicone implants promoting a thickness reduction of peri-implant fibrous capsule, upregulating IL-10 expression, and reducing the production of both IL-1\u3b2 and IL-6. These results underline the potential application of MC2625 and MC2780 in inflammation-related diseases