Potential drivers of the atopic march - unraveling the skin-lung crosstalk

Atopic diseases such as atopic dermatitis (AD), food allergy, allergic rhinitis, and allergic asthma are not only an economical burden to the healthcare system but also a highly physiological and psychological burden for patients. Similar pathophysiological patterns like increased IgE plasma concentration as well as Th2-dominant inflammation characterize these diseases. By now, it is well-accepted that AD is the “entry point“ for a successive development of further atopic diseases during the first 10 years of life - a process known as the atopic march. However, the underlying mechanisms are still not fully understood and by far not all responsible factors have been identified; a drawback resulting in poor options for prevention. In this thesis, a human-based two-organ co-culture of skin disease equivalents, mimicking hallmarks of AD, and healthy bronchial epithelial equivalents was established for studying the pathophysiological crosstalk between skin and bronchi. Already a short co-cultivation period of six days induced a clear hyperproliferative phenotype with elevated mucus-secretion and an increase of inflammatory markers as determined on gene and protein level. Consequently, this led to the suggestion that either epithelial or dermal factors might play a role due to missing immune cells in this co-culture. Secretome of skin disease equivalents as well as proteome and transcriptome analysis patient-derived fibroblasts revealed significant changes of extracellular matrix (ECM)-related genes and proteins. In an exemplary study, the effect of five ECM-related compounds (complement factor C3, fibronectin, syndecan-4 (SD-4), cluster of differentiation 44 (CD44), and thrombospondin-1 (TSP-1)) was tested on bronchial epithelial equivalents and naïve activated CD4+ T cells. For all compounds, an asthma-like inflammation was induced in bronchial epithelial equivalents. Furthermore, a compound-specific polarization of naïve activated CD4+ T cells into different Th subsets was observed. In addition to the human-based in vitro co-culture model, three out of five ECM-related compounds (SD-4, CD44, and TSP-1) were tested in vivo in healthy BALB/c mice in the context of a translational study. In line with the in vitro experiments, a polarization of murine CD4+ T cells towards Th1, Th17, and Th22 subtypes was observed in a first low-dose approach. Interestingly, no histological changes in skin and lung were observed. However, treated mice had an increased size and weight of spleen. Taken together, the established human-based two-organ co-culture system not only enables investigation of the crosstalk between healthy skin and bronchus, but also of the pathophysiological communication in the context of the atopic march by using diseased skin equivalents. The results of this thesis demonstrate the importance of ECM in the complex pathophysiology of the atopic march - a hitherto underestimated feature. In this regard, further research is needed to finally assess new targets for the prevention of the atopic march

La construction d’un projet dans un cadre territorial périurbain : la Plaine du Var

Posant la question du territoire, comme espace de projet, l’étude présentée souligne le décalage entre l’urbain (réalité factuelle) et l’urbanité (objectif à atteindre) ; elle développe l’» attitude de projet » selon une argumentation en trois points. D’une part, les conditions changent lorsque l’on passe de la condition classique de projet architectural à celle de projet territorial, qui démultiplie l’échelle spatiale et l’éventail des acteurs en position de maîtrise d’ouvrage, modifiant en profondeur l’élaboration et l’application du projet. Par ailleurs, les connaissances préalables nécessaires : bilan et diagnostic de l’existant, ne constituent pas en eux-mêmes un projet. Enfin, pour « faire projet », une idée, une projection dans l’avenir sont nécessaires. Un début de réponse est donné, avec le compte-rendu d’une expérimentation en cours : le projet d’aménagement de la Plaine du Var, action à long terme d’échelle intercommunale dont la mise en œuvre est amorcée.By raising the question of territory as a space for project planning, our study emphasizes the discrepancy between urban as a fact and urbanity as an aim ; it develops the “project planner’s attitude” in three parts. First, conditions change a lot when passing from a classical architectural scheme to a territorial project. The spatial scale and the range of action for the project managers are increased, considerably modifying the way the project gets worked out. Otherwise, a diagnosis and a statement of the existing conditions cannot be called a project. Finally, to have a project worthy of the name you need an idea and a forecast in the future as a prerequisite. We give a tentative answer with an experiment in progress : the development project of the Var flat valley, a long-term intermunicipal scheme, just in its beginning stage

A comparison of multivariate and univariate time series approaches to modelling and forecasting emergency department demand in Western Australia

Objective: To develop multivariate vector-ARMA (VARMA) forecast models for predicting emergency department (ED) demand in Western Australia (WA) and compare them to the benchmark univariate autoregressive moving average (ARMA) and Winters’ models. Methods: Seven-year monthly WA state-wide public hospital ED presentation data from 2006/07 to 2012/13 were modelled. Graphical and VARMA modelling methods were used for descriptive analysis and model fitting. The VARMA models were compared to the benchmark univariate ARMA and Winters’ models to determine their accuracy to predict ED demand. The best models were evaluated by using error correction methods for accuracy. Results: Descriptive analysis of all the dependent variables showed an increasing pattern of ED use with seasonal trends over time. The VARMA models provided a more precise and accurate forecast with smaller confidence intervals and better measures of accuracy in predicting ED demand in WA than the ARMA and Winters’ method. Conclusion: VARMA models are a reliable forecasting method to predict ED demand for strategic planning and resource allocation. While the ARMA models are a closely competing alternative, they under-estimated future ED demand

Rare mutations in SQSTM1 modify susceptibility to frontotemporal lobar degeneration

Mutations in the gene coding for Sequestosome 1 (SQSTM1) have been genetically associated with amyotrophic lateral sclerosis (ALS) and Paget disease of bone. In the present study, we analyzed the SQSTM1 coding sequence for mutations in an extended cohort of 1,808 patients with frontotemporal lobar degeneration (FTLD), ascertained within the European Early-Onset Dementia consortium. As control dataset, we sequenced 1,625 European control individuals and analyzed whole-exome sequence data of 2,274 German individuals (total n = 3,899). Association of rare SQSTM1 mutations was calculated in a meta-analysis of 4,332 FTLD and 10,240 control alleles. We identified 25 coding variants in FTLD patients of which 10 have not been described. Fifteen mutations were absent in the control individuals (carrier frequency < 0.00026) whilst the others were rare in both patients and control individuals. When pooling all variants with a minor allele frequency < 0.01, an overall frequency of 3.2 % was calculated in patients. Rare variant association analysis between patients and controls showed no difference over the whole protein, but suggested that rare mutations clustering in the UBA domain of SQSTM1 may influence disease susceptibility by doubling the risk for FTLD (RR = 2.18 [95 % CI 1.24-3.85]; corrected p value = 0.042). Detailed histopathology demonstrated that mutations in SQSTM1 associate with widespread neuronal and glial phospho-TDP-43 pathology. With this study, we provide further evidence for a putative role of rare mutations in SQSTM1 in the genetic etiology of FTLD and showed that, comparable to other FTLD/ALS genes, SQSTM1 mutations are associated with TDP-43 pathology

Replication of EPHA1 and CD33 associations with late-onset Alzheimer's disease: a multi-centre case-control study

<p>Abstract</p> <p>Background</p> <p>A recently published genome-wide association study (GWAS) of late-onset Alzheimer's disease (LOAD) revealed genome-wide significant association of variants in or near <it>MS4A4A, CD2AP, EPHA1 </it>and <it>CD33</it>. Meta-analyses of this and a previously published GWAS revealed significant association at <it>ABCA7 </it>and <it>MS4A</it>, independent evidence for association of <it>CD2AP, CD33 </it>and <it>EPHA1 </it>and an opposing yet significant association of a variant near <it>ARID5B</it>. In this study, we genotyped five variants (in or near <it>CD2AP, EPHA1, ARID5B</it>, and <it>CD33</it>) in a large (2,634 LOAD, 4,201 controls), independent dataset comprising six case-control series from the USA and Europe. We performed meta-analyses of the association of these variants with LOAD and tested for association using logistic regression adjusted by age-at-diagnosis, gender, and <it>APOE ε4 </it>dosage.</p> <p>Results</p> <p>We found no significant evidence of series heterogeneity. Associations with LOAD were successfully replicated for <it>EPHA1 </it>(rs11767557; OR = 0.87, p = 5 × 10^-4) and <it>CD33 </it>(rs3865444; OR = 0.92, p = 0.049), with odds ratios comparable to those previously reported. Although the two <it>ARID5B </it>variants (rs2588969 and rs494288) showed significant association with LOAD in meta-analysis of our dataset (p = 0.046 and 0.008, respectively), the associations did not survive adjustment for covariates (p = 0.30 and 0.11, respectively). We had insufficient evidence in our data to support the association of the <it>CD2AP </it>variant (rs9349407, p = 0.56).</p> <p>Conclusions</p> <p>Our data overwhelmingly support the association of <it>EPHA1 </it>and <it>CD33 </it>variants with LOAD risk: addition of our data to the results previously reported (total n > 42,000) increased the strength of evidence for these variants, providing impressive p-values of 2.1 × 10^-15(<it>EPHA1</it>) and 1.8 × 10^-13(<it>CD33</it>).</p

Hierarchical spectral clustering reveals brain size and shape changes in asymptomatic carriers of C9orf72

Traditional methods for detecting asymptomatic brain changes in neurodegenerative diseases such as Alzheimer\u27s disease or frontotemporal degeneration typically evaluate changes in volume at a predefined level of granularity, e.g. voxel-wise or in a priori defined cortical volumes of interest. Here, we apply a method based on hierarchical spectral clustering, a graph-based partitioning technique. Our method uses multiple levels of segmentation for detecting changes in a data-driven, unbiased, comprehensive manner within a standard statistical framework. Furthermore, spectral clustering allows for detection of changes in shape along with changes in size. We performed tensor-based morphometry to detect changes in the Genetic Frontotemporal dementia Initiative asymptomatic and symptomatic frontotemporal degeneration mutation carriers using hierarchical spectral clustering and compared the outcome to that obtained with a more conventional voxel-wise tensor- and voxel-based morphometric analysis. In the symptomatic groups, the hierarchical spectral clustering-based method yielded results that were largely in line with those obtained with the voxel-wise approach. In asymptomatic C9orf72 expansion carriers, spectral clustering detected changes in size in medial temporal cortex that voxel-wise methods could only detect in the symptomatic phase. Furthermore, in the asymptomatic and the symptomatic phases, the spectral clustering approach detected changes in shape in the premotor cortex in C9orf72. In summary, the present study shows the merit of hierarchical spectral clustering for data-driven segmentation and detection of structural changes in the symptomatic and asymptomatic stages of monogenic frontotemporal degeneration

TBK1 mutation spectrum in an extended European patient cohort with frontotemporal dementia and amyotrophic lateral sclerosis

We investigated the mutation spectrum of the TANK-Binding Kinase 1 (TBK1) gene and its associated phenotypic spectrum by exonic resequencing of TBK1 in a cohort of 2,538 patients with frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), or FTD plus ALS, ascertained within the European Early-Onset Dementia Consortium. We assessed pathogenicity of predicted protein-truncating mutations by measuring loss of RNA expression. Functional effect of in-frame amino acid deletions and missense mutations was further explored in vivo on protein level and in vitro by an NFB-induced luciferase reporter assay and measuring phosphorylated TBK1. The protein-truncating mutations led to the loss of transcript through nonsense-mediated mRNA decay. For the in-frame amino acid deletions, we demonstrated loss of TBK1 or phosphorylated TBK1 protein. An important fraction of the missense mutations compromised NFB activation indicating that at least some functions of TBK1 are lost. Although missense mutations were also present in controls, over three times more mutations affecting TBK1 functioning were found in the mutation fraction observed in patients only, suggesting high-risk alleles (P = 0.03). Total mutation frequency for confirmed TBK1 LoF mutations in the European cohort was 0.7%, with frequencies in the clinical subgroups of 0.4% in FTD, 1.3% in ALS, and 3.6% in FTD-ALS