Conceptual Modeling Applied to Genomics: Challenges Faced in Data Loading

Todays genomic domain evolves around insecurity: too many imprecise concepts, too much information to be properly managed. Considering that conceptualization is the most exclusive human characteristic, it makes full sense to try to conceptualize the principles that guide the essence of why humans are as we are. This question can of course be generalized to any species, but we are especially interested in this work in showing how conceptual modeling is strictly required to understand the ''execution model'' that human beings ''implement''. The main issue is to defend the idea that only by having an in-depth knowledge of the Conceptual Model that is associated to the Human Genome, can this Human Genome properly be understood. This kind of Model-Driven perspective of the Human Genome opens challenging possibilities, by looking at the individuals as implementation of that Conceptual Model, where different values associated to different modeling primitives will explain the diversity among individuals and the potential, unexpected variations together with their unwanted effects in terms of illnesses. This work focuses on the challenges faced in loading data from conventional resources into Information Systems created according to the above mentioned conceptual modeling approach. The work reports on various loading efforts, problems encountered and the solutions to these problems. Also, a strong argument is made about why conventional methods to solve the so called `data chaos¿ problems associated to the genomics domain so often fail to meet the demands.Van Der Kroon ., M. (2011). Conceptual Modeling Applied to Genomics: Challenges Faced in Data Loading. http://hdl.handle.net/10251/16993Archivo delegad

Comparative Genomics of Parastagonospora and Pyrenophora species

The advancing technology and tools available for the important agricultural pathogens P. nodorum and P. avenaria pathosystems are leveraged here for intra-species comparison as well as comparisons to other related species. The comparisons have yielded insight into the evolutionary history of pathogen and host, insights into the mechanisms of genome evolution, and prediction of genes integral to fungal pathogenicity

Atassie eredodegenerative ad esordio precoce: descrizione del pattern di alterazione patologica mediante neuroimaging avanzato e studio neuropsicologico per la definizione di indicatori paraclinici utili al monitoraggio dell'evoluzione o alla verifica di efficacia del trattamento.

Background/aims Friedreich's ataxia (FRDA) is an autosomal-recessive neurodegenrative conditions caused by a GAA repeat triplet. There are no quantitative objective biomarkers to show reliable correlations with progression rate and disease severity. Methods or Materials or Case Report Twenty-one early-onset molecularly-defined FRDA patients underwent clinical-neuropsychological assessment. A subgroup of patients (n=17) and age-matched healthy controls (HC) underwent a neuroimaging study protocol on a 3T-MRI (DTI, fMRI). Non-parametric voxel-based permutations were performed on the WM-maps regions-of-interest (ROI), via a general linear model (GLM) (p<0.05). An fMRI sequence was acquired during a simple block-design finger-tapping task. Results All patients were homozygous for GAA expansion (mean GAA1 653.7±221), but one with a point-mutaion/170GAA. Age-at-onset: 10.6±4.6yrs, age-at-assessment 26.9±10.3yrs, disease-duration 16.3±8.8yrs. Deficit on executive and attentive functions were ob-served. We found altered FA and MD parameters, through voxel- and ROI-based analysis, in the cerebellar peduncles; corticospinal, lemnisceal systems, major commissural fibres, thalamic and optic radiations. A finger-tapping task demonstrated significantly higher cerebellar-cortex activation (lobule V, IV) in HC compared to FRDA. Conclusion We report our clinical and neuroimaging experience of a FRDA cohort in order to explore WM con-nectivity and motor dysfunction. DTI changes in selected areas and BOLD-signal in the cerebellar, ipsilateral cerebellar-cortex in response to a simple motor task show strong intergroup-discriminating power and may prove to be useful paraclinical disease markers. A longitudinal study is needed to investigate and validate the sensitivity of these indicators

Differential evolution of non-coding DNA across eukaryotes and its close relationship with complex multicellularity on Earth

Here, I elaborate on the hypothesis that complex multicellularity (CM, sensu Knoll) is a major evolutionary transition (sensu Szathmary), which has convergently evolved a few times in Eukarya only: within red and brown algae, plants, animals, and fungi. Paradoxically, CM seems to correlate with the expansion of non-coding DNA (ncDNA) in the genome rather than with genome size or the total number of genes. Thus, I investigated the correlation between genome and organismal complexities across 461 eukaryotes under a phylogenetically controlled framework. To that end, I introduce the first formal definitions and criteria to distinguish ‘unicellularity’, ‘simple’ (SM) and ‘complex’ multicellularity. Rather than using the limited available estimations of unique cell types, the 461 species were classified according to our criteria by reviewing their life cycle and body plan development from literature. Then, I investigated the evolutionary association between genome size and 35 genome-wide features (introns and exons from protein-coding genes, repeats and intergenic regions) describing the coding and ncDNA complexities of the 461 genomes. To that end, I developed ‘GenomeContent’, a program that systematically retrieves massive multidimensional datasets from gene annotations and calculates over 100 genome-wide statistics. R-scripts coupled to parallel computing were created to calculate >260,000 phylogenetic controlled pairwise correlations. As previously reported, both repetitive and non-repetitive DNA are found to be scaling strongly and positively with genome size across most eukaryotic lineages. Contrasting previous studies, I demonstrate that changes in the length and repeat composition of introns are only weakly or moderately associated with changes in genome size at the global phylogenetic scale, while changes in intron abundance (within and across genes) are either not or only very weakly associated with changes in genome size. Our evolutionary correlations are robust to: different phylogenetic regression methods, uncertainties in the tree of eukaryotes, variations in genome size estimates, and randomly reduced datasets. Then, I investigated the correlation between the 35 genome-wide features and the cellular complexity of the 461 eukaryotes with phylogenetic Principal Component Analyses. Our results endorse a genetic distinction between SM and CM in Archaeplastida and Metazoa, but not so clearly in Fungi. Remarkably, complex multicellular organisms and their closest ancestral relatives are characterized by high intron-richness, regardless of genome size. Finally, I argue why and how a vast expansion of non-coding RNA (ncRNA) regulators rather than of novel protein regulators can promote the emergence of CM in Eukarya. As a proof of concept, I co-developed a novel ‘ceRNA-motif pipeline’ for the prediction of “competing endogenous” ncRNAs (ceRNAs) that regulate microRNAs in plants. We identified three candidate ceRNAs motifs: MIM166, MIM171 and MIM159/319, which were found to be conserved across land plants and be potentially involved in diverse developmental processes and stress responses. Collectively, the findings of this dissertation support our hypothesis that CM on Earth is a major evolutionary transition promoted by the expansion of two major ncDNA classes, introns and regulatory ncRNAs, which might have boosted the irreversible commitment of cell types in certain lineages by canalizing the timing and kinetics of the eukaryotic transcriptome.:Cover page Abstract Acknowledgements Index 1. The structure of this thesis 1.1. Structure of this PhD dissertation 1.2. Publications of this PhD dissertation 1.3. Computational infrastructure and resources 1.4. Disclosure of financial support and information use 1.5. Acknowledgements 1.6. Author contributions and use of impersonal and personal pronouns 2. Biological background 2.1. The complexity of the eukaryotic genome 2.2. The problem of counting and defining “genes” in eukaryotes 2.3. The “function” concept for genes and “dark matter” 2.4. Increases of organismal complexity on Earth through multicellularity 2.5. Multicellularity is a “fitness transition” in individuality 2.6. The complexity of cell differentiation in multicellularity 3. Technical background 3.1. The Phylogenetic Comparative Method (PCM) 3.2. RNA secondary structure prediction 3.3. Some standards for genome and gene annotation 4. What is in a eukaryotic genome? GenomeContent provides a good answer 4.1. Background 4.2. Motivation: an interoperable tool for data retrieval of gene annotations 4.3. Methods 4.4. Results 4.5. Discussion 5. The evolutionary correlation between genome size and ncDNA 5.1. Background 5.2. Motivation: estimating the relationship between genome size and ncDNA 5.3. Methods 5.4. Results 5.5. Discussion 6. The relationship between non-coding DNA and Complex Multicellularity 6.1. Background 6.2. Motivation: How to define and measure complex multicellularity across eukaryotes? 6.3. Methods 6.4. Results 6.5. Discussion 7. The ceRNA motif pipeline: regulation of microRNAs by target mimics 7.1. Background 7.2. A revisited protocol for the computational analysis of Target Mimics 7.3. Motivation: a novel pipeline for ceRNA motif discovery 7.4. Methods 7.5. Results 7.6. Discussion 8. Conclusions and outlook 8.1. Contributions and lessons for the bioinformatics of large-scale comparative analyses 8.2. Intron features are evolutionarily decoupled among themselves and from genome size throughout Eukarya 8.3. “Complex multicellularity” is a major evolutionary transition 8.4. Role of RNA throughout the evolution of life and complex multicellularity on Earth 9. Supplementary Data Bibliography Curriculum Scientiae Selbständigkeitserklärung (declaration of authorship

High Performance Computing for DNA Sequence Alignment and Assembly

Recent advances in DNA sequencing technology have dramatically increased the scale and scope of DNA sequencing. These data are used for a wide variety of important biological analyzes, including genome sequencing, comparative genomics, transcriptome analysis, and personalized medicine but are complicated by the volume and complexity of the data involved. Given the massive size of these datasets, computational biology must draw on the advances of high performance computing. Two fundamental computations in computational biology are read alignment and genome assembly. Read alignment maps short DNA sequences to a reference genome to discover conserved and polymorphic regions of the genome. Genome assembly computes the sequence of a genome from many short DNA sequences. Both computations benefit from recent advances in high performance computing to efficiently process the huge datasets involved, including using highly parallel graphics processing units (GPUs) as high performance desktop processors, and using the MapReduce framework coupled with cloud computing to parallelize computation to large compute grids. This dissertation demonstrates how these technologies can be used to accelerate these computations by orders of magnitude, and have the potential to make otherwise infeasible computations practical

Novel Algorithm Development for ‘NextGeneration’ Sequencing Data Analysis

In recent years, the decreasing cost of ‘Next generation’ sequencing has spawned numerous applications for interrogating whole genomes and transcriptomes in research, diagnostic and forensic settings. While the innovations in sequencing have been explosive, the development of scalable and robust bioinformatics software and algorithms for the analysis of new types of data generated by these technologies have struggled to keep up. As a result, large volumes of NGS data available in public repositories are severely underutilised, despite providing a rich resource for data mining applications. Indeed, the bottleneck in genome and transcriptome sequencing experiments has shifted from data generation to bioinformatics analysis and interpretation. This thesis focuses on development of novel bioinformatics software to bridge the gap between data availability and interpretation. The work is split between two core topics – computational prioritisation/identification of disease gene variants and identification of RNA N6 -adenosine Methylation from sequencing data. The first chapter briefly discusses the emergence and establishment of NGS technology as a core tool in biology and its current applications and perspectives. Chapter 2 introduces the problem of variant prioritisation in the context of Mendelian disease, where tens of thousands of potential candidates are generated by a typical sequencing experiment. Novel software developed for candidate gene prioritisation is described that utilises data mining of tissue-specific gene expression profiles (Chapter 3). The second part of chapter investigates an alternative approach to candidate variant prioritisation by leveraging functional and phenotypic descriptions of genes and diseases from multiple biomedical domain ontologies (Chapter 4). Chapter 5 discusses N6 AdenosineMethylation, a recently re-discovered posttranscriptional modification of RNA. The core of the chapter describes novel software developed for transcriptome-wide detection of this epitranscriptomic mark from sequencing data. Chapter 6 presents a case study application of the software, reporting the previously uncharacterised RNA methylome of Kaposi’s Sarcoma Herpes Virus. The chapter further discusses a putative novel N6-methyl-adenosine -RNA binding protein and its possible roles in the progression of viral infection

Structural auditing methodologies for controlled terminologies

Several auditing methodologies for large controlled terminologies are developed. These are applied to the Unified Medical Language System XXXX and the National Cancer Institute Thesaurus (NCIT). Structural auditing methodologies are based on the structural aspects such as IS-A hierarchy relationships groups of concepts assigned to semantic types and groups of relationships defined for concepts. Structurally uniform groups of concepts tend to be semantically uniform. Structural auditing methodologies focus on concepts with unlikely or rare configuration. These concepts have a high likelihood for errors. One of the methodologies is based on comparing hierarchical relationships between the META and SN, two major knowledge sources of the UMLS. In general, a correspondence between them is expected since the SN hierarchical relationships should abstract the META hierarchical relationships. It may indicate an error when a mismatch occurs. The UMLS SN has 135 categories called semantic types. However, in spite of its medium size, the SN has limited use for comprehension purposes because it cannot be easily represented in a pictorial form, it has many (about 7,000) relationships. Therefore, a higher-level abstraction for the SN called a metaschema, is constructed. Its nodes are meta-semantic types, each representing a connected group of semantic types of the SN. One of the auditing methodologies is based on a kind of metaschema called a cohesive metaschema. The focus is placed on concepts of intersections of meta-semantic types. As is shown, such concepts have high likelihood for errors. Another auditing methodology is based on dividing the NCIT into areas according to the roles of its concepts. Moreover, each multi-rooted area is further divided into pareas that are singly rooted. Each p-area contains a group of structurally and semantically uniform concepts. These groups, as well as two derived abstraction networks called taxonomies, help in focusing on concepts with potential errors. With genomic research being at the forefront of bioscience, this auditing methodology is applied to the Gene hierarchy as well as the Biological Process hierarchy of the NCIT, since processes are very important for gene information. The results support the hypothesis that the occurrence of errors is related to the size of p-areas. Errors are more frequent for small p-areas

Dissemination and visualisation of biological data

With the recent advent of various waves of technological advances, the amount of biological data being generated has exploded. As a consequence of this data deluge, new challenges have emerged in the field of biological data management. In order to maximize the knowledge extracted from the huge amount of biological data produced it is of great importance for the research community that data dissemination and visualisation challenges are tackled. Opening and sharing our data and working collaboratively will benefit the scientific community as a whole and to move towards that end, new developements, tools and techniques are needed. Nowadays, many small research groups are capable of producing important and interesting datasets. The release of those datasets can greatly increase their scientific value. In addition, the development of new data analysis algorithms greatly benefits from the availability of a big corpus of annotated datasets for training and testing purposes, giving new and better algorithms to biomedical sciences in return. None of these would be feasible without large amounts of biological data made freely and publicly available. Dissemination The Distributed Annotation System (DAS) is a protocol designed to publish and integrate annotations on biological entities in a distributed way. DAS is structured as a client-server system where the client retrieves data from one or more servers and to further process and visualise. Nowadays, setting up a DAS server imposes some requirements not met by many research groups. With the aim of removing the hassle of setting up a DAS server, a new software platform has been developed: easyDAS. easyDAS is a hosted platform to automatically create DAS servers. Using a simple web interface the user can upload a data file, describe its contents and a new DAS server will be automatically created and data will be publicly available to DAS clients. Visualisation One of the most broadly used visualization paradigms for genomic data are genomic browsers. A genomic browser is capable of displaying different sets of features positioned relative to a sequence. It is possible to explore the sequence and the features by moving around and zooming in and out. When this project was started, in 2007, all major genome browsers offered quite an static experience. It was possible to browse and explore data, but is was done through a set of buttons to the genome a certain amount of bases to left or right or zooming in and out. From an architectural point of view, all web-based genome browsers were very similar: they all had a relatively thin clien-side part in charge of showing images and big backend servers taking care of everything else. Every change in the display parameters made by the user triggered a request to the server, impacting the perceived responsiveness. We created a new prototype genome browser called GenExp, an interactive web-based browser with canvas based client side data rendering. It offers fluid direct interaction with the genome representation and it's possible to use the mouse drag it and use the mouse wheel to change the zoom level. GenExp offers also some quite unique features, such as its multi-window capabilities that allow a user to create an arbitrary number of independent or linked genome windows and its ability to save and share browsing sessions. GenExp is a DAS client and all data is retrieved from DAS sources. It is possible to add any available DAS data source including all data in Ensembl, UCSC and even the custom ones created with easyDAS. In addition, we developed a javascript DAS client library, jsDAS. jsDAS is a complete DAS client library that will take care of everything DAS related in a javascript application. jsDAS is javascript library agnostic and can be used to add DAS capabilities to any web application. All software developed in this thesis is freely available under an open source license.Les recents millores tecnològiques han portat a una explosió en la quantitat de dades biològiques que es generen i a l'aparició de nous reptes en el camp de la gestió de les dades biològiques. Per a maximitzar el coneixement que podem extreure d'aquestes ingents quantitats de dades cal que solucionem el problemes associats al seu anàlisis, i en particular a la seva disseminació i visualització. La compartició d'aquestes dades de manera lliure i gratuïta pot beneficiar en gran mesura a la comunitat científica i a la societat en general, però per a fer-ho calen noves eines i tècniques. Actualment, molts grups són capaços de generar grans conjunts de dades i la seva publicació en pot incrementar molt el valor científic. A més, la disponibilitat de grans conjunts de dades és necessària per al desenvolupament de nous algorismes d'anàlisis. És important, doncs, que les dades biològiques que es generen siguin accessibles de manera senzilla, estandaritzada i lliure. Disseminació El Sistema d'Anotació Distribuïda (DAS) és un protocol dissenyat per a la publicació i integració d'anotacions sobre entitats biològiques de manera distribuïda. DAS segueix una esquema de client-servidor, on el client obté dades d'un o més servidors per a combinar-les, processar-les o visualitzar-les. Avui dia, però, crear un servidor DAS necessita uns coneixements i infraestructures que van més enllà dels recursos de molts grups de recerca. Per això, hem creat easyDAS, una plataforma per a la creació automàtica de servidors DAS. Amb easyDAS un usuari pot crear un servidor DAS a través d'una senzilla interfície web i amb només alguns clics. Visualització Els navegadors genomics són un dels paradigmes de de visualització de dades genòmiques més usats i permet veure conjunts de dades posicionades al llarg d'una seqüència. Movent-se al llarg d'aquesta seqüència és possibles explorar aquestes dades. Quan aquest projecte va començar, l'any 2007, tots els grans navegadors genomics oferien una interactivitat limitada basada en l'ús de botons. Des d'un punt de vista d'arquitectura tots els navegadors basats en web eren molt semblants: un client senzill encarregat d'ensenyar les imatges i un servidor complex encarregat d'obtenir les dades, processar-les i generar les imatges. Així, cada canvi en els paràmetres de visualització requeria una nova petició al servidor, impactant molt negativament en la velocitat de resposta percebuda. Vam crear un prototip de navegador genòmic anomenat GenExp. És un navegador interactiu basat en web que fa servir canvas per a dibuixar en client i que ofereix la possibilitatd e manipulació directa de la respresentació del genoma. GenExp té a més algunes característiques úniques com la possibilitat de crear multiples finestres de visualització o la possibilitat de guardar i compartir sessions de navegació. A més, com que és un client DAS pot integrar les dades de qualsevol servidor DAS com els d'Ensembl, UCSC o fins i tot aquells creats amb easyDAS. A més, hem desenvolupat jsDAS, la primera llibreria de client DAS completa escrita en javascript. jsDAS es pot integrar en qualsevol aplicació DAS per a dotar-la de la possibilitat d'accedir a dades de servidors DAS. Tot el programari desenvolupat en el marc d'aquesta tesis està lliurement disponible i sota una llicència de codi lliure