Discovering lesser known molecular players and mechanistic patterns in Alzheimer's disease using an integrative disease modelling approach

Convergence of exponentially advancing technologies is driving medical research with life changing discoveries. On the contrary, repeated failures of high-profile drugs to battle Alzheimer's disease (AD) has made it one of the least successful therapeutic area. This failure pattern has provoked researchers to grapple with their beliefs about Alzheimer's aetiology. Thus, growing realisation that Amyloid-β and tau are not 'the' but rather 'one of the' factors necessitates the reassessment of pre-existing data to add new perspectives. To enable a holistic view of the disease, integrative modelling approaches are emerging as a powerful technique. Combining data at different scales and modes could considerably increase the predictive power of the integrative model by filling biological knowledge gaps. However, the reliability of the derived hypotheses largely depends on the completeness, quality, consistency, and context-specificity of the data. Thus, there is a need for agile methods and approaches that efficiently interrogate and utilise existing public data. This thesis presents the development of novel approaches and methods that address intrinsic issues of data integration and analysis in AD research. It aims to prioritise lesser-known AD candidates using highly curated and precise knowledge derived from integrated data. Here much of the emphasis is put on quality, reliability, and context-specificity. This thesis work showcases the benefit of integrating well-curated and disease-specific heterogeneous data in a semantic web-based framework for mining actionable knowledge. Furthermore, it introduces to the challenges encountered while harvesting information from literature and transcriptomic resources. State-of-the-art text-mining methodology is developed to extract miRNAs and its regulatory role in diseases and genes from the biomedical literature. To enable meta-analysis of biologically related transcriptomic data, a highly-curated metadata database has been developed, which explicates annotations specific to human and animal models. Finally, to corroborate common mechanistic patterns — embedded with novel candidates — across large-scale AD transcriptomic data, a new approach to generate gene regulatory networks has been developed. The work presented here has demonstrated its capability in identifying testable mechanistic hypotheses containing previously unknown or emerging knowledge from public data in two major publicly funded projects for Alzheimer's, Parkinson's and Epilepsy diseases

RNA CoMPASS: RNA Comprehensive Multi-Processor Analysis System for Sequencing

The main theme of this dissertation is to develop a distributed computational pipeline for processing next-generation RNA sequencing (RNA-seq) data. RNA-seq experiments generate hundreds of millions of short reads for each DNA/RNA sample. There are many existing bioinformatics tools developed for the analysis and visualization of this data, but very large studies present computational and organizational challenges that are difficult to overcome manually. We designed a comprehensive pipeline for the analysis of RNA sequencing which leverages many existing tools and parallel computing technology to facilitate the analysis of extremely large studies. RNA CoMPASS provides a web-based graphical user interface and distributed computational pipeline including endogenous transcriptome quantification and additionally the investigation of exogenous sequences

RNA CoMPASS: RNA Comprehensive Multi-Processor Analysis System for Sequencing

The main theme of this dissertation is to develop a distributed computational pipeline for processing next-generation RNA sequencing (RNA-seq) data. RNA-seq experiments generate hundreds of millions of short reads for each DNA/RNA sample. There are many existing bioinformatics tools developed for the analysis and visualization of this data, but very large studies present computational and organizational challenges that are difficult to overcome manually. We designed a comprehensive pipeline for the analysis of RNA sequencing which leverages many existing tools and parallel computing technology to facilitate the analysis of extremely large studies. RNA CoMPASS provides a web-based graphical user interface and distributed computational pipeline including endogenous transcriptome quantification and additionally the investigation of exogenous sequences

The Role of bZIP Transcription Factors in Green Plant Evolution: Adaptive Features Emerging from Four Founder Genes

BACKGROUND: Transcription factors of the basic leucine zipper (bZIP) family control important processes in all eukaryotes. In plants, bZIPs are regulators of many central developmental and physiological processes including photomorphogenesis, leaf and seed formation, energy homeostasis, and abiotic and biotic stress responses. Here we performed a comprehensive phylogenetic analysis of bZIP genes from algae, mosses, ferns, gymnosperms and angiosperms. METHODOLOGY/PRINCIPAL FINDINGS: We identified 13 groups of bZIP homologues in angiosperms, three more than known before, that represent 34 Possible Groups of Orthologues (PoGOs). The 34 PoGOs may correspond to the complete set of ancestral angiosperm bZIP genes that participated in the diversification of flowering plants. Homologous genes dedicated to seed-related processes and ABA-mediated stress responses originated in the common ancestor of seed plants, and three groups of homologues emerged in the angiosperm lineage, of which one group plays a role in optimizing the use of energy. CONCLUSIONS/SIGNIFICANCE: Our data suggest that the ancestor of green plants possessed four bZIP genes functionally involved in oxidative stress and unfolded protein responses that are bZIP-mediated processes in all eukaryotes, but also in light-dependent regulations. The four founder genes amplified and diverged significantly, generating traits that benefited the colonization of new environments

Joint learning from multiple information sources for biological problems

Thanks to technological advancements, more and more biological data havebeen generated in recent years. Data availability offers unprecedented opportunities to look at the same problem from multiple aspects. It also unveils a more global view of the problem that takes into account the intricated inter-play between the involved molecules/entities. Nevertheless, biological datasets are biased, limited in quantity, and contain many false-positive samples. Such challenges often drastically downgrade the performance of a predictive model on unseen data and, thus, limit its applicability in real biological studies. Human learning is a multi-stage process in which we usually start with simple things. Through the accumulated knowledge over time, our cognition ability extends to more complex concepts. Children learn to speak simple words before being able to formulate sentences. Similarly, being able to speak correct sentences supports our learning to speak correct and meaningful paragraphs, etc. Generally, knowledge acquired from related learning tasks would help boost our learning capability in the current task. Motivated by such a phenomenon, in this thesis, we study supervised machine learning models for bioinformatics problems that can improve their performance through exploiting multiple related knowledge sources. More specifically, we concern with ways to enrich the supervised models’ knowledge base with publicly available related data to enhance the computational models’ prediction performance. Our work shares commonality with existing works in multimodal learning, multi-task learning, and transfer learning. Nevertheless, there are certain differences in some cases. Besides the proposed architectures, we present large-scale experiment setups with consensus evaluation metrics along with the creation and release of large datasets to showcase our approaches’ superiority. Moreover, we add case studies with detailed analyses in which we place no simplified assumptions to demonstrate the systems’ utilities in realistic application scenarios. Finally, we develop and make available an easy-to-use website for non-expert users to query the model’s generated prediction results to facilitate field experts’ assessments and adaptation. We believe that our work serves as one of the first steps in bridging the gap between “Computer Science” and “Biology” that will open a new era of fruitful collaboration between computer scientists and biological field experts

Integrative bioinformatics applications for complex human disease contexts

This thesis presents new methods for the analysis of high-throughput data from modern sources in the context of complex human diseases, at the example of a bioinformatics analysis workflow. New measurement techniques improve the resolution with which cellular and molecular processes can be monitored. While RNA sequencing (RNA-seq) measures mRNA expression, single-cell RNA-seq (scRNA-seq) resolves this on a per-cell basis. Long-read sequencing is increasingly used in genomics. With imaging mass spectrometry (IMS) the protein level in tissues is measured spatially resolved. All these techniques induce specific challenges, which need to be addressed with new computational methods. Collecting knowledge with contextual annotations is important for integrative data analyses. Such knowledge is available through large literature repositories, from which information, such as miRNA-gene interactions, can be extracted using text mining methods. After aggregating this information in new databases, specific questions can be answered with traceable evidence. The combination of experimental data with these databases offers new possibilities for data integrative methods and for answering questions relevant for complex human diseases. Several data sources are made available, such as literature for text mining miRNA-gene interactions (Chapter 2), next- and third-generation sequencing data for genomics and transcriptomics (Chapters 4.1, 5), and IMS for spatially resolved proteomics (Chapter 4.4). For these data sources new methods for information extraction and pre-processing are developed. For instance, third-generation sequencing runs can be monitored and evaluated using the poreSTAT and sequ-into methods. The integrative (down-stream) analyses make use of these (heterogeneous) data sources. The cPred method (Chapter 4.2) for cell type prediction from scRNA-seq data was successfully applied in the context of the SARS-CoV-2 pandemic. The robust differential expression (DE) analysis pipeline RoDE (Chapter 6.1) contains a large set of methods for (differential) data analysis, reporting and visualization of RNA-seq data. Topics of accessibility of bioinformatics software are discussed along practical applications (Chapter 3). The developed miRNA-gene interaction database gives valuable insights into atherosclerosis-relevant processes and serves as regulatory network for the prediction of active miRNA regulators in RoDE (Chapter 6.1). The cPred predictions, RoDE results, scRNA-seq and IMS data are unified as input for the 3D-index Aorta3D (Chapter 6.2), which makes atherosclerosis related datasets browsable. Finally, the scRNA-seq analysis with subsequent cPred cell type prediction, and the robust analysis of bulk-RNA-seq datasets, led to novel insights into COVID-19. Taken all discussed methods together, the integrative analysis methods for complex human disease contexts have been improved at essential positions.Die Dissertation beschreibt Methoden zur Prozessierung von aktuellen Hochdurchsatzdaten, sowie Verfahren zu deren weiterer integrativen Analyse. Diese findet Anwendung vor allem im Kontext von komplexen menschlichen Krankheiten. Neue Messtechniken erlauben eine detailliertere Beobachtung biomedizinischer Prozesse. Mit RNA-Sequenzierung (RNA-seq) wird mRNA-Expression gemessen, mit Hilfe von moderner single-cell-RNA-seq (scRNA-seq) sogar für (sehr viele) einzelne Zellen. Long-Read-Sequenzierung wird zunehmend zur Sequenzierung ganzer Genome eingesetzt. Mittels bildgebender Massenspektrometrie (IMS) können Proteine in Geweben räumlich aufgelöst quantifiziert werden. Diese Techniken bringen spezifische Herausforderungen mit sich, die mit neuen bioinformatischen Methoden angegangen werden müssen. Für die integrative Datenanalyse ist auch die Gewinnung von geeignetem Kontextwissen wichtig. Wissenschaftliche Erkenntnisse werden in Artikeln veröffentlicht, die über große Literaturdatenbanken zugänglich sind. Mittels Textmining können daraus Informationen extrahiert werden, z.B. miRNA-Gen-Interaktionen, die in eigenen Datenbank aggregiert werden um spezifische Fragen mit nachvollziehbaren Belegen zu beantworten. In Kombination mit experimentellen Daten bieten sich so neue Möglichkeiten für integrative Methoden. Durch die Extraktion von Rohdaten und deren Vorprozessierung werden mehrere Datenquellen erschlossen, wie z.B. Literatur für Textmining von miRNA-Gen-Interaktionen (Kapitel 2), Long-Read- und RNA-seq-Daten für Genomics und Transcriptomics (Kapitel 4.2, 5) und IMS für Protein-Messungen (Kapitel 4.4). So dienen z.B. die poreSTAT und sequ-into Methoden der Vorprozessierung und Auswertung von Long-Read-Sequenzierungen. In der integrativen (down-stream) Analyse werden diese (heterogenen) Datenquellen verwendet. Für die Bestimmung von Zelltypen in scRNA-seq-Experimenten wurde die cPred-Methode (Kapitel 4.2) erfolgreich im Kontext der SARS-CoV-2-Pandemie eingesetzt. Auch die robuste Pipeline RoDE fand dort Anwendung, die viele Methoden zur (differentiellen) Datenanalyse, zum Reporting und zur Visualisierung bereitstellt (Kapitel 6.1). Themen der Benutzbarkeit von (bioinformatischer) Software werden an Hand von praktischen Anwendungen diskutiert (Kapitel 3). Die entwickelte miRNA-Gen-Interaktionsdatenbank gibt wertvolle Einblicke in Atherosklerose-relevante Prozesse und dient als regulatorisches Netzwerk für die Vorhersage von aktiven miRNA-Regulatoren in RoDE (Kapitel 6.1). Die cPred-Methode, RoDE-Ergebnisse, scRNA-seq- und IMS-Daten werden im 3D-Index Aorta3D (Kapitel 6.2) zusammengeführt, der relevante Datensätze durchsuchbar macht. Die diskutierten Methoden führen zu erheblichen Verbesserungen für die integrative Datenanalyse in komplexen menschlichen Krankheitskontexten

Bayesian nonparametric clusterings in relational and high-dimensional settings with applications in bioinformatics.

Recent advances in high throughput methodologies offer researchers the ability to understand complex systems via high dimensional and multi-relational data. One example is the realm of molecular biology where disparate data (such as gene sequence, gene expression, and interaction information) are available for various snapshots of biological systems. This type of high dimensional and multirelational data allows for unprecedented detailed analysis, but also presents challenges in accounting for all the variability. High dimensional data often has a multitude of underlying relationships, each represented by a separate clustering structure, where the number of structures is typically unknown a priori. To address the challenges faced by traditional clustering methods on high dimensional and multirelational data, we developed three feature selection and cross-clustering methods: 1) infinite relational model with feature selection (FIRM) which incorporates the rich information of multirelational data; 2) Bayesian Hierarchical Cross-Clustering (BHCC), a deterministic approximation to Cross Dirichlet Process mixture (CDPM) and to cross-clustering; and 3) randomized approximation (RBHCC), based on a truncated hierarchy. An extension of BHCC, Bayesian Congruence Measuring (BCM), is proposed to measure incongruence between genes and to identify sets of congruent loci with identical evolutionary histories. We adapt our BHCC algorithm to the inference of BCM, where the intended structure of each view (congruent loci) represents consistent evolutionary processes. We consider an application of FIRM on categorizing mRNA and microRNA. The model uses latent structures to encode the expression pattern and the gene ontology annotations. We also apply FIRM to recover the categories of ligands and proteins, and to predict unknown drug-target interactions, where latent categorization structure encodes drug-target interaction, chemical compound similarity, and amino acid sequence similarity. BHCC and RBHCC are shown to have improved predictive performance (both in terms of cluster membership and missing value prediction) compared to traditional clustering methods. Our results suggest that these novel approaches to integrating multi-relational information have a promising future in the biological sciences where incorporating data related to varying features is often regarded as a daunting task

Single-Molecule Imaging Reveals that Argonaute Re-Shapes the Properties of its Nucleic Acid Guides: A Dissertation

Small RNA silencing pathways regulate development, viral defense, and genomic integrity in all kingdoms of life. An Argonaute (Ago) protein, guided by a tightly bound, small RNA or DNA, lies at the core of these pathways. Argonaute uses its small RNA or DNA to find its target sequences, which it either cleaves or stably binds, acting as a binding scaffold for other proteins. We used Co-localization Single-Molecule Spectroscopy (CoSMoS) to analyze target binding and cleavage by Ago and its guide. We find that both eukaryotic and prokaryotic Argonaute proteins re-shape the fundamental properties of RNA:RNA, RNA:DNA, and DNA:DNA hybridization: a small RNA or DNA bound to Argonaute as a guide no longer follows the well-established rules by which oligonucleotides find, bind, and dissociate from complementary nucleic acid sequences. Counter to the rules of nucleic acid hybridization alone, we find that mouse AGO2 and its guide bind to microRNA targets 17,000 times tighter than the guide without Argonaute. Moreover, AGO2 can distinguish between microRNA-like targets that make seven base pairs with the guide and the products of cleavage, which bind via nine base pairs: AGO2 leaves the cleavage products faster, even though they pair more extensively. This thesis presents a detailed kinetic interrogation of microRNA and RNA interference pathways. We discovered sub-domains within the previously defined functional domains created by Argonaute and its bound DNA or RNA guide. These sub-domains have features that no longer conform to the well-established properties of unbound oligonucleotides. It is by re-writing the rules for nucleic acid hybridization that Argonautes allow oligonucleotides to serve as specificity determinants with thermodynamic and kinetic properties more typical of RNA-binding proteins than that of RNA or DNA. Taken altogether, these studies further our understanding about the biology of small RNA silencing pathways and may serve to guide future work related to all RNA-guided endonucleases