A Novel Graph-Theoretical Approach for Identifying Inter-correlations and Functional Pathways in Microbiome Data

The human microbiome represents a vastly complex ecosystem that is tightly linked to our development. Microbiomes play critical roles in nutrient uptake, immune system development, and vitamin production. The evolution of human microbiomes continues to impact the overall health and quality of life of all humans. For example, despite the ability of microbiota to restrict pathogen invasion, pathogens have evolved tougher due to changes in diets, host environments, and use of antibiotics and other drugs. Such evolution has impacted the organization and composition of the microbial community which, in turn, has influenced susceptibility to and severity of the different type of infections. In fact, the interactions between elements of microbiome significantly shape its host microbial community. Hence, recent metagenomics- based studies of microbiomes has been focusing on how to characterize the composition of species in the microbiome and their co-occurrence patterns. In this study, we propose a comprehensive graph-theoretic framework that integrates microbe-microbe and microbe-host interactions. The study uses microbiome data from Crohn’s disease patients and healthy individuals in a Korean population to develop and test an integrated bioinformatics pipeline. We utilize the integrated pipeline to characterize the taxonomic and metabolic pathway composition in both groups. We show that different groups of bacteria are significantly associated with various phenotypes related to metabolic pathways in patient samples as compared to healthy samples. The obtained results also reveal that microbial elements extracted from within the highly correlated group among Crohn’s disease patients are closely associated with the metabolic mechanisms that have been linked to Crohn’s disease

Disentangling ecological networks in marine microbes

There is a myriad of microorganisms on Earth contributing to global biogeochemical cycles, and their interactions are considered pivotal for ecosystem function. Previous studies have already determined relationships between a limited number of microorganisms. Yet, we still need to understand a large number of interactions to increase our knowledge of complex microbiomes. This is challenging because of the vast number of possible interactions. Thus, microbial interactions still remain barely known to date. Networks are a great tool to handle the vast number of microorganisms and their connections, explore potential microbial interactions, and elucidate patterns of microbial ecosystems. This thesis locates at the intersection of network inference and network analysis. The presented methodology aims to support and advance marine microbial investigations by reducing noise and elucidating patterns in inferred association networks for subsequent biological down-stream analyses. This thesis’s main contribution to marine microbial interactions studies is the development of the program EnDED (Environmentally-Driven Edge Detection), a computational framework to identify environmentally-driven associations inside microbial association networks, inferred from omics datasets. We applied the methodology to a model marine microbial ecosystem at the Blanes Bay Microbial Observatory (BBMO) in the North-Western Mediterranean Sea (ten years of monthly sampling). We also applied the methodology to a dataset compilation covering six global-ocean regions from the surface (3 m) to the deep ocean (down to 4539 m). Thus, our methodology provided a step towards studying the marine microbial distribution in space via the horizontal (ocean regions) and vertical (water column) axes.Hi ha una infinitat de microorganismes a la Terra que contribueixen als cicles biogeoquímics mundials i les seves interaccions es consideren fonamentals pel funcionament dels ecosistemes. Estudis previs ja han determinat les relacions entre un nombre limitat de microorganismes. Tot i això, encara hem d’entendre un gran nombre d’interaccions per augmentar el nostre coneixement dels microbiomes complexos. Això és un repte a causa del gran nombre d'interaccions possibles. Per això, les interaccions microbianes encara són poc conegudes fins ara. Les xarxes són una gran eina per tractar el gran nombre de microorganismes i les seves connexions, explorar interaccions microbianes potencials i dilucidar patrons d’ecosistemes microbians. Aquesta tesi es situa a la intersecció de la inferència de xarxes i l’anàlisi de la xarxes. La metodologia presentada té com a objectiu donar suport i avançar en investigacions microbianes marines reduint el soroll i dilucidant patrons en xarxes d’associació inferides per a posteriors anàlisis biològiques. La principal contribució d’aquesta tesi als estudis d’interaccions microbianes marines és el desenvolupament del programa EnDED (Environmentally-Driven Edge Detection), un marc computacional per identificar associacions impulsades pel medi ambient dins de xarxes d’associació microbiana, inferides a partir de conjunts de dades òmics. S’ha aplicat la metodologia a un model d’ecosistema microbià marí a l’Observatori Microbià de la Badia de Blanes (BBMO) al mar Mediterrani nord-occidental (deu anys de mostreig mensual). També s’ha la metodologia a una recopilació de dades que cobreix sis regions oceàniques globals des de la superfície (3 m) fins a l'oceà profund (fins a 4539 m).Hay una gran cantidad de microorganismos en la Tierra que contribuyen a los ciclos biogeoquímicos globales, y sus interacciones se consideran fundamentales para la función del ecosistema. Estudios previos ya han determinado relaciones entre un número limitado de microorganismos. Sin embargo, todavía necesitamos comprender una gran cantidad de interacciones para aumentar nuestro conocimiento de los microbiomas más complejos. Esto representa un gran desafío debido a la gran cantidad de posibles interacciones. Por lo tanto, las interacciones microbianas son aun poco conocidas. Las redes representan una gran herramienta para analizar la gran cantidad de microorganismos y sus conexiones, explorar posibles interacciones y dilucidar patrones en ecosistemas microbianos. Esta tesis se ubica en la intersección entre la inferencia de redes y el análisis de redes. La metodología presentada tiene como objetivo avanzar las investigaciones sobre interacciones microbianas marinas mediante la reducción del ruido en las inferencias de redes y elucidar patrones en redes de asociación permitiendo análisis biológicos posteriores. La principal contribución de esta tesis a los estudios de interacciones microbianas marinas es el desarrollo del programa EnDED (Environmentally-Driven Edge Detection), un marco computacional para identificar asociaciones generadas por el medio ambiente en redes de asociaciones microbianas, inferidas a partir de datos ómicos. Aplicamos la metodología a un modelo de ecosistema microbiano marino en el Observatorio Microbiano de la Bahía de Blanes (BBMO) en el Mar Mediterráneo Noroccidental (diez años de muestreo mensual). También, aplicamos la metodología a una compilación de conjuntos de datos que cubren seis regiones oceánicas globales desde la superficie (3 m) hasta las profundidades del océano (hasta 4539 m). Por lo tanto, nuestra metodología significa un paso adelante hacia de los patrones temporales microbianos marinos y el estudio de la distribución microbiana marina en el espacio a través de los ejes horizontal (regiones oceánicas) y vertical (columna de agua). Para llegar a hipótesis de interacción precisas, es importante determinar, cuantificar y eliminar las asociaciones generadas por el medio ambiente en las redes de asociaciones microbianas marinas. Además, nuestros resultados subrayaron la necesidad de estudiar la naturaleza dinámica de las redes, en contraste con el uso de redes estáticas únicas agregadas en el tiempo o el espacio. Nuestras nuevas metodologías pueden ser utilizadas por una amplia gama de investigadores que investigan redes e interacciones en diversos microbiomas.Postprint (published version

Integration and visualisation of clinical-omics datasets for medical knowledge discovery

In recent decades, the rise of various omics fields has flooded life sciences with unprecedented amounts of high-throughput data, which have transformed the way biomedical research is conducted. This trend will only intensify in the coming decades, as the cost of data acquisition will continue to decrease. Therefore, there is a pressing need to find novel ways to turn this ocean of raw data into waves of information and finally distil those into drops of translational medical knowledge. This is particularly challenging because of the incredible richness of these datasets, the humbling complexity of biological systems and the growing abundance of clinical metadata, which makes the integration of disparate data sources even more difficult. Data integration has proven to be a promising avenue for knowledge discovery in biomedical research. Multi-omics studies allow us to examine a biological problem through different lenses using more than one analytical platform. These studies not only present tremendous opportunities for the deep and systematic understanding of health and disease, but they also pose new statistical and computational challenges. The work presented in this thesis aims to alleviate this problem with a novel pipeline for omics data integration. Modern omics datasets are extremely feature rich and in multi-omics studies this complexity is compounded by a second or even third dataset. However, many of these features might be completely irrelevant to the studied biological problem or redundant in the context of others. Therefore, in this thesis, clinical metadata driven feature selection is proposed as a viable option for narrowing down the focus of analyses in biomedical research. Our visual cortex has been fine-tuned through millions of years to become an outstanding pattern recognition machine. To leverage this incredible resource of the human brain, we need to develop advanced visualisation software that enables researchers to explore these vast biological datasets through illuminating charts and interactivity. Accordingly, a substantial portion of this PhD was dedicated to implementing truly novel visualisation methods for multi-omics studies.Open Acces

iVikodak—A Platform and Standard Workflow for Inferring, Analyzing, Comparing, and Visualizing the Functional Potential of Microbial Communities

Background: The objectives of any metagenomic study typically include identification of resident microbes and their relative proportions (taxonomic analysis), profiling functional diversity (functional analysis), and comparing the identified microbes and functions with available metadata (comparative metagenomics). Given the advantage of cost-effectiveness and convenient data-size, amplicon-based sequencing has remained the technology of choice for exploring phylogenetic diversity of an environment. A recent school of thought, employing the existing genome annotation information for inferring functional capacity of an identified microbiome community, has given a promising alternative to Whole Genome Shotgun sequencing for functional analysis. Although a handful of tools are currently available for function inference, their scope, functionality and utility has essentially remained limited. Need for a comprehensive framework that expands upon the existing scope and enables a standardized workflow for function inference, analysis, and visualization, is therefore felt.Methods: We present iVikodak, a multi-modular web-platform that hosts a logically inter-connected repertoire of functional inference and analysis tools, coupled with a comprehensive visualization interface. iVikodak is equipped with microbial co-inhabitance pattern driven published algorithms along with multiple updated databases of various curated microbe-function maps. It also features an advanced task management and result sharing system through introduction of personalized and portable dashboards.Results: In addition to inferring functions from 16S rRNA gene data, iVikodak enables (a) an in-depth analysis of specific functions of interest (b) identification of microbes contributing to various functions (c) microbial interaction patterns through function-driven correlation networks, and (d) simultaneous functional comparison between multiple microbial communities. We have bench-marked iVikodak through multiple case studies and comparisons with existing state of art. We also introduce the concept of a public repository which provides a first of its kind community-driven framework for scientific data analytics, collaboration and sharing in this area of microbiome research.Conclusion: Developed using modern design and task management practices, iVikodak provides a multi-modular, yet inter-operable, one-stop framework, that intends to simplify the entire approach toward inferred function analysis. It is anticipated to serve as a significant value addition to the existing space of functional metagenomics.iVikodak web-server may be freely accessed at https://web.rniapps.net/iVikodak/

Emerging Priorities for Microbiome Research

Microbiome research has increased dramatically in recent years, driven by advances in technology and significant reductions in the cost of analysis. Such research has unlocked a wealth of data, which has yielded tremendous insight into the nature of the microbial communities, including their interactions and effects, both within a host and in an external environment as part of an ecological community. Understanding the role of microbiota, including their dynamic interactions with their hosts and other microbes, can enable the engineering of new diagnostic techniques and interventional strategies that can be used in a diverse spectrum of fields, spanning from ecology and agriculture to medicine and from forensics to exobiology. From June 19–23 in 2017, the NIH and NSF jointly held an Innovation Lab on Quantitative Approaches to Biomedical Data Science Challenges in our Understanding of the Microbiome. This review is inspired by some of the topics that arose as priority areas from this unique, interactive workshop. The goal of this review is to summarize the Innovation Lab’s findings by introducing the reader to emerging challenges, exciting potential, and current directions in microbiome research. The review is broken into five key topic areas: (1) interactions between microbes and the human body, (2) evolution and ecology of microbes, including the role played by the environment and microbe-microbe interactions, (3) analytical and mathematical methods currently used in microbiome research, (4) leveraging knowledge of microbial composition and interactions to develop engineering solutions, and (5) interventional approaches and engineered microbiota that may be enabled by selectively altering microbial composition. As such, this review seeks to arm the reader with a broad understanding of the priorities and challenges in microbiome research today and provide inspiration for future investigation and multi-disciplinary collaboration

Resolving Drivers of Microbial Community Structure in The Dry Valleys of Antarctica

Understanding the processes that underlie patterns of microbial distribution is fundamental to the field of microbial ecology, but extremely challenging given the complexity of natural systems. Antarctica’s ice-free regions possess unique ecosystems of simple trophic structure, shaped by the harsh environmental conditions that typify the continent. The Dry Valleys comprise the largest of these regions and have one of the simplest food webs on the planet, making them a tractable system to comprehensively define relationships that influence microbial distributions at the landscape scale. The New Zealand Terrestrial Antarctic Biocomplexity Survey (nzTABS) was aimed at identifying factors that control, and can predict, biological distributions in the Dry Valleys. As part of nzTABS, the goal of the research presented in this thesis was to elucidate the factors that influence bacterial community structure in Dry Valley soils. This study explored how topographic, physicochemical, and spatial variation influence bacterial diversity and community structure across a Dry Valley landscape. Bacterial communities were characterized in 471 soil samples using automated ribosomal intergenic spacer analysis (ARISA). Diversity and community composition were most strongly related to variation in physicochemical soil properties, though significant relationships with topographic and spatial variables were also observed. This identified, for the first time, the influence of environmental variables on bacterial diversity and community composition across the landscape, and presents a structural equation model identifying those relationships. The phylogenetic diversity of bacterial communities in Dry Valley soils was also examined. High-throughput sequencing of 16S rRNA gene amplicons was used to analyze bacterial communities in 177 soil samples. This work identified significant relationships between the relative abundances of bacterial taxa and both abiotic and biotic variables, though these relationships explained only a small amount of community variation collectively. The relative abundances of several bacterial taxa were, however, significantly coupled to one another, suggesting that interactions between bacterial taxa may influence community compositions. Lastly, the bacterial composition of aerosols above the Dry Valleys was examined. High-throughput sequencing of 16S rRNA gene amplicons was used to analyze two aerosol samples collected in the Miers Valley, and their compositions were compared to those of previously characterized aerosols and soils from across the continent. Bacteria present in the aerosols were found to be distinct from those of local soils; instead, aerosol compositions were more similar to those of air samples reported from elsewhere on the planet. Importantly, these findings suggest that local redistribution of Dry Valley soil bacteria through atmospheric processes may be largely restricted to periods when high winds mobilize soil particles and associated biota. This study provides novel insights into the microbial ecology of the Dry Valleys. Despite the relative simplicity of the ecosystem, the factors that influence bacterial distributions within the Dry Valleys appear to be highly complex, and include a combination of abiotic and biotic drivers. Continued research will help to disentangle relationships that influence microbial community compositions in Antarctica’s ice-free ecosystems, and will improve understanding of processes that influence microbial community assembly globally

On Computable Protein Functions

Proteins are biological machines that perform the majority of functions necessary for life. Nature has evolved many different proteins, each of which perform a subset of an organism’s functional repertoire. One aim of biology is to solve the sparse high dimensional problem of annotating all proteins with their true functions. Experimental characterisation remains the gold standard for assigning function, but is a major bottleneck due to resource scarcity. In this thesis, we develop a variety of computational methods to predict protein function, reduce the functional search space for proteins, and guide the design of experimental studies. Our methods take two distinct approaches: protein-centric methods that predict the functions of a given protein, and function-centric methods that predict which proteins perform a given function. We applied our methods to help solve a number of open problems in biology. First, we identified new proteins involved in the progression of Alzheimer’s disease using proteomics data of brains from a fly model of the disease. Second, we predicted novel plastic hydrolase enzymes in a large data set of 1.1 billion protein sequences from metagenomes. Finally, we optimised a neural network method that extracts a small number of informative features from protein networks, which we used to predict functions of fission yeast proteins

Aging and Health

Aging is a major risk factor for chronic diseases, which in turn can provide information about the aging of a biological system. This publication serves as an introduction to systems biology and its application to biological aging. Key pathways and processes that impinge on aging are reviewed, and how they contribute to health and disease during aging is discussed. The evolution of this situation is analyzed, and the consequences for the study of genetic effects on aging are presented. Epigenetic programming of aging, as a continuation of development, creates an interface between the genome and the environment. New research into the gut microbiome describes how this interface may operate in practice with marked consequences for a variety of disorders. This analysis is bolstered by a view of the aging organism as a whole, with conclusions about the mechanisms underlying resilience of the organism to change, and is expanded with a discussion of circadian rhythms in aging

Aging and Health

Aging is a major risk factor for chronic diseases, which in turn can provide information about the aging of a biological system. This publication serves as an introduction to systems biology and its application to biological aging. Key pathways and processes that impinge on aging are reviewed, and how they contribute to health and disease during aging is discussed. The evolution of this situation is analyzed, and the consequences for the study of genetic effects on aging are presented. Epigenetic programming of aging, as a continuation of development, creates an interface between the genome and the environment. New research into the gut microbiome describes how this interface may operate in practice with marked consequences for a variety of disorders. This analysis is bolstered by a view of the aging organism as a whole, with conclusions about the mechanisms underlying resilience of the organism to change, and is expanded with a discussion of circadian rhythms in aging