Machine learning and data-parallel processing for viral metagenomics

More than 2 million cancer cases around the world each year are caused by viruses. In addition, there are epidemiological indications that other cancer-associated viruses may also exist. However, the identification of highly divergent and yet unknown viruses in human biospecimens is one of the biggest challenges in bio- informatics. Modern-day Next Generation Sequencing (NGS) technologies can be used to directly sequence biospecimens from clinical cohorts with unprecedented speed and depth. These technologies are able to generate billions of bases with rapidly decreasing cost but current bioinformatics tools are inefficient to effectively process these massive datasets. Thus, the objective of this thesis was to facilitate both the detection of highly divergent viruses among generated sequences as well as large-scale analysis of human metagenomic datasets. To re-analyze human sample-derived sequences that were classified as being of “unknown” origin by conventional alignment-based methods, we used a meth- odology based on profile Hidden Markov Models (HMM) which can capture evolutionary changes by using multiple sequence alignments. We thus identified 510 sequences that were classified as distantly related to viruses. Many of these sequences were homologs to large viruses such as Herpesviridae and Mimiviridae but some of them were also related to small circular viruses such as Circoviridae. We found that bioinformatics analysis using viral profile HMM is capable of extending the classification of previously unknown sequences and consequently the detection of viruses in biospecimens from humans. Different organisms use synonymous codons differently to encode the same amino acids. To investigate whether codon usage bias could predict the presence of virus in metagenomic sequencing data originating from human samples, we trained Random Forest and Artificial Neural Networks based on Relative Synonymous Codon Usage (RSCU) frequency. Our analysis showed that machine learning tech- niques based on RSCU could identify putative viral sequences with area under the ROC curve of 0.79 and provide important information for taxonomic classification. For identification of viral genomes among raw metagenomic sequences, we devel- oped the tool ViraMiner, a deep learning-based method which uses Convolutional Neural Networks with two convolutional branches. Using 300 base-pair length sequences, ViraMiner achieved 0.923 area under the ROC curve which is con- siderably improved performance in comparison with previous machine learning methods for virus sequence classification. The proposed architecture, to the best of our knowledge, is the first deep learning tool which can detect viral genomes on raw metagenomic sequences originating from a variety of human samples. To enable large-scale analysis of massive metagenomic sequencing data we used Apache Hadoop and Apache Spark to develop ViraPipe, a scalable parallel bio- informatics pipeline for viral metagenomics. Comparing ViraPipe (executed on 23 nodes) with the sequential pipeline (executed on a single node) was 11 times faster in the metagenome analysis. The new distributed workflow contains several standard bioinformatics tools and can scale to terabytes of data by accessing more computer power from the nodes. To analyze terabytes of RNA-seq data originating from head and neck squamous cell carcinoma samples, we used our parallel bioinformatics pipeline ViraPipe and the most recent version of the HPV sequence database. We detected transcription of HPV viral oncogenes in 92/500 cancers. HPV 16 was the most important HPV type, followed by HPV 33 as the second most common infection. If these cancers are indeed caused by HPV, we estimated that vaccination might prevent about 36 000 head and neck cancer cases in the United States every year. In conclusion, the work in this thesis improves the prospects for biomedical researchers to classify the sequence contents of ultra-deep datasets, conduct large- scale analysis of metagenome studies, and detect presence of viral genomes in human biospecimens. Hopefully, this work will contribute to our understanding of biodiversity of viruses in humans which in turn can help exploring infectious causes of human disease

The International Virus Bioinformatics Meeting 2023

The 2023 International Virus Bioinformatics Meeting was held in Valencia, Spain, from 24–26 May 2023, attracting approximately 180 participants worldwide. The primary objective of the conference was to establish a dynamic scientific environment conducive to discussion, collaboration, and the generation of novel research ideas. As the first in-person event following the SARS-CoV-2 pandemic, the meeting facilitated highly interactive exchanges among attendees. It served as a pivotal gathering for gaining insights into the current status of virus bioinformatics research and engaging with leading researchers and emerging scientists. The event comprised eight invited talks, 19 contributed talks, and 74 poster presentations across eleven sessions spanning three days. Topics covered included machine learning, bacteriophages, virus discovery, virus classification, virus visualization, viral infection, viromics, molecular epidemiology, phylodynamic analysis, RNA viruses, viral sequence analysis, viral surveillance, and metagenomics. This report provides rewritten abstracts of the presentations, a summary of the key research findings, and highlights shared during the meeting

The Promises and Pitfalls of Machine Learning for Detecting Viruses in Aquatic Metagenomes

Tools allowing for the identification of viral sequences in host-associated and environmental metagenomes allows for a better understanding of the genetics and ecology of viruses and their hosts. Recently, new approaches using machine learning methods to distinguish viral from bacterial signal using k-mer sequence signatures were published for identifying viral contigs in metagenomes. The promise of these content-based approaches is the ability to discover new viruses, with no or few known relatives. In this perspective paper, we examine the use of the content-based machine learning tool VirFinder for the identification of viral sequences in aquatic metagenomes and explore the possibility of using ecosystem-focused models targeted to marine metagenomes. We discuss the impact of the training set composition on the tool performance and the current limitation for the retrieval of low abundance viral sequences in metagenomes. We identify potential biases that could arise from machine learning approaches for viral hunting in real-world datasets and suggest possible avenues to overcome them.National Science Foundation [1640775]Open access journal.This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Convolutional Neural Network Applied to SARS-CoV-2 Sequence Classification

COVID-19, the illness caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus belonging to the Coronaviridade family, a single-strand positive-sense RNA genome, has been spreading around the world and has been declared a pandemic by the World Health Organization. On 17 January 2022, there were more than 329 million cases, with more than 5.5 million deaths. Although COVID-19 has a low mortality rate, its high capacities for contamination, spread, and mutation worry the authorities, especially after the emergence of the Omicron variant, which has a high transmission capacity and can more easily contaminate even vaccinated people. Such outbreaks require elucidation of the taxonomic classification and origin of the virus (SARS-CoV-2) from the genomic sequence for strategic planning, containment, and treatment of the disease. Thus, this work proposes a high-accuracy technique to classify viruses and other organisms from a genome sequence using a deep learning convolutional neural network (CNN). Unlike the other literature, the proposed approach does not limit the length of the genome sequence. The results show that the novel proposal accurately distinguishes SARS-CoV-2 from the sequences of other viruses. The results were obtained from 1557 instances of SARS-CoV-2 from the National Center for Biotechnology Information (NCBI) and 14,684 different viruses from the Virus-Host DB. As a CNN has several changeable parameters, the tests were performed with forty-eight different architectures; the best of these had an accuracy of 91.94 +/- 2.62% in classifying viruses into their realms correctly, in addition to 100% accuracy in classifying SARS-CoV-2 into its respective realm, Riboviria. For the subsequent classifications (family, genera, and subgenus), this accuracy increased, which shows that the proposed architecture may be viable in the classification of the virus that causes COVID-19.Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)High-Performance Computing Center at UFRN(NPAD/UFRN

What the Phage: a scalable workflow for the identification and analysis of phage sequences

Phages are among the most abundant and diverse biological entities on earth. Phage prediction from sequence data is a crucial first step to understanding their impact on the environment. A variety of bacteriophage prediction tools have been developed over the years. They differ in algorithmic approach, results, and ease of use. We, therefore, developed "What the Phage"(WtP), an easy-to-use and parallel multitool approach for phage prediction combined with an annotation and classification downstream strategy, thus supporting the user's decision-making process by summarizing the results of the different prediction tools in charts and tables. WtP is reproducible and scales to thousands of datasets through a workflow manager (Nextflow). WtP is freely available under a GPL-3.0 license (https://github.com/replikation/What_the_Phage)