FILER: a framework for harmonizing and querying large-scale functional genomics knowledge

ABSTRACT Querying massive functional genomic and annotation data collections, linking and summarizing the query results across data sources/data types are important steps in high-throughput genomic and genetic analytical workflows. However, these steps are made difficult by the heterogeneity and breadth of data sources, experimental assays, biological conditions/tissues/cell types and file formats. FILER (FunctIonaL gEnomics Repository) is a framework for querying large-scale genomics knowledge with a large, curated integrated catalog of harmonized functional genomic and annotation data coupled with a scalable genomic search and querying interface. FILER uniquely provides: (i) streamlined access to >50 000 harmonized, annotated genomic datasets across >20 integrated data sources, >1100 tissues/cell types and >20 experimental assays; (ii) a scalable genomic querying interface; and (iii) ability to analyze and annotate user’s experimental data. This rich resource spans >17 billion GRCh37/hg19 and GRCh38/hg38 genomic records. Our benchmark querying 7 × 109 hg19 FILER records shows FILER is highly scalable, with a sub-linear 32-fold increase in querying time when increasing the number of queries 1000-fold from 1000 to 1 000 000 intervals. Together, these features facilitate reproducible research and streamline integrating/querying large-scale genomic data within analyses/workflows. FILER can be deployed on cloud or local servers (https://bitbucket.org/wanglab-upenn/FILER) for integration with custom pipelines and is freely available (https://lisanwanglab.org/FILER).

ATP6V1B2 and IFI27 and their intrinsic functional genomic characteristics associated with SARS-CoV-2

Genes functionally associated with SARS-CoV-2 and genes functionally related to COVID-19 disease can be different, whose distinction will become the first essential step for successfully fighting against the COVID-19 pandemic. Unfortunately, this first step has not been completed in all biological and medical research. Using a newly developed max-competing logistic classifier, two genes, ATP6V1B2 and IFI27, stand out to be critical in transcriptional response to SARS-CoV-2 with differential expressions derived from NP/OP swab PCR. This finding is evidenced by combining these two genes with one another gene in predicting disease status to achieve better-indicating power than existing classifiers with the same number of genes. In addition, combining these two genes with three other genes to form a five-gene classifier outperforms existing classifiers with ten or more genes. With their exceptional predicting power, these two genes can be critical in fighting against the COVID-19 pandemic as a new focus and direction. Comparing the functional effects of these genes with a five-gene classifier with 100% accuracy identified and tested from blood samples in the literature, genes and their transcriptional response and functional effects to SARS-CoV-2 and genes and their functional signature patterns to COVID-19 antibody are significantly different, which can be interpreted as the former is the point of a phenomenon, and the latter is the essence of the disease. Such significant findings can help explore the causal and pathological clue between SARS-CoV-2 and COVID-19 disease and fight against the disease with more targeted vaccines, antiviral drugs, and therapies.

Molecular Paleobiology of the Echinoderm Skeleton

Molecular paleobiology provides a promising avenue to merge data from deep time, molecular biology and genomics, gaining insights into the evolutionary process at multiple levels. The echinoderm skeleton is a model for molecular paleobioloogical studies. I begin with an overview of the skeletogenic process in echinoderms, as well as a discussion of what gene regulatory networks are, and why they are of interest to paleobiologists. I then highlight recent advances in the evolution of the echinoderm skeleton from both paleobiological and molecular/functional genomic perspectives, highlighting examples where diverse approaches provide complementary insight and discussing potential of this field of research.

A deep learning framework for predicting human essential genes from population and functional genomic data

Being able to predict essential genes intolerant to loss-of-function (LOF) mutations can dramatically improve our ability to identify genes associated with genetic disorders. Numerous computational methods have recently been developed to predict human essential genes from population genomic data; however, the existing methods have limited power in pinpointing short essential genes due to the sparsity of polymorphisms in the human genome. Here we present an evolution-based deep learning model, DeepLOF, which integrates population and functional genomic data to improve gene essentiality prediction. Compared to previous methods, DeepLOF shows unmatched performance in predicting ClinGen haploinsufficient genes, mouse essential genes, and essential genes in human cell lines. Furthermore, DeepLOF discovers 109 potentially essential genes that are too short to be identified by previous methods. Altogether, DeepLOF is a powerful computational method to aid in the discovery of essential genes.

Classifications of Iris L. genus at the biological and molecular levels as a basis for modern phylogenetic studies

Aim. To analyze and summarize literature data on the state of classification and taxonomy of Iris genus plants of Iridaceae family.Results and discussion. The genus Iris L. is one of the largest species of Iridaceae Juss family, which includes 1,800 species of 80 genera. Despite the fact that plants are very common as ornamental plants, there is currently no single classification for both horticultural and wild species of Iris genus. Various classifications and systems of irises have been proposed, the main difference between them is related to the understanding of the genus scope. The taxonomy of Iris genus underwent dramatic changes in the last century, especially in recent decades after the introduction of molecular methods. This paper deals with the analysis of current literature data on the existing classifications of Iris genus plants, the correspondence between taxonomic studies and the results of recent molecular studies. The data presented summarize the traditional classifications according to Rodionenko and Matthew, and also describe modern achievements in phylogenetic studies of this genus using functional genomic studies, involving molecular markers in irises. Conclusions. The data analyzed and summarized in the article can be useful for solving issues of taxonomy, phylogeny, genetics and practical problems of selection of plants of Iris genus; in addition, they will contribute to a more appropriate use of the Iris raw material in practical pharmacy.

A multi-layer functional genomic analysis to understand noncoding genetic variation in lipids

A major challenge of genome-wide association studies (GWAS) is to translate phenotypic associations into biological insights. Here, we integrate a large GWAS on blood lipids involving 1.6 million individuals from five ancestries with a wide array of functional genomic datasets to discover regulatory mechanisms underlying lipid associations. We first prioritize lipid-associated genes with expression quantitative trait locus (eQTL) colocalizations, and then add chromatin interaction data to narrow the search for functional genes. Polygenic enrichment analysis across 697 annotations from a host of tissues and cell types confirms the central role of the liver in lipid levels, and highlights the selective enrichment of adipose-specific chromatin marks in high-density lipoprotein cholesterol and triglycerides. Overlapping transcription factor (TF) binding sites with lipid-associated loci identifies TFs relevant in lipid biology. In addition, we present an integrative framework to prioritize causal variants at GWAS loci, producing a comprehensive list of candidate causal genes and variants with multiple layers of functional evidence. Two prioritized genes, CREBRF and RRBP1, show convergent evidence across functional datasets supporting their roles in lipid biology.

Rationally designed mariner vectors for functional genomic analysis of Actinobacillus pleuropneumoniae and other Pasteurellaceae species by transposon-directed insertion-site sequencing (TraDIS)

Comprehensive identification of conditionally essential genes requires efficient tools for generating high-density transposon libraries that, ideally, can be analysed using next-generation sequencing methods such as Transposon Directed Insertion-site Sequencing (TraDIS). The Himar1 (mariner) transposon is ideal for generating near-saturating mutant libraries, especially in AT-rich chromosomes, as the requirement for integration is a TA dinucleotide, and this transposon has been used for mutagenesis of a wide variety of bacteria. However, plasmids for mariner delivery do not necessarily work well in all bacteria. In particular, there are limited tools for functional genomic analysis of Pasteurellaceae species of major veterinary importance, such as swine and cattle pathogens, Actinobacillus pleuropneumoniae and Pasteurella multocida, respectively. Here, we developed plasmids, pTsodCPC9 and pTlacPC9 (differing only in the promoter driving expression of the transposase gene), that allow delivery of mariner into both these pathogens, but which should also be applicable to a wider range of bacteria. Using the pTlacPC9 vector, we have generated, for the first time, saturating mariner mutant libraries in both A. pleuropneumoniae and P. multocida that showed a near random distribution of insertions around the respective chromosomes as detected by TraDIS. A preliminary screen of 5000 mutants each identified 8 and 14 genes, respectively, that are required for growth under anaerobic conditions. Future high-throughput screening of the generated libraries will facilitate identification of mutants required for growth under different conditions, including in vivo, highlighting key virulence factors and pathways that can be exploited for development of novel therapeutics and vaccines.

Olorofim and the azoles are antagonistic in A. fumigatus and functional genomic screens reveal mechanisms of cross resistance.

Aspergillosis, in its various manifestations, is a major cause of morbidity and mortality. Very few classes of antifungal have been approved for clinical use to treat these diseases and resistance to the first line therapeutics is increasing. A new class of antifungals, the orotomides, are currently in development with the first compound in this class olorofim in late-stage clinical trials. In this study, we characterise a network of genes that govern olorofim response in A. fumigatus. We reveal that the number of transcription factors that regulate olorofim susceptibility are far fewer than we have previously observed for the azoles and the change in sensitivity observed in these isolates is less extreme. Intriguingly, loss of function in two higher order transcriptional regulators, HapB a member of the heterotrimeric HapB/C/E (CBC) complex or the regulator of nitrogen metabolic genes AreA, leads to cross resistance to both the azoles and olorofim. However, a clinical azole resistant isolate with a point mutation in HapE (hapEP88L) retains sensitivity to olorofim. Our transcriptomic analysis suggests that altered sensitivity to olorofim may emerge via modification of genes involved in the production of pyrimidine biosynthetic precursors. Finally, we also show that the action of the azoles are antagonistic to olorofim in vitro.

Olorofim and the azoles are antagonistic in Aspergillus fumigatus and functional genomic screens reveal mechanisms of cross resistance.

Aspergillosis, in its various manifestations, is a major cause of morbidity and mortality. Very few classes of antifungal have been approved for clinical use to treat these diseases and resistance to the first line therapeutics is increasing. A new class of antifungals, the orotomides, are currently in development with the first compound in this class olorofim in late-stage clinical trials. In this study, we characterise a network of genes that govern olorofim response in A. fumigatus. We reveal that the number of transcription factors that regulate olorofim susceptibility are far fewer than we have previously observed for the azoles and the change in sensitivity observed in these isolates is less extreme. Intriguingly, loss of function in two higher order transcriptional regulators, HapB a member of the heterotrimeric HapB/C/E (CBC) complex or the regulator of nitrogen metabolic genes AreA, leads to cross resistance to both the azoles and olorofim. However, a clinical azole resistant isolate with a point mutation in HapE (hapEP88L) retains sensitivity to olorofim. Our transcriptomic analysis suggests that altered sensitivity to olorofim may emerge via modification of genes involved in the production of pyrimidine biosynthetic precursors. Finally, we also show that the action of the azoles are antagonistic to olorofim in vitro.