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Shotgun transcriptome, spatial omics, and isothermal profiling of SARS-CoV-2 infection reveals unique host responses, viral diversification, and drug interactions.

Daniel Butler | Christopher Mozsary | Cem Meydan | Jonathan Foox | Joel Rosiene | Alon Shaiber | David Danko | Ebrahim Afshinnekoo | Matthew MacKay | Fritz J Sedlazeck | Nikolay A Ivanov | Maria Sierra | Diana Pohle | Michael Zietz | Undina Gisladottir | Vijendra Ramlall | Evan T Sholle | Edward J Schenck | Craig D Westover | Ciaran Hassan | Krista Ryon | Benjamin Young | Chandrima Bhattacharya | Dianna L Ng | Andrea C Granados | Yale A Santos | Venice Servellita | Scot Federman | Phyllis Ruggiero | Arkarachai Fungtammasan | Chen-Shan Chin | Nathaniel M Pearson | Bradley W Langhorst | Nathan A Tanner | Youngmi Kim | Jason W Reeves | Tyler D Hether | Sarah E Warren | Michael Bailey | Justyna Gawrys | Dmitry Meleshko | Dong Xu | Mara Couto-Rodriguez | Dorottya Nagy-Szakal | Joseph Barrows | Heather Wells | Niamh B O'Hara | Jeffrey A Rosenfeld | Ying Chen | Peter A D Steel | Amos J Shemesh | Jenny Xiang | Jean Thierry-Mieg | Danielle Thierry-Mieg | Angelika Iftner | Daniela Bezdan | Elizabeth Sanchez | Thomas R Campion | John Sipley | Lin Cong | Arryn Craney | Priya Velu | Ari M Melnick | Sagi Shapira | Iman Hajirasouliha | Alain Borczuk | Thomas Iftner | Mirella Salvatore | Massimo Loda | Lars F Westblade | Melissa Cushing | Shixiu Wu | Shawn Levy | Charles Chiu | Robert E Schwartz | Nicholas Tatonetti | Hanna Rennert | Marcin Imielinski | Christopher E Mason
Nature communications | 2021

In less than nine months, the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) killed over a million people, including >25,000 in New York City (NYC) alone. The COVID-19 pandemic caused by SARS-CoV-2 highlights clinical needs to detect infection, track strain evolution, and identify biomarkers of disease course. To address these challenges, we designed a fast (30-minute) colorimetric test (LAMP) for SARS-CoV-2 infection from naso/oropharyngeal swabs and a large-scale shotgun metatranscriptomics platform (total-RNA-seq) for host, viral, and microbial profiling. We applied these methods to clinical specimens gathered from 669 patients in New York City during the first two months of the outbreak, yielding a broad molecular portrait of the emerging COVID-19 disease. We find significant enrichment of a NYC-distinctive clade of the virus (20C), as well as host responses in interferon, ACE, hematological, and olfaction pathways. In addition, we use 50,821 patient records to find that renin-angiotensin-aldosterone system inhibitors have a protective effect for severe COVID-19 outcomes, unlike similar drugs. Finally, spatial transcriptomic data from COVID-19 patient autopsy tissues reveal distinct ACE2 expression loci, with macrophage and neutrophil infiltration in the lungs. These findings can inform public health and may help develop and drive SARS-CoV-2 diagnostic, prevention, and treatment strategies.

Pubmed ID: 33712587 RIS Download

Associated grants

  • Agency: NIMH NIH HHS, United States
    Id: R01 MH117406
  • Agency: NLM NIH HHS, United States
    Id: T15 LM007079
  • Agency: NCATS NIH HHS, United States
    Id: TL1 TR002386
  • Agency: NIAID NIH HHS, United States
    Id: U19 AI144297
  • Agency: NIGMS NIH HHS, United States
    Id: R35 GM138152
  • Agency: NCATS NIH HHS, United States
    Id: UL1 TR002384
  • Agency: NIDDK NIH HHS, United States
    Id: R01 DK121072
  • Agency: NIGMS NIH HHS, United States
    Id: R35 GM131905
  • Agency: NCATS NIH HHS, United States
    Id: UL1 TR000457
  • Agency: NIBIB NIH HHS, United States
    Id: R25 EB020393
  • Agency: NIAID NIH HHS, United States
    Id: R01 AI151059

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This is a list of tools and resources that we have found mentioned in this publication.


LAMP (tool)

RRID:SCR_001740

Software for linkage and association modeling in pedigrees that uses a maximum likelihood model to extract information on genetic linkage and association from samples of unrelated individuals, sib pairs, trios and larger pedigrees (Li et al, 2005; Li et al, 2006). It provides estimates of genetic model parameters and powerful tests of association in settings where population stratification is not a concern.

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MUlti SImulation Coordinator (tool)

RRID:SCR_001756

Software that allows large scale neuron simulators to communicate during runtime. It allows exchange of data among parallel applications in a cluster environment, interconnects large-scale neuronal network simulators with each other or with other tools, participates in multi-simulations, and is continuously developed and extended. Three simulators currently have MUSIC interfaces: Moose, NEURON and NEST. Three applications execute in parallel while exchanging data via MUSIC. The software interface promotes interoperability by allowing models written for different simulators to be simulated together in a larger system. It enables re-usability of models or tools by providing a standard interface. As data are distributed over a number of processors, it is non-trivial to coordinate data transfer so that it reaches the correct destination at the correct time. Current and future simulators can make use of MUSIC - compliant general purpose tools and participate in multi-simulations, for example when: * Different parts of a complex nervous system model are optimally implemented in different simulators, and need to communicate with each other. * Post-processing of generated data is needed, where the amounts of data are too large for intermediate storage, and requires the simulator to pass the data directly to the post-processing module. A standard interface enables straight-forward independent third-party development and community sharing of interoperable software tools for parallel processing. * Library and utilities are written in C++, uses MPI. * It is possible to add a MUSIC interface to existing simulators. * Works independently, no assumptions are made about other applications to facilitate development of general purpose tools. * Performance Data transport with high bandwidth and low latency.

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Ensembl (tool)

RRID:SCR_002344

Collection of genome databases for vertebrates and other eukaryotic species with DNA and protein sequence search capabilities. Used to automatically annotate genome, integrate this annotation with other available biological data and make data publicly available via web. Ensembl tools include BLAST, BLAT, BioMart and the Variant Effect Predictor (VEP) for all supported species.

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scikit-learn (tool)

RRID:SCR_002577

scikit-learn: machine learning in Python

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GenBank (tool)

RRID:SCR_002760

NIH genetic sequence database that provides annotated collection of all publicly available DNA sequences for almost 280 000 formally described species (Jan 2014) .These sequences are obtained primarily through submissions from individual laboratories and batch submissions from large-scale sequencing projects, including whole-genome shotgun (WGS) and environmental sampling projects. Most submissions are made using web-based BankIt or standalone Sequin programs, and GenBank staff assigns accession numbers upon data receipt. It is part of International Nucleotide Sequence Database Collaboration and daily data exchange with European Nucleotide Archive (ENA) and DNA Data Bank of Japan (DDBJ) ensures worldwide coverage. GenBank is accessible through NCBI Entrez retrieval system, which integrates data from major DNA and protein sequence databases along with taxonomy, genome, mapping, protein structure and domain information, and biomedical journal literature via PubMed. BLAST provides sequence similarity searches of GenBank and other sequence databases. Complete bimonthly releases and daily updates of GenBank database are available by FTP.

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Gene Set Enrichment Analysis (tool)

RRID:SCR_003199

Software package for interpreting gene expression data. Used for interpretation of a large-scale experiment by identifying pathways and processes.

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RefSeq (tool)

RRID:SCR_003496

Collection of curated, non-redundant genomic DNA, transcript RNA, and protein sequences produced by NCBI. Provides a reference for genome annotation, gene identification and characterization, mutation and polymorphism analysis, expression studies, and comparative analyses. Accessed through the Nucleotide and Protein databases.

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STAR (tool)

RRID:SCR_004463

Software performing alignment of high-throughput RNA-seq data. Aligns RNA-seq reads to reference genome using uncompressed suffix arrays.

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RSeQC (tool)

RRID:SCR_005275

Software package to comprehensively evaluate different aspects of RNA-seq experiments, such as sequence quality, GC bias, polymerase chain reaction bias, nucleotide composition bias, sequencing depth, strand specificity, coverage uniformity and read distribution over the genome structure. RSeQC takes both SAM and BAM files as input, which can be produced by most RNA-seq mapping tools as well as BED files, which are widely used for gene models.

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Bioconductor (tool)

RRID:SCR_006442

Software repository for R packages related to analysis and comprehension of high throughput genomic data. Uses separate set of commands for installation of packages. Software project based on R programming language that provides tools for analysis and comprehension of high throughput genomic data.

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Promega (tool)

RRID:SCR_006724

An Antibody supplier

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QIAGEN (tool)

RRID:SCR_008539

A commercial organization which provides assay technologies to isolate DNA, RNA, and proteins from any biological sample. Assay technologies are then used to make specific target biomolecules, such as the DNA of a specific virus, visible for subsequent analysis.

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Ingenuity Pathway Analysis (tool)

RRID:SCR_008653

A web-based software application that enables users to analyze, integrate, and understand data derived from gene expression, microRNA, and SNP microarrays, metabolomics, proteomics, and RNA-Seq experiments, and small-scale experiments that generate gene and chemical lists. Users can search for targeted information on genes, proteins, chemicals, and drugs, and build interactive models of experimental systems. IPA allows exploration of molecular, chemical, gene, protein and miRNA interactions, creation of custom molecular pathways, and the ability to view and modify metabolic, signaling, and toxicological canonical pathways. In addition to the networks and pathways that can be created, IPA can provide multiple layering of additional information, such as drugs, disease genes, expression data, cellular functions and processes, or a researchers own genes or chemicals of interest.

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Subread (tool)

RRID:SCR_009803

Software package for high-performance read alignment, quantification and mutation discovery.General purpose read aligner which can be used to map both genomic DNA-seq reads and RNA-seq reads. Subread aligner as fast, accurate and scalable read mapping by seed-and-vote.These programs were also implemented in Bioconductor R package Rsubread.

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edgeR (tool)

RRID:SCR_012802

Bioconductor software package for Empirical analysis of Digital Gene Expression data in R. Used for differential expression analysis of RNA-seq and digital gene expression data with biological replication.

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Abcam (tool)

RRID:SCR_012931

A commercial antibody supplier which supplies primary and secondary antibodies, biochemicals, proteins, peptides, lysates, immunoassays and other kits.

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New England Biolabs (tool)

RRID:SCR_013517

An Antibody supplier

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FastQC (tool)

RRID:SCR_014583

Quality control software that perform checks on raw sequence data coming from high throughput sequencing pipelines. This software also provides a modular set of analyses which can give a quick impression of the quality of the data prior to further analysis.

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ggplot2 (tool)

RRID:SCR_014601

Open source software package for statistical programming language R to create plots based on grammar of graphics. Used for data visualization to break up graphs into semantic components such as scales and layers.

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FactoMineR (tool)

RRID:SCR_014602

Software R package for multivariate analysis which takes into account different types of data structure. Data can be organized in groups of variable, groups of individuals, or into hierarchy of variables.

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GENCODE (tool)

RRID:SCR_014966

Human and mouse genome annotation project which aims to identify all gene features in the human genome using computational analysis, manual annotation, and experimental validation.

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MultiQC (tool)

RRID:SCR_014982

Data aggregate that compiles results from bioinformatics analyses across multiple samples into a single report. It is written in Python.

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DESeq2 (tool)

RRID:SCR_015687

Software package for differential gene expression analysis based on the negative binomial distribution. Used for analyzing RNA-seq data for differential analysis of count data, using shrinkage estimation for dispersions and fold changes to improve stability and interpretability of estimates.

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rsamtools (tool)

RRID:SCR_024257

Software R package provides interface to the 'samtools', 'bcftools', and 'tabix' utilities for manipulating Sequence Alignment Map, FASTA, binary variant call and compressed indexed tab-delimited files.

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