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Joint profiling of chromatin accessibility and CAR-T integration site analysis at population and single-cell levels.

Proceedings of the National Academy of Sciences of the United States of America | 2020

Chimeric antigen receptor (CAR)-T immunotherapy has yielded impressive results in several B cell malignancies, establishing itself as a powerful means to redirect the natural properties of T lymphocytes. In this strategy, the T cell genome is modified by the integration of lentiviral vectors encoding CAR that direct tumor cell killing. However, this therapeutic approach is often limited by the extent of CAR-T cell expansion in vivo. A major outstanding question is whether or not CAR-T integration itself enhances the proliferative competence of individual T cells by rewiring their regulatory landscape. To address this question, it is critical to define the identity of an individual CAR-T cell and simultaneously chart where the CAR-T vector integrates into the genome. Here, we report the development of a method called EpiVIA (https://github.com/VahediLab/epiVIA) for the joint profiling of the chromatin accessibility and lentiviral integration site analysis at the population and single-cell levels. We validate our technique in clonal cells with previously defined integration sites and further demonstrate the ability to measure lentiviral integration sites and chromatin accessibility of host and viral genomes at the single-cell resolution in CAR-T cells. We anticipate that EpiVIA will enable the single-cell deconstruction of gene regulation during CAR-T therapy, leading to the discovery of cellular factors associated with durable treatment.

Pubmed ID: 32094195 RIS Download

Associated grants

  • Agency: NCI NIH HHS, United States
    Id: P01 CA214278
  • Agency: NCI NIH HHS, United States
    Id: R01 CA241762
  • Agency: NHLBI NIH HHS, United States
    Id: R01 HL145754
  • Agency: NIA NIH HHS, United States
    Id: U01 AG066100

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


ChIP-seq (tool)

RRID:SCR_001237

Set of software modules for performing common ChIP-seq data analysis tasks across the whole genome, including positional correlation analysis, peak detection, and genome partitioning into signal-rich and signal-poor regions. The tools are designed to be simple, fast and highly modular. Each program carries out a well defined data processing procedure that can potentially fit into a pipeline framework. ChIP-Seq is also freely available on a Web interface.

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SAMTOOLS (tool)

RRID:SCR_002105

Original SAMTOOLS package has been split into three separate repositories including Samtools, BCFtools and HTSlib. Samtools for manipulating next generation sequencing data used for reading, writing, editing, indexing,viewing nucleotide alignments in SAM,BAM,CRAM format. BCFtools used for reading, writing BCF2,VCF, gVCF files and calling, filtering, summarising SNP and short indel sequence variants. HTSlib used for reading, writing high throughput sequencing data.

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NCBI Epigenomics (tool)

RRID:SCR_006151

THIS RESOURCE IS NO LONGER IN SERVICE, documented on January 19, 2022.

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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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GenoList genome browser (tool)

RRID:SCR_007683

GenoList is a resource for microbial genomes. It provides links to databases that give information on specific microbes. A link to the Genolist itself also incorporates many microbial genomes and search and analysis tools. Of the links provided on the GenoList homepage, species included are: Bacillus subtilis Escherichia coli Mycobacterium tuberculosis Mycobacterium leprae Mycobacterium bovis Mycobacterium ulcerans Listeria monocytogenes Legionella pneumophila Helicobacter pylori Mycoplasma pulmonis Synechocystis Anabaena Staphylococcus aureus Streptococcus pneumoniae Streptococcus agalactiae Photorhabdus luminescens Candida albicans Bacillus subtilis Escherichia coli Mycobacterium tuberculosis Mycobacterium leprae Mycobacterium bovis Mycobacterium ulcerans Listeria monocytogenes Legionella pneumophila Helicobacter pylori Mycoplasma pulmonis Synechocystis Anabaena Staphylococcus aureus Streptococcus pneumoniae Streptococcus agalactiae Photorhabdus luminescens Candida albicans

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MatPlotLib (tool)

RRID:SCR_008624

Python 2D plotting library which produces publication quality figures in variety of hardcopy formats and interactive environments across platforms. Used in python scripts, web application servers, and six graphical user interface toolkits. Used to generate plots, histograms, power spectra, bar charts, error charts, scatter plots.

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Homer2 (tool)

RRID:SCR_009586

Software matlab scripts used for analyzing fNIRS data to obtain estimates and maps of brain activation. Graphical user interface (GUI) for visualization and analysis of functional near-infrared spectroscopy (fNIRS) data.

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Circos (tool)

RRID:SCR_011798

A software package for visualizing data and information. It visualizes data in a circular layout - this makes Circos ideal for exploring relationships between objects or positions.

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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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Atac (tool)

RRID:SCR_015980

Alignment analysis software tool for comparative mapping between two genome assemblies or between two different genomes. It can cache intermediate results to speed a comparisons of multiple sequences.

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liftOver (tool)

RRID:SCR_018160

Web tool to convert genome coordinates and genome annotation files between assemblies. Used to translate genomic coordinates from one assembly version into another and retrieves putative orthologous regions in other species using UCSC chained and netted alignments.

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HEK293T (tool)

RRID:CVCL_0063

Cell line HEK293T is a Transformed cell line with a species of origin Homo sapiens (Human)

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