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Using CRISPR genome screens and dual transcriptome analyses to dissect host-pathogen interactions in tuberculosis

Periodic Reporting for period 1 - Host-TB (Using CRISPR genome screens and dual transcriptome analyses to dissect host-pathogen interactions in tuberculosis)

Reporting period: 2019-06-01 to 2020-11-30

With more than 10 million cases annually, tuberculosis (TB) remains a global health problem. TB epidemic is exacerbated by the spread of multidrug-resistant TB. Host-directed therapies (HDTs) can improve immune mechanisms by augmenting the ability of host cells to kill M. tuberculosis (Mtb) or by modulating the immune response to prevent excessive inflammation, cell death and tissue damage. Progress with HDT development has been slowed down by the limited understanding of host-pathogen interactions during Mtb infection. Screens of the whole human genome can identify novel genes involved in the immune responses to Mtb infection and susceptibility to TB. Previously, we successfully used genome-wide association studies to identify human genes associated with susceptibility to TB. Here, we will for the first time use the groundbreaking CRISPR technology to screen the human genome in macrophages infected with Mtb and discover genes that are critically involved in host-pathogen interactions. Then, we will comprehensively characterise pathways that mediate impacts of these genes on both the human macrophage and the intracellular Mtb bacilli using dual transcriptome analyses and high-throughput microscopy assays. This novel approach will dissect crucial mechanisms of host-pathogen interaction during Mtb infection and will point to new targets for HDTs of TB.
We performed first screens of human genome in CRISPR-edited macrophages after their infection with M. tuberculosis. These screens revealed several genes that affect viability of human macrophages during infection.
Novel human genes that affect susceptibility of macrophages to M. tuberculosis infection have been identified using CRISPR-Cas9 screens of the human genome. These genes will be validated in the follow-up experiments. Their interaction with virulence factors of M. tuberculosis will be also studied. Proteins encoded by these genes may become novel targets for host-directed therapy of tuberculosis.
Human macrophages engulfing mycobacteria