CRISPR Correction of the GBA Mutation in Human-Induced Pluripotent Stem Cells Restores Normal Function to Gaucher Macrophages and Increases Their Susceptibility to Mycobacterium tuberculosis

Ramalingam, S and Kumar, A and Krug, S and Mohan, H and Rao, DN and Bishai, WR and Chandrasegaran, S (2023) CRISPR Correction of the GBA Mutation in Human-Induced Pluripotent Stem Cells Restores Normal Function to Gaucher Macrophages and Increases Their Susceptibility to Mycobacterium tuberculosis. [Preprint]

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Abstract

Gaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the β-glucocerebrosidase (GCase) GBA gene, which result in macrophage dysfunction. CRISPR (clustered regularly interspaced short palindromic repeats) editing of the homozygous L444P (1448T�C) GBA mutation in type 2 GD (GBA-/-) human-induced pluripotent stem cells (hiPSCs) yielded both heterozygous (GBA+/-) and homozygous (GBA+/+) isogenic lines. Macrophages derived from GBA-/-, GBA+/- and GBA+/+ hiPSCs showed that GBA mutation correction restores normal macrophage functions: GCase activity, motility, and phagocytosis. Furthermore, infection of GBA-/-, GBA+/- and GBA+/+ macrophages with the Mycobacterium tuberculosis H37Rv strain showed that impaired mobility and phagocytic activity were correlated with reduced levels of bacterial engulfment and replication suggesting that GD may be protective against tuberculosis. © 2023 The Author(s). Published by Oxford University Press on behalf of Infectious Diseases Society of America. All rights reserved.

Item Type:	Preprint
Publication:	Journal of Infectious Diseases
Publisher:	Oxford University Press
Additional Information:	The copyright for this article belongs to authors.
Keywords:	gamma interferon; glucosylceramidase; interleukin 1beta; interleukin 6; tumor necrosis factor; glucosylceramidase, Article; bacterial growth; cell activation; cell migration; cell motility; clustered regularly interspaced short palindromic repeat; colony forming unit; controlled study; CRISPR-CAS9 system; fibroblast; Gaucher disease; Gaucher disease type 2; gene mutation; genetic susceptibility; genotype; human; human cell; immune response; induced pluripotent stem cell; karyotyping; lysosome storage disease; macrophage function; monocyte; Mycobacterium tuberculosis; necrosis; nonhuman; phagocytosis; phenotype; tuberculosis; clustered regularly interspaced short palindromic repeat; genetics; macrophage; metabolism; mutation, Clustered Regularly Interspaced Short Palindromic Repeats; Gaucher Disease; Glucosylceramidase; Humans; Induced Pluripotent Stem Cells; Macrophages; Mutation; Mycobacterium tuberculosis
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	14 Dec 2024 00:20
Last Modified:	14 Dec 2024 00:20
URI:	http://eprints.iisc.ac.in/id/eprint/85429

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