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Engineering an inhibitor resistant human CSF1R variant for microglia replacement [InVivo_RNA]

ABSTRACT: Engineering an inhibitor resistant human CSF1R variant for microglia replacement [InVivo_RNA]

PROVIDER: PRJNA909243 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR22548600_1.fastq.gz	Fastqsanger.gz
	SRR22548600_2.fastq.gz	Fastqsanger.gz
	SRR22548601_1.fastq.gz	Fastqsanger.gz
	SRR22548601_2.fastq.gz	Fastqsanger.gz
	SRR22548602_1.fastq.gz	Fastqsanger.gz

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Engineering an inhibitor resistant human CSF1R variant for microglia replacement

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Engineering an inhibitor resistant human CSF1R variant for microglia replacement [InVitro_RNA]

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Engineering an inhibitor resistant human CSF1R variant for microglia replacement [InVitro_RNA]

Project description:iPSC-derived microglia expressing the wildtype, G795A, or G795C CSF1R variants were generated in vitro and treated with either DMSO or Plexxicon 5622 in order to compare the gene expression differences exising between genotypes at baseline or following CSF1R inhibtion.

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Engineering an inhibitor resistant human CSF1R variant for microglia replacement [InVivo_RNA]

Project description:iPSC-derived hematopoietic progenitor cells expressing either the wildtype or G795A CSF1R variant were xenotransplanted into neonatal mice and subsequently aged for two months to compare in vivo gene expression differences between the two CSF1R genotypes.

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Engineering an inhibitor-resistant human CSF1R variant for microglia replacement.

Project description:Hematopoietic stem cell transplantation (HSCT) can replace endogenous microglia with circulation-derived macrophages but has high mortality. To mitigate the risks of HSCT and expand the potential for microglia replacement, we engineered an inhibitor-resistant CSF1R that enables robust microglia replacement. A glycine to alanine substitution at position 795 of human CSF1R (G795A) confers resistance to multiple CSF1R inhibitors, including PLX3397 and PLX5622. Biochemical and cell-based assays show no discernable gain or loss of function. G795A- but not wildtype-CSF1R expressing macrophages efficiently engraft the brain of PLX3397-treated mice and persist after cessation of inhibitor treatment. To gauge translational potential, we CRISPR engineered human-induced pluripotent stem cell-derived microglia (iMG) to express G795A. Xenotransplantation studies demonstrate that G795A-iMG exhibit nearly identical gene expression to wildtype iMG, respond to inflammatory stimuli, and progressively expand in the presence of PLX3397, replacing endogenous microglia to fully occupy the brain. In sum, we engineered a human CSF1R variant that enables nontoxic, cell type, and tissue-specific replacement of microglia.

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Engineering an inhibitor resistant human CSF1R variant for microglia replacement

Project description:This SuperSeries is composed of the SubSeries listed below.

2022-12-21 | GSE220215 | GEO

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Project description:Microglia are important immune cells in the central nervous system (CNS). Dysfunctions of gene-deficient microglia contribute to the development and progression of multiple CNS diseases. Microglia replacement by nonself cells has been proposed to treat microglia associated disorders. However, most of attempts are failed due to low replacement efficiencies, such as with the traditional bone marrow transplantation approach. In this study, we develop three efficient strategies for microglia replacement: microglia replacement by bone marrow transplantation (mrBMT), microglia replacement by peripheral blood (mrPB) and microglia replacement by microglia transplantation (mrMT). mrBMT and mrPB allow microglia-like cells to efficiently replace resident microglia in the whole CNS. On the other hand, mrMT achieves microglia replacement in brain regions of interest. In summary, the present study offers effective tactics for microglia replacement with diverse application scenarios, which potentially opens up a window on treating microglia-associated CNS disorders.

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