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Serine-threonine Kinase, ROCK2, Regulates Germinal Center B Cell Positioning and Cholesterol Biosynthesis

ABSTRACT: Serine-threonine Kinase, ROCK2, Regulates Germinal Center B Cell Positioning and Cholesterol Biosynthesis

PROVIDER: PRJNA593194 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10575201_1.fastq.gz	Fastqsanger.gz
	SRR10575201_2.fastq.gz	Fastqsanger.gz
	SRR10575202_1.fastq.gz	Fastqsanger.gz
	SRR10575202_2.fastq.gz	Fastqsanger.gz
	SRR10575203_1.fastq.gz	Fastqsanger.gz

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Serine-threonine Kinase, ROCK2, Regulates Germinal Center B Cell Positioning and Cholesterol Biosynthesis

Project description:Splenic follicular B cells (FoB; B220+GL7-CD38hiCD23+) and Germinal Center B cells (GCB; B220+GL7+CD38lo) were FACS sorted from WT or CD23-Cre.Rock2 mice after 7d post-immunization with NP-CGG. RNA-seq was then employed to compare the transcriptome of GCBs from the indicated mice. Here we show that loss of ROCK2 leads to altered gene expression in GCBs. We also employed ATAC-seq analysis and found that differentially expressed genes from the ROCK2-deficient GC B cells contained loci that were enriched for IRF binding sites.

2020-05-22 | GSE141328 | GEO

Serine-threonine kinase ROCK2 regulates germinal center B cell positioning and cholesterol biosynthesis.

Project description:Germinal center (GC) responses require B cells to respond to a dynamic set of intercellular and microenvironmental signals that instruct B cell positioning, differentiation, and metabolic reprogramming. RHO-associated coiled-coil-containing protein kinase 2 (ROCK2), a serine-threonine kinase that can be therapeutically targeted by ROCK inhibitors or statins, is a key downstream effector of RHOA GTPases. Although RHOA-mediated pathways are emerging as critical regulators of GC responses, the role of ROCK2 in B cells is unknown. Here, we found that ROCK2 was activated in response to key T cell signals like CD40 and IL-21 and that it regulated GC formation and maintenance. RNA-Seq analyses revealed that ROCK2 controlled a unique transcriptional program in GC B cells that promoted optimal GC polarization and cholesterol biosynthesis. ROCK2 regulated this program by restraining AKT activation and subsequently enhancing FOXO1 activity. ATAC-Seq (assay for transposase-accessible chromatin with high-throughput sequencing) and biochemical analyses revealed that the effects of ROCK2 on cholesterol biosynthesis were instead mediated via a novel mechanism. ROCK2 directly phosphorylated interferon regulatory factor 8 (IRF8), a crucial mediator of GC responses, and promoted its interaction with sterol regulatory element-binding transcription factor 2 (SREBP2) at key regulatory regions controlling the expression of cholesterol biosynthetic enzymes, resulting in optimal recruitment of SREBP2 at these sites. These findings thus uncover ROCK2 as a multifaceted and therapeutically targetable regulator of GC responses.

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