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Gene expression analysis of human sickle cell disease (SCD) and healthy donor (HD) CD34+ hematopoietic stem/progenitor cells (HSPCs) treated with CRISPR/Cas9 targeting the HBG promoters or untreated samples

ABSTRACT: Gene expression analysis of human sickle cell disease (SCD) and healthy donor (HD) CD34+ hematopoietic stem/progenitor cells (HSPCs) treated with CRISPR/Cas9 targeting the HBG promoters or untreated samples

PROVIDER: PRJNA1045560 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR26955600_1.fastq.gz	Fastqsanger.gz
	SRR26955600_2.fastq.gz	Fastqsanger.gz
	SRR26955601_1.fastq.gz	Fastqsanger.gz
	SRR26955601_2.fastq.gz	Fastqsanger.gz
	SRR26955602_1.fastq.gz	Fastqsanger.gz

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Project description:Pregnancy in Sickle Cell Disease (SCD) women is associated to increased risk of clinical and obstetrical complications. Placentas from SCD pregnancies can present increased abnormal findings, which may lead to placental insufficiency, favoring adverse perinatal outcome. These placental abnormalities are well known and reported, however little is known about the molecular mechanisms, such as epigenetics. Thus, our aim was to evaluate the DNA methylation profile in placentas from women with SCD (HbSS and HbSC genotypes), compared to uncomplicated controls (HbAA). We included in this study 11 pregnant women with HbSS, 11 with HbSC and 21 with HbAA genotypes. Illumina Methylation EPIC BeadChip was used to assess the whole placental DNA methylation. Pyrosequencing was used for array data validation and qRT-PCR was applied for gene expression analysis. Our results showed high frequency of hypermethylated CpGs sites in HbSS and HbSC groups with 73.5% and 76.2% respectively, when compared with the control group. Differentially methylated regions (DMRs) also showed an increased hypermethylation status for the HbSS (89%) and HbSC (86%) groups, when compared with the control group methylation data. DMRs were selected for methylation validation (4 DMRs-HbSS and 3 DMRs the HbSC groups) and after analyses three were validated in the HbSS group, and none in the HbSC group. The gene expression analysis showed differential expression for the PTGFR (-2.97-fold) and GPR56 (3.0-fold) genes in the HbSS group, and for the SPOCK1 (-2.40-fold) and ADCY4 (1.80-fold) genes in the HbSC group. Taken together, these data strongly suggest that SCD (HbSS and HbSC genotypes) can alter placental DNA methylation and lead to gene expression changes. These changes possibly contribute to abnormal placental development and could impact in the clinical course, especially for the fetus, possibly leading to increased risk of abortion, fetal growth restriction (FGR), stillbirth, small for gestational age newborns and prematurity.

Dataset Information

Gene expression analysis of human sickle cell disease (SCD) and healthy donor (HD) CD34+ hematopoietic stem/progenitor cells (HSPCs) treated with CRISPR/Cas9 targeting the HBG promoters or untreated samples

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