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Next Generation Sequencing Analysis of human hematopoietic stem/progenitor cells (HSPCs) differentiated from pluripotent stem cells

ABSTRACT: Next Generation Sequencing Analysis of human hematopoietic stem/progenitor cells (HSPCs) differentiated from pluripotent stem cells

PROVIDER: PRJNA464429 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR7130980_1.fastq.gz	Fastqsanger.gz
	SRR7130980_2.fastq.gz	Fastqsanger.gz
	SRR7130990_1.fastq.gz	Fastqsanger.gz
	SRR7130990_2.fastq.gz	Fastqsanger.gz
	SRR7131000_1.fastq.gz	Fastqsanger.gz

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Next Generation Sequencing Analysis of human hematopoietic stem/progenitor cells (HSPCs) differentiated from pluripotent stem cells

Project description:Purpose: global gene expression profiling of H1 and iPSC derived HSPCs. Methods: SMART-seq2 amplified polyA RNA from differentiation day 8 CD31+CD34+CD43+ and CD31+CD34+CD43- cells, and day 5 CD31+CD34+ cells cultured in medium with or without insulin supplement Results: Modulation of key cell signaling pathways lead to highly efficient HSPC differentiation from human pluripotent stem cells. Conclusions: Highly efficient HSPC differentiation from human pluripotent stem cells in a monolayer, chemically defined culture system.

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Project description:Haematopoietic stem and progenitor cells (HSPCs) in the foetus and adult possess distinct molecular landscapes that regulate cell fate and change their susceptibility to initiation and progression of haematopoietic malignancies. The proteomic programs that govern these differences remain elusive. In this study, we have utilized a mass spectrometry-based quantitative proteomics approach to comprehensively describe and compare the proteome of foetal and adult HSPCs. We found that the proteome of foetal HSPCs is relatively simple, characterized by proteins involved in cell cycle and cell proliferation, while their adult counterparts are defined by a larger set of proteins that are involved in more diverse cellular processes. These adult characteristics include an arsenal of proteins important for viral and bacterial defence, as well as protection against ROS-induced protein oxidation. Our further analyses of Type I interferon signalling shows that foetal HSPCs are sensitive to Interferon a (IFNa), which impairs their production of mature lymphoid cells, whereas stimulation with IFNa to the pregnant mother enhances the production of early progenitors from foetal HSCs. Our results provide new and important insights into the molecular landscape of foetal and adult haematopoiesis that advance our understanding of normal and malignant haematopoiesis during foetal and adult life.

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Project description:Purpose: Illumina next-generation sequencing (NGS) has been used to interrogate the transcriptome profiling (bulk RNA-seq) of primary human HSPCs in the presence and absence of RIOK2. Primary human hematopoietic stem and progenitor cells (HSPCs) isolated from 3 different donors were genome edited to obtain knockdown (KD) and knockout (KO) of RIOK2. The genome edited HSPCs were then differentiated for 48 hours and their total RNA was isolated to perform cDNA synthesis and bulk RNA sequencing. The overall goal of this study was to investigate the global alterations in gene expressions with dose-dependent loss of RIOK2 in primary human HSPCs, that would expand our understanding of RIOK2-dependent transcriptomic changes involved in hematopoietic differentiation.

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