Browse
Submit Data
Databases
API
Help

Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

39 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

RNA sequencing comparing SCN CSF3R-d715 RUNX1-D171N (RHD) treated with the HDAC inhibitor MS275, octyl-(R)-2hydroxyglutarate (2HG) or solvent control (DMSO)

ABSTRACT: Effects of TET2 on repressing inflammation have been assigned to its methylcytosine dioxygenase activity, but also to its ability to recruit HDAC-mediated repressor activity to pro-inflammatory genes. To discriminate between the two above-proposed functions of TET2, we asked how inhibition of its enzymatic activity, by administration of octyl-(R)-2hydroxyglutarate (2HG), and its ability to recruit HDAC-activity, by administration of the HDAC-inhibitor MS275 (Entinostat), affected the transcriptional profile of CSF3R-d715 RUNX1-D171N (RHD) expressing SCN-iPSC derived CD34+CD45+ HPCs. HPCs were generated from CRISPR-Cas9 genome edited CSF3R-d715 SCN-derived iPSCs with the STEMdiff™ Hematopoietic Kit (STEMCELL Technologies). The RUNX1-D171N (RHD) mutation was lentivirally introduced to the hematopoietic induction cultures 60 hours before harvesting the floating cells. DMSO (solvent control), 0.1 mM Octyl-(R)-2HG (Sigma-Aldrich) or 2 µM MS275 (Santa Cruz) was added for 16 hours to the hematopoietic induction cultures before harvesting the floating cells. Floating cells were harvested at Day 12 of the hematopoietic induction protocol. CD34+CD45+ HPCs were FACS sorted in TRIzol and RNA was isolated according to the manufacturer’s protocol. SMARTer Ultra Low Input RNA kit for sequencing (Clontech, v4 Cat# 634891) was used to generate cDNA. Sequencing libraries were generated using TruSeq Nano DNA Sample Preparation kits (Illumina, Cat# 20015964), according to the low sample protocol and paired-end sequenced on a Novaseq 6000 (Illumina).

INSTRUMENT(S): Illumina NovaSeq 6000

ORGANISM(S): Homo sapiens

SUBMITTER: Patricia Olofsen

PROVIDER: E-MTAB-9374 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Json Xml

Similar Datasets

RNA sequencing comparing the effect of the introduction of RUNX1-D171N (RHD) in control- or SCN -CSF3R-d715 hematopoietic progenitor cells (HPCs)

Project description:To address how CSF3R and RUNX1 mutations affect hematopoiesis in a severe congenital neutropenia (SCN) background we used SCN patient, or control, derived induced pluripotent stem cells (iPSC). HPCs were generated from CRISPR-Cas9 genome edited CSF3R-d715 SCN or control-derived iPSCs with the STEMdiff™ Hematopoietic Kit (STEMCELL Technologies). The RUNX1-D171N (RHD) mutation, or an empty vector control, was lentivirally introduced to the hematopoietic induction cultures 60 hours before harvesting the floating cells. Floating cells were harvested at Day 12 of the hematopoietic induction protocol. CD34+CD45+ HPCs were FACS sorted in TRIzol and RNA was isolated according to the manufacturer’s protocol. SMARTer Ultra Low Input RNA kit for sequencing (Clontech, v4 Cat# 634891) was used to generate cDNA. Sequencing libraries were generated using TruSeq Nano DNA Sample Preparation kits (Illumina, Cat# 20015964), according to the low sample protocol and paired-end sequenced on a HiSeq 2500 or Novaseq 6000 (both Illumina).

2020-08-24 | E-MTAB-9375 | biostudies-arrayexpress

RNA sequencing comparing the effect of the introduction of CSF3R-d715 in control- or SCN- hematopoietic progenitor cells (HPCs)

Project description:To address how CSF3R mutations affect hematopoiesis in a severe congenital neutropenia (SCN) background we used SCN patient, or control, derived induced pluripotent stem cells (iPSC). HPCs were generated from CRISPR-Cas9 genome edited CSF3R-d715 SCN or control-derived iPSCs with the STEMdiff™ Hematopoietic Kit (STEMCELL Technologies). Floating cells were harvested at Day 12 of the hematopoietic induction protocol. CD34+CD45+ HPCs were FACS sorted in TRIzol and RNA was isolated according to the manufacturer’s protocol. SMARTer Ultra Low Input RNA kit for sequencing (Clontech, v4 Cat# 634891) was used to generate cDNA. Sequencing libraries were generated using TruSeq Nano DNA Sample Preparation kits (Illumina, Cat# 20015964), according to the low sample protocol and paired-end sequenced on a HiSeq 2500 or Novaseq 6000 (both Illumina).

2021-04-01 | E-MTAB-10288 | biostudies-arrayexpress

RNA sequencing comparing the effect of the introduction of RUNX1-D171N in Csf3r-d715 mice bone marrow cells

Project description:To address how Csf3r and RUNX1 mutations in combination with CSF3 administration affect hematopoiesis in vitro, we performed hematopoietic expansion cultures in LODISH stem cell expansion medium. Lineage-negative Csfr-d715 BM cell cells were retrovirally transduced with the patient specific RUNX1-D171N (RHD) mutant or an empty vector control and subsequently treated with puromycin to select for the transduced cells. Lineage- c-Kit+ (LK) cells were FACS sorted in TRIzol 2, 5 and 9 days after puromycin selection and subsequently used for RNA isolation according to the manufacturer’s protocol. SMARTer Ultra Low Input RNA kit for sequencing (Clontech, v4 Cat# 634891) was used to generate cDNA. Sequencing libraries were generated using TruSeq Nano DNA Sample Preparation kits (Illumina, Cat# 20015964), according to the low sample protocol and paired-end sequenced on a HiSeq 2500 (Illumina).

2020-08-24 | E-MTAB-9373 | biostudies-arrayexpress

Genome-wide mapping of the transcription factor GABP-alpha binding in human CD34+CD133+ hematopoietic progenitor cells.

Project description:We used ChIP-Seq to map GABP-alpha binding sites in human hematopoietic progenitor cells (HPCs). Coupled with functional assays using GABP-alpha deficient mouse model and bioinformatics analysis, we systematically determined a transcriptional module controlled by GABP in HPCs. Examination of the role of GABP in hematopoietic stem cells

2010-12-10 | E-GEOD-24933 | biostudies-arrayexpress

DNA sequencing comparing different stages of leukemic progression in a mouse model of severe congenital neutropenia (SCN)

Project description:To address how Csf3r and RUNX1 mutations in combination with CSF3 administration affect hematopoiesis in vivo, we performed serial transplantation experiments. Lineage-negative Csfr-d715 BM cell cells were lentivirally transduced with the patient specific RUNX1-D171N (RHD) mutant or an empty vector control. Primary recipients showed a pre-leukemic condition characterised by myeloblasts in the peripheral blood, but not overt AML. Upon serial transplantation, one of the Csfr-d715/RUNX1-D171N mutant mice, treated with CSF3, could repopulate secondary and tertiary recipients. Whole exome sequencing on these mice were performed to investigate whether these mice acquired an additional mutation. Enzymatically fragmented genomic DNA was used to construct sample libraries following the SeqCap EZ HyperPlusCap workflow User’s Guide version 1.0 (Roche). Unique, dual index adapters (Integrated DNA technologies) were used for ligation. After ligation of adapters and an amplification step, exome target sequences were captured using in-solution oligonucleotide baits (SeqCap EZ Developer Library mm9_exome_L2R_D02). Amplified captured sample libraries were paired-end sequenced on the HiSeq 2500 platform (Illumina).

2020-08-24 | E-MTAB-9376 | biostudies-arrayexpress

Loss of BTG1 function promotes ETV6-RUNX1-mediated leukemic transformation by upregulation of BCL6

Project description:Analysis of gene signatures in WT+Ctrl vs WT+ETV6-RUNX1, Btg1-/- and Btg1-/-+ETV6-RUNX1 in cKit+Ter119- fetal liver-derived hematopoietic progenitor cells (FL-HPCs). The Btg1-/-+ETV6-RUNX1 FL-HPCs display a strong increase in proliferation compared to WT+ETV6-RUNX1. Total RNA otained from WT+Ctrl, WT+ETV6-RUNX1, Btg1-/-+Ctrl and Btg1-/-+ETV6-RUNX1 FL-HPCs cells that were cultured for 12 days in expansion medium.

2014-08-22 | E-GEOD-60589 | biostudies-arrayexpress

RNA sequencing comparing different stages of leukemic progression in a mouse model of severe congenital neutropenia (SCN)

Project description:To address how Csf3r and RUNX1 mutations in combination with CSF3 administration affect hematopoiesis in vivo, we performed serial transplantation experiments. Lineage-negative Csfr-d715 BM cell cells were lentivirally transduced with the patient specific RUNX1-D171N (RHD) mutant or an empty vector control. An additionaly acquired mutation in Cxxc4 in mouse 29 transformed the pre-leukemic condition, characterized by excess pheripheral lineage- c-kit+ (LK) myeloblasts, into acute myeloid leukemia (AML). LK blasts were FACS sorted in TriZol and RNA was isolated according to the manufacturer’s protocol. SMARTer Ultra Low Input RNA kit for sequencing (Clontech, v4 Cat# 634891) was used to generate cDNA. Sequencing libraries were generated using TruSeq Nano DNA Sample Preparation kits (Illumina, Cat# 20015964), according to the low sample protocol and paired-end sequenced on a HiSeq 2500 (Illumina).

2020-08-24 | E-MTAB-9377 | biostudies-arrayexpress

LIM domain binding protein 1 regulates a transcriptional program essential for hematopoietic stem cell maintenance

Project description:We used ChIP-Seq to map Ldb1, Scl and Gata2 binding sites in mouse hematopoietic progenitor cells (HPCs). Together with functional studies using conventional and conditional Ldb1 deficient mouse models and bioinformatics analysis, we systematically determined a transcriptional program controlled by Ldb1 complexes in HSC maintenance. To evaluate the role of Ldb1 in hematopoietic stem cell maintenance.

2011-01-04 | E-GEOD-26031 | biostudies-arrayexpress

Expression data from murine Hemophagocytes (HPCs) versus resting splenic macrophages

Project description:Hemophagocytes are activated macrophages seen morphologically to have engulfed other hematopoietic cells. Their function is unknown. Attempts to induce these cells in vitro or purify them ex vivo have been unsuccessful. We isolated HPCs and resting splenic macrophages by laser capture microdissection. RNA was isolated from captured cells and microarray performed to compare the activation state of HPCs versus resting splenic macrophages. We treated WT mice with PBS or CpG1826 and an IL-10receptor blocking antibody. Cells were captured from splenic touch preps on PEN-coated slides, RNA isolated by WT-Ovation One-Direct Amplification System and assayed on Affymetrix mouse gene ST 1.0 chips.

2013-12-31 | E-GEOD-47430 | biostudies-arrayexpress

Expression data from human CD34+ HPC subpopulations isolated from umbilical cord blood (Haddad et al. Blood 104:3918, 2004)

Project description:We used microarrays to analyze the gene expression profile of CD34+CD45RA+CD7+, CD34+CD45RA+CD10+CD19- and CD34+CD45+CD7-CD10-CD19- HPCs isolated from umbilical cord blood CD34+CD45RA+CD7+(CD10-) and CD34+CD45RA+CD10+(CD7-CD19-) HPCs correspond respectively to prothymocytes and early pre-proB precursors. CD34+CD45RA+CD7-CD10-CD19- HPCs correspond to lympho-granulo-macrophagic precursors The corresponding populations were sorted from total CD34+ HPCs isolated from 2 or 3 individual donors

2011-05-25 | E-GEOD-29522 | biostudies-arrayexpress

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data