Transcriptome data of hematopoietic stem cells of mice under ionizing radiation
Ontology highlight
ABSTRACT: Analysis of hematopoietic stem cells (HSC, Lineage-Sca-1+c-Kit+Flt3–CD34–). In order to better experiment, we treated mice with total body irradiation and local irradiation respectively. Next, the HSC were purified from the bone marrow of 8 weeks WT mice (Ctrl), total body radiation mice (IR), irradiated legs of locally irradiated mice (L_IR) and the other leg which unirradiated of locally irradiated mice (Ab_IR). Results provide the injury effect of IR on HSC under different conditions.
Project description:Analysis of the RNA-seq data performed in IR vs NIR hematopoietic stem cells show the loss of the TNF_via_NFKB signature. We showed that the loss of this signature could be associated with hematopoietic stem cell gene deregulation. To validate this association, we tested if TNFa treatment before irradiation was able to prevent IR-effect on HSC gene deregulation. For this purpose, we treated mice with TNFa 1h before irradiation (IR_TNF) and performed RNA-seq experiments on hematopoietic stem cells 1 month after irradiation and compared them to hematopoietic stem cells sorted from non irradiated mice (NIR) and from non-treated irradiated mice (IR).
Project description:using transplantation of transgenic knock-in JAK2V617F hematopoietic cells into a single irradiated leg, we show development of Polycythemia Vera (PV) from a single anatomical site in immuno-competent mice. Mutant cells colonizing the non-irradiated leg efficiently induce PV. We compared transcriptomic profiles of sorted JAK2V617F HSPC and their reciprocal endogenous WT HSPC in the non-irradiated and irradiated legs from recipient mice transplanted with JAK2V617F hematopoietic cells.
Project description:Ionizing radiation (IR) has long been associated with reduced hematopoietic function and increased malignancies, although the mechanisms behind this relationship remain poorly understood. The carcinogenic effect of IR has been commonly attributed to the direct induction of DNA damage. We demonstrate that IR exposure results in long-term, somatically heritable, cell-intrinsic reductions in HSC self-renewal that is mediated by C/EBP? and reversed by Notch, both of which are associated with human leukemias. Remarkably, restoration of HSC self-renewal prevents selection for C/EBP? loss of function in previously irradiated HSC pools. We propose that environmental insults prompt HSC to initiate a program limiting their self-renewal to prevent damaged HSC from contributing to hematopoiesis. This "programmed mediocrity" is advantageous for the localized insults animals have evolved to deal with, but becomes tumor promoting when the entire HSC compartment is damaged, such as during total body irradiation, by increasing selective pressure for adaptive oncogenic mutations Examination of mRNA levels in in vitro and in vivo Hematopoietic Stem Cell that exposed to IR Ionizing radiation (IR) or control. Each group has three replicates.
Project description:Irradiation of the K-rasLA1 mouse model with a fractionated dose of 1.0Gy 56Fe- particles increases the incidence of invasive carcinoma compared to unirradiated controls or those irradiated with an acute dose. Microarray profiling was perfromed on whole lungs from K-rasLA1 mice in order to determine global expression changes in the lung following radiation exposure. RNA was extracted from K-rasLA1 lungs from unirradiated control animals or those irradiated with a fractionated or acute dose of 1.0Gy 56Fe- particles 70 days post-irradiation when lungs are still histologically indistiguishable and only contain benign lesions. 129S2 K-rasLA1 mice were either irradiated with an acute (1.0Gy x1; n = 15) or fractionated (0.2Gy x5; n=13) dose of 1.0Gy 56Fe- particles or left unirradiated (n=13). Animals were euthanized 70 days post-irradiation and RNA was extracted from the whole left lung of each animal.
Project description:Ionizing radiation (IR) has long been associated with reduced hematopoietic function and increased malignancies, although the mechanisms behind this relationship remain poorly understood. The carcinogenic effect of IR has been commonly attributed to the direct induction of DNA damage. We demonstrate that IR exposure results in long-term, somatically heritable, cell-intrinsic reductions in HSC self-renewal that is mediated by C/EBPα and reversed by Notch, both of which are associated with human leukemias. Remarkably, restoration of HSC self-renewal prevents selection for C/EBPα loss of function in previously irradiated HSC pools. We propose that environmental insults prompt HSC to initiate a program limiting their self-renewal to prevent damaged HSC from contributing to hematopoiesis. This "programmed mediocrity" is advantageous for the localized insults animals have evolved to deal with, but becomes tumor promoting when the entire HSC compartment is damaged, such as during total body irradiation, by increasing selective pressure for adaptive oncogenic mutations
Project description:Comprehensive gene expression analysis in BM-resident stromal cells was performed for an overview of BM environmental change caused by total body irradiation (TBI). Total RNA samples collected from BM-resident stromal cells with or without TBI were subjected to high sensitivity DNA microarray assays Three-condition experiment: Unirradiated, 1 day after TBI and 3 days after TBI. Bone marrow stromal cells were obtained from C57BL/6 mice (n = 6) either non-irradiated or after 9.5 Gy irradiation at indicated times.
Project description:A 2 Gy-total body irradiation (TBI) of WT mice induces a second wave of ROS and a second wave of apoptosis in hematopoietic stem cells starting from 6 days after irradiation. In CD169DTR/+ mice, which are depleted in CD169+ resident macrophages after a diphtheria toxin injection, these two waves do not occur and a total recovery of the LT-HSC pool is observed, contrarily to the LT-HSC pool from WT mice, which is decreased. Our goal is to understand why these secondary waves of apoptosis and ROS occur in LT-HSC from irradiated WT mice and not in LT-HSC from irradiated mice depleted in resident macrophages, allowing a recovery of the LT-HSC pool. We used microarrays to compare the global gene expression in LT-HSC isolated from WT and CD169DTR/+ mice 5 days after irradiation, i.e. just before both secondary waves of ROS and apoptosis occurred in WT mice.
Project description:Irradiation of the K-rasLA1 mouse model with a fractionated dose of 1.0Gy 56Fe- particles increases the incidence of invasive carcinoma compared to unirradiated controls or those irradiated with an acute dose. Microarray profiling was perfromed on whole lungs from K-rasLA1 mice in order to determine global expression changes in the lung following radiation exposure. RNA was extracted from K-rasLA1 lungs from unirradiated control animals or those irradiated with a fractionated or acute dose of 1.0Gy 56Fe- particles 70 days post-irradiation when lungs are still histologically indistiguishable and only contain benign lesions.
Project description:Analysis of MHCII-high (MHCII-hi) and MHCII-low (MHCII-lo) hematopoietic stem cells (Lineage-Sca-1+c-Kit+CD150+Flt3-CD48-). The two population of hematopoietic stem cell (HSC) were purified from the bone marrow of mice at 3 months post 5 Gy total body irradiation (IR). Results provide insight into the role of MHCII in HSC.
Project description:Analysis of the RNA-seq data performed in IR vs NIR hematopoietic stem cells show the loss of the TNF_via_NFKB signature. We showed that the loss of this signature could be associated with H3K9me3 loss at specific retrotransposable elements . To validate this association, we tested if TNFa treatment before irradiation was able to prevent IR-effect on H3K9me3 loss at retrotransposable elements. For this purpose, we treated mice with TNFa 1h before irradiation (IR_TNF) and performed H3K9me3 cut&tag experiments on hematopoietic stem cells 1 month after irradiation and compared them to hematopoietic stem cells sorted from non irradiated mice (NIR) and from non-treated irradiated mice (IR).