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Project description:Endothelial Map2k1 mutation affects vasculogenesis: RNA sequencing analysis in a conditional murine model
| PRJNA940423 | ENA
Project description:Somatic activating MAP2K1 mutations in endothelial cells (ECs) cause extracranial arteriovenous malformation (AVM). We previously reported the generation of a mouse line allowing inducible expression of constitutively active MAP2K1 (p.K57N) from the Rosa locus (R26GT-Map2k1-GFP/+) and showed, using Tg-Cdh5CreER, that EC expression of mutant MAP2K1 is sufficient for the development of vascular malformations in the brain, ear, and intestines. To gain further insight into the mechanism by which mutant MAP2K1 drives AVM development, we induced MAP2K1 (p.K57N) expression in ECs of postnatal-day-1 pups (P1) and investigated the changes in gene expression in P9 brain ECs by RNA-seq. We found that over-expression of MAP2K1 altered the transcript abundance of > 1600 genes. Several genes had > 20-fold changes between MAP2K1 expressing and wild-type ECs; the highest were Col15a1 (39-fold) and Itgb3 (24-fold). Increased expression of COL15A1 in R26GT-Map2k1-GFP/+; Tg-Cdh5CreER+/- brain ECs was validated by immunostaining. Ontology showed that differentially expressed genes were involved in processes important for vasculogenesis (e.g., cell migration, adhesion, extracellular matrix organization, tube formation, angiogenesis). Understanding how these genes and pathways contribute to AVM formation will help identify targets for therapeutic intervention.
| S-EPMC10329712 | biostudies-literature
Project description:We previously identified somatic activating KRAS mutations in a majority of human arteriovenous malformations (AVMs), using whole exome sequencing, which were enhanced in AVM endothelial cell fractions. We have now performed whole genome sequencing on AVM endothelial and non-endothelial cell fractions, as well as paired blood samples, in order to identify further somatic mutations.
| EGAS00001006719 | EGA
Project description:The mammalian genome contains two ERK/MAP kinase kinase genes, Map2k1 and Map2k2, encoding dual-specificity kinases responsible for ERK/MAP kinase activation. Loss of Map2k1 function in mouse causes embryonic lethality due to placental defects, while Map2k2 mutant mice survive with a normal lifespan. The Map2k1 mutation interferes with the growth of the labyrinthine region of the placenta and its vascularization. Map2k1+/-Map2k2+/- embryos also die during gestation of underdevelopment of the labyrinth. In Map2k1+/-Map2k2+/- mutants, the vascularization of the labyrinth is reduced and defects in SynT-II formation lead to the accumulation of multinucleated trophoblasts giant cells (MTG). Deletion of both Map2k1 alleles in allantoïs-derived tissues in Map2k1+/ Map2k2+/- placenta (Map2k1flox/-Map2k2+/-Tg+/Sox2Cre) increases the penetrance and the expressivity of the MTG placental phenotype. Microarray analysis with RNA extracted from placentas of E12.5 wt, Map2k1+/-Map2k2+/- and Map2k1flox/-Map2k2+/-Tg+/Sox2Cre embryos was performed to evaluate the molecular impact of the loss of Map2k alleles on placenta development. Total RNA was isolated from placentas of E12.5 Map2k1+/+Map2k2+/+(control), from E12.5 Map2k1+/-Map2k2+/- embryos (experimental) and from E12.5 and Map2k1flox/-Map2k2+/-Tg+/Sox2Cre embryos (experimental). Four specimens were analyzed per genotype.
2014-08-01 | E-GEOD-51644 | biostudies-arrayexpress
Project description:The mammalian genome contains two ERK/MAP kinase kinase genes, Map2k1 and Map2k2, encoding dual-specificity kinases responsible for ERK/MAP kinase activation. Loss of Map2k1 function in mouse causes embryonic lethality due to placental defects, while Map2k2 mutant mice survive with a normal lifespan. The Map2k1 mutation interferes with the growth of the labyrinthine region of the placenta and its vascularization. Map2k1+/-Map2k2+/- embryos also die during gestation of underdevelopment of the labyrinth. In Map2k1+/-Map2k2+/- mutants, the vascularization of the labyrinth is reduced and defects in SynT-II formation lead to the accumulation of multinucleated trophoblasts giant cells (MTG). Deletion of both Map2k1 alleles in allantoïs-derived tissues in Map2k1+/ Map2k2+/- placenta (Map2k1flox/-Map2k2+/-Tg+/Sox2Cre) increases the penetrance and the expressivity of the MTG placental phenotype. Microarray analysis with RNA extracted from placentas of E12.5 wt, Map2k1+/-Map2k2+/- and Map2k1flox/-Map2k2+/-Tg+/Sox2Cre embryos was performed to evaluate the molecular impact of the loss of Map2k alleles on placenta development.
2014-08-01 | GSE51644 | GEO
Project description:Background: Cancers result from accumulation of somatic mutations and their properties are thought to reflect the sum of these mutations. However, little is known about the consequences of altering the order of mutation acquisition. Methods: Mutation order was determined in myeloproliferative neoplasm patients by genotyping hematopoietic colonies or next generation sequencing. Stem and progenitor cells were isolated to study the effect of mutation order on mature and immature hematopoietic cells. Results: Age of presentation, acquisition of JAK2V617F homozygosity and the balance of immature progenitors were all influenced by mutation order. Compared to TET2-first patients, JAK2-first patients had an increased likelihood of presenting with polycythemia vera than essential thrombocythemia, an increased risk of thrombosis and an increased sensitivity of JAK2-mutant progenitors to ruxolitinib in vitro. In studies of single hematopoietic stem and progenitor cells (HSPCs), mutation order influenced the proliferative response to JAK2V617F and the capacity of double-mutant HSPCs to generate colony-forming cells. Moreover the HSPC compartment was dominated by TET2 single-mutant cells in TET2-first patients but by JAK2/TET2 double-mutant cells in JAK2-first patients. Prior mutation of TET2 altered the transcriptional consequences of JAK2V617F in a cell-intrinsic manner, and prevented JAK2V617F from up-regulating genes associated with proliferation. These data demonstrate that mutation order influences progenitor proliferation and terminal cell expansion, thus influencing clinical presentation, thrombosis risk and progenitor response to targeted therapy. Conclusions: The order in which JAK2 and TET2 mutations are acquired influences clinical features, stem/progenitor cell biology and clonal evolution in patients with myeloproliferative neoplasms.
2014-11-17 | E-MTAB-3086 | biostudies-arrayexpress
Project description:Interplay between non-cancerous cells (immune, fibroblasts, mesenchymal stromal cells (MSC), and endothelial cells (EC)) has been identified as vital in driving tumor progression. As studying such interactions in vivo is challenging, ex vivo systems that can recapitulate in vivo scenarios can aid in unraveling the factors impacting tumorigenesis and metastasis. Using the synthetic tumor microenvironment mimics (STEMs)-a spheroid system composed of breast cancer cells (BCC) with defined human MSC and EC fractions, here we show that EC organization into vascular structures is BC phenotype dependent, and independent of ERα expression in epithelial cancer cells, and involves MSC-mediated Notch1 signaling. In a 3D-bioprinted model system to mimic local invasion, MDA STEMs collectively respond to serum gradient and form invading cell clusters. STEMs grown on chick chorioallantoic membrane undergo local invasion to form CAM tumors that can anastomose with host vasculature and bear the typical hallmarks of human BC and this process requires both EC and MSC. This study provides a framework for developing well-defined in vitro systems, including patient-derived xenografts that recapitulate in vivo events, to investigate heterotypic cell interactions in tumors, to identify factors promoting tumor metastasis-related events, and possibly drug screening in the context of personalized medicine.
| S-EPMC9701219 | biostudies-literature
Project description:To identify potential biological targets of the TGFβ pathway involved in AVM formation, we performed RNA-seq on endothelial cells (ECs) isolated from a Smad4 inducible, EC specific knockout (Smad4-iECKO; Smad4f/f;Cdh5-CreERT2) mouse model that develops retinal AVMs.
2018-12-18 | GSE116230 | GEO
Project description:PurposeThe purpose of this study was to establish a murine model of endothelial keratoplasty.MethodsEndothelial keratoplasty (EK) was performed using C57BL/6 donor and BALB/c recipient mice. The central endothelium and Descemet membrane were removed from the recipient cornea, and a 1.5-mm posterior lamellar donor graft was made adherent to the recipient cornea with a small amount of viscoelastic. Mice were followed through slitlamp microscopy postoperatively, and OCT was used to assess the cornea and anterior chamber and measure central corneal thickness. Histology and immunohistochemistry were performed to confirm graft adherence and endothelial cell morphology.ResultsSuccessfully attached EK grafts were visualized in all transplanted animals. Histology and immunostaining confirmed proper graft orientation and adherence, as well as the presence of donor endothelium on transplanted grafts. We observed maximal corneal edema in all animals at day 1 postoperatively which gradually subsided. EK graft survival was 97% at 8 weeks.ConclusionsIn this study, we describe a novel murine model for EK which we anticipate will enable detailed investigation into the cellular and molecular mechanisms involved in EK pathobiology.
| S-EPMC9805546 | biostudies-literature
Project description:Endothelial cell late G1 arrest induced by Palbociclib modulates the expression of genes regulating arterio-venous identity and prevents AVM development induced by Alk1 genetic deletion.
2023-12-30 | GSE244923 | GEO