5 research outputs found
Nanog-Tcf15 axis during exit from naïve pluripotency
Pluripotent cells have the dual abilities to self-renewal and to differentiate
into all three germ layers. Pluripotent cells can be isolated from two different stages
of mouse embryogenesis. Embryonic stem cells (ESCs) are isolated from the inner
cell mass (ICM) of the pre-implantation embryo and are considered to be in a naïve
state. On the other hand, cells isolated from epiblast of the post-implantation embryo
are referred as epiblast stem cells (EpiSC) and are representative of primed
pluripotency. ESCs and EpiSCs are distinct from each other in terms of the
morphology, the gene regulatory network and the signalling pathways regulating
self-renewal. Under certain conditions, ESCs and EpiSCs can be transitioned into
each other. However, the mechanism that regulates this transition from naïve to
primed pluripotent state remains to be solved.
Nanog, Oct4 and Sox2 form the core gene regulatory network of
pluripotency. Additionally, the Id protein family is also important in the maintenance
of pluripotency in ESCs. Id proteins function by inhibiting the activity of pro-differentiation
factors. Tcf15 is identified as one of the targets of Id inhibition in
ESCs. Moreover, Tcf15 has been identified as a repression target of Nanog.
In this study, to understand the function of Tcf15, the expression of Tcf15
was characterized in differentiating ESCs. The transient upregulation of Tcf15
mRNA and protein was detected at early stages of differentiation before lineage
commitment. Furthermore, Tcf15 protein was heterogeneously expressed in
differentiating cells. Mutually exclusive expression of Nanog and Tcf15 proteins
were demonstrated in both self-renewing and differentiating ESCs.
Further characterization of the effect of Nanog on Tcf15 transcription showed
that Tcf15 pre-mRNA was downregulated within 20 minute of Nanog induction. A
Nanog binding site was identified at +32kb relative to the Tcf15 transcription start
site (TSS). Initially, Nanog binding at this region was confirmed by performing
ChIP-PCR experiments. Then, this Nanog binding region was further analysed for its
enhancer activity related to the Tcf15 gene. Deletion of the Nanog binding region
using CRISPR-Cas9 confirmed that this region acts as Tcf15 enhancer; it was shown
that this region was required for the activation of Tcf15 transcription during
differentiation.
Tcf15 induction experiments were performed in order to the check whether
Tcf15 affects Nanog transcription. The results indicate that Nanog is not a direct
target of Tcf15, but Tcf15 contributes indirectly to the repression of Nanog.
Additional analysis with the Tcf15 enhancer deletion cells showed that Tcf15
is not required for efficient downregulation of naïve markers and the upregulation of
primed markers. However, the genes related to the regulation of adhesion properties
of cells such as Zyc, Itga3 were induced with lower efficiency in the absence of
Tcf15 compared to the wild type cells.
In summary, I investigated the reciprocal regulation of Tcf15 and Nanog and
the role of Tcf15 for the differentiation. My results suggest that Tcf15 is expressed
in the cells that have initiated differentiation but are not lineage-committed.
Additionally, Tcf15 can contribute to the regulation of adhesion related genes in
order to help the epithelisation of the cells required during the differentiation from
naïve to the primed pluripotent state. As a conclusion, Nanog is proposed to help to
prevent certain aspects of ESCs differentiation by repressing Tcf15
Immunization status in chronic obstructive pulmonary disease: A multicenter study from Turkey
intepe, yavuz selim/0000-0002-5697-5291; Erol, Serhat/0000-0003-1645-7761; Korkmaz, Celalettin/0000-0001-8602-0368; Sertogullarindan, Bunyamin/0000-0002-1478-1990; Kasapoglu, Umut Sabri/0000-0003-2869-9872; Bulbul, Yilmaz/0000-0002-8488-3650WOS: 000455908800007PubMed: 30745939OBJECTIVE: The purpose of this study is to detect the prevalence and the factors associated with influenza and pneumococcal vaccination and outcomes of vaccination during 2013-2014 season in patients with chronic obstructive pulmonary disease (COPD) in Turkey. METHODS: This was a multicenter retrospective cohort study performed in 53 different centers in Turkey. RESULTS: During the study period, 4968 patients were included. COPD was staged as GOLD 1-2-3-4 in 9.0%, 42.8%, 35.0%, and 13.2% of the patients, respectively. Influenza vaccination rate in the previous year was 37.9%; and pneumococcus vaccination rate, at least once during in a life time, was 13.3%. Patients with older age, higher level of education, more severe COPD, and comorbidities, ex-smokers, and patients residing in urban areas had higher rates of influenza vaccination. Multivariate logistic regression analysis showed that advanced age, higher education levels, presence of comorbidities, higher COPD stages, and exacerbation rates were associated with both influenza and pneumococcal vaccination. The number of annual physician/outpatient visits and hospitalizations due to COPD exacerbation was 2.73 +/- 2.85 and 0.92 +/- 1.58 per year, respectively. Patients with older age, lower education levels, more severe COPD, comorbid diseases, and lower body mass index and patients who are male and are residing in rural areas and vaccinated for influenza had significantly higher rates of COPD exacerbation. CONCLUSIONS: The rates of influenza and pneumococcal vaccination in COPD patients were quite low, and the number of annual physician/outpatient visits and hospitalizations due to COPD exacerbation was high in Turkey. Advanced age, higher education levels, comorbidities, and higher COPD stages were associated with both influenza and pneumococcal vaccination
MPP8 is essential for sustaining self-renewal of ground-state pluripotent stem cells
Deciphering the mechanisms that control the pluripotent ground state is key for understanding embryonic development. Nonetheless, the epigenetic regulation of ground-state mouse embryonic stem cells (mESCs) is not fully understood. Here, we identify the epigenetic protein MPP8 as being essential for ground-state pluripotency. Its depletion leads to cell cycle arrest and spontaneous differentiation. MPP8 has been suggested to repress LINE1 elements by recruiting the human silencing hub (HUSH) complex to H3K9me3-rich regions. Unexpectedly, we find that LINE1 elements are efficiently repressed by MPP8 lacking the chromodomain, while the unannotated C-terminus is essential for its function. Moreover, we show that SETDB1 recruits MPP8 to its genomic target loci, whereas transcriptional repression of LINE1 elements is maintained without retaining H3K9me3 levels. Taken together, our findings demonstrate that MPP8 protects the DNA-hypomethylated pluripotent ground state through its association with the HUSH core complex, however, independently of detectable chromatin binding and maintenance of H3K9me3
TAK1 inhibition leads to RIPK1-dependent apoptosis in immune-activated cancers
Poor survival and lack of treatment response in glioblastoma (GBM) is attributed to the persistence of glioma stem cells (GSCs). To identify novel therapeutic approaches, we performed CRISPR/Cas9 knockout screens and discovered TGFβ activated kinase (TAK1) as a selective survival factor in a significant fraction of GSCs. Loss of TAK1 kinase activity results in RIPK1-dependent apoptosis via Caspase-8/FADD complex activation, dependent on autocrine TNFα ligand production and constitutive TNFR signaling. We identify a transcriptional signature associated with immune activation and the mesenchymal GBM subtype to be a characteristic of cancer cells sensitive to TAK1 perturbation and employ this signature to accurately predict sensitivity to the TAK1 kinase inhibitor HS-276. In addition, exposure to pro-inflammatory cytokines IFN gamma and TNFα can sensitize resistant GSCs to TAK1 inhibition. Our findings reveal dependency on TAK1 kinase activity as a novel vulnerability in immune-activated cancers, including mesenchymal GBMs that can be exploited therapeutically