Grainyhead-like 2 inhibits the coactivator p300, suppressing tubulogenesis and the epithelial–mesenchymal transition

Developmental morphogenesis and tumor progression require a transient or stable breakdown of epithelial junctional complexes to permit programmed migration, invasion, and anoikis resistance, characteristics endowed by the epithelial–mesenchymal transition (EMT). The epithelial master-regulatory transcription factor Grainyhead-like 2 (GRHL2) suppresses and reverses EMT, causing a mesenchymal–epithelial transition to the default epithelial phenotype. Here we investigated the role of GRHL2 in tubulogenesis of Madin–Darby canine kidney cells, a process requiring transient, partial EMT. GRHL2 was required for cystogenesis, but it suppressed tubulogenesis in response to hepatocyte growth factor. Surprisingly, GRHL2 suppressed this process by inhibiting the histone acetyltransferase coactivator p300, preventing the induction of matrix metalloproteases and other p300-dependent genes required for tubulogenesis. A 13–amino acid region of GRHL2 was necessary for inhibition of p300, suppression of tubulogenesis, and interference with EMT. The results demonstrate that p300 is required for partial or complete EMT occurring in tubulogenesis or tumor progression and that GRHL2 suppresses EMT in both contexts through inhibition of p300

Steps towards the hyperfine splitting measurement of the muonic hydrogen ground state: pulsed muon beam and detection system characterization

The high precision measurement of the hyperfine splitting of the muonic-hydrogen atom ground state with pulsed and intense muon beam requires careful technological choices both in the construction of a gas target and of the detectors. In June 2014, the pressurized gas target of the FAMU experiment was exposed to the low energy pulsed muon beam at the RIKEN RAL muon facility. The objectives of the test were the characterization of the target, the hodoscope and the X-ray detectors. The apparatus consisted of a beam hodoscope and X-rays detectors made with high purity Germanium and Lanthanum Bromide crystals. In this paper the experimental setup is described and the results of the detector characterization are presented.Comment: 22 pages, 14 figures, published and open access on JINS

Regulation of anti-apoptotic signaling by Kruppel-like factors 4 and 5 mediates lapatinib resistance in breast cancer

The Kruppel-like transcription factors (KLFs) 4 and 5 (KLF4/5) are coexpressed in mouse embryonic stem cells, where they function redundantly to maintain pluripotency. In mammary carcinoma, KLF4/5 can each impact the malignant phenotype, but potential linkages to drug resistance remain unclear. In primary human breast cancers, we observed a positive correlation between KLF4/5 transcript abundance, particularly in the human epidermal growth factor receptor 2 (HER2)-enriched subtype. Furthermore, KLF4/5 protein was rapidly upregulated in human breast cancer cells following treatment with the HER2/epidermal growth factor receptor inhibitor, lapatinib. In addition, we observed a positive correlation between these factors in the primary tumors of genetically engineered mouse models (GEMMs). In particular, the levels of both factors were enriched in the basal-like tumors of the C3(1) TAg (SV40 large T antigen transgenic mice under control of the C3(1)/prostatein promoter) GEMM. Using tumor cells derived from this model as well as human breast cancer cells, suppression of KLF4 and/or KLF5 sensitized HER2-overexpressing cells to lapatinib. Indicating cooperativity, greater effects were observed when both genes were depleted. KLF4/5-deficient cells had reduced basal mRNA and protein levels of the anti-apoptotic factors myeloid cell leukemia 1 (MCL1) and B-cell lymphoma-extra large (BCL-XL). Moreover, MCL1 was upregulated by lapatinib in a KLF4/5-dependent manner, and enforced expression of MCL1 in KLF4/5-deficient cells restored drug resistance. In addition, combined suppression of KLF4/5 in cultured tumor cells additively inhibited anchorage-independent growth, resistance to anoikis and tumor formation in immunocompromised mice. Consistent with their cooperative role in drug resistance and other malignant properties, KLF4/5 levels selectively stratified human HER2-enriched breast cancer by distant metastasis-free survival. These results identify KLF4 and KLF5 as cooperating protumorigenic factors and critical participants in resistance to lapatinib, furthering the rationale for combining anti-MCL1/BCL-XL inhibitors with conventional HER2-targeted therapies

Experimental determination of the energy dependence of the rate of the muon transfer reaction from muonic hydrogen to oxygen for collision energies up to 0.1 eV

We report the first experimental determination of the collision-energy dependence of the muon transfer rate from the ground state of muonic hydrogen to oxygen at near-thermal energies. A sharp increase by nearly an order of magnitude in the energy range 0 - 70 meV was found that is not observed in other gases. The results set a reliable reference for quantum-mechanical calculations of low-energy processes with exotic atoms, and provide firm ground for the measurement of the hyperfine splitting in muonic hydrogen and the determination of the Zemach radius of the proton by the FAMU collaboration.Comment: 30 pages, 10 figure

First measurement of the temperature dependence of muon transfer rate from muonic hydrogen atoms to oxygen

We report the first measurement of the temperature dependence of muon transfer rate from muonic hydrogen atoms to oxygen between 100 and 300 K. Data were obtained from the X-ray spectra of delayed events in a gaseous target, made of a H2/O2 mixture, exposed to a muon beam. This work sets constraints on theoretical models of muon transfer and is of fundamental importance for the measurement of the hyperfine splitting of muonic hydrogen ground state as proposed by the FAMU collaboration

First measurement of the temperature dependence of muon transfer rate from muonic hydrogen atoms to oxygen

We report the first measurement of the temperature dependence of muon transfer rate from muonic hydrogen atoms to oxygen between 100 and 300 K. Data were obtained from the X-ray spectra of delayed events in a gaseous target, made of a H2/O2 mixture, exposed to a muon beam. This work sets constraints on theoretical models of muon transfer and is of fundamental importance for the measurement of the hyperfine splitting of muonic hydrogen ground state as proposed by the FAMU collaboration

Differential reprogramming of breast cancer subtypes in 3D cultures and implications for sensitivity to targeted therapy

Screening for effective candidate drugs for breast cancer has shifted from two-dimensional (2D) to three-dimensional (3D) cultures. Here we systematically compared the transcriptomes of these different culture conditions by RNAseq of 14 BC cell lines cultured in both 2D and 3D conditions. All 3D BC cell cultures demonstrated increased mitochondrial metabolism and downregulated cell cycle programs. Luminal BC cells in 3D demonstrated overall limited reprogramming. 3D basal B BC cells showed increased expression of extracellular matrix (ECM) interaction genes, which coincides with an invasive phenotype not observed in other BC cells. Genes downregulated in 3D were associated with metastatic disease progression in BC patients, including cyclin dependent kinases and aurora kinases. Furthermore, the overall correlation of the cell line transcriptome to the BC patient transcriptome was increased in 3D cultures for all TNBC cell lines. To define the most optimal culture conditions to study the oncogenic pathway of interest, an open source bioinformatics strategy was established.Toxicolog

FAMU: study of the energy dependent transfer rate \u39b \u3bcp \u2192 \u3bcO

The main goal of the FAMU experiment is the measurement of the hyperfine splitting (hfs) in the 1S state of muonic hydrogen \u394Ehfs (\u3bc - p)1S. The physical process behind this experiment is the following: \u3bcp are formed in a mixture of hydrogen and a higher-Z gas. When absorbing a photon at resonance-energy \u394Ehfs 48 0.182 eV, in subsequent collisions with the surrounding H 2 molecules, the \u3bcp is quickly de-excited and accelerated by ~ 2/3 of the excitation energy. The observable is the time distribution of the K-lines X-rays emitted from the \u3bcZ formed by muon transfer (\u3bcp) + Z \u2192 (\u3bcZ)* + p, a reaction whose rate depends on the \u3bcp kinetic energy. The maximal response, to the tuned laser wavelength, of the time distribution of X-ray from K-lines of the (\u3bcZ)* cascade indicate the resonance. During the preparatory phase of the FAMU experiment, several measurements have been performed both to validate the methodology and to prepare the best configuration of target and detectors for the spectroscopic measurement. We present here the crucial study of the energy dependence of the transfer rate from muonic hydrogen to oxygen (\u39b \u3bcp \u2192 \u3bc0 ), precisely measured for the first time

First FAMU observation of muon transfer from \u3bcp atoms to higher-Z elements

Abstract: The FAMU experiment aims to accurately measure the hyperfine splitting of the ground state of the muonic hydrogen atom. A measurement of the transfer rate of muons from hydrogen to heavier gases is necessary for this purpose. In June 2014, within a preliminary experiment, a pressurized gas-target was exposed to the pulsed low-energy muon beam at the RIKEN RAL muon facility (Rutherford Appleton Laboratory, U.K.). The main goal of the test was the characterization of both the noise induced by the pulsed beam and the X-ray detectors. The apparatus, to some extent rudimental, has served admirably to this task. Technical results have been published that prove the validity of the choices made and pave the way for the next steps. This paper presents the results of physical relevance of measurements of the muon transfer rate to carbon dioxide, oxygen, and argon from non-thermalized excited \u3bcp atoms. The analysis methodology and the approach to the systematics errors are useful for the subsequent study of the transfer rate as function of the kinetic energy of the \u3bcp currently under way