The transcription factor ATF5: role in cellular differentiation, stress responses, and cancer.

Activating transcription factor 5 (ATF5) is a cellular prosurvival transcription factor within the basic leucine zipper (bZip) family that is involved in cellular differentiation and promotes cellular adaptation to stress. Recent studies have characterized the oncogenic role of ATF5 in the development of several different types of cancer, notably glioblastoma. Preclinical assessment of a systemically deliverable dominant-negative ATF5 (dnATF5) biologic has found that targeting ATF5 results in tumor regression and tumor growth inhibition of glioblastoma xenografts in mouse models. In this review, we comprehensively and critically detail the current scientific literature on ATF5 in the context of cellular differentiation, survival, and response to stressors in normal tissues. Furthermore, we will discuss how the prosurvival role of ATF5 aides in cancer development, followed by current advances in targeting ATF5 using dominant-negative biologics, and perspectives on future research

Dominant-negative ATF5 rapidly depletes survivin in tumor cells.

Survivin (BIRC5, product of the BIRC5 gene) is highly expressed in many tumor types and has been widely identified as a potential target for cancer therapy. However, effective anti-survivin drugs remain to be developed. Here we report that both vector-delivered and cell-penetrating dominant-negative (dn) forms of the transcription factor ATF5 that promote selective death of cancer cells in vitro and in vivo cause survivin depletion in tumor cell lines of varying origins. dn-ATF5 decreases levels of both survivin mRNA and protein. The depletion of survivin protein appears to be driven at least in part by enhanced proteasomal turnover and depletion of the deubiquitinase USP9X. Survivin loss is rapid and precedes the onset of cell death triggered by dn-ATF5. Although survivin downregulation is sufficient to drive tumor cell death, survivin over-expression does not rescue cancer cells from dn-ATF5-promoted apoptosis. This indicates that dn-ATF5 kills malignant cells by multiple mechanisms that include, but are not limited to, survivin depletion. Cell-penetrating forms of dn-ATF5 are currently being developed for potential therapeutic use and the present findings suggest that they may pose an advantage over treatments that target only survivin

Chemoenzymatic synthesis of isotopically labelled folates

Dihydrofolate reductase (DHFR) is a key enzyme in cellular anabolism. It catalyses the reduction of 7,8-dihydrofolate (H2F) to 5,6,7,8-tetrahydrofolate (H4F) via hydride transfer from the C4 position of NADPH to the C6 position of H2F accompanied with protonation at the N5 position of H2F. Due to the importance of DHFR as an anticancer and antimicrobial target, the catalytic mechanism of DHFR has long been the focus of intense research. Kinetic isotope effect (KIE) measurements can provide insight into the mechanism of DHFR catalysis and guide the rational design of novel anti-DHFR drugs. However,because of a lack of a practical strategy to introduce heavy atoms (15N, 13C) into H2F, current research is mostly restrained to the study of hydrogen isotope effects. In this thesis, a fourteen step, one-pot chemoenzymatic synthesis of labelled H2F is reported. The flexibility of this synthetic approach enables the production of various isotopically enriched H2Fs from simple starting materials such as D-glucose. The labelled substrates were used to measure, for the first time, heavy atom KIEs and to derive information about the transition state of the chemical step during DHFR catalysis. This methodology is widely applicable to other biochemically important substrates and cofactors and it can be used for a wide variety of in vitro and in vivo investigations

Christine Jacobs-Wagner: Drawing the bacterial organizational chart

Jacobs-Wagner has been at the forefront of a revolution in bacterial cell biology

Suppression of planar cell polarity signaling and migration in glioblastoma by Nrdp1-mediated Dvl polyubiquitination.

The lethality of the aggressive brain tumor glioblastoma multiforme (GBM) results in part from its strong propensity to invade surrounding normal brain tissue. Although oncogenic drivers such as epidermal growth factor receptor activation and Phosphatase and Tensin homolog inactivation are thought to promote the motility and invasiveness of GBM cells via phosphatidylinostitol 3-kinase activation, other unexplored mechanisms may also contribute to malignancy. Here we demonstrate that several components of the planar cell polarity (PCP) arm of non-canonical Wnt signaling including VANGL1, VANGL2 and FZD7 are transcriptionally upregulated in glioma and correlate with poorer patient outcome. Knockdown of the core PCP pathway component VANGL1 suppresses the motility of GBM cell lines, pointing to an important mechanistic role for this pathway in glioblastoma malignancy. We further observe that restoration of Nrdp1, a RING finger type E3 ubiquitin ligase whose suppression in GBM also correlates with poor prognosis, reduces GBM cell migration and invasiveness by suppressing PCP signaling. Our observations indicate that Nrdp1 physically interacts with the Vangl1 and Vangl2 proteins to mediate the K63-linked polyubiquitination of the Dishevelled, Egl-10 and Pleckstrin (DEP) domain of the Wnt pathway protein Dishevelled (Dvl). Ubiquitination hinders Dvl binding to phosphatidic acid, an interaction necessary for efficient Dvl recruitment to the plasma membrane upon Wnt stimulation of Fzd receptor and for the propagation of downstream signals. We conclude that the PCP pathway contributes significantly to the motility and hence the invasiveness of GBM cells, and that Nrdp1 acts as a negative regulator of PCP signaling by inhibiting Dvl through a novel polyubiquitination mechanism. We propose that the upregulation of core PCP components, together with the loss of the key negative regulator Nrdp1, act coordinately to promote GBM invasiveness and malignancy

Optimization of Ni-Based WC/Co/Cr Composite Coatings Produced by Multilayer Laser Cladding

As a surface coating technique, laser cladding (LC) has been developed for improving wear, corrosion, and fatigue properties of mechanical components. The main advantage of this process is the capability of introducing hard particles such as SiC, TiC, and WC as reinforcements in the metallic matrix such as Ni-based alloy, Co-based alloy, and Fe-based alloy to form ceramic-metal composite coatings, which have very high hardness and good wear resistance. In this paper, Ni-based alloy (Colmonoy 227-F) and Tungsten Carbides/Cobalt/Chromium (WC/Co/Cr) composite coatings were fabricated by the multilayer laser cladding technique (MLC). An optimization procedure was implemented to obtain the combination of process parameters that minimizes the porosity and produces good adhesion to a stainless steel substrate. The optimization procedure was worked out with a mathematical model that was supported by an experimental analysis, which studied the shape of the clad track generated by melting coaxially fed powders with a laser. Microstructural and microhardness analysis completed the set of test performed on the coatings