Insulin Receptor Substrate-2 (IRS-2): A Novel Hypoxia-Responsive Gene in Breast Cancer: A Dissertation

Breast cancer is the most common malignancy among women in the U.S. While many successful treatments exist for primary breast cancer, very few are available for patients with metastatic disease. The purpose of this study was to understand the role of Insulin Receptor Subtrate-2 (IRS-2) in breast cancer metastasis. IRS-2 belongs to the IRS family of cytoplasmic adaptor proteins that mediate signaling from cell surface receptors, many of which have been implicated in cancer. Although the IRS proteins are highly homologous in structure and have some complementary functions, growing evidence supports that the IRS proteins have unique roles in cancer. IRS-1 has been shown to promote tumor cell proliferation, while IRS-2 has been positively associated with cancer cell invasion, glycolysis and tumor metastasis. In the current work, we identified IRS-2 as a novel hypoxia-responsive gene in breast carcinoma cells. In contrast, IRS-1 expression does not increase in response to hypoxia, supporting the notion of their non-overlapping functions. Hypoxia promotes the adaptation and resistance of cancer cells to chemo- and radiation therapy, and also promotes tumor cell survival, invasion and metastasis by selecting for aggressive tumor cells that can survive under stressful low oxygen conditions. We have shown that IRS-2 upregulation in response to hypoxia promotes Akt signaling and tumor cell viability and invasion. We identified a cell context-dependent role for Hypoxia Inducible Factor (HIF) in the regulation of IRS-2 expression in hypoxia, with HIF-2 playing a more dominant role than HIF-1. We also demonstrate that binding of Snail, a regulator of the EMT, to the IRS-2 promoter keeps the chromatin in an open conformation that is permissive for HIF-dependent transcription of IRS-2 in hypoxia. IRS-2 is not upregulated by hypoxia in well-differentiated epithelial-like carcinoma cells that do not express Snail, implicating IRS-2 gene expression as part of the EMT programming. In summary, we have identified an endogenous mechanism by which cancer cells can shift the balance of IRS-1 and IRS-2 to favor IRS-2 expression and function, which promotes survival, invasion, and ultimately metastasis. Understanding the mechanism of IRS-2 regulation by hypoxia may reveal new therapeutic targets for metastatic breast cancer

Expression and function of the insulin receptor substrate proteins in cancer

The Insulin Receptor Substrate (IRS) proteins are cytoplasmic adaptor proteins that function as essential signaling intermediates downstream of activated cell surface receptors, many of which have been implicated in cancer. The IRS proteins do not contain any intrinsic kinase activity, but rather serve as scaffolds to organize signaling complexes and initiate intracellular signaling pathways. As common intermediates of multiple receptors that can influence tumor progression, the IRS proteins are positioned to play a pivotal role in regulating the response of tumor cells to many different microenvironmental stimuli. Limited studies on IRS expression in human tumors and studies on IRS function in human tumor cell lines and in mouse models have provided clues to the potential function of these adaptor proteins in human cancer. A general theme arises from these studies; IRS-1 and IRS-4 are most often associated with tumor growth and proliferation and IRS-2 is most often associated with tumor motility and invasion. In this review, we discuss the mechanisms by which IRS expression and function are regulated and how the IRS proteins contribute to tumor initiation and progression

Single-crystal silver nanowires: Preparation and Surface-enhanced Raman Scattering (SERS) property

Ordered Ag nanowire arrays with high aspect ratio and high density self-supporting Ag nanowire patterns were successfully prepared using potentiostatic electrodeposition within the confined nanochannels of a commercial porous anodic aluminium oxide (AAO) template. X-ray diffraction and selected area electron diffraction analysis show that the as-synthesized samples have preferred (220) orientation. Transmission electron microscopy and scanning electron microscopy investigation reveal that large-area and ordered Ag nanowire arrays with smooth surface and uniform diameter were synthesized. Surface-enhanced Raman Scattering (SERS) spectra show that the Ag nanowire arrays as substrates have high SERS activity.Comment: 5 pages, 4 figure

Chemical composition and structural changes of porous templates obtained by anodising aluminium in phosphoric acid electrolyte

Ordered anodic aluminium oxide (AAO) filmswere first prepared by anodising in a phosphoric acid electrolyte and then studied extensively and characterised by field emission gun-scanning electron microscopy (FEG-SEM), X-ray diffraction, Raman and infrared spectroscopy at a macroscopic scale. These analyses showed that the as-prepared AAO film is in fact amorphous, partially hydrated and that its initial global chemical composition can be described, in agreement with previous works, as: Al2O3, 0.186AlPO4 · 0.005H2O. Additional analyses (thermogravimetric analysis, differential thermal analysis and FEG-SEM) showed geometrical changes of the film structure at different scales, explained by various steps of dehydration and allotropic transformations of the resulting crystallised alumina. However, because their structure remains unchanged up to 900 ◦C, the phosphoric templates appear to be particularly suitable for applications or processes atmedium or high temperatures, such as the preparation of carbon nanotubes or oxide rods

Reliable Pb(Zr,Ti)O3-based thin film piezoelectric micromirrors for space-applications

Although Pb(Zr,Ti)O3 (PZT) piezoelectric thin films are finding widespread applications on Earth, it is yet unclear if they are suitable for space-related applications. In space, their long-term reliability is a significant concern due to the difficulties of repairing and replacing malfunctioning devices. In this work, PZT thin film micromirrors for compact interferometric 3D imaging systems have been exposed to operating conditions encountered on a space mission and tested according to criteria set by the European Space Agency. Thermal cycling in vacuum, sinusoidal and random mechanical vibrations, and -radiation with and without bias did not degrade key functional device properties of the micromirror such as angular deflection, resonance frequency, polarization, and permittivity. Apart from -radiation, stressing the devices enhanced their large-signal angular deflection and improved their electrical lifetime compared to pristine devices. Their dielectric and ferroelectric characteristics remained comparable to that of a lab-scale environment. Simultaneously applying a 10 V field-down bias while -radiating the micromirrors changed the capacitance-field and polarization-field characteristics and enhanced the electrical imprint. After stress-testing, the median time-to-failure in moderate acceleration conditions of 150 kV/cm and 175 °C ranged from 1.95 to 2.64 h, close to 2.11 h as measured for a reference group. All actuator membranes had shorter electrical lifetimes, smaller voltage acceleration factors, and smaller activation energies, ranging from 2.56 to 2.88 V−1 and 1.03 to 1.09 eV, than simple bonding pads. This work is a device-level report covering a full set of space-relevant tests demonstrating that PZT-based thin film piezomicroelectromechanical systems technology is space-ready.acceptedVersio

Supported Molten Metal Membranes for Hydrogen Separation

We describe here our results on the feasibility of a novel dense metal membrane for hydrogen separation: Supported Molten Metal Membrane, or SMMM.1 The goal in this work was to develop these new membranes based on supporting thin films of low-melting, non- precious group metals, e.g., tin (Sn), indium (In), gallium (Ga), or their alloys, to provide a flux and selectivity of hydrogen that rivals the conventional but substantially more expensive palladium (Pd) or Pd alloy membranes, which are susceptible to poisoning by the many species in the coal-derived syngas, and further possess inadequate stability and limited operating temperature range. The novelty of the technology presented numerous challenges during the course of this project, however, mainly in the selection of appropriate supports, and in the fabrication of a stable membrane. While the wetting instability of the SMMM remains an issue, we did develop an adequate understanding of the interaction between molten metal films with porous supports that we were able to find appropriate supports. Thus, our preliminary results indicate that the Ga/SiC SMMM at 550 ºC has a permeance that is an order of magnitude higher than that of Pd, and exceeds the 2015 DOE target. To make practical SMM membranes, however, further improving the stability of the molten metal membrane is the next goal. For this, it is important to better understand the change in molten metal surface tension and contact angle as a function of temperature and gas-phase composition. A thermodynamic theory was, thus, developed, that is not only able to explain this change in the liquid-gas surface tension, but also the change in the solid-liquid surface tension as well as the contact angle. This fundamental understanding has allowed us to determine design characteristics to maintain stability in the face of changing gas composition. These designs are being developed. For further progress, it is also important to understand the nature of solution and permeation process in these molten metal membranes. For this, a comprehensive microkinetic model was developed for hydrogen permeation in dense metal membranes, and tested against data for Pd membrane over a broad range of temperatures.3 It is planned to obtain theoretical and experimental estimates of the parameters to corroborate the model against mental results for SMMM

LIM kinase inhibitors disrupt mitotic microtubule organization and impair tumor cell proliferation

The actin and microtubule cytoskeletons are critically important for cancer cell proliferation, and drugs that target microtubules are widely-used cancer therapies. However, their utility is compromised by toxicities due to dose and exposure. To overcome these issues, we characterized how inhibition of the actin and microtubule cytoskeleton regulatory LIM kinases could be used in drug combinations to increase efficacy. A previously-described LIMK inhibitor (LIMKi) induced dose-dependent microtubule alterations that resulted in significant mitotic defects, and increased the cytotoxic potency of microtubule polymerization inhibitors. By combining LIMKi with 366 compounds from the GSK Published Kinase Inhibitor Set, effective combinations were identified with kinase inhibitors including EGFR, p38 and Raf. These findings encouraged a drug discovery effort that led to development of CRT0105446 and CRT0105950, which potently block LIMK1 and LIMK2 activity in vitro, and inhibit cofilin phosphorylation and increase αTubulin acetylation in cells. CRT0105446 and CRT0105950 were screened against 656 cancer cell lines, and rhabdomyosarcoma, neuroblastoma and kidney cancer cells were identified as significantly sensitive to both LIMK inhibitors. These large-scale screens have identified effective LIMK inhibitor drug combinations and sensitive cancer types. In addition, the LIMK inhibitory compounds CRT0105446 and CRT0105950 will enable further development of LIMK-targeted cancer therapy

Caspase-resistant ROCK1 expression prolongs survival of Eµ-Myc B cell lymphoma mice

Apoptosis is characterized by membrane blebbing and apoptotic body formation. Caspase cleavage of ROCK1 generates an active fragment that promotes actin-myosin mediated contraction and membrane blebbing during apoptosis. Expression of caspase-resistant non-cleavable ROCK1 (Rock1 NC) prolonged survival of mice that rapidly develop B cell lymphomas due to Eµ-Myc transgene expression. Eµ-Myc; Rock1 NC mice had significantly fewer bone marrow cells relative to Eµ-Myc mice expressing wild-type ROCK1 (Rock1 WT), which was associated with altered cell cycle profiles. Circulating macrophage numbers were lower in Eµ-Myc; Rock1 NC mice, but there were higher levels of bone marrow macrophages, consistent with spontaneous cell death in Eµ-Myc; Rock1 NC mice bone marrows being more inflammatory. Rock1 WT recipient mice transplanted with pre-neoplastic Eµ-Myc; Rock1 NC bone marrow cells survived longer than mice transplanted with Eµ-Myc; Rock1 WT cells, indicating that the survival benefit was intrinsic to the Eµ-Myc; Rock1 NC bone marrow cells. The results suggest that the apoptotic death of Eµ-Myc; Rock1 NC cells generates a proliferation-suppressive microenvironment in bone marrows that reduces cell numbers and prolongs B cell lymphoma mouse survival

The production of hydrogen through the use of a 77 wt% Pd 23 wt% Ag membrane water gas shift reactor

Hydrogen as an energy carrier has the potential to decarbonize the energy sector. This work presents the application of a palladium-silver (PdeAg) membrane-based reactor. The membrane reactor which is made from PdeAg film supported by porous stainless steel (PSS) is evaluated for the production of hydrogen and the potential replacement of the current two-stage Water-Gas Shift (WGS) reaction by a single stage reaction. The permeability of a 20 mmPdeAg membrane reactor was examined at 320° C, 380° C and 430° C. The effect of continuous hydrogen exposure on the PdeAg membrane at high temperature and low temperature was examined to investigate the thermal stability and durability of the membrane. During continuous operation to determine thermal stability, the membrane reactor exhibited stable hydrogen permeation at 320° C for 120 h and unstable hydrogen permeation at 430° C was observed. For the WGS reaction, the reactor was loaded with Ferrochrome catalyst. The membrane showed the ability to produce high purity hydrogen, with a CO conversion and an H2 recovery of 84% and 88%, respectively. The membrane suffered from hydrogen embrittlement due to desorption and adsorption of hydrogen on the membrane surface. SEM analysis revealed cracks that occurred on the surface of the membrane after hydrogen exposure. XRD analysis revealed lattice expansion after hydrogen loading which suggests the occurrence of phase change from a-phase to the more brittle b-phase.DHE