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Excel spreadsheet of NUAK1 inhibitor induced phospho-peptide alterations

Supplementary Methods

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Exploiting oxidative stress has recently emerged as a plausible strategy for treatment of human cancer, and antioxidant defenses are implicated in resistance to chemotherapy and radiotherapy. Targeted suppression of antioxidant defenses could thus broadly improve therapeutic outcomes. Here, we identify the AMPK-related kinase NUAK1 as a key component of the antioxidant stress response pathway and reveal a specific requirement for this role of NUAK1 in colorectal cancer. We show that NUAK1 is activated by oxidative stress and that this activation is required to facilitate nuclear import of the antioxidant master regulator NRF2: Activation of NUAK1 coordinates PP1β inhibition with AKT activation in order to suppress GSK3β-dependent inhibition of NRF2 nuclear import. Deletion of NUAK1 suppresses formation of colorectal tumors, whereas acute depletion of NUAK1 induces regression of preexisting autochthonous tumors. Importantly, elevated expression of NUAK1 in human colorectal cancer is associated with more aggressive disease and reduced overall survival. Significance: This work identifies NUAK1 as a key facilitator of the adaptive antioxidant response that is associated with aggressive disease and worse outcome in human colorectal cancer. Our data suggest that transient NUAK1 inhibition may provide a safe and effective means for treatment of human colorectal cancer via disruption of intrinsic antioxidant defenses. Cancer Discov; 8(5); 632–47. ©2018 AACR. This article is highlighted in the In This Issue feature, p. 517

Objectives: Little is known about the impact of HIV infection on biological ageing in sub-Saharan Africa. The study aimed to assess biological ageing in South African HIV-infected adults and HIV-seronegative individuals using two validated biomarkers, telomere length and CDKN2A expression (a mediator of cellular senescence). Design: A case–control study. Methods: Two hundred and thirty-six HIV-infected adults aged at least 30 years and 250 age and sex frequency matched HIV-seronegative individuals were recruited from clinics in township communities in Cape Town. Biological ageing was evaluated by measurement of telomere length and CDKN2A expression in peripheral blood leukocytes. Results: The median ages of the HIV-infected and HIV-seronegative participants were 39 and 40 years, respectively. Among HIV-infected participants, 87.1% were receiving antiretroviral therapy (ART), their median CD4+ cell count was 468 cells/μl and 84.3% had undetectable viral load. Both biomarkers were validated against chronological age in HIV-seronegative individuals. Telomere length was significantly shorter in HIV-infected individuals than in HIV-seronegative individuals (mean relative T/S ratio ±SE:0.91 ± 0.007 vs. 1.07 ± 0.008, P

The discovery that the 5'AMP-activated protein kinase (AMPK) serves to link the tumour suppressors LKB1 and the tuberous sclerosis complex and functions to slow macromolecular synthesis through attenuation of the mechanistic target of rapamycin complex 1 revealed a role for AMPK in tumour suppression. On the other hand, the well-recognized role of AMPK in maintaining ATP homeostasis, through suppression of anabolism and promotion of catabolism, as well as the role of AMPK in neutralizing reactive oxygen species, via maintenance of NADPH-dependent reductive capacity, point to tumour-protective roles in the context of metabolic stress, which is a key feature of many solid tumours. A growing number of studies thus suggest a duality of functions for AMPK that are either pro- or anti-cancer, depending upon context. Importantly, AMPK is composed of three subunits, and multiple isoforms exist for all three, allowing for different permutations to assemble and the potential for specific AMPK complexes to regulate distinct cellular processes. Moreover, certain subunits of the AMPK complex are frequently overexpressed in a spectrum of human cancer types, suggesting an outright oncogenic function for specific AMPK complexes. Adding complexity to this picture, the catalytic AMPK alpha subunits belong to a family of 14 kinases that can all be activated by LKB1 and studies are beginning to reveal a similar duality of roles in cancer for other members of the AMPK-related kinase family.

The thermochemistry of oxo transfer from coordinated nitrite in the dinitro(5,10,15,20-tetrakis(o-pivalamidophenyl)porphinato)iron(III) anion, ion-paired with the tetrapropylammonium ion, {[Fe(III)TpivPP(NO(2))(2)](-)Pr(4)N(+)}, has been evaluated in acetonitrile solution. This oxo-transfer half-reaction of {[Fe(III)TpivPP(NO(2))(2)](-)Pr(4)N(+)} has been assessed on the basis of the determination of the E(1/2) = +0.54 V vs SHE for the reversible [Fe(II/III)TpivPP(NO)(NO(2))](-)(/0) couple and the measurement of the formation constants for the association of NO and NO(2)(-) with the mononitroiron(III) porphyrin derivative. The formation constant for nitric oxide association, K(NO), has the value (1.21 +/- 0.08) x 10(3). The stability constant, K(2), for association of a second nitro ligand in 0.0100 M tetrapropylammonium perchlorate medium has been estimated as 2.18 x 10(3). The oxo-transfer half-reaction free energy, DeltaG degrees ((X/XO)), for addition of oxygen to [Fe(II)TpivPP(NO)(NO(2))](-) to form {[Fe(III)TpivPP(NO(2))(2)](-)Pr(4)N(+)} has been found to be -50 kJ/mol.

The New York Bight extends seaward some 80 to 100 miles (ca. 129 to 161 km) from the Long Island and New Jersey shorelines to the edge of the continental shelf. Over 14 × 10 6 m 3 of sewage sludge, dredge spoils, acid wastes, and cellar dirt are discharged into this area each year. Large populations of Bacillus sp. resistant to 20 μg of mercury per ml were observed in Bight sediments contaminated by these wastes. Resistant Bacillus populations were much greater in sediments containing high concentrations of Hg and other heavy metals than in sediments from areas further offshore where dumping has never been practiced and where heavy-metal concentrations were found to be low. Ampicillin resistance due mainly to β-lactamase production was significantly ( P < 0.001) more frequent in Bacillus strains from sediments near the sewage sludge dump site than in similar Bacillus populations from control sediments. Bacillus strains with combined ampicillin and Hg resistances were almost six times as frequent at the sludge dump site as in control sediments. This observation suggests that genes for Hg resistance and β-lactamase production are simultaneously selected for in Bacillus and that heavy-metal contamination of an ecosystem can result in a selection pressure for antibiotic resistance in bacteria in that system. Also, Hg resistance was frequently linked with other heavy-metal resistances and, in a substantial proportion of Bacillus strains, involved reduction to volatile metallic Hg (Hg°).

We undertook this study to determine the impact of urbanization on the microbial content of waters of a major recreational area of Lake Champlain. We followed changes in the numbers of total coliforms, fecal coliforms, fecal streptococci, and ascertained resistance of coliforms to heavy metals and antibiotics. We determined the distribution of these bacterial indicators of pollution by examining water and sediment samples. We also surveyed changes in physical parameters such as pH, temperature, turbidity, and wind direction to evaluate their effect on the examined aquatic microbial population. The level of all standard microbial indices of pollution frequently exceeded acceptable levels for recreational waters. Sediment samples indicated prolonged deposition of microbial pollutants, but efforts to demonstrate their release from sediment by dislocation and resuspension were inconclusive. We confirmed the presence of a population of heavy-metal and antibiotic-resistant coliforms isolated from harbor waters. Eighty-four percent of the total coliforms isolated from harbor water samples were resistant to one or more antibiotics and 96% were resistant to two or more heavy metals. The presence of these antibiotic resistant properties in coliforms, as well as elevated fecal coliform and fecal streptococci counts in both water and sediment samples, suggests to us that the bay is polluted by human fecal matter. This pollution probably emanates from south harbor sources and affects the shores of the major northern bathing area. It constitutes a potential hazard to public health because of the geophysical nature of the harbor and the location of the pollution source.