Regulation of pH by Carbonic Anhydrase 9 Mediates Survival of Pancreatic Cancer Cells With Activated KRAS in Response to Hypoxia.

Background & Aims Most pancreatic ductal adenocarcinomas (PDACs) express an activated form of KRAS, become hypoxic and dysplastic, and are refractory to chemo and radiation therapies. To survive in the hypoxic environment, PDAC cells upregulate enzymes and transporters involved in pH regulation, including the extracellular facing carbonic anhydrase 9 (CA9). We evaluated the effect of blocking CA9, in combination with administration of gemcitabine, in mouse models of pancreatic cancer. Methods We knocked down expression of KRAS in human (PK-8 and PK-1) PDAC cells with small hairpin RNAs. Human and mouse (KrasG12D/Pdx1-Cre/Tp53/RosaYFP) PDAC cells were incubated with inhibitors of MEK (trametinib) or extracellular signal-regulated kinase (ERK), and some cells were cultured under hypoxic conditions. We measured levels and stability of the hypoxia-inducible factor 1 subunit alpha (HIF1A), endothelial PAS domain 1 protein (EPAS1, also called HIF2A), CA9, solute carrier family 16 member 4 (SLC16A4, also called MCT4), and SLC2A1 (also called GLUT1) by immunoblot analyses. We analyzed intracellular pH (pHi) and extracellular metabolic flux. We knocked down expression of CA9 in PDAC cells, or inhibited CA9 with SLC-0111, incubated them with gemcitabine, and assessed pHi, metabolic flux, and cytotoxicity under normoxic and hypoxic conditions. Cells were also injected into either immune-compromised or immune-competent mice and growth of xenograft tumors was assessed. Tumor fragments derived from patients with PDAC were surgically ligated to the pancreas of mice and the growth of tumors was assessed. We performed tissue microarray analyses of 205 human PDAC samples to measure levels of CA9 and associated expression of genes that regulate hypoxia with outcomes of patients using the Cancer Genome Atlas database. Results Under hypoxic conditions, PDAC cells had increased levels of HIF1A and HIF2A, upregulated expression of CA9, and activated glycolysis. Knockdown of KRAS in PDAC cells, or incubation with trametinib, reduced the posttranscriptional stabilization of HIF1A and HIF2A, upregulation of CA9, pHi, and glycolysis in response to hypoxia. CA9 was expressed by 66% of PDAC samples analyzed; high expression of genes associated with metabolic adaptation to hypoxia, including CA9, correlated with significantly reduced survival times of patients. Knockdown or pharmacologic inhibition of CA9 in PDAC cells significantly reduced pHi in cells under hypoxic conditions, decreased gemcitabine-induced glycolysis, and increased their sensitivity to gemcitabine. PDAC cells with knockdown of CA9 formed smaller xenograft tumors in mice, and injection of gemcitabine inhibited tumor growth and significantly increased survival times of mice. In mice with xenograft tumors grown from human PDAC cells, oral administration of SLC-0111 and injection of gemcitabine increased intratumor acidosis and increased cell death. These tumors, and tumors grown from PDAC patient-derived tumor fragments, grew more slowly than xenograft tumors in mice given control agents, resulting in longer survival times. In KrasG12D/Pdx1-Cre/Tp53/RosaYFP genetically modified mice, oral administration of SLC-0111 and injection of gemcitabine reduced numbers of B cells in tumors. Conclusions In response to hypoxia, PDAC cells that express activated KRAS increase expression of CA9, via stabilization of HIF1A and HIF2A, to regulate pH and glycolysis. Disruption of this pathway slows growth of PDAC xenograft tumors in mice and might be developed for treatment of pancreatic cancer

The importance of language for language development: Linguistic determinism in the 1980s

The semantic and syntactic functions of verbs are the major aspects of linguistic complexity that contribute to the cognitive requirements for learning language between two and three years of age. Several contrastive categories of verbs emerged from our studies with action/state as the largest and most general. Contrastive subcategories of action verbs were locative/nonlocative action, durative/nondurative action, and completive/noncompletive action. The subcategories of state verbs were volitional/epistemic/notice/communication states. The psychological and linguistic validity of these semantic categories rests on their being coextensive with major grammatical developments and/or their sequential development

Impacts of air pollutants from rural Chinese households under the rapid residential energy transition

Rural residential energy consumption in China is experiencing a rapid transition towards clean energy, nevertheless, solid fuel combustion remains an important emission source. Here we quantitatively evaluate the contribution of rural residential emissions to PM2.5 (particulate matter with an aerodynamic diameter less than 2.5 μm) and the impacts on health and climate. The clean energy transitions result in remarkable reductions in the contributions to ambient PM2.5, avoiding 130,000 (90,000-160,000) premature deaths associated with PM2.5 exposure. The climate forcing associated with this sector declines from 0.057 ± 0.016 W/m2 in 1992 to 0.031 ± 0.008 W/m2 in 2012. Despite this, the large remaining quantities of solid fuels still contributed 14 ± 10 μg/m3 to population-weighted PM2.5 in 2012, which comprises 21 ± 14% of the overall population-weighted PM2.5 from all sources. Rural residential emissions affect not only rural but urban air quality, and the impacts are highly seasonal and location dependent

The evolution of fetal protection policies

This article examines the evolution of fetal protection policies (FPPs) by detailing their historical legacy and a range of contemporary social forces that have contributed to their maintenance. It is based on a case study of the 1977 U.S. Department of Labor, Occupational Safety and Health Administration (OSHA) hearings to revise the industrial lead standard, the 1991 U.S. Supreme Court decision that such policies are unconstitutional ( United Auto Workers v. Johnson Controls , 1991), and the case law preceding that decision. A primary issue is the notion that women and fetuses are disproportionately susceptible to lead. This study reveals the ways in which this belief is framed, disputed, and appropriated by various parties to the fetal protection policy debate. Implications of this case study for family health policy are also discussed.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/44656/1/10834_2006_Article_BF02353687.pd

Empty nest, cohort, and employment in the well-being of midlife women

Whether the empty-nest experience has positive or negative consequences for women's well-being at midlife may depend on their historical cohort membership and employment status. In this study, it was posited that the empty nest was likely to be a negative experience among the particular cohort of women (Cohort II) who reached adulthood during the period of strong societal emphasis on women's maternal role known as the feminine mystique, would be experienced positively among the earlier cohort (Cohort I) who as young adults were encouraged to enter the labor force during World War II. Analyses of covariance tested the relationships among empty-nest status, cohort membership, and employment status, and three measures of psychological well-being, adjusted for age, education, and marital status. The results of this study show that cohort and employment each have important independent associations with women's well-being at midlife, but that the experience of the empty nest depends on these two factors, especially cohort experiences.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/45589/1/11199_2004_Article_BF00287990.pd

A deadly double dose for India's poor

Cooking with solid fuels is a major source of early death and ill health among lower-income Indians. The same group also suffers disproportionately from air pollution generated by other, more general sources