Faculty Bibliography

BACKGROUND: Posttranslational histone modifications (PTHMs) are altered by arsenic, an environmental carcinogen. PTHMs are also influenced by nutritional methyl donors involved in one-carbon metabolism (OCM), which may protect against epigenetic dysregulation. METHODS: We measured global levels of three PTHMs, which are dysregulated in cancers (H3K36me2, H3K36me3, H3K79me2), in peripheral blood mononuclear cells (PBMCs) from 324 participants enrolled in the Folic Acid and Creatine Trial, a randomized trial in arsenic-exposed Bangladeshi adults. Sex-specific associations between several blood OCM indices (folate, vitamin B12, choline, betaine, homocysteine) and PTHMs were examined at baseline using regression models, adjusted for multiple tests by controlling for the false discovery rate (PFDR). We also evaluated the effects of FA supplementation (400 microg/day for 12 weeks), compared with placebo, on PTHMs. RESULTS: Associations between choline and H3K36me2 and between vitamin B12 and H3K79me2, differed significantly by sex (Pdiff < 0.01 and < 0.05, respectively). Among men, plasma choline was positively associated with H3K36me2 (PFDR < 0.05), and among women, plasma vitamin B12 was positively associated with H3K79me2 (PFDR < 0.01). FA supplementation did not alter any of the PTHMs examined (PFDR = 0.80). CONCLUSIONS: OCM indices may influence PTHMs in a sex-dependent manner, and FA supplementation, at this dose and duration, does not alter PTHMs in PBMCs. IMPACT: This is the first study to examine the influences of OCM indices on PTHMs in a population that may have increased susceptibility to cancer development due to widespread exposure to arsenic-contaminated drinking water and a high prevalence of hyperhomocysteinemia.

Both nickel and cadmium compounds have been established as group I carcinogens for several decades. Despite over-whelming evidence of these compounds' carcinogenicity in humans, the specific underlying molecular mechanisms that govern metal induced cellular transformation remain unclear. In this study, we found that there were slightly different effects on decreased SLBP mRNA and protein as well as increased polyA H3.1 in our nickel exposed cells. This suggested that nickel and arsenic have similar effects on canonical histone mRNA transcription and translation. We also saw that the depletion of SLBP protein was reversed by inhibiting the proteosome. Finally, we showed that inhibiting the SLBP mRNA and protein levels were rescued by epigenetic modifiers suggesting that nickel's effects on SLBP may be mediated via epigenetic mechanisms. Taken together these results suggest a similar mechanism by which both arsenic and nickel may exert their carcinogenic effects.

This study reports levels and profiles of polycyclic aromatic hydrocarbons (PAHs) in dust samples collected from three different microenvironments (cars, air conditioner (AC) filters and household floor dust) of Jeddah, Saudi Arabia (KSA) and Kuwait. To the best of our knowledge, this is first study reporting PAHs in indoor microenvironments of KSA, which makes these findings important. Benzo(b)fluoranthene (BbF), benzo(a)pyrene (BaP), phenanthrene (Phe), and pyrene (Pyr) were found to be the major chemicals in dust samples from all selected microenvironments. SigmaPAHs occurred at median concentrations (ng/g) of 3450, 2200, and 2650 in Saudi AC filter, car and household floor dust, respectively. The median levels (ng/g) of SigmaPAHs in Kuwaiti car (950) and household floor (1675) dust samples were lower than Saudi dust. The PAHs profile in Saudi dust was dominated by high molecular weight (HMW) (4-5 ring) PAHs while in Kuwaiti dust 3 ring PAHs have marked contribution. BaP equivalent, a marker for carcinogenic PAHs, was high in Saudi household floor and AC filter dust with median levels (ng/g) of 370 and 455, respectively. Different exposure scenarios, using 5th percentile, median, mean, and 95th percentile levels, were estimated for adults and toddlers. For Saudi and Kuwaiti toddlers worst exposure scenario of SigmaPAHs was calculated at 175 and 85ng/kg body weight/day (ng/kgbw/d), respectively. For Saudi toddlers, the calculated worst exposure scenarios for carcinogenic BaP (27.7) and BbF (29.3ng/kgbw/d) was 2-4 times higher than Kuwaiti toddlers. This study is based on small number of samples which necessitate more detailed studies for better understanding of dynamics of PAHs in the indoor environments of this region. Nevertheless, our finding supports the ongoing exposure of organic pollutants to population that accumulates indoor.

BACKGROUND: Exposure to inorganic arsenic is associated with numerous adverse health outcomes, with susceptibility differing by sex. While evidence from in vitro studies suggests that arsenic alters post-translational histone modifications (PTHMs), evidence in humans is limited. OBJECTIVES: The objectives were to determine: 1) if arsenic exposure is associated with global (%) levels of PTHMs: H3K36me2, H3K36me3, and H3K79me2 in a sex-dependent manner and 2) if %PTHMs are stable when arsenic exposure is reduced. METHODS: We examined associations between arsenic, measured in blood and urine, and %PTHMs in peripheral blood mononuclear cells from 317 participants enrolled in the Bangladesh Folic Acid and Creatine Trial (FACT). We also examined the stability of %PTHMs after the use of arsenic-removal water filters (n = 60). RESULTS: Associations between natural log-transformed (ln) urinary arsenic, adjusted for creatinine (uAsCr), and %H3K36me2 differed significantly between men and women (p = 0.01). Ln-uAsCr was positively associated with %H3K36me2 in men (beta = 0.12; 95% CI: 0.01, 0.23, p = 0.03), but was negatively associated with %H3K36me2 in women (beta = -0.05; 95% CI: -0.12, 0.02, p = 0.19). The patterns of associations with blood arsenic were similar. On average, water filter use was also associated with reductions in %H3K36me2 (p < 0.01), but this did not differ significantly by sex. Arsenic was not significantly associated with %H3K36me3 or %H3K79me2 in men or women. CONCLUSIONS: Arsenic exposure was associated with %H3K36me2 in a sex-specific manner, but was not associated with %H3K36me3 or %H3K79me2. Additional studies are needed to assess changes in %H3K36me2 after arsenic removal.

Oxygen levels range from 2-9% in vivo. Atmospheric O2 levels (21%) are known to induce cell proliferation defects and cellular senescence in primary cell cultures. However, the mechanistic basis of the deleterious effects of higher O2 levels is not fully understood. On the other hand, immortalized cells including cancer cell lines, which evade cellular senescence are normally cultured at 21% O2 and the effects of higher O2 on these cells are understudied. Here we addressed this problem by culturing immortalized human bronchial epithelial (BEAS-2B) cells at ambient atmospheric, 21% O2 and lower, 10% O2 . Our results show increased inflammatory response at 21% O2 but not at 10% O2 . We found higher RelA binding at the NF-kappaB1/RelA target gene promoters as well as upregulation of several pro-inflammatory cytokines in cells cultured at 21% O2 . RelA knockdown prevented the upregulation of the pro-inflammatory cytokines at 21% O2 , suggesting NF-kappaB1/RelA as a major mediator of inflammatory response in cells cultured at 21% O2 . Interestingly, unlike the 21% O2 cultured cells, exposure of 10% O2 cultured cells to H2 O2 did not elicit inflammatory response, suggesting increased ability to tolerate oxidative stress in cells cultured at lower O2 levels

Oxygen (O2) levels range from 2-9% in vivo. However, cell culture experiments are performed at atmospheric O2 levels (21%). Oxidative stress due to generation of reactive oxygen species (ROS) in cells cultured at higher than physiological levels is implicated in multitude of deleterious effects including DNA damage, genomic instability and senescence. In addition, oxidative stress activates redox sensitive transcription factors related to inflammatory signaling and apoptotic signaling. Furthermore, several chromatin-modifying enzymes are affected by ROS, potentially impacting epigenetic regulation of gene expression. While primary cells are cultured at lower O2 levels due to their inability to grow at higher O2, the immortalized cells, which display no such apparent growth difficulties, are typically cultured at 21% O2. This review will provide an overview of issues associated with increased oxygen levels in in vitro cell culture and point out the benefits of using lower levels of oxygen tension even for immortalized cells.

The special AT-rich sequence-binding protein 2 (SATB2) is a protein that binds to the nuclear matrix attachment region of the cell and regulates gene expression by altering chromatin structure. In our previous study, we reported that SATB2 gene expression was induced in human bronchial epithelial BEAS-2B cells transformed by arsenic, chromium, nickel and vanadium. In this study, we show that ectopic expression of SATB2 in the normal human bronchial epithelial cell-line BEAS-2B increased anchorage-independent growth and cell migration, meanwhile, shRNA-mediated knockdown of SATB2 significantly decreased anchorage-independent growth in Ni transformed BEAS-2B cells. RNA sequencing analyses of SATB2 regulated genes revealed the enrichment of those involved in cytoskeleton, cell adhesion and cell-movement pathways. Our evidence supports the hypothesis that SATB2 plays an important role in BEAS-2B cell transformation.

Ultraviolet radiation (UVR) from sunlight is the primary effector of skin DNA damage. Chromatin remodeling and histone post-translational modification (PTM) are critical factors in repairing DNA damage and maintaining genomic integrity, however, the dynamic changes of histone marks in response to solar UVR are not well characterized. Here we report global changes in histone PTMs induced by solar simulated UVR (ssUVR). A decrease in lysine acetylation of histones H3 and H4, particularly at positions of H3 lysine 9, lysine 56, H4 lysine 5, and lysine 16, was found in human keratinocytes exposed to ssUVR. These acetylation changes were highly associated with ssUVR in a dose-dependent and time-specific manner. Interestingly, H4K16ac, a mark that is crucial for higher order chromatin structure, exhibited a persistent reduction by ssUVR that was transmitted through multiple cell divisions. In addition, the enzymatic activities of histone acetyltransferases were significantly reduced in irradiated cells, which may account for decreased global acetylation. Moreover, depletion of histone deacetylase SIRT1 in keratinocytes rescued ssUVR-induced H4K16 hypoacetylation. These results indicate that ssUVR affects both HDAC and HAT activities, leading to reduced histone acetylation.