DNA-Based Epigenetic Changes in Recurrent and Tamoxifen-Resistant Breast Cancer

Roughly two-thirds of all breast cancers are Estrogen Receptor a (ER)-positive and can be treated with an anti-estrogen such as Tamoxifen, however resistance occurs in 33% of women who take the drug for more than 5 years. In addition to this acquired antiestrogen resistance, de novo- or intrinsic-resistance occurs primarily in ER-negative tumors but also occasionally in ER-positive tumors. Aberrant DNA promoter methylation, a major epigenetic mechanism by which gene expression is altered in cancer, is thought to play a role in this resistance. To date, few studies have examined promoter methylation and Tamoxifen resistance in breast cancer. Of the studies conducted, one detected drug-specific promoter methylation and gene expression profiles in an ER-positive, Tamoxifen-selected MCF-7 derivative cell line. However, studies using both ER-positive and –negative, Tamoxifen-selected cell lines have not been described until now. To develop an understanding of Tamoxifen-resistance and identify novel pathways and targets of aberrant methylation, I first analyzed two Tamoxifen-resistant clones of MCF-7, one that retained expression of ER (TMX2-11) and one that lost expression of the gene (TMX2-28) after 6-months of Tamoxifen treatment, by Illumina HumanMethylation450 BeadChip (HM450BC). I found that prolonged treatment with Tamoxifen induced hypermethylation and hypomethylation throughout the genome. Compared to MCF-7, the ER-positive line, TMX2-11 had 4,000 hypermethylated sites, while the ER-negative line, TMX2-28 had over 33,000. Analysis of CpG sites in both TMX2-11 and TMX2-28 revealed that the two Tamoxifen-selected lines share 3,000 hypermethylated CpG sites with 21% of those sites being located in the promoter region. Promoter methylation and expression of two genes, MAGED1 and ZNF350, in both Tamoxifen-resistant cell lines demonstrated cell line-specific responses to treatment with 5-aza-2’deoxycitidine (5-Aza). Sixteen additional genes involved in signal transduction, cell adhesion, transcriptional repression, inflammatory response, cell proliferation and hormone response were chosen for further analysis based on their shared hypermethylation or their reduced expression in TMX2-28 as detected in a previously completed expression array. Five genes, RORA, THBS1, CAV2, TGFβ2, and BMP2 had decreased expression in TMX2-28, but not TMX2-11 as compared to MCF-7, and 5-Aza increased expression of the genes. This indicates that Tamoxifen is affecting a set of genes similarly in both the ER-positive and -negative breast cancer cell lines, however overall methylation changes are more pronounced in the ER-negative line. Our data as well as others suggest that DNA methylation may be contributing to Tamoxifen-resistance. I hypothesized that both ER-positive and ER-negative second human breast tumors occurring after anti-estrogen treatment would be hypermethylated. I characterized the methylation profiles of 70 human breast tumor samples using the HM450BC. These data confirm previous findings that ER-positive breast tumors have more hypermethylated CpG sites than ER-negative tumors. Stratification of the tumors by ER-positive first and second tumor sets shows that methylation is greater in first tumors.. Additionally, I saw that first tumors from ipsilateral pairs had higher methylation than the second tumors; in contrast, second tumors from contralateral pairs had higher methylation than in the first tumor. These data, together with the fact that tumor progression is associated with an increase in methylation, are consistent with the prediction that ipsilateral, not contralateral, tumors are more likely to be a true recurrence. Pathway analysis was conducted to provide insight into biomarkers associated with tumors that recur. Two pathways, ‘homophilic cell adhesion via plasma membrane adhesion molecules’ and ‘cell fate commitment’, were selected for further analysis. ER-positive first tumors that recurred as either ER-positive or ER-negative compared with non-recurrent tumors shared hypermethylated genes in the homophilic cell adhesion pathway. ER-positive first tumors that recurred as ER-negative compared with ER-positive first tumors that recurred as ER-positive were associated with a unique set of hypermethylated genes in the cell fate commitment pathway. To examine the association of methylation changes in my tumor data set with breast cancer patient survival data, Kaplan-Meier plots were created using TGCA breast cancer data available online. Expression of the genes only hypermethylated in each individual comparison group in the homophilic cell adhesion pathway was linked to overall survival. These data suggest that the genes hypermethylated only in ER-positive tumors recurring as ER-negative are a potential signature for poor survival. The underlying mechanisms of anti-estrogen resistance are poorly understood. Variable responses to breast cancer therapy highlights the need for biomarkers that can effectively guide treatment. The findings presented here underscore the potential use of breast tumor stratification based on methylation biomarkers in guiding treatment

Sav1 Loss Induces Senescence and Stat3 Activation Coinciding with Tubulointerstitial Fibrosis

Tubulointerstitial fibrosis (TIF) is recognized as a final phenotypic manifestation in the transition from chronic kidney disease (CKD) to end-stage renal disease (ESRD). Here we show that conditional inactivation of Sav1 in the mouse renal epithelium resulted in upregulated expression of profibrotic genes and TIF. Loss of Sav1 induced Stat3 activation and a senescence-associated secretory phenotype (SASP) that coincided with the development of tubulointerstitial fibrosis. Treatment of mice with the YAP inhibitor verteporfin (VP) inhibited activation of genes associated with senescence, SASPs, and activation of Stat3 as well as impeded the development of fibrosis. Collectively, our studies offer novel insights into molecular events that are linked to fibrosis development from Sav1 loss and implicate VP as a potential pharmacological inhibitor to treat patients at risk for developing CKD and TIF

Communication Strategies for Connecting OneVCU

This project analyzes and makes improvement suggestions for communication to VCU faculty and staff. The COVID-19 crisis has brought to light a variety of communication needs. Faculty and staff are feeling overwhelmed by a flood of emails, while at the same time are missing key information. We have assessed the current communication infrastructure, and together with the key VCU communicators have identified improved processes and policies

Slow progressors to type 1 diabetes lose islet autoantibodies over time, have few islet antigen-specific CD8+ T cells and exhibit a distinct CD95hi B cell phenotype

ims/hypothesis The aim of this study was to characterise islet autoantibody profiles and immune cell phenotypes in slow progressors to type 1 diabetes. Methods Immunological variables were compared across peripheral blood samples obtained from slow progressors to type 1 diabetes, individuals with newly diagnosed or long-standing type 1 diabetes, and healthy individuals. Polychromatic flow cytometry was used to characterise the phenotypic attributes of B and T cells. Islet autoantigen-specific B cells were quantified using an enzyme-linked immunospot (ELISpot) assay and islet autoantigen-specific CD8+ T cells were quantified using peptide–HLA class I tetramers. Radioimmunoassays were used to detect islet autoantibodies. Sera were assayed for various chemokines, cytokines and soluble receptors via ELISAs. Results Islet autoantibodies were lost over time in slow progressors. Various B cell subsets expressed higher levels of CD95 in slow progressors, especially after polyclonal stimulation, compared with the corresponding B cell subsets in healthy donors (p < 0.05). The phenotypic characteristics of CD4+ and CD8+ T cells were similar in slow progressors and healthy donors. Lower frequencies of CD4+ T cells with a central memory phenotype (CD27int, CD127+, CD95int) were observed in slow progressors compared with healthy donors (mean percentage of total CD4+ T cells was 3.00% in slow progressors vs 4.67% in healthy donors, p < 0.05). Autoreactive B cell responses to proinsulin were detected at higher frequencies in slow progressors compared with healthy donors (median no. of spots was 0 in healthy donors vs 24.34 in slow progressors, p < 0.05) in an ELISpot assay. Islet autoantigen-specific CD8+ T cell responses were largely absent in slow progressors and healthy donors. Serum levels of DcR3, the decoy receptor for CD95L, were elevated in slow progressors compared with healthy donors (median was 1087 pg/ml in slow progressors vs 651 pg/ml in healthy donors, p = 0.06). Conclusions/interpretation In this study, we found that slow progression to type 1 diabetes was associated with a loss of islet autoantibodies and a distinct B cell phenotype, consistent with enhanced apoptotic regulation of peripheral autoreactivity via CD95. These phenotypic changes warrant further studies in larger cohorts to determine their functional implications

Physical activity across the curriculum: year one process evaluation results

<p>Abstract</p> <p>Background</p> <p>Physical Activity Across the Curriculum (PAAC) is a 3-year elementary school-based intervention to determine if increased amounts of moderate intensity physical activity performed in the classroom will diminish gains in body mass index (BMI). It is a cluster-randomized, controlled trial, involving 4905 children (2505 intervention, 2400 control).</p> <p>Methods</p> <p>We collected both qualitative and quantitative process evaluation data from 24 schools (14 intervention and 10 control), which included tracking teacher training issues, challenges and barriers to effective implementation of PAAC lessons, initial and continual use of program specified activities, and potential competing factors, which might contaminate or lessen program effects.</p> <p>Results</p> <p>Overall teacher attendance at training sessions showed exceptional reach. Teachers incorporated active lessons on most days, resulting in significantly greater student physical activity levels compared to controls (p < 0.0001). Enjoyment ratings for classroom-based lessons were also higher for intervention students. Competing factors, which might influence program results, were not carried out at intervention or control schools or were judged to be minimal.</p> <p>Conclusion</p> <p>In the first year of the PAAC intervention, process evaluation results were instrumental in identifying successes and challenges faced by teachers when trying to modify existing academic lessons to incorporate physical activity.</p

Environmental attitudes associated with large-scale cultural differences, not local environmental conflicts

Species and ecosystems are under constant pressure from a rapidly-growing human population. Human tolerance of carnivores, including the willingness to live in areas with these predators, is key to the success of large carnivore conservation. In the Scandinavian Peninsula, large carnivore populations conflict with human activity; low tolerance among local people may lead to illegal hunting. A survey of 2521 Scandinavian respondents to measure environmental value orientation, using the new environmental paradigm (NEP) scale and attitudes toward large carnivores, revealed attitudes towards the presence of carnivores were not related to carnivore abundance. Nor was there a significant relationship between environmental value orientation and personal experiences with loss of domestic sheep or hunting dogs. Environmental values were mainly explained by country differences; Swedes had a more ecocentric value orientation than Norwegians. Significantly more Norwegians (45 %) than Swedes (19 %) responded that there were too many carnivores in their country. Historic differences in how government is perceived between Norway and Sweden may result in different attitudes towards illegal hunting and towards carnivores. Specifically, Norwegians may hold a more anthropocentric view, based on a suspicion of central authorities, whereas Swedes may hold a more ecocentric view

Physical Activity Across the Curriculum (PAAC): a randomized controlled trial to promote physical activity and diminish overweight and obesity in elementary school children

Objective Physical Activity Across the Curriculum (PAAC) was a three-year cluster randomized controlled trial to promote physical activity and diminish increases in overweight and obesity in elementary school children. Methods Twenty-four elementary schools were cluster randomized to the PAAC intervention or served as control. All children in grades two and three were followed to grades four and five. PAAC promoted 90 minutes/wk of moderate to vigorous intensity physically active academic lessons delivered by classroom teachers. BMI was the primary outcome, daily PA and academic achievement were secondary outcomes. Results The three-year change in BMI for PAAC was 2.0 ± 1.9 and control 1.9 ± 1.9, respectively (NS). However, change in BMI from baseline to three years was significantly influenced by exposure to PAAC. Schools with ≥75 minutes of PAAC/wk showed significantly less increase in BMI at three years compared to schools that had <75 minutes of PAAC (1.8 ± 1.8 vs. 2.4 ± 2.0, p=0.02). PAAC schools had significantly greater changes in daily PA and academic achievement scores. Conclusions The PAAC approach may promote daily PA and academic achievement in elementary school children. Additionally, 75 minutes of PAAC activities may attenuate increases in BMI

word~river literary review (2009)

wordriver is a literary journal dedicated to the poetry, short fiction and creative nonfiction of adjuncts and part-time instructors teaching in our universities, colleges, and community colleges. Our premier issue was published in Spring 2009. We are always looking for work that demonstrates the creativity and craft of adjunct/part-time instructors in English and other disciplines. We reserve first publication rights and onetime anthology publication rights for all work published. We define adjunct instructors as anyone teaching part-time or full-time under a semester or yearly contract, nationwide and in any discipline. Graduate students teaching under part-time contracts during the summer or who have used up their teaching assistant time and are teaching with adjunct contracts for the remainder of their graduate program also are eligible.https://digitalscholarship.unlv.edu/word_river/1002/thumbnail.jp