Regulation of CCL2 expression in human vascular endothelial cells by a neighboring divergently transcribed long noncoding RNA.

Atherosclerosis is a chronic inflammatory disease that is driven, in part, by activation of vascular endothelial cells (ECs). In response to inflammatory stimuli, the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway orchestrates the expression of a network of EC genes that contribute to monocyte recruitment and diapedesis across the endothelium. Although many long noncoding RNAs (lncRNAs) are dysregulated in atherosclerosis, they remain poorly characterized, especially in the context of human vascular inflammation. Prior studies have illustrated that lncRNAs can regulate their neighboring protein-coding genes via interaction with protein complexes. We therefore identified and characterized neighboring interleukin-1ß (IL-1ß)-regulated messenger RNA (mRNA)-lncRNA pairs in ECs. We found these pairs to be highly correlated in expression, especially when located within the same chromatin territory. Additionally, these pairs were predominantly divergently transcribed and shared common gene regulatory elements, characterized by active histone marks and NF-κB binding. Further analysis was performed on lncRNA-CCL2, which is transcribed divergently to the gene, CCL2, encoding a proatherosclerotic chemokine. LncRNA-CCL2 and CCL2 showed coordinate up-regulation in response to inflammatory stimuli, and their expression was correlated in unstable symptomatic human atherosclerotic plaques. Knock-down experiments revealed that lncRNA-CCL2 positively regulated CCL2 mRNA levels in multiple primary ECs and EC cell lines. This regulation appeared to involve the interaction of lncRNA-CCL2 with RNA binding proteins, including HNRNPU and IGF2BP2. Hence, our approach has uncovered a network of neighboring mRNA-lncRNA pairs in the setting of inflammation and identified the function of an lncRNA, lncRNA-CCL2, which may contribute to atherogenesis in humans.

Dynamic regulation of VEGF-inducible genes by an ERK/ERG/p300 transcriptional network.

The transcriptional pathways activated downstream of vascular endothelial growth factor (VEGF) signaling during angiogenesis remain incompletely characterized. By assessing the signals responsible for induction of the Notch ligand delta-like 4 (DLL4) in endothelial cells, we find that activation of the MAPK/ERK pathway mirrors the rapid and dynamic induction of DLL4 transcription and that this pathway is required for DLL4 expression. Furthermore, VEGF/ERK signaling induces phosphorylation and activation of the ETS transcription factor ERG, a prerequisite for DLL4 induction. Transcription of DLL4 coincides with dynamic ERG-dependent recruitment of the transcriptional co-activator p300. Genome-wide gene expression profiling identified a network of VEGF-responsive and ERG-dependent genes, and ERG chromatin immunoprecipitation (ChIP)-seq revealed the presence of conserved ERG-bound putative enhancer elements near these target genes. Functional experiments performed in vitro and in vivo confirm that this network of genes requires ERK, ERG and p300 activity. Finally, genome-editing and transgenic approaches demonstrate that a highly conserved ERG-bound enhancer located upstream of HLX (which encodes a transcription factor implicated in sprouting angiogenesis) is required for its VEGF-mediated induction. Collectively, these findings elucidate a novel transcriptional pathway contributing to VEGF-dependent angiogenesis.

Improving the health assistant in nursing employment model through entry to practice nursing student perceptions: a cross-sectional study.

Background: A model employing entry to practice nursing students as health assistants in nursing (HANs) was developed to support the nursing workforce.Objective: This research examines the perceived benefits of being employed as a HAN to graduate entry to practice nursing students.Design: Cross-sectional studyMethods: A web-based 33-item questionnaire was open to entry to practice nursing students employed as a HAN. Categorical measures were summarised using descriptive statistics. Qualitative comments were analysed thematically.Results: Of the 39 enrolled participants, 38 (97.4%) completed the study. Fifteen students (39.5%) commenced a graduate year in 2019, 12 (31.6%) commenced a graduate year in 2020 and 11 (28.9%) will commence a graduate year in 2021. The participants viewed HAN employment as an opportunity to further develop as nurses and suggested that the HAN scope of practice expand to include their existing nursing scope of practice, thereby optimising their learning experience.Conclusion: The HAN model is viewed as a valuable model of employment by entry to practice nursing students; however, the HAN scope of practice is viewed as too limited and not conducive to self-development. This may impact on the retention of HAN employees as they transition to registered graduate nurse employment.Impact statement: Graduate entry to practice nursing students offer direction on how to improve the Health Assistants in Nursing employment model.

Advancing Nursing Informatics Through Clinical Placements: Pilot Study.

Work integrated learning in the space of nursing informatics is a new concept in Australian nursing curriculum. This study examined nursing students' experiences in the pilot nursing informatics clinical placement centered on electronic medical records, their use in patient care and clinical decision making. Students completed reflective diaries of their learning during the four-week placement. Data was explored by thematic analysis. Emergent themes included: importance of adequate training in using EMR; impact of EMR on nursing workflow and patient care; shaping future career choices; forming rewarding relationships; and potential for improvements. These themes will be used to enhance teaching and learning opportunities as this pilot placement evolves into permeant part of the nursing curriculum.

Conserved regulatory logic at accessible and inaccessible chromatin during the acute inflammatory response in mammals.

The regulatory elements controlling gene expression during acute inflammation are not fully elucidated. Here we report the identification of a set of NF-κB-bound elements and common chromatin landscapes underlying the acute inflammatory response across cell-types and mammalian species. Using primary vascular endothelial cells (human/mouse/bovine) treated with the pro-inflammatory cytokine, Tumor Necrosis Factor-α, we identify extensive (~30%) conserved orthologous binding of NF-κB to accessible, as well as nucleosome-occluded chromatin. Regions with the highest NF-κB occupancy pre-stimulation show dramatic increases in NF-κB binding and chromatin accessibility post-stimulation. These 'pre-bound' regions are typically conserved (~56%), contain multiple NF-κB motifs, are utilized by diverse cell types, and overlap rare non-coding mutations and common genetic variation associated with both inflammatory and cardiovascular phenotypes. Genetic ablation of conserved, 'pre-bound' NF-κB regions within the super-enhancer associated with the chemokine-encoding CCL2 gene and elsewhere supports the functional relevance of these elements.