Protein kinase a-regulated assembly of a MEF2·HDAC4 repressor complex controls c-jun expression in vascular smooth muscle cells

Vascular smooth muscle cells (VSMCs) maintain the ability to modulate their phenotype in response to changing environmental stimuli. This phenotype modulation plays a critical role in the development of most vascular disease states. In these studies, stimulation of cultured vascular smooth muscle cells with platelet-derived growth factor resulted in marked induction of c-jun expression, which was attenuated by protein kinase Cδ and calcium/calmodulin-dependent protein kinase inhibition. Given that these signaling pathways have been shown to relieve the repressive effects of class II histone deacetylases (HDACs) on myocyte enhancer factor (MEF) 2 proteins, we ectopically expressed HDAC4 and observed repression of c-jun expression. Congruently, suppression of HDAC4 by RNA interference resulted in enhanced c-jun expression. Consistent with these findings, mutation of the MEF2 cis-element in the c-jun promoter resulted in promoter activation during quiescent conditions, suggesting that the MEF2 cis-element functions as a repressor in this context. Furthermore, we demonstrate that protein kinase A attenuates c-Jun expression by promoting the formation of a MEF2-HDAC4 repressor complex by inhibiting salt-inducible kinase 1. Finally, we document a physical interaction between c-Jun and myocardin, and we document that forced expression of c-Jun represses the ability of myocardin to activate smooth muscle gene expression. Thus, MEF2 and HDAC4 act to repress c-Jun expression in quiescent VSMCs, protein kinase A enhances this repression, and platelet-derived growth factor derepresses c-Jun expression through calcium/calmodulin-dependent protein kinases and novel protein kinase Cs. Regulation of this molecular "switch" on the c-jun promoter may thus prove critical for toggling between the activated and quiescent VSMC phenotypes

In vitro and in vivo mRNA delivery using lipid-enveloped pHresponsive polymer nanoparticles

Biodegradable core−shell structured nanoparticles with a poly(β-amino ester) (PBAE) core enveloped by a phospholipid bilayer shell were developed for in vivo mRNA delivery with a view toward delivery of mRNA-based vaccines. The pH-responsive PBAE component was chosen to promote endosome disruption, while the lipid surface layer was selected to minimize toxicity of the polycation core. Messenger RNA was efficiently adsorbed via electrostatic interactions onto the surface of these net positively charged nanoparticles. In vitro, mRNA-loaded particle uptake by dendritic cells led to mRNA delivery into the cytosol with low cytotoxicity, followed by translation of the encoded protein in these difficult-to-transfect cells at a frequency of 30%. Particles loaded with mRNA administered intranasally (i.n.) in mice led to the expression of the reporter protein luciferase in vivo as soon as 6 h after administration, a time point when naked mRNA given i.n. showed no expression. At later time points, luciferase expression was detected in naked mRNA-treated mice, but this group showed a wide variation in levels of transfection, compared to particle-treated mice. This system may thus be promising for noninvasive delivery of mRNA-based vaccines.United States. Dept. of Defense (Institute for Soldier Nanotechnology, contract W911NF-07-D-0004)Ragon Institute of MGH, MIT and HarvardSingapore. Agency for Science, Technology and ResearchHoward Hughes Medical Institute (Investigator

Productive margins, regulating for engagement: Seeing regulation differently 2013-2018

'Seeing regulation differently’ is one of seven projects under the ‘Productive Margins: Regulating for Engagement’ programme of research, co-produced between academics and community organisations in South Wales and Bristol. ‘Seeing regulation differently’ was a cross-programme project, starting later than many of the other projects (2016-2018) and drawing on discussions at project and programme level, exploring how regulation is seen and talked about by community organisations, researchers and community participants. The data collected were from two elements included in this project: 1. A year-long artists residency with artists Close and Remote (Sept 2016 - Sept 2017); 2. Discussions about regulation within the programme, including workshops with academics from several universities, specialising in regulation theory. Information about the regulation project including the residency can be found on the Productive Margins website and will be the subject of a book to be published in December 2018 by Policy Press. ‘Seeing regulation differently’ is one of seven projects under the ‘Productive Margins: Regulating for Engagement’ programme of research, co-produced between academics and community organisations in South Wales and Bristol. ‘Seeing regulation differently’ was a cross-programme project, starting later than many of the other projects (2016-2018) and drawing on discussions at project and programme level, exploring how regulation is seen and talked about by community organisations, researchers and community participants. </p

Productive margins, regulation for engagement: Isolation and loneliness of older people 2015-2016

This project co-produced research exploring the isolation and loneliness of older people (ILOP) in an area of South Bristol (Greater Bedminster) in the UK. Data was collected through (1) A postcard writing activity with older people (2) Interviews with older people (3) A focus group with people who worked in community services/activities for older people (voluntary and statutory) and (4) A focus group with members of the community research group. Apart from (1), data is not stored as there was no consent to share. The research was part of the Productive Margins: Regulating for Engagement programme.Busy streets, laughter, the sound of children playing - but what lies behind closed doors in our communities, especially for those in later life, who may feel lonely and isolated.</p

Productive margins, regulation for engagement: Who decides what's in my fridge 2015-2016

Data was collected through two surveys that were undertaken in Knowle West, and interviews that were carried out with working group members (Academic and community organisation staff on the research team) and community participants. For the purpose of storage, data has been anonymised only when requested with some names replaced and no surnames included. Much of the ground work was carried out in community-based workshops. Who Decides What’s in my Fridge was a co-produced research project exploring how people experienced the regulation of their food habits. The project was a collaboration between the University of Bristol and three community organisations in Bristol; Knowle West Media Centre (KWMC) in Knowle West, Single Parent Action Network (SPAN) in Easton, and the Community Kitchen at Coexist in Stokes Croft. A researcher was based in the community working with groups and individuals from the nearby areas of two of the partner organisations involved; a group of local residents in Knowle West (working with KWMC) and a group of Somali women in Easton (working with SPAN). The researcher also worked with a group of eight ‘Junior Digital Producers’ that were employed by KWMC to collaborate on the project. The group received research training and the data sets were collected by the group members with the researcher between October 2015 and January 2016. </p

Productive margins, regulating for engagement: Women and data futures 2017

Workshops data, consisting of notes made by the University of Bristol researcher of workshops held in each locality. Topics included data privacy, understanding of personal and big data, the role of digital data in participants’ neighbourhoods, and how big data companies use personal data. 'Women and Data Futures’ was a joint project and knowledge exchange between two community organisations: the Knowle West Media Centre (KWMC) in Bristol and the 3Gs Community Development Centre (3Gs) in Merthyr Tydfil, South Wales. Both organisations were involved as partners in the Productive Margins: Regulating for Engagement programme. This project aimed to explore perceptions of local women about the use of the personal data they put online (such as on Facebook and Snapchat), and how they could take some control over their personal information online. Data was collected through six workshops in each location between February and March 2017, and a workshop with both groups together in May 2017. Further information including two podcasts, one based on sound recordings from the workshops, and the other from a public symposium on data held in October 2017, are available on the KWMC website.</p