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

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

Productive margins, regulating for engagement: Life chances, low income families in urban areas 2015-2016

‘Life Chances’ is a co-produced research project exploring life on a low income for families with children, and the regulatory systems they encounter in their everyday lives. The project was co-designed by academics from Bristol and Cardiff Universities, artists Close and Remote, and two community organisations: Single Parent Action Network in Bristol and South Riverside Community Development Centre in Cardiff. The project also aimed to co-produce the research with community participants in both areas. Data was collected through 11 workshops in each location, a joint visit to Chepstow castle and two workshops with both groups together, between December 2015 and June 2016. In addition, 13 individual interviews were undertaken by the Research Associate between January and May 2016. An evaluation session with both groups also took place in July 2016. The data stored are from workshops and interviews undertaken between December 2015 and November 2016. This project is part of the Productive Margins: Regulating for Engagement programme, and concerns concerns life on a low income for families with children in two urban areas in England and Wales. Using creative methods, it involved collaboration between academics, two community organisations and artists.</p

Productive margins, regulating for engagement: Life chances, low income families in urban areas 2015-2016

‘Life Chances’ is a co-produced research project exploring life on a low income for families with children, and the regulatory systems they encounter in their everyday lives. The project was co-designed by academics from Bristol and Cardiff Universities, artists Close and Remote, and two community organisations: Single Parent Action Network in Bristol and South Riverside Community Development Centre in Cardiff. The project also aimed to co-produce the research with community participants in both areas. Data was collected through 11 workshops in each location, a joint visit to Chepstow castle and two workshops with both groups together, between December 2015 and June 2016. In addition, 13 individual interviews were undertaken by the Research Associate between January and May 2016. An evaluation session with both groups also took place in July 2016. The data stored are from workshops and interviews undertaken between December 2015 and November 2016. This project is part of the Productive Margins: Regulating for Engagement programme, and concerns concerns life on a low income for families with children in two urban areas in England and Wales. Using creative methods, it involved collaboration between academics, two community organisations and artists.</p

Replication through partnership: the evolution of partnerships between community land trusts and housing associations in England

© 2016 Informa UK Limited, trading as Taylor & Francis Group. Community land trusts (CLTs) have emerged as an innovative way of addressing the affordable housing crisis in England, as they seek to control and own housing to ensure lasting affordability and to democratically manage assets through voluntarism and community ownership structures. However, there can be difficulties that impede their progress, including legitimacy as new forms of housing organisation, access to finance, and voluntary capacity. CLTs have increasingly begun to partner with housing associations to overcome these issues, combining community leadership with professional expertise and experience. While partnerships may be critiqued for standardising community initiatives or for marrying contrasting institutional logics, housing association support has led directly to the growth of the CLT sector and created new frameworks in which communities can pursue local goals. This paper reports on empirical research into the constitution and effectiveness of partnerships, and considers their implications for future community-led housing development