Untying a nanoscale knotted polymer structure to linear chains for efficient gene delivery in vitro and to the brain

The purpose of this study was to develop a platform transfection technology, for applications in the brain, which could transfect astrocytes without requiring cell specific functionalization and without the common cause of toxicity through high charge density. Here we show that a simple and scalable preparation technique can be used to produce a “knot” structured cationic polymer, where single growing chains can crosslink together via disulphide intramolecular crosslinks (internal cyclizations). This well-defined knot structure can thus “untie” under reducing conditions, showing a more favorable transfection profile for astrocytes compared to 25 kDa-PEI (48-fold), SuperFect® (39-fold) and Lipofectamine®2000 (18-fold) whilst maintaining neural cell viability at over 80% after four days of culture. The high transfection/lack of toxicity of this knot structured polymer in vitro, combined with its ability to mediate luciferase transgene expression in the adult rat brain, demonstrates its use as a platform transfection technology which should be investigated further for neurodegenerative disease therapies

Untying a nanoscale knotted polymer structure to linear chains for efficient gene delivery in vitro and to the brain

The purpose of this study was to develop a platform transfection technology, for applications in the brain, which could transfect astrocytes without requiring cell specific functionalization and without the common cause of toxicity through high charge density. Here we show that a simple and scalable preparation technique can be used to produce a "knot" structured cationic polymer, where single growing chains can crosslink together via disulphide intramolecular crosslinks (internal cyclizations). This well-defined knot structure can thus "untie" under reducing conditions, showing a more favorable transfection profile for astrocytes comp-red to 25 kDa-PEI (48-fold), SuperFect (R) (39-fold) and Lipofectamine (R) 2000 (18-fold) whilst maintaining neural cell viability at over 80% after four days of culture. The high transfection/lack of toxicity of this knot structured polymer in vitro, combined with its ability to mediate luciferase transgene expression in the adult rat brain, demonstrates its use as - platform transfection technology which should be investigated further for neurodegenerative disease therapies

Gdnf gene delivery via a 2-(dimethylamino)ethyl methacrylate based cyclized knot polymer for neuronal cell applications

Nonviral genetic therapeutic intervention strategies for neurological disorders hold great promise, but a lack of vector efficacy, coupled with vector toxicity, continue to hinder progress. Here we report the application of a newly developed class of polymer, distinctly different from conventional branched polymers, as a transfection agent for the delivery of glial cell line derived neurotrophic factor (GDNF) encoding gene. This new 2-(dimethylamino)ethyl methacrylate (DMAEMA) based cyclized knot polymer was studied for neuronal cell transfection applications, in comparison to branched polyethyleneimine (PEI). While showing a similar transfection profile over multiple cell types, the cyclized knot polymer showed far lower toxicity. In addition, transfection of Neu7 astrocytes with the GDNF encoding gene was able to cause neurite outgrowth when cocultured with dorsal root ganglia (DRGs). The cyclized knot polymer assessed here (PD-E 8%PEG), synthesized via a simple one-pot reaction, was shown to have great potential for neuronal gene therapy applications

Gdnf gene delivery via a 2-(dimethylamino)ethyl methacrylate based cyclized knot polymer for neuronal cell applications

Nonviral genetic therapeutic intervention strategies for neurological disorders hold great promise, but a lack of vector efficacy, coupled with vector toxicity, continue to hinder progress. Here we report the application of a newly developed class of polymer, distinctly different from conventional branched polymers, as a transfection agent for the delivery of glial cell line derived neurotrophic factor (GDNF) encoding gene. This new 2-(dimethylamino)ethyl methacrylate (DMAEMA) based cyclized knot polymer was studied for neuronal cell transfection applications, in comparison to branched polyethyleneimine (PEI). While showing a similar transfection profile over multiple cell types, the cyclized knot polymer showed far lower toxicity. In addition, transfection of Neu7 astrocytes with the GDNF encoding gene was able to cause neurite outgrowth when cocultured with dorsal root ganglia (DRGs). The cyclized knot polymer assessed here (PD-E 8%PEG), synthesized via a simple one-pot reaction, was shown to have great potential for neuronal gene therapy applications

ESPON on the road: bringing closer ESPON evidence for decision making: ECP Transnational Networking Activity Final Report

The ESPONontheRoad project was a Transnational Networking Activity (TNA) of nineteen ESPON Contact Points with the aim to bring ESPON results closer for decision-making and thus increase the capitalisation of the ESPON Programme. During the relatively short project lifetime of one year, participating ECPs brought ESPON closer to the local and regional level, and to citizens in physical and virtual forms as well. The project built a bridge between the issues on a local level and scientific evidence on EU territorial development policy themes. After taking stock of the most recent policy issues in each national context, ECPs formed transnational working groups to have a common understanding of the messages coming from ESPON results. These working groups elaborated their issues for their target groups more in details and designed the most appropriate and efficient form of communication with target groups. In this way ESPON results were put into the macroregional context of West, South, North and Central-Eastern areas, and both the content-related and the organisational tasks were organised in a balanced way

ESPON on the road - Bringing closer ESPON evidence for decision making

The ESPONontheRoad project was a Transnational Networking Activity (TNA) of nineteen ESPON Contact Points (ECPs) with the aim to bring ESPON results closer for decision-making and thus increase the capitalisation of the ESPON Programme. During a year, participating ECPs brought ESPON closer to the local and regional level, and to citizens in physical and virtual forms. The project built a bridge between the issues on a local level and scientific evidence on EU territorial development policy themes. After taking stock of the most recent policy issues in each national context, ECPs formed transnational working groups to have a common understanding of the messages coming from ESPON results. These working groups designed the most appropriate and efficient form of communication for their target groups. In this way ESPON results were put into the macroregional context of West, South, North and Central-Eastern areas, and both the content-related and the organisational tasks were organised in a balanced way. The activity report summaries the goals of the project, presents how they were implemented and what are the lessons learnt. It concludes with recommendations for the future

ESPON on the road: bringing closer ESPON evidence for decision making: ECP Transnational Networking Activity Final Report

The ESPONontheRoad project was a Transnational Networking Activity (TNA) of nineteen ESPON Contact Points with the aim to bring ESPON results closer for decision-making and thus increase the capitalisation of the ESPON Programme. During the relatively short project lifetime of one year, participating ECPs brought ESPON closer to the local and regional level, and to citizens in physical and virtual forms as well. The project built a bridge between the issues on a local level and scientific evidence on EU territorial development policy themes. After taking stock of the most recent policy issues in each national context, ECPs formed transnational working groups to have a common understanding of the messages coming from ESPON results. These working groups elaborated their issues for their target groups more in details and designed the most appropriate and efficient form of communication with target groups. In this way ESPON results were put into the macroregional context of West, South, North and Central-Eastern areas, and both the content-related and the organisational tasks were organised in a balanced way