Final Report Summary - SITRANSPORTER (Identification of molecular transporters that facilitate siRNA skin delivery)
Identification of molecular transporters that facilitate siRNA skin delivery
Robyn Hickerson PhD, Marie Curie Fellow and Independent Investigator
School of Life Sciences, University of Dundee
Final progress report by Professor Irwin McLean FRS FRSE FMedSci
The main aims of this project were (i) to develop a rapid, highly efficient method of coupling small molecule chemical groups to nucleic acids in order to create libraries of chemical-coupled oligonucleotides suitable for identification of cellular penetration enhancing chemicals via high-throughput screening; and (ii) to develop an improved ex vivo organ culture system for mammalian skin in order to facilitate high-throughput drug and nucleic acid screening on living skin.
Both of these aims were achieved. Specifically, a high-throughput chemical coupling method was developed to allow thousands of chemicals to be covalently linked to nucleic acids with close to 100% efficiency and importantly, in a single-tube reaction. Using this new system, a number of high-throughput screens were performed to explore the delivery of short-interfering RNA molecules into a variety of cell lines presenting cell types found in the skin (keratinocytes, fibroblasts and melanocytes). Although no uptake-enhancing chemical structures were identified in these small-scale screens of a few thousand compounds, the protocols are now in place for carrying out larger screens. Furthermore, the technology developed is readily adaptable for screens involving not only siRNA but other forms of nucleic acid therapy, such as antisense oligonucleotides (ASOs). In addition, the technology is in place to explore uptake enhancement chemistry in relation to siRNAs or ASOs delivery into other cell and tissue types. Thus, this Marie Curie project has led to development of an enabling technology to support future oligonucleotide therapeutics research.
Secondly, an advanced organ culture system was developed for mammalian skin to act as a platform for drug/oligonucleotide delivery and bio-distribution studies in living skin. The system developed is able to keep skin alive longer and in a more metabolically active state, that more closely mirrors the viability and gene/protein expression characteristics of the in vivo situation. This ex vivo skin culture system is a major development that has a myriad of applications across the entire field of dermatology, both in the context of academia and industry and is the subject of two patents filed by The University of Dundee, with further IP currently being generated. This aspect of the work is currently the central focus of major grant applications and commercialization activity, all of which is being led by the Marie Curie Fellow, Dr Robyn Hickerson.
Overall, this Marie Curie Fellowship allowed the recruitment and retention of a top-flight US-based scientist with a wealth of industrial and academic experience. The cutting edge research outputs and intellectual property she has produced during the tenure of the Fellowship are set to attract major grant and industrial funding that will consolidate the Fellow’s career in the short-term future and allow her establish herself as a fully independent Principal Investigator in Europe.
Robyn Hickerson PhD, Marie Curie Fellow and Independent Investigator
School of Life Sciences, University of Dundee
Final progress report by Professor Irwin McLean FRS FRSE FMedSci
The main aims of this project were (i) to develop a rapid, highly efficient method of coupling small molecule chemical groups to nucleic acids in order to create libraries of chemical-coupled oligonucleotides suitable for identification of cellular penetration enhancing chemicals via high-throughput screening; and (ii) to develop an improved ex vivo organ culture system for mammalian skin in order to facilitate high-throughput drug and nucleic acid screening on living skin.
Both of these aims were achieved. Specifically, a high-throughput chemical coupling method was developed to allow thousands of chemicals to be covalently linked to nucleic acids with close to 100% efficiency and importantly, in a single-tube reaction. Using this new system, a number of high-throughput screens were performed to explore the delivery of short-interfering RNA molecules into a variety of cell lines presenting cell types found in the skin (keratinocytes, fibroblasts and melanocytes). Although no uptake-enhancing chemical structures were identified in these small-scale screens of a few thousand compounds, the protocols are now in place for carrying out larger screens. Furthermore, the technology developed is readily adaptable for screens involving not only siRNA but other forms of nucleic acid therapy, such as antisense oligonucleotides (ASOs). In addition, the technology is in place to explore uptake enhancement chemistry in relation to siRNAs or ASOs delivery into other cell and tissue types. Thus, this Marie Curie project has led to development of an enabling technology to support future oligonucleotide therapeutics research.
Secondly, an advanced organ culture system was developed for mammalian skin to act as a platform for drug/oligonucleotide delivery and bio-distribution studies in living skin. The system developed is able to keep skin alive longer and in a more metabolically active state, that more closely mirrors the viability and gene/protein expression characteristics of the in vivo situation. This ex vivo skin culture system is a major development that has a myriad of applications across the entire field of dermatology, both in the context of academia and industry and is the subject of two patents filed by The University of Dundee, with further IP currently being generated. This aspect of the work is currently the central focus of major grant applications and commercialization activity, all of which is being led by the Marie Curie Fellow, Dr Robyn Hickerson.
Overall, this Marie Curie Fellowship allowed the recruitment and retention of a top-flight US-based scientist with a wealth of industrial and academic experience. The cutting edge research outputs and intellectual property she has produced during the tenure of the Fellowship are set to attract major grant and industrial funding that will consolidate the Fellow’s career in the short-term future and allow her establish herself as a fully independent Principal Investigator in Europe.