OPTIMISED DELIVERY OF THERAPEUTIC siRNA INTO HUMAN SKIN

Lead Research Organisation: University of Dundee
Department Name: Unlisted

Abstract

RNAi is a new form of therapy that allows faulty genes to be switched off in a wide range of genetic diseases and in cancer. The barrier to applying this revolutionary new form of medicine is how to deliver it into the body, since the molecules involved are larger than normal drugs and so cells require some help in taking this material up so it can have its beneficial effect. The main aim of this project is to revolutionise the delivery of these new therapeutics into the skin, as a test organ system. Skin has the advantage of being on the outside of the body for easy access, and excess human skin from cosmetic surgery (breast reductions and tummy tucks) is readily available for experimentation to optimise delivery of RNAi molecules. In pursuit of this, we will develop an innovative new tool combining chemistry, ultrasound and arrays of microscopic painless needles for fast, effective and reliable delivery into skin at high coverage rates. Once optimised for skin diseases, this hand-held device will be readily adapted for use on internal organs using keyhole surgery methods, so that the potential applications of this device across all branches of medicine are huge. The team assembled in Dundee brings together leading molecular biologists with engineers, supported by further international collaborators with diverse skills across many different fields, in order to piece together elements of technology in ways that have never been done before. The estimated market for solving the therapy delivery problem being tackled here is in the order of 10s of billions of dollars per year.

Technical Summary

RNA-interference (RNAi) has revolutionised biomedical sciences as a research tool and has enormous potential for treatment a host of genetic diseases and cancer. In 2004, the projected market for this new class of therapeutics was estimated at $25 billion in the USA alone. The major limiting factor preventing RNAi technology from clinical application is effective delivery. The skin is an attractive target organ for delivery development due its accessibility but the stratum corneum, the impermeable barrier layer at the epidermal surface, is difficult to cross. Here we have assembled a multidisciplinary team consisting of two molecular biologists with expertise in genetic skin disease and an physicist highly active in design of microfabrication of new medical devices for ultrasonic delivery of molecules (sonoporation), all based in a new translational research division within the 5* research environment at the University of Dundee. The team is supported by collaborators that include a clinical dermatologist, the CEO of a siRNA biopharma start-up and the leading experts in use of sonoporation for skin delivery and in use of microneedles for trans-stratum corneum delivery. The overarching objective for this project is to revolutionise the delivery of molecular therapeutics across the stratum corneum and into the underlying living tissue. In pursuit of this, we will develop an innovative new tool offering the critical capability for fast, effective and reliable delivery at coverage rates approaching square cm/sec, and all in a pain-free format. This will be achieved by marrying the latest cutting edge findings drawn from ultrasound driven micro-fluid mechanics, with state-of-the-art RNAi biotechnology. Here, we make a quantum leap in procedures by coupling the initial penetrative passage through the stratum corneum with an active mode of subsequent dispersal into the viable dermis using ultrasound activated microbubbles that contain the siRNA. Delivery testing will involve use of human skin biopsy material and an animal model. This hybrid technology has not been attempted before but holds great promise. Due to the activity building around RNAi therapeutics in the biotech industry, a window of opportunity to develop the first effective means of widespread RNAi delivery. If successful in skin, this technology could be readily adapted for laporascopic use in other organ systems. Truly multidisciplinary, high-risk, high potential return research of this nature is difficult to fund from a single source such as EPSRC, MRC etc and so the Milstein scheme is an ideal funding stream for this project.

Publications

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publication icon
Gruber R (2012) An appraisal of oral retinoids in the treatment of pachyonychia congenita. in Journal of the American Academy of Dermatology

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Hegde V (2014) In vivo gene silencing following non-invasive siRNA delivery into the skin using a novel topical formulation. in Journal of controlled release : official journal of the Controlled Release Society

 
Description European Dermatology Forum
Geographic Reach Europe 
Policy Influence Type Citation in other policy documents
 
Description Clinical Fellowship
Amount £82,258 (GBP)
Organisation Dystrophic Epidermolysis Bullosa Research Association (DEBRA) 
Sector Charity/Non Profit
Country Austria
Start 10/2015 
End 10/2016
 
Description Industrial collaboration
Amount £200,000 (GBP)
Organisation Wave Life Sciences 
Sector Private
Country Unknown
Start 09/2015 
End 08/2017
 
Description MRC Programme Grant
Amount £1,426,740 (GBP)
Funding ID G0802780 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 03/2010 
End 02/2015
 
Description New Investigator Award
Amount £548,800 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 07/2014 
End 06/2017
 
Description Project Grant
Amount £38,500 (GBP)
Organisation Dystrophic Epidermolysis Bullosa Research Association (DEBRA) 
Sector Charity/Non Profit
Country Austria
Start 08/2015 
End 07/2016
 
Description Project Grant
Amount £169,773 (GBP)
Funding ID 1450/1 
Organisation Fight for Sight 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2013 
End 09/2016
 
Description Wellcome Trust Strategic Award
Amount £5,869,875 (GBP)
Funding ID 098439/Z/12/Z 
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 08/2012 
End 07/2017
 
Title K12 humanized knock-in mouse 
Description Knock-in mouse expressing human KRT12 gene carrying a dominant-negative mutation under control of the murine Krt12 gene. 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Provided To Others? No  
Impact Manuscript at advanced stage, submission planned Q4 of 2014. 
 
Title K12-luciferase knock-in mouse 
Description Expresses luciferase reporter gene under the control of the murine Krt12 gene. Carries multiple human therapy target sequences. 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Provided To Others? No  
Impact Manuscript at advanced stage, for submission Q4 of 2014. 
 
Title K9 humanized knock-in mouse 
Description Knock-in mouse carrying a dominant-negative mutation in the human KRT9 gene. 
Type Of Material Model of mechanisms or symptoms - human 
Provided To Others? No  
Impact Publication planned 2015. 
 
Title K9 knockout mouse 
Description Knockout mouse for the palm/sole specific keratin gene Krt9 encoding the K9 protein. 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Provided To Others? No  
Impact Paper published in 2013 (on-line) and 2014 (print): Fu DJ, Thomson C, Lunny DP, Dopping-Hepenstal PJ, McGrath JA, Smith FJD, McLean WHI and Leslie Pedrioli DM (2014) Keratin 9 is Required for the Structural Integrity and Terminal Differentiation of Palmoplantar Epidermis. J Invest Dermatol 134:754-763 [Epub ahead of print Aug 20, 2013] (PubMed ID: 23962810) 
 
Title K9-luciferase reporter gene mouse 
Description Knock-in mouse expressing luciferase under the control of the murine Krt9 gene. Carries multiple therapy target sequences. 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Provided To Others? No  
Impact Publication planned 2015. 
 
Description Roslin 
Organisation University of Edinburgh
Department The Roslin Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution Developing systems for ex vivo culture of skin.
Collaborator Contribution Provision of discarded pig skin following ova harvesting.
Impact This collaboration is helping us optimise an ex vivo skin culture system. No final outcome yet.
Start Year 2015
 
Description WaVe 
Organisation WaVe Biosciences
Country United States 
Sector Private 
PI Contribution We established an informal collaboration with WaVe Biosciences in 2014 to test their proprietary "stereopure" antisense oligonucleotides using in vitro and in vivo model systems we have available.
Collaborator Contribution None as yet but we anticipate obtaining "stereopure" oligonucleotides for testing in 2015.
Impact None as yet.
Start Year 2014
 
Description Consultant Geneticist on Channel 4's "Embarrassing Bodies" 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Consultant on Channel 4's "Embarrassing Bodies" in relation to an undiagnosed patient with pachyonychia congenita. Following this mass-media appearance, more than 50 new undiagnosed families with pachyonychia congenita or related conditions contacted the research group and were given genetic testing and patient support. The programme has been repeated in the UK and other nations worldwide, and each time, further undiagnosed families have come forth and been given genetic testing, contact with patient groups and other support. Many famililes contacted in this way have attended patient support meetings organised by the group.
Year(s) Of Engagement Activity 2012,2013,2014
 
Description DEBRA Annual Conference, Weybridge, UK 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Type Of Presentation Workshop Facilitator
Geographic Reach National
Primary Audience Patients, carers and/or patient groups
Results and Impact More than 100 patients with the inherited skin blistering disorder epidermolysis bullosa simplex (EBS) attended 1-day meetings in Weybridge, South of England, and Staffordshire, South of England, to learn about the molecular basis of their disease and hear about progress being made in therapy development.

Owing to the success of this meeting and the preceding one in Scotland, a follow-up meeting was organised to spread this information to patients in the north of England in November 2012. These EBS patient meetings will now be an annual event.
Year(s) Of Engagement Activity 2012,2015
 
Description DEBRA UK research leadership visit 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Patients, carers and/or patient groups
Results and Impact Visits from DEBRA UK Research Managers and lead fundraisers to laboratories, with updates on current research and future planning.
Year(s) Of Engagement Activity 2015,2016
 
Description Open Doors Day, College of Life Sciences, Dundee 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Attended College of Life Sciences Open Doors Day where scientists explain their research to a few hundred members of the general public.
Year(s) Of Engagement Activity 2011
 
Description PC Project America Patient support meeting 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact About 100 patients and family members affected by the skin disease pachyonychia congenita attended a 2 day workshop in North America, co-organised by members of the Dundee research group. These meetings are run every 2 years in the USA or Canada for the benefit of patients from the Americas.

Very positive feedback received from patients.
Year(s) Of Engagement Activity 2007,2009,2011,2013
 
Description PC Project European Patient Support Meeting 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Type Of Presentation Workshop Facilitator
Geographic Reach International
Primary Audience Patients, carers and/or patient groups
Results and Impact More than 100 patients and family members affected by the skin disease pachyonychia congenita attended a 2 day meeting in Edinburgh organised by the Dundee research group.

Very positive feedback received from patients and their families. This is now a biannual event, with the next date set for Oct 2014.
Year(s) Of Engagement Activity 2006,2008,2010,2012,2014
 
Description PC Project European Patient Support Meeting 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact About 100 patients and family members affected by the genetic skin disease pachyonychia congenita from all over Europe attended a 2 day meeting organised by this research group. This allowed close interaction of patients, family members, clinicians and scientists. This meeting is run every 2 years by the Dundee group and was hosted in Dundee in 2006, 2008 and in Edinburgh 2010, 2012, 2014.

Very positive feedback received from the patients and other attendees.
Year(s) Of Engagement Activity 2006,2008,2010,2012,2014
 
Description PC Project France Patient support meeting 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact About 50 French patients and family members affected by the skin disease pachyonychia congenita attended a 2 day French language workshop organised by members of the Dundee group. This will be a biannual meeting from now on, with the next one scheduled for 2012.

Very positive feedback obtained from the families. This is to be a bi-annual event, next meeting 2016.
Year(s) Of Engagement Activity 2010,2012,2014
 
Description Pachyonychia congenita patient support, Hefei, China 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact About 20 patients with pachyonychia congenita plus their attendant family members, clinicians and scientists, all from mainland China, attended the first patient support meeting for this disease in the Far East.

Positive feedbak received from Chinese patients, many of whom had never had a firm diagnosis or any real understanding of their disease. A number of patients were given genetic diagnosis as a result of this meeting and some misdiagnosed cases correctly reassigned.
Year(s) Of Engagement Activity 2008
 
Description Press release about election to The Royal Society 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Press release about election to The Royal Society:
http://www.dundee.ac.uk/news/2014/professor-irwin-mclean-elected-to-royal-society.php
Several press articles and local radio coverage
Year(s) Of Engagement Activity 2014
URL http://www.dundee.ac.uk/news/2014/professor-irwin-mclean-elected-to-royal-society.php
 
Description Press release regarding Buchanan Medal 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Press release on Buchannan Medal of the Royal Society, local and national press coverage
http://www.dundee.ac.uk/news/2015/professor-irwin-mclean-awarded-buchanan-medal-by-royal-society.php
Year(s) Of Engagement Activity 2015
URL http://www.dundee.ac.uk/news/2015/professor-irwin-mclean-awarded-buchanan-medal-by-royal-society.php
 
Description Press release, Wellcome Trust Strategic Award to Dundee Dermatology and Genetic Medicine 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Media interviews about the Wellcome Trust's major funding boost for dermatology research at the University of Dundee
e.g. http://www.bbc.co.uk/news/uk-scotland-tayside-central-18883942
Year(s) Of Engagement Activity 2012
 
Description Public Lecture 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact 65 people attended the public lecture.

Invited to do a follow-up lecture in 2014
Year(s) Of Engagement Activity 2013
URL http://www.cafesciencedundee.co.uk/?p=1235
 
Description Public Lecture (British Association for the Advancement of Science) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact "Are you thick skinned enough? Dermatology Discoveries in Dundee". Audience of about 60 people, all from the Tayside region general public. 60 min lecture plus 30 min discussion.
Year(s) Of Engagement Activity 2017
 
Description Public Lecture, Comber Rotary Club, County Down, Northern Ireland. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Public talk hosted by a Rotary Club. "Studying inherited skin disease through thick and thin". Audience of about 30-40 Rotarians and interested public. 30 min lecture plus 15 min discussion.
Year(s) Of Engagement Activity 2017
 
Description Public Talk, South West Scotland 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact By direct invitation, a 45 min presentation was given to about 100 members of the public involved in fund-raising for skin disease charities, particularly relating to the filaggrin gene in ichthyosis, common dry skin and eczema/allergy, as well as outlining our efforts to develop therapies based on this gene. Lengthy discussion followed - about 15 min formal and 2 hours informal.

Small financial donation to the lab was received from this group.
Year(s) Of Engagement Activity 2010
 
Description Scottish EBS Patient Support Meeting 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Participants in your research and patient groups
Results and Impact About 100 patients and family members with the inherited skin blistering disease epidermolysis bullosa simplex (EBS) attended a 1-day meeting to learn about the molecular basis of their disease and progress being made in therapy development.

The success of this meeting led to organisation of bigger meeting in the south of England (May 2012) and north of England (Nov 2012), with help from the EB charity, DEBRA.
Year(s) Of Engagement Activity 2011
 
Description UK (North) Epidermolysis bullosa simplex patient support meeting 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Type Of Presentation Workshop Facilitator
Geographic Reach National
Primary Audience Patients, carers and/or patient groups
Results and Impact More than 100 patients with the inherited skin blistering disorder epidermolysis bullosa simplex (EBS) attended 1-day meetings in Manchester, and in Dundee to learn about the molecular basis of their disease and hear about progress being made in therapy development.

These EBS patient meetings will now be an annual event, with 3 being held (Scotland, North of England, South of England) to cover the UK and allow patients to travel to the nearest venue.
Year(s) Of Engagement Activity 2012,2015
 
Description UK (South) Epidermolysis bullosa simplex patient support meeting 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Participants in your research and patient groups
Results and Impact About 100 patients with epidermolysis bullosa simplex (EBS) and their carers/families attended a 1 day meeting in Reading, co-organised by the Dundee group and the patient support organisation DEBRA. The purpose of this meeting was to update these individuals on the state of the art in relation to understanding the molecular basis of their disease and importantly, what therapies are in development.

None as yet.
Year(s) Of Engagement Activity 2014
 
Description Undergraduate Open Day 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Undergraduate students
Results and Impact Undergraduate Open Day, School of Life Sciences
Year(s) Of Engagement Activity 2015