Cell labelling for in vivo use and in process quality control
Lead Research Organisation:
Queen Mary University of London
Department Name: Blizard Institute of Cell and Molecular
Abstract
The clinical component of the project is a single clinical trial of the efficacy of spray cultured allogeneic keratinocytes in acceleration of re-epithelialisation of life-threatening burn injury [and other non-burn large area skin loss] split-thickness
skin graft donor sites based on a novel cell labelling system.
Whilst cultured keratinocyte allogeneic cells do not survive transplantation longterm, they appear to have a useful clinical role in both acute and chronic wound healing. In the former, they accelerate epithelialisation of partial-thickness cutaneous
wounds, and in the latter, they appear to convert the wound phenotype to a more acute one. These effects are mediated by the release of a wide range of cytokines and growth factors in an orchestrated way. The process of allograft cell loss is not
classical, has been described as creeping substitution and is poorly understood. The wound distribution of sprayed cells, their attachment and survival, and their subsequent migration have been almost impossible to follow in the absence of an in
vivo label [we have used sex-mismatching and a Y-probe in the past]. The effects can be seen clinically, but in a very subjective way as shortened "time to healing". A cell label will allow an assessment of the relative contribution of allogeneic
and autologous keratinocytes to accelerated restoration of epidermal barrier function over time.
This clinical study will establish the efficacy of spray cultured allogeneic keratinoyte adjuvant therapy at a new level of objectivity, and at the same time introduce a novel cell labelling system to the clinical environment. The population of lifethreatening
burn injury [>30% TBSA] has been chosen for a variety of reasons. This is the population for whom cultured keratinocyte stem cell technology has historically applied, since timely wound closure limits sepsis related mortality. Large
donor sites are created, and the patients return to theatre regularly, often daily, for further surgery or large dressing changes. This will allow longitudinal retrieval of punch biopsy material following spray application.
Three arms will be included, each represented by a 1% TBSA donor site wound:
- control donor site wound, managed in a standard way with Jelonet dressings,
- spray cultured allogeneic keratinocyte treated donor site wound,
- spray cultured allogeneic keratinocyte including cell label treated wound.
The wounds will be followed by serial clinical evaluation and standardised photography at dressing changes. 6mm punch biopsies will be retrieved from each site at T0, immediately following creation of the donor site, and two further time points
over the following two weeks - T48hrs and T5days ideally, but for ethical reasons at times otherwise when general anaesthetic procedures are required. Biopsies will be bisected and snap frozen or fixed for evaluation.
All patients will be adult, and recruited only with informed consent. A power calculation will be performed on statistical dvice, and a patient number of between 6 and 10 is currently anticipated. The work will be conducted by a Surgical research fellow toward a higher degree, MDRes or PhD under PI supervision.
The aspiration is for a detection system that will demonstrate the distribution, survival and migration of labelled allogeneic cells macroscopically. Tissue will be processed for detection of the cell label, for morphological evidence of foreign body
reaction or other, for the relative contribution of allo- vs autograft to volume, and for markers of keratinocyte migration, proliferation, differentiation and apoptosis.
skin graft donor sites based on a novel cell labelling system.
Whilst cultured keratinocyte allogeneic cells do not survive transplantation longterm, they appear to have a useful clinical role in both acute and chronic wound healing. In the former, they accelerate epithelialisation of partial-thickness cutaneous
wounds, and in the latter, they appear to convert the wound phenotype to a more acute one. These effects are mediated by the release of a wide range of cytokines and growth factors in an orchestrated way. The process of allograft cell loss is not
classical, has been described as creeping substitution and is poorly understood. The wound distribution of sprayed cells, their attachment and survival, and their subsequent migration have been almost impossible to follow in the absence of an in
vivo label [we have used sex-mismatching and a Y-probe in the past]. The effects can be seen clinically, but in a very subjective way as shortened "time to healing". A cell label will allow an assessment of the relative contribution of allogeneic
and autologous keratinocytes to accelerated restoration of epidermal barrier function over time.
This clinical study will establish the efficacy of spray cultured allogeneic keratinoyte adjuvant therapy at a new level of objectivity, and at the same time introduce a novel cell labelling system to the clinical environment. The population of lifethreatening
burn injury [>30% TBSA] has been chosen for a variety of reasons. This is the population for whom cultured keratinocyte stem cell technology has historically applied, since timely wound closure limits sepsis related mortality. Large
donor sites are created, and the patients return to theatre regularly, often daily, for further surgery or large dressing changes. This will allow longitudinal retrieval of punch biopsy material following spray application.
Three arms will be included, each represented by a 1% TBSA donor site wound:
- control donor site wound, managed in a standard way with Jelonet dressings,
- spray cultured allogeneic keratinocyte treated donor site wound,
- spray cultured allogeneic keratinocyte including cell label treated wound.
The wounds will be followed by serial clinical evaluation and standardised photography at dressing changes. 6mm punch biopsies will be retrieved from each site at T0, immediately following creation of the donor site, and two further time points
over the following two weeks - T48hrs and T5days ideally, but for ethical reasons at times otherwise when general anaesthetic procedures are required. Biopsies will be bisected and snap frozen or fixed for evaluation.
All patients will be adult, and recruited only with informed consent. A power calculation will be performed on statistical dvice, and a patient number of between 6 and 10 is currently anticipated. The work will be conducted by a Surgical research fellow toward a higher degree, MDRes or PhD under PI supervision.
The aspiration is for a detection system that will demonstrate the distribution, survival and migration of labelled allogeneic cells macroscopically. Tissue will be processed for detection of the cell label, for morphological evidence of foreign body
reaction or other, for the relative contribution of allo- vs autograft to volume, and for markers of keratinocyte migration, proliferation, differentiation and apoptosis.
Planned Impact
The goal of the clinical translational group is to demonstrate the in vivo efficacy and relevance of the novel cell labelling system.
Academic learning and dissemination
Research output will be disseminated locally, Nationally, and Internationally through presentation and publication. The group hosts the Southeast of England Burn Injury Research Network [BIRN] - www.burnshealing.com, and the PI is one of only three HEFCE funded academics in Plastic Surgery in the Southeast able to provide a platform for such projects and their dissemination, and the only one Nationally with a background in burn care.
Research collaborations
Within the Blizard Institute, the Cutaneous Research Centre skin engineering group contributes to institute-wide themes including a somatic stem cell theme, collaborating with other clinical academic groups focussed on other body systems.
The Cutaneous Centre holds a CR-UK programme, and many collaborations within European networks: Geneskin, Therapeuskin and UVI-EPI-CA. The PI is developing a unique Burns & Plastic Surgery research base at the Postgraduate
Medical Institute of Anglia Ruskin University, Chelmsford.
Commercial aspects
Demonstration of in vivo efficacy of the cell label will improve the efficiency of such cultured keratinocyte technology, lower the cost, and increase clinical take up. Commercial development will be achieved through sharing in house results with
third party contacts, in the first instance Roslin Cellab, and subsequently to other companies eg Geron and GE Healthcare. New partners will either have regenerative products on the market/in development, or a significant market position in supplying tools and products to the regenerative market.
Dialogue with users
The main current end user of cultured keratinocyte technology is the burn surgeon. That technology has received a very mixed reception over the years. The clear demonstration of efficacy of the keratinocyte system using the cell label would
appeal to the end user. The clinical group is well placed to disseminate that.
Publicity for research
The group publishes in the skin engineering field. Its profile is extended internationally by the distance learning teaching programme in burn care - Postgraduate Diploma in Burn Care - it delivers as far afield as the Gaza strip. It run a parallel programme in scar research. Further education programmes in microsurgery simulation and the principles of safe aesthetic practice and their associated advertisements, clinical events and seminars attract a significant commercial and public interest.
Academic learning and dissemination
Research output will be disseminated locally, Nationally, and Internationally through presentation and publication. The group hosts the Southeast of England Burn Injury Research Network [BIRN] - www.burnshealing.com, and the PI is one of only three HEFCE funded academics in Plastic Surgery in the Southeast able to provide a platform for such projects and their dissemination, and the only one Nationally with a background in burn care.
Research collaborations
Within the Blizard Institute, the Cutaneous Research Centre skin engineering group contributes to institute-wide themes including a somatic stem cell theme, collaborating with other clinical academic groups focussed on other body systems.
The Cutaneous Centre holds a CR-UK programme, and many collaborations within European networks: Geneskin, Therapeuskin and UVI-EPI-CA. The PI is developing a unique Burns & Plastic Surgery research base at the Postgraduate
Medical Institute of Anglia Ruskin University, Chelmsford.
Commercial aspects
Demonstration of in vivo efficacy of the cell label will improve the efficiency of such cultured keratinocyte technology, lower the cost, and increase clinical take up. Commercial development will be achieved through sharing in house results with
third party contacts, in the first instance Roslin Cellab, and subsequently to other companies eg Geron and GE Healthcare. New partners will either have regenerative products on the market/in development, or a significant market position in supplying tools and products to the regenerative market.
Dialogue with users
The main current end user of cultured keratinocyte technology is the burn surgeon. That technology has received a very mixed reception over the years. The clear demonstration of efficacy of the keratinocyte system using the cell label would
appeal to the end user. The clinical group is well placed to disseminate that.
Publicity for research
The group publishes in the skin engineering field. Its profile is extended internationally by the distance learning teaching programme in burn care - Postgraduate Diploma in Burn Care - it delivers as far afield as the Gaza strip. It run a parallel programme in scar research. Further education programmes in microsurgery simulation and the principles of safe aesthetic practice and their associated advertisements, clinical events and seminars attract a significant commercial and public interest.
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| Simon Myers (Principal Investigator) |