Using key models for developing stem cell-based therapies for aganglionic gut disorders

Lead Research Organisation: University College London
Department Name: Institute of Child Health

Abstract

Life threatening developmental disorders of the gut enteric nervous system (ENS) such as Hirschsprung?s disease (HSCR) occur commonly and are characterised by absent (aganglionic) nerve cells in the gut wall. Current surgical treatments for HSCR are unsatisfactory due to persistent complications and are best classed as palliative therapies rather than definitive cures. The overall research goal is to develop curative therapies for these disorders by testing whether specialised stem cells isolated from the gut (ENS stem cells) are capable of restoring missing ENS following transplantation into, and restoring function of, affected gut. To date, the applicants? research has laid the groundwork for progression to the critical step of testing such therapeutic strategies in key animal models that closely resemble HSCR, prior to clinical trials in patients. This work includes the recent identification of a novel source of ENS stem cells from the gut lining (mucosa), which provides a safer, more accessible and reproducible source of stem cells. ENS stem cells isolated from the gut mucosa of post-natal animals and humans (including from the normal? segment of HSCR gut) have been shown to restore components of the ENS when transplanted into recipient aganglionc gut grown in the laboratory. In order to translate these findings into an effective cell replacement therapy in humans, the proposed research aims to study whether gut mucosal ENS stem cells can be safely and efficiently delivered into the gut of live animals suffering HSCR-like disorders. The research will study the best methods to deliver ENS stem cells to the affected gut and optimise their ability to colonise the gut, form ENS and integrate with other components of the gut musculature to restore function. In addition to access to the best available mouse models of HSCR, the applicants are well placed to carry out this research given their track record in the field of ENS development, stem cell biology and transplantation, and ultimately expertise in the clinical management of children suffering ENS disorders.

Technical Summary

Current surgical treatments for common life threatening enteric nervous system (ENS) disorders such as Hirschsprung?s disease (HSCR) are unsatisfactory and palliative. The overall aim of the proposed research is to develop more effective and curative therapies utilising ENS stem cells. The applicants have an international track record in the field of ENS development and ENS stem cell biology and have already addressed key prerequisites for progressing the research most effectively towards definitive trials of ENS stem cells in humans. These include the isolation, harvesting, and characterisation of ENS stem cells from post-natal mouse and human gut (including from normal gut and the ganglionated portion of HSCR gut) and their effective transplantation into in vitro models of gut aganglionosis. The assessment of transplantation success in these in vitro studies, however, has been limited by a number of practical issues and the research needs to progress to definitive in vivo research employing robust animal models of HSCR. This research proposal aims to utilise both an established and well characterised mouse models of HSCR (monoisoformic Ret51) as well as a transgenic mouse line (Wnt1Cre;ROSA26YFPstop) which allows accurate and permanent lineage tracing of transplanted neural crest-derived ENS stem cells. The work will also use ENS stem cells isolated from gut mucosa as a novel, more accessible and reproducible source of cells. The applicants propose to utilise protocols routinely employed by the research teams to isolate and expand ENS stem cells obtained from wild-type or HSCR-like postnatal animals, and from gut mucosal biopsies from human patients. These will be transplanted in vivo into both embryonic and post-natal aganglionic gut segments of Ret51 mice. Transplant success will be assessed by characterising the ability of the transplanted ENS stem cells to colonise the aganglionic gut, generate an anatomically mature and integrated ENS, and ultimately to effect functional rescue of the dysmotile gut.

Publications

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Burns AJ (2009) Development of the enteric nervous system: bringing together cells, signals and genes. in Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society

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Burns AJ (2014) Neural stem cell therapies for enteric nervous system disorders. in Nature reviews. Gastroenterology & hepatology

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Cooper JE (2017) In vivo transplantation of fetal human gut-derived enteric neural crest cells. in Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society

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Fiori E (2017) Letter regarding 'Covered versus uncovered metal stents for malignant gastric outlet obstruction: Systematic review and meta-analysis'. in Digestive endoscopy : official journal of the Japan Gastroenterological Endoscopy Society

 
Description White paper for enteric neural stem cell therapy
Geographic Reach Multiple continents/international 
Policy Influence Type Membership of a guideline committee
 
Description development of Academy of Paediatric Gastroenterology
Geographic Reach National 
Policy Influence Type Influenced training of practitioners or researchers
Impact Improvement of clinical skills and knowledge in turn improving the care of patients cared for in the secondary and tertiary medical setting and referral to more specialist units
 
Description European Commission Horizon 2020
Amount € 5,990,464 (EUR)
Organisation European Commission 
Sector Public
Country European Union (EU)
Start 01/2016 
End 12/2021
 
Description Great Ormond Street Hospital Children's Charity Clinical Research Starter Grant
Amount £135,204 (GBP)
Organisation Great Ormond Street Hospital Children's Charity (GOSHCC) 
Sector Charity/Non Profit
Country United Kingdom
Start 11/2014 
End 05/2016
 
Description UEG Week 2012 Basic Science Travel Grant
Amount € 1,000 (EUR)
Funding ID UEGW12-2002 
Organisation United European Gastroenterology 
Sector Charity/Non Profit
Country Austria
Start 10/2012 
End 10/2012
 
Description UEG Week 2013 Basic Science Travel Grant
Amount € 1,000 (EUR)
Organisation United European Gastroenterology 
Sector Charity/Non Profit
Country Austria
Start 10/2013 
End 10/2013
 
Title Gut Physiology Lab 
Description Intention was to develop capability to measure physiology of transplanted cells and of the recipient tissue they were transplanted into 
Type Of Material Improvements to research infrastructure 
Year Produced 2015 
Provided To Others? Yes  
Impact Better ability to gauge the success and functional outcomes of enteric neural stem cell transplantation 
 
Title Neuromuscular Disorders Patient Database 
Description Development of a detailed database of all patients sufffeing from gut neuromuscular disoders that are beibg treated within the clinical service 
Type Of Material Model of mechanisms or symptoms - human 
Provided To Others? No  
Impact This will allow better characterisation of patient phenotypes and chanractersitics and allow better transplation of current research 
 
Title Sox10CreERT2 (X3) 
Description Transgenic mouse lines that express Cre recombinase under the control of the Sox10 promoter 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2011 
Provided To Others? Yes  
Impact Understanding the role of enteric progenitor and glial cells in the development and regeneration of the enteric nervous system 
 
Description Comparative pathogenetic mechanisms of enteric neuropathies in animal models and humans 
Organisation Ohio State University
Department Department of Pediatrics, Gastroenterology/Nutrition Unit
Country United States 
Sector Academic/University 
PI Contribution Initiation of network and contribute expertise on animal models of disease and their relationship to clinical conditions. Development of innovative therapies such as stem cells
Collaborator Contribution Intellectual input into the pathogenesis of enteric neuropathies namely geneticsIntellectual input into the pathogenesis of enteric neuropathies namely viral aetiologies and neurodegenerationIntellectual input into the clinical presentation and outcomes of enteric neuropathies
Impact agreement to have interactive meetings and apply for additional funding
Start Year 2011
 
Description Comparative pathogenetic mechanisms of enteric neuropathies in animal models and humans 
Organisation University Medical Center Gronigen
Country Netherlands 
Sector Hospitals 
PI Contribution Initiation of network and contribute expertise on animal models of disease and their relationship to clinical conditions. Development of innovative therapies such as stem cells
Collaborator Contribution Intellectual input into the pathogenesis of enteric neuropathies namely geneticsIntellectual input into the pathogenesis of enteric neuropathies namely viral aetiologies and neurodegenerationIntellectual input into the clinical presentation and outcomes of enteric neuropathies
Impact agreement to have interactive meetings and apply for additional funding
Start Year 2011
 
Description Comparative pathogenetic mechanisms of enteric neuropathies in animal models and humans 
Organisation University of Amsterdam
Department Academic Medical Center, Pediatric Gastroenterology
Country Netherlands 
Sector Academic/University 
PI Contribution Initiation of network and contribute expertise on animal models of disease and their relationship to clinical conditions. Development of innovative therapies such as stem cells
Collaborator Contribution Intellectual input into the pathogenesis of enteric neuropathies namely geneticsIntellectual input into the pathogenesis of enteric neuropathies namely viral aetiologies and neurodegenerationIntellectual input into the clinical presentation and outcomes of enteric neuropathies
Impact agreement to have interactive meetings and apply for additional funding
Start Year 2011
 
Description Comparative pathogenetic mechanisms of enteric neuropathies in animal models and humans 
Organisation University of Bologna
Department Department of Gastroenterology
Country Italy 
Sector Academic/University 
PI Contribution Initiation of network and contribute expertise on animal models of disease and their relationship to clinical conditions. Development of innovative therapies such as stem cells
Collaborator Contribution Intellectual input into the pathogenesis of enteric neuropathies namely geneticsIntellectual input into the pathogenesis of enteric neuropathies namely viral aetiologies and neurodegenerationIntellectual input into the clinical presentation and outcomes of enteric neuropathies
Impact agreement to have interactive meetings and apply for additional funding
Start Year 2011
 
Description Gastrointestinal Tissue Engineering 
Organisation Francis Crick Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution My research team provides expertise related to re-innervation (with neural crest derived cells) of engineered segments of gastrointestinal tract. The team is able to harvest progenitors of these cells from the gastrointestinal tract, culture and propagate them and ultimately use them in models of disease or engineered segments of intestine.
Collaborator Contribution There are multiple partners based in these institutes each addressing a specific aspect of tissue engineering or dealing with a particular constituent cell type of the gastrointestinal tract. The work is brought together in a European Union Programme Grant (Horizon 2020) - PI Professor Paolo De Coppi
Impact None as yet
Start Year 2015
 
Description Gastrointestinal Tissue Engineering 
Organisation Royal Netherlands Academy of Arts and Sciences
Department Hubrecht Institute
Country Netherlands 
Sector Academic/University 
PI Contribution My research team provides expertise related to re-innervation (with neural crest derived cells) of engineered segments of gastrointestinal tract. The team is able to harvest progenitors of these cells from the gastrointestinal tract, culture and propagate them and ultimately use them in models of disease or engineered segments of intestine.
Collaborator Contribution There are multiple partners based in these institutes each addressing a specific aspect of tissue engineering or dealing with a particular constituent cell type of the gastrointestinal tract. The work is brought together in a European Union Programme Grant (Horizon 2020) - PI Professor Paolo De Coppi
Impact None as yet
Start Year 2015
 
Description Gastrointestinal Tissue Engineering 
Organisation Swiss Federal Institute of Technology in Lausanne (EPFL)
Country Switzerland 
Sector Public 
PI Contribution My research team provides expertise related to re-innervation (with neural crest derived cells) of engineered segments of gastrointestinal tract. The team is able to harvest progenitors of these cells from the gastrointestinal tract, culture and propagate them and ultimately use them in models of disease or engineered segments of intestine.
Collaborator Contribution There are multiple partners based in these institutes each addressing a specific aspect of tissue engineering or dealing with a particular constituent cell type of the gastrointestinal tract. The work is brought together in a European Union Programme Grant (Horizon 2020) - PI Professor Paolo De Coppi
Impact None as yet
Start Year 2015
 
Description Gastrointestinal Tissue Engineering 
Organisation University College London
Department Institute of Child Health
Country United Kingdom 
Sector Academic/University 
PI Contribution My research team provides expertise related to re-innervation (with neural crest derived cells) of engineered segments of gastrointestinal tract. The team is able to harvest progenitors of these cells from the gastrointestinal tract, culture and propagate them and ultimately use them in models of disease or engineered segments of intestine.
Collaborator Contribution There are multiple partners based in these institutes each addressing a specific aspect of tissue engineering or dealing with a particular constituent cell type of the gastrointestinal tract. The work is brought together in a European Union Programme Grant (Horizon 2020) - PI Professor Paolo De Coppi
Impact None as yet
Start Year 2015
 
Description Gastrointestinal Tissue Engineering 
Organisation University of Cambridge
Department Cambridge Stem Cell Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution My research team provides expertise related to re-innervation (with neural crest derived cells) of engineered segments of gastrointestinal tract. The team is able to harvest progenitors of these cells from the gastrointestinal tract, culture and propagate them and ultimately use them in models of disease or engineered segments of intestine.
Collaborator Contribution There are multiple partners based in these institutes each addressing a specific aspect of tissue engineering or dealing with a particular constituent cell type of the gastrointestinal tract. The work is brought together in a European Union Programme Grant (Horizon 2020) - PI Professor Paolo De Coppi
Impact None as yet
Start Year 2015
 
Description Gastrointestinal Tissue Engineering 
Organisation University of Copenhagen
Department Biotech Research and Innovation Center (BRIC)
Country Denmark 
Sector Academic/University 
PI Contribution My research team provides expertise related to re-innervation (with neural crest derived cells) of engineered segments of gastrointestinal tract. The team is able to harvest progenitors of these cells from the gastrointestinal tract, culture and propagate them and ultimately use them in models of disease or engineered segments of intestine.
Collaborator Contribution There are multiple partners based in these institutes each addressing a specific aspect of tissue engineering or dealing with a particular constituent cell type of the gastrointestinal tract. The work is brought together in a European Union Programme Grant (Horizon 2020) - PI Professor Paolo De Coppi
Impact None as yet
Start Year 2015
 
Description Gastrointestinal Tissue Engineering 
Organisation University of Edinburgh
Department The Roslin Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution My research team provides expertise related to re-innervation (with neural crest derived cells) of engineered segments of gastrointestinal tract. The team is able to harvest progenitors of these cells from the gastrointestinal tract, culture and propagate them and ultimately use them in models of disease or engineered segments of intestine.
Collaborator Contribution There are multiple partners based in these institutes each addressing a specific aspect of tissue engineering or dealing with a particular constituent cell type of the gastrointestinal tract. The work is brought together in a European Union Programme Grant (Horizon 2020) - PI Professor Paolo De Coppi
Impact None as yet
Start Year 2015
 
Description Gut Physiology 
Organisation University Hospitals Leuven
Department Department of Gastroenterology Louvain
Country Belgium 
Sector Academic/University 
PI Contribution Transplantation of enteric neural stem cells into postnatal gut including models of aganglionosis
Collaborator Contribution Assessement of gut neuromuscular physiology in transplanted tissues
Impact Increased understanding of the functional integrity and integration of transplanted enteric neural stem cells. Presentations at international meetings (described in appropriate section)
Start Year 2012
 
Description The genetics of Hirschsprung's Disease and other enteric neuropathies 
Organisation Erasmus University Rotterdam
Department Biomedical Genetics
Country Netherlands 
Sector Academic/University 
PI Contribution Generation of enteric neural cell progenitors derived from the gut to examine gene expression
Collaborator Contribution Intellectual input including providing a better understanding of the genes involved in formation of the enteric nervous system and disorders arising from defective formation e.g. Hirschsprung's disease. These will allow the research to be best informed of the best possible strategies for enteric neural stem cell therapies including exploring the possibility of genetic rescue - a major focus of the research
Impact Collaborative grants published peer reviewed paper - PMID 20977903
Start Year 2012
 
Description The genetics of Hirschsprung's Disease and other enteric neuropathies 
Organisation University of Groningen
Department Department of Genetics
Country Netherlands 
Sector Academic/University 
PI Contribution Generation of enteric neural cell progenitors derived from the gut to examine gene expression
Collaborator Contribution Intellectual input including providing a better understanding of the genes involved in formation of the enteric nervous system and disorders arising from defective formation e.g. Hirschsprung's disease. These will allow the research to be best informed of the best possible strategies for enteric neural stem cell therapies including exploring the possibility of genetic rescue - a major focus of the research
Impact Collaborative grants published peer reviewed paper - PMID 20977903
Start Year 2012
 
Description Understanding potential aetiologies of enteric neuropathies 
Organisation University of Bologna
Department Department of Gastroenterology
Country Italy 
Sector Academic/University 
PI Contribution Provision of gut derived enteric neural cells (neurones and glia) to provide a test bed for potential aetiologies for enteric disease e.g. neuropathic viruses
Collaborator Contribution Intellectual ideas and development of the project
Impact no outputs as yet - still early in collaboration
Start Year 2010
 
Description Parents network meeting in Hospital 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact 30+ parents of patients attended a parent network event. There was education and discussion on the research and how parents may have a role in supporting such initiatives that are designed for health improvement

organisation of larger parent and patient directed 'information' events to talk about medical conditions especially complex ones or those with relatively poor prognosis at present and how research and innovation may improve outcomes
Year(s) Of Engagement Activity 2010,2011,2012