Pre-clinical safety studies of erythrocyte encapsulated thymidine phosphorylase

Lead Research Organisation: St George's University of London
Department Name: Clinical Developmental Sciences

Abstract

MNGIE (Mitochondrial neurogastrointestinal encephalomyopathy) is a life-threatening inherited metabolic disorder caused by a defect in the gene coding for the enzyme thymidine phosphorylase, resulting in affected individuals producing little or no active enzyme. Thymidine phosphorylase is essential for the normal metabolism of DNA metabolites, and in its absence, these metabolites accumulate in the body producing toxic effects on the nervous system and skeletal muscle, causing gastrointestinal dysmotility (e.g. vomiting and anorexia), neuropathy (nerve damage leading to, for example, loss of sensation and abnormal eye movements) and severe muscle weakness. MNGIE is relentlessly progressive with patients dying at an average reported age of 38 years. No specific treatment is currently available. The research team at St. George?s University of London are world leaders in developing the red blood cell as a vehicle for carrying therapeutic proteins in the blood. In the current study they are investigating the effectiveness of using patient?s own red blood cells to carry the missing thymidine phosphorylase in the circulation. The red blood cells provide a protected environment in which the enzyme can function. The encapsulated enzyme reduces the levels of toxic metabolites in the blood, thus relieving the nervous system and muscle of their toxic effects. The aim of this proposal is to conduct pre-clinical safety studies in two animal species and to establish a manufacturing process for the production of clinical grade thymidine phosphorylase; the data obtained will support their application to the UK and USA regulatory bodies to extend this work into a phase II/III clinical trials programme. Results from these studies will be made available to patient support groups and charities specialising in rare inherited metabolic disorders.

Technical Summary

Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a fatal inherited metabolic disorder caused by a deficiency of thymidine phosphorylase, leading to a pathological accumulation of thymidine and deoxyuridine. MNGIE is relentlessly progressive with patients dying at an average age of 37.6 years. There is no recognised treatment. The encapsulation of therapeutic enzymes within autologous erythrocytes is an approach for enzyme replacement therapy which is employed clinically by our group. Encapsulation within the erythrocyte has the advantage of maintaining enzyme activity within the circulation for the life span of the erythrocyte and preventing the formation of inactivating antibodies against the enzyme. This approach is applicable to disorders where pathologically elevated plasma metabolites are able to permeate the erythrocyte membrane. Preliminary data from our clinical pilot studies of erythrocyte encapsulated thymidine phosphorylase show a reduction in plasma thymidine. The aim of this proposal is to obtain regulatory pre-clinical toxicity data from two animal species, and to establish a GMP-compliant manufacturing pathway for recombinant thymidine phosphorylase for clinical use. Both will make a significant contribution to our clinical trial applications to the regulatory bodies, the MHRA and the FDA, from which approval will be sought to develop this investigational therapy into a clinical trials programme.

Publications

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Godfrin Y (2012) International seminar on the red blood cells as vehicles for drugs. in Expert opinion on biological therapy

 
Title In2Science video 
Description In2Science student describing the project she was working on during her work experience at St George's University of London 
Type Of Art Film/Video/Animation 
Year Produced 2018 
Impact Student received In2Science first price for video 
URL https://www.youtube.com/watch?v=W40uDO38ljo&t=11s
 
Description External Expert Advisor to the Department of Health, Gene Therapy Advisory Committee
Geographic Reach National 
Policy Influence Type Participation in a advisory committee
Impact Provided coments on gene therapy proposal for a clinical trial
 
Description iCPS Rare Diseases EU Roundtable
Geographic Reach Europe 
Policy Influence Type Gave evidence to a government review
URL http://rarediseases.parlicentre.org/
 
Description Higher Education Funding Council for Britain
Amount £14,280 (GBP)
Organisation St George's University of London 
Sector Academic/University
Country United Kingdom
Start 01/2011 
End 01/2012
 
Description Innovation Award
Amount £18,019 (GBP)
Organisation St George's University of London 
Sector Academic/University
Country United Kingdom
Start 11/2018 
End 06/2019
 
Description Molecular and Clinical Sciences Research Institute Funding
Amount £6,274 (GBP)
Organisation St George's University 
Sector Academic/University
Country Grenada
Start 05/2017 
End 11/2017
 
Description Molecular and Clinical Sciences Research Institute Funding (IPA software)
Amount £9,542 (GBP)
Organisation St George's University 
Sector Academic/University
Country Grenada
Start 12/2017 
End 11/2018
 
Description PhD studentship
Amount £112,000 (GBP)
Organisation Purine Metabolic Patients’ Association (PUMPA) 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2012 
End 01/2016
 
Description Pilot study Extended expression profiling study of Cerebral Organoids
Amount £4,787 (GBP)
Organisation Purine Metabolic Patients’ Association (PUMPA) 
Sector Charity/Non Profit
Country United Kingdom
Start 05/2017 
End 09/2017
 
Description Pilot study Extended miRNA expression profiling study
Amount £10,000 (GBP)
Organisation Purine Metabolic Patients’ Association (PUMPA) 
Sector Charity/Non Profit
Country United Kingdom
Start 06/2017 
End 12/2017
 
Description Postdoctoral salary fund
Amount £22,754 (GBP)
Organisation Orphan Technologies 
Sector Private
Country Switzerland
Start 04/2018 
End 09/2018
 
Description Principal Investigator award
Amount £76,680 (GBP)
Organisation United Mitochondrial Disease Foundation 
Sector Charity/Non Profit
Country United States
Start 06/2008 
End 06/2011
 
Description Research Funding
Amount £37,000 (GBP)
Funding ID LILY-2017.18 (APPLICATION 3 - BAX) 
Organisation The Lily Foundation 
Sector Charity/Non Profit
Country United Kingdom
Start 04/2018 
End 03/2020
 
Description South of England Specialised Commissioning group
Amount £200,000 (GBP)
Organisation Northamptonshire Healthcare NHS Foundation Trust 
Department South Central Specialised Commissioning Group
Sector Charity/Non Profit
Country United Kingdom
Start 02/2013 
End 06/2013
 
Description Specialised Equipment
Amount £40,000 (GBP)
Organisation Purine Metabolic Patients’ Association (PUMPA) 
Sector Charity/Non Profit
Country United Kingdom
Start 12/2012 
End 12/2014
 
Description Specialised Patient Care
Amount £999,420 (GBP)
Funding ID WEF010812 
Organisation Mid Hampshire Primary Care Trust 
Start 08/2012 
End 07/2017
 
Description Specialised Patient care
Amount £957,271 (GBP)
Organisation Northamptonshire Healthcare NHS Foundation Trust 
Department South Central Specialised Commissioning Group
Sector Charity/Non Profit
Country United Kingdom
Start 02/2008 
End 02/2012
 
Description The development of knock-out cell culture models for investigating the underlying molecular mechanisms that contribute to the neuronal aspects of Mitochondrial Neurogastrointestinal Encephalomyopathy
Amount £142,133 (GBP)
Organisation Purine Metabolic Patients’ Association (PUMPA) 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2014 
End 03/2018
 
Title GLP immunoassay 
Description Validated GLP-compliant immunoassay for the detection of anti-thymidine phosphorylase antibodies for use in a clinical trial 
Type Of Material Technology assay or reagent 
Year Produced 2013 
Provided To Others? No  
Impact Reliable and accurate assay for monitoring antibodies in the clinical application of thymidine phosphorylase. Orphan Drug designation granted by FDA and EMA for therapeutic approach Data supports Clinical trial application to MHRA 
 
Title GLP immunoassay for EE-TP clinical trial 
Description Validated immunoassay for the measurement of human anti-thymidine phosphorylase antibodies in patients treated with EE-TP 
Type Of Material Biological samples 
Year Produced 2018 
Provided To Others? Yes  
Impact Supporting documentation for clinical trial of EE-TP 
URL https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6170929/
 
Title GLP toxicity data 
Description GLP-compliant toxicity data from two animal species to support a clinical trial application to the regulatory bodies 
Type Of Material Biological samples 
Year Produced 2014 
Provided To Others? Yes  
Impact Orphan Drug designation approval by FDA and EMA for therapeutic approach Production of a Clinical Trial Application package Expression of collaboration interest from four biotechnology Companies 
 
Title GMP -TP 
Description Production of three batches of GMP thymidine phosphorylase for a clinical trial of EE-TP for the treatment of MNGIE 
Type Of Material Technology assay or reagent 
Year Produced 2017 
Provided To Others? No  
Impact Three validation batches have provided data to support IMPD and pre-submission briefing package to EMA 
 
Title Validated HPLC assay 
Description Assay validated for the measurement of EE-TP to support QP release in Clinical trial 
Type Of Material Technology assay or reagent 
Year Produced 2014 
Provided To Others? Yes  
Impact Quantification of doses given to patients in compassionate use 
 
Title Validated immunoassay 
Description Immunoassay for detection of human antibodies against thymidine phosphorylase 
Type Of Material Biological samples 
Year Produced 2016 
Provided To Others? No  
Impact Support Clinical trial 
 
Title iPSC of MNGIE and healthy controls 
Description We have developed iPSCs of MNGIE and healthy controls to further the understanding of the molecular mechanisms that underlie MNGIE. Cerebral organoids have been created 
Type Of Material Cell line 
Year Produced 2017 
Provided To Others? Yes  
Impact Data obtained will support the submission of a PhD thesis. Model to replace mouse double knock-out 
URL http://www.nmd-journal.com/article/S0960-8966(17)30270-5/fulltext
 
Description Bioinformatics collaboration 
Organisation University of Lisbon
Country Portugal 
Sector Academic/University 
PI Contribution Sharing data for bioinformatic analysis
Collaborator Contribution Training in the use of bioinformatic databases, and pathway analysis
Impact miRNA- gene target networks Enrichment analysis of miRNA targets PhD thesis
Start Year 2015
 
Description Characterisation of biomarkers for monitoring disease progression and treatment efficacy in MNGIE 
Organisation St George's University of London
Department Department of Basic Medical Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution Analysed preliminary data and wrote/submitted grant application,
Collaborator Contribution Provided guidance on the appropriate techniques to be employed in project
Impact Funding for a PhD studentship
Start Year 2011
 
Description Collaboration with University of Salamanca 
Organisation University of Salamanca
Country Spain 
Sector Academic/University 
PI Contribution Provided training and experience with handling human erythrocytes and training using the Principal Investigator's loading technique
Collaborator Contribution Two PhD students worked in Principal Investigator's laboratory for six months each to exchange animal/human experience with red cells as drug carriers.
Impact Award of two PhDs by the University of Salamanca. For one student, the Principal Investigator was the First Vocal at the PhD Tribunal. One publication: Pub med reference number18674740
Start Year 2006
 
Description Commercial partnership 
Organisation Orphan Technologies
Country Switzerland 
Sector Private 
PI Contribution Intellectual knowledge, expertise in EE-TP manufacture, knowledge of disease (MNGIE)
Collaborator Contribution Expertise in regulatory issues, manufacturing to GMP, clinical trial activities
Impact Recent submission of DPFS/DCS MRC application
Start Year 2012
 
Description MNGIE Consensus group 
Organisation University of Bologna
Country Italy 
Sector Academic/University 
PI Contribution Contributed to an International Systematic Review and Evidence Mapping on the state-of-knowledge of MNGIE.
Collaborator Contribution Contributed to an International Systematic Review and Evidence Mapping on the state-of-knowledge of MNGIE.
Impact Mapping review
Start Year 2018
 
Description MNGIE partnership 
Organisation Ege University
Department Faculty of Medicine
Country Turkey 
Sector Academic/University 
PI Contribution Sharing knowledge , collaborative project
Collaborator Contribution Sharing knowledge, access to patient samples
Impact Sample collection. PI site for clinical trial
Start Year 2018
 
Description MNGIE partnership 
Organisation Vall d' Hebron Research Institute
Country Spain 
Sector Academic/University 
PI Contribution Sharing of knowledge, involvement in clinical study
Collaborator Contribution Sharing of knowledge, access to patients for clinical trial
Impact Clinical trial site identified
Start Year 2019
 
Description MNGIE: An Italian network for MNGIE epidemiology, molecular mechanisms and enzyme replacement therapy by stem cell transplant 
Organisation University Hospital Spedali Civili
Department Clinical Neurology
Country Italy 
Sector Academic/University 
PI Contribution Sharing data obtained from the treatment of patients with enzyme replacement therapy with thymidine phosphorylase
Collaborator Contribution The research team has diagnosed two patients with MNGIE via this collaboration, providing the research team with two potential patients for the Clinical trial they are intending to initiate. Plasma and urine samples have been donated to the research team's Biomarker study.Supply of plasma and urine samples for Research team's Biomarker study
Impact A network of Italian clinicians (neurologists, gastroenterologists) and European researchers has been established, who have experience in MNGIE. The aim is to share information in order to better understand the frequency, natural history and clinical aspects of this complex and rare disease, and to share protocols for both pre-treatment and follow-up studies.
Start Year 2011
 
Description MNGIE: An Italian network for MNGIE epidemiology, molecular mechanisms and enzyme replacement therapy by stem cell transplant 
Organisation University of Verona
Department Department of Neurological Sciences and Vision
Country Italy 
Sector Academic/University 
PI Contribution Sharing data obtained from the treatment of patients with enzyme replacement therapy with thymidine phosphorylase
Collaborator Contribution The research team has diagnosed two patients with MNGIE via this collaboration, providing the research team with two potential patients for the Clinical trial they are intending to initiate. Plasma and urine samples have been donated to the research team's Biomarker study.Supply of plasma and urine samples for Research team's Biomarker study
Impact A network of Italian clinicians (neurologists, gastroenterologists) and European researchers has been established, who have experience in MNGIE. The aim is to share information in order to better understand the frequency, natural history and clinical aspects of this complex and rare disease, and to share protocols for both pre-treatment and follow-up studies.
Start Year 2011
 
Description MiRNA profiling in MNGIE 
Organisation Integrated University Hospital Verona
PI Contribution miRNA profiling and bioinformatic analysis of of plasma samples
Collaborator Contribution Patient samples
Impact Poster presentation at the Annual Neuromuscular Translational Research Conference 2017. Oral presentation at the 17th Symposium of the Purine and Pyrimidine Society conference and prize awarded for best presentation. Successful funding from the Lily Foundation to continue study.
Start Year 2017
 
Title EE-TP therapy for MNGIE disease 
Description Enzyme replacement therapy for a rare inherited metabolic disease. Enzyme is encapsulated in patients red blood cells. 
IP Reference GB1116767.3 
Protection Patent application published
Year Protection Granted 2012
Licensed Yes
Impact Increased awareness in patient and clinician community. Requests for compassionate use. International Health authorities funding compassionate therapy.
 
Title TREATMENT FOR MITROCHONDRIAL NEURO-GASTROINTESTINAL ENCEPHALOMYOPATHY (MNGIE) 
Description The invention provides a method of treating MNGIEt, comprising administering to the patient autologous erythrocytes that contain thymidine phosphorylase 
IP Reference WO2013045885 
Protection Patent granted
Year Protection Granted 2019
Licensed Yes
Impact Clinical trial of EE-TP approved by MHRA
 
Title Treatment for mitrochondrial neurogastrointestinal Encephalomyopathy (MNGIE) 
Description New European Patent Application No. 12756551.3 National Phase of PCT/GB2012/052157 
IP Reference EP2760459 
Protection Patent application published
Year Protection Granted 2014
Licensed Yes
Impact Clinical trial funding
 
Title Treatment for mitrochondrial neurogastrointestinal encephalomyopathy (mngie) 
Description a method of treating mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) in a patient, comprising administering to the patient autologous erythrocytes that contain thymidine phosphorylase and are free of animal proteins other than proteins derived from the patient. 
IP Reference WO2013045885 
Protection Patent granted
Year Protection Granted 2018
Licensed Yes
Impact Clinical trial of EE-TP approval by MHRA
 
Title Erythrocyte encapsulated thymidine phosphorylase using an automated manufacturing process 
Description Erythrocyte encapsulated thymidine phosphorylase manufactured using an automated GMP manufacturing process has been employed in the compassionate use setting. 
Type Therapeutic Intervention - Cellular and gene therapies
Current Stage Of Development Early clinical assessment
Year Development Stage Completed 2018
Development Status Under active development/distribution
Impact Other impacts include supporting data for our clinical trial protocol and regulatory package development 
 
Title Erythrocyte encapsulated thymidine phosphorylase 
Description A red cell loading device has been validated for the encapsulation of thymidine phosphorylase into erythrocytes. Its use has been applied in the compassionate treatment setting prior to moving into a clinical trial which is due to start mid 2018. 
Type Therapeutic Intervention - Medical Devices
Current Stage Of Development Refinement. Clinical
Year Development Stage Completed 2017
Development Status Under active development/distribution
Impact Validation data has supported the development of our study protocol, and regulatory package 
 
Title Erythrocyte encapsulated thymidine phosphorylase 
Description Erythrocyte encapsulated thymidine phosphorylase for the treatment of mitochondrial neurogastrointestinal encephalomyopathy. Received MHRA and ethical approval December 2018 
Type Therapeutic Intervention - Cellular and gene therapies
Current Stage Of Development Early clinical assessment
Year Development Stage Completed 2018
Development Status Under active development/distribution
Impact Application to a compassionate use setting 
 
Title GMP Working Cell bank for thymidine phosphorylase 
Description The product under development is erythrocyte encapsulated thymidine phosphorylase (EE-TP) for the treatment of MNGIE. The thymidine phosphorylase developed is a recombinant enzyme and to date thre engineering batches have been produced for clinical trial use. Master and working cell banks have been created. 
Type Therapeutic Intervention - Cellular and gene therapies
Current Stage Of Development Early clinical assessment
Year Development Stage Completed 2016
Development Status Under active development/distribution
Impact The product has been employed in the compassionate use setting and has eliminated previously observed immune reactions that were observed using a less purified version 
 
Title GMP thymidine phosphorylase 
Description The manufacture of two GMP batches of thymidine phosphorylase for clinical trial use (Q3 2018) 
Type Therapeutic Intervention - Cellular and gene therapies
Current Stage Of Development Early clinical assessment
Year Development Stage Completed 2018
Development Status Under active development/distribution
Impact This material will be used in our clinical trial of erythrocyte encapsulated thymidine phosphorylase which is due to start in mid 2018 
 
Title GMP thymidine phosphorylase 
Description The product under development is erythrocyte encapsulated thymidine phosphorylase (EE-TP) for the treatment of MNGIE. The thymidine phosphorylase developed is a recombinant enzyme and to date three engineering batches have been produced for clinical trial use. Master and working cell banks have been created. The development is funded by the MRC. 
Type Therapeutic Intervention - Cellular and gene therapies
Current Stage Of Development Early clinical assessment
Year Development Stage Completed 2016
Development Status Under active development/distribution
Impact Documentation from manufacture will support clinical trial application 
 
Title Thymidine phosphorylase 
Description The production of a pre GMP recombinant E. coli thymidine phosphorylase. The production has been adapted to processes which are compatible for GMP production. Modifications in the production process were made to minimise hazardous materials, exclude the use of animal-derived products and reduce endotoxin levels. 
Type Therapeutic Intervention - Drug
Current Stage Of Development Refinement. Clinical
Year Development Stage Completed 2012
Development Status Actively seeking support
Impact The development of a refined preparation of thymidine phosphorylase has provided the investigators with an clinical grade enzyme; this has resulted in an amelioration of the mild adverse reactions previously observed on infusion in the investigators compassionate use of this therapy. The availibility of Master Manufacturing Formula documentation to support a clinical trial application to the MHRA. 
 
Description Article 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? Yes
Type Of Presentation Paper Presentation
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Global dissemination resource for the wider scientific, technology and research communities

Contacted by patients with MNGIE for update on availability of therapy in the USA and Turkey
Year(s) Of Engagement Activity 2012
URL http://www.research-europe.com
 
Description How to achieve funding support from the MRC 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact Provided a presentation on the experiences of a MRC grant holder
Year(s) Of Engagement Activity 2017
 
Description In2Science student 
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 Schools
Results and Impact Hosted an In2Science student for one week. The charity aims are to empower students from disadvantaged backgrounds to achieve their potential and progress to STEM and research careers through high quality work placements and careers guidance. Impact included mentoring, provision of university application guidance and insight into a STEM career.
Year(s) Of Engagement Activity 2018
URL https://www.youtube.com/watch?v=W40uDO38ljo&t=11s
 
Description Interview one with Lily Foundation 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Patients, carers and/or patient groups
Results and Impact Interview with the Lily Foundation to talk about funding received to support a biomarker study for patients with a rare disease.
Year(s) Of Engagement Activity 2018
URL https://www.thelilyfoundation.org.uk/news/outsmarting-syndrome/
 
Description Interview two with Lily Foundation 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Patients, carers and/or patient groups
Results and Impact Interview with Lily Foundation to talk about regulatory approval and ethics approval obtained for a clinical trial
Year(s) Of Engagement Activity 2018
URL https://www.thelilyfoundation.org.uk/news/enzyme-replacement-therapy-mngie-approved-clinical-trials-...
 
Description Patient/clinician seminar 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Patients, carers and/or patient groups
Results and Impact 40 individuals (health professionals, researchers, patient carers and diagnostic laboratory staff) attended a seminar on raising the awareness of MNGIE.

Summary of seminar sent out to patients and carers via Newsletter.

Participating clinician of the Association of British Neurologist entered details of clinical study to monthly electronic newsletter sent out by British Neurological Surveillance Unit.
Year(s) Of Engagement Activity 2010
 
Description Rare Disease Day 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact International MNGIE Consortium Meeting and Innsbruck Rare Disease Day Symposium 2016

Monday, February 29 2016, Innsbruck
Presentation of recent / interesting issues:
Discussions on Liver cirrhosis in MNGIE, liver transplantation and gene therapy using an AAV vector
Year(s) Of Engagement Activity 2016
 
Description Rare Disease Day 2019 raising awareness 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Set up a stand to mark Rare Disease Day. Researchers asked passers-by to try their luck at a lighthearted quiz about rare diseases with the chance to win a cupcake or a temporary tattoo. They also took the opportunity to explain rare diseases to the public, staff and students.
Year(s) Of Engagement Activity 2019
URL https://www.sgul.ac.uk/news/news-archive/genetic-therapies-in-the-spotlight-for-rare-disease-day
 
Description Rare Disease Day article 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Article in Guardian Newspaper Rare Disease supplement 28th February covering clinical trial of enzyme replacement therapy. Purpose was to engage with the Rare Disease community. Received six telephone calls from patients and/or families with regard to participation in clinical trial.

https://www.healthawareness.co.uk/rare-diseases/enzyme-replacement-therapy-approved-for-uk-clinical-trials/
Year(s) Of Engagement Activity 2019
URL https://issuu.com/mediaplanetuk/docs/rare_diseases_pages