Engineering human artificial chromosomes containing the dystrophin locus for autologous cell therapy of Duchenne Muscular Dystrophy.

Lead Research Organisation: University College London
Department Name: Cell and Developmental Biology

Abstract

Duchenne muscular dystrophy (DMD) is the most common and severe form of muscular dystrophy (1/3500 males). It affects skeletal and cardiac muscles, leading to progressive loss of movements, confinement to a wheelchair and finally to total paralysis and assisted ventilation. Death occurs usually in the third decade of life because of cardiac and/or respiratory failure. Improved medical assistance has increased the lifespan of patients but still there is no efficacious therapy and steroids are the only palliative treatment available. Among several novel strategies that are entering clinical experimentation, our group focused on cell therapy, based upon intra-arterial infusion of mesoangioblasts (MABs), stem/progenitor cells associated to blood vessels that showed efficacy in dystrophic mice and dogs. Although a phase I/II clinical trial based upon allogeneic transplantation of mesoangioblasts is currently running, gene therapy of autologous cells would be preferable (no immune suppression and no need of HLA-matched donor) but the large size of the dystrophin gene (2.4Mb) hampers the use of viral vectors. Our group pioneered the use of human artificial chromosomes (HACs) containing the whole dystrophin locus for muscular dystrophy (DYS-HAC), showing efficacy of this strategy in dystrophic mouse cells. Based on the above, we now plan to develop a definitive strategy that would allow translation to human cells and boosting the therapeutic effect.
First, we will insert into the DYS-HAC an "immortalizing cassette" containing genes that allow bypassing senescence (telomerase and Bm1) and a suicide gene as a safety device. Preliminary results confirm that this cassette works in human cells. The cassette is floxed and thus can be excised from the HAC before cells are infused into patients.The DYS-HAC will also contain an inducible MyoD (i.e linked to the Estrogen Receptor: MyoD-ER), a myogenic master gene that will allow induction of muscle differentiation (which is variable among different patients).
The optimal number of copies of dystrophin to include will be determined to achieve the optimal level of expression for a single genetically corrected nucleus to compensate for the muscle fibre resident nuclei that cannot synthesize it. All the subsequent generations of HACs will be transferred in human dystrophic mesoangioblasts that will be then challenged for their ability to repair dystrophic muscle and ameliorate the disease.
The PI is a leader in the field of cell therapy; his expertise, and that of his colleagues, will guarantee a high probability of success for this ambitious but realistic project, whose results may lead to rapid clinical translation and, at the same time, pave the way for other monogenic diseases, characterized by mutations of a very large gene.

Technical Summary

The aim of this project is to develop a novel cell therapy for Duchenne Muscular Dystrophy (DMD) based upon transplantation of autologous mesoangioblasts (MABs; vessel associated progenitors) engineered with a human artificial chromosome (HAC) vector containing the entire 2.4 megabases of the human dystrophin locus (DYS-HAC). Specifically we will insert into the DYS-HAC:
a) Floxed immortalizing sequences (human telomerase reverse transcriptase (hTERT), Bmi-1 and the suicide gene Herpex Simplex Virus Thymidine Kinase (HSV-TK). These will allow us to extend, reversibly, the lifespan of primary human MABs, excision of the floxed cassette by a non-integrating lentiviral vector expressing the Cre recombinase and negative selection of cells where the cassette has not been excised;
b) An inducible MyoD (MyoD-ER) where the myogenic factor is linked to the Estrogen Receptor, to induce myogenesis at will, since mesoangioblasts vary in their myogenic potency among different patients.
c) Two or more full-length human dystrophin cDNAs to compensate for gene dosage insufficiency deriving from fusion of relatively few genetically corrected cells in a fiber containing a large majority of genetically defective nuclei. Transfer of different HACs, each with a different selection marker, will allow us to determine the number of copies required for optimal protein production.
Subsequent generations of HACs, progressively incorporating the above mentioned functions, will be built and transferred into MABs from DMD patients. Proliferation, differentiation, transformation/tumorigenicity of cells will be tested in vitro and in vivo (scid/mdx dystrophic and immune-deficient mice) before and after excision of floxed immortalizing sequences (by a non-integrating lentiviral vector). Engraftment, dystrophin production, amelioration of the pathology and functional recovery will be tested in transplanted mice. This project will set the stage for a definitive cell therapy of DMD.

Planned Impact

Although cardiovascular and neurological diseases remain the most widespread and disabling in the Western world, musculoskeletal impairments are among the most frequent causes of disability, encompassing more than 150 diseases and affecting every age and socioeconomic group. The classic example is muscular dystrophy, which can appear from infancy to adulthood and affects over 300,000 European citizens. This group of diseases alone poses serious social and economic problems for each national health system, considering the cost for repeated hospitalizations, home assistance, physiotherapy, surgery, medical devices for a devastating disease that takes now decades from diagnosis to death with a progressively deterioration of life quality.
Our project aims to demonstrate safety and efficacy for rapid clinical translation of this cell therapy strategy for Duchenne muscular dystrophy, though restricted to patients in a paediatric age and whose mutation cannot be treated by the simpler exon-skipping strategy. We aim to achieve a significant stabilization/amelioration of clinical symptoms. Importantly, even a partial effect would produce an enormous benefit for patients.
Together with patients, their families would benefit of the success of this project, not only in terms of emotional impact but also in practical terms, due to a reduction of the daily heavy burden related to continuous assistance to patients who are non ambulant and, later, totally unable to move.
The National Health Service would also indirectly benefit, not only for the implementation of available therapies, but also in economic terms. Although cell therapies are expensive and patient-tailored, they would still be cheaper that the total costs for the NHS, related to diagnosis, medical care, surgery, physiotherapy, drugs, medical devices that are required for decades.

Publications

10 25 50

 
Description Faculty, EUREKA Summerschool on Translational Medicine, Utrecht (NL), Jul 2018
Geographic Reach Multiple continents/international 
Policy Influence Type Influenced training of practitioners or researchers
Impact Faculty, EUREKA Summerschool on Translational Medicine, Utrecht (NL), Jul 2018
URL https://eurekainstitute.org/utrecht-summerschool-course-2017-2/
 
Description Muscular Dystrophy UK research strategy reviewing panel
Geographic Reach Europe 
Policy Influence Type Participation in a advisory committee
 
Description Presentation to general paediatricians
Geographic Reach Local/Municipal/Regional 
Policy Influence Type Influenced training of practitioners or researchers
Impact Two seminars were given in UK hospitals, including a district general hospital. Target audience: secondary care paediatricians. Positive feedback were given following this event, as clinicians felt they knew much more on the new therapeutic options for muscular dystrophy.
 
Description Teaching session for clinicians on new therapies for muscular dystrophies
Geographic Reach Local/Municipal/Regional 
Policy Influence Type Influenced training of practitioners or researchers
Impact Teaching session for clinicians on new therapies for muscular dystrophies (North Middlesex Hospital, London, UK: paediatric department)
 
Description UCL Co-lead, London Stem Cell Network
Geographic Reach Local/Municipal/Regional 
Policy Influence Type Influenced training of practitioners or researchers
Impact Established of the first London-wide stem cell network, with first symposium oversubscribed and highly successful.
URL https://lscn.crick.ac.uk
 
Description UCL Co-lead, London Stem Cell Network
Geographic Reach National 
Policy Influence Type Participation in a advisory committee
Impact One of the leads of the largest stem cell network in London. Its first inaugural event was oversubscribed within a few days with more than 400 participants.
URL https://lscn.crick.ac.uk
 
Description Collaborative research grant
Amount € 398,826 (EUR)
Funding ID 201440.30.31.32 
Organisation La Marató de TV3 Foundation 
Sector Charity/Non Profit
Country Spain
Start 07/2015 
End 06/2018
 
Description ERC Starting Grant "HISTOID"
Amount € 1,500,000 (EUR)
Funding ID 759108 
Organisation European Research Council (ERC) 
Sector Public
Country European Union (EU)
Start 09/2018 
End 08/2023
 
Description European Research Council Starting Grant
Amount € 1,499,738 (EUR)
Funding ID HISTOID 
Organisation European Research Council (ERC) 
Sector Public
Country European Union (EU)
Start 06/2018 
End 05/2023
 
Description Muscular Dystrophy Campaign PhD Studentship
Amount £114,326 (GBP)
Funding ID RA4/3023/1 
Organisation Muscular Dystrophy UK 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2015 
End 12/2018
 
Description New Frontier Science
Amount $364,623 (USD)
Organisation Takeda Pharmaceutical Company 
Sector Private
Country Global
Start 02/2014 
End 07/2016
 
Description PhD Studenship
Amount £77,314 (GBP)
Funding ID BB/J014567/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 06/2014 
End 09/2017
 
Description PluriMes EC FP7 consortium
Amount € 6,000,000 (EUR)
Funding ID 602423 
Organisation European Commission 
Department Seventh Framework Programme (FP7)
Sector Public
Country European Union (EU)
Start 02/2014 
End 01/2018
 
Description Research Grant
Amount £46,000 (GBP)
Organisation Duchenne Parent Project Onlus Italy 
Sector Charity/Non Profit
Country Italy
Start 08/2015 
End 02/2017
 
Description Understanding motor neuron and muscle disease using human stem cells
Amount £73,500 (GBP)
Funding ID Main PI: Dr Rickie Patani (Co-I: Dr Tedesco) 
Organisation Rosetrees Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2018 
End 09/2021
 
Title A Highly Efficient Method of Microcell Production for Chromosome Transfer 
Description Microcell-mediated chromosome transfer (MMCT) is a method that enables the transfer of chromosomes from donor to recipient cells. It involves fusion of microcells (small particles of nuclear and plasma membrane containing one or a few chromosomes) with cells, and the following selection of cell hybrids. MMCT can be used to study in-vitro, or in-vivo outcomes of the transferred chromosome with a range of cell lines. We have developed a method to overcome current obstacles that are limiting MMCT utilisation. This protocol results in a cost-effective microcell manufacturing procedure that is amenable to scale-up and diffuse microcell-based technologies such as HAC-transfer for gene therapy. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact We obtained a decrease in cost and time complemented by a significant increase in the number of microcells extracted, eliminating the requirement of a super-centrifuge machine, setting the basis to scale-up microcell production for chromosome transfer. 
 
Title A new HAC containing an inducible MyoD (MyoD-ER) 
Description A tamoxifen inducible MyoD was inserted inside the new HAC in order to induce muscle differentiation upon tamoxifen administration. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact It is expected that this reagent will facilitate muscle differentiation of cells harboring the novel HAC 
 
Title A novel DYS-HAC, devoid of immunogenic proteins (e-GFP and HSTK) with floxed resistance genes 
Description A new intermediate HAC has been developed after removal of HSTK and GFP that are strongly immunogenic. In addition, resistance genes (also potentially immunogenic but needed for selection) have been floxed so that they can be excised when needed 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact The new vectors will be instrumental towards the building of the final HAC, aim of this project. 
 
Title A novel method for direct microcell production for HAC transfer from DT40 cells 
Description We have developed a new method that allows adhesion of chicken DT40 cells (normally growing in suspension) for direct production of microcells. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact Microcells could be then directly used to target cells for HAC transfer. This method significantly accelerate HAC transfer, avoiding the intermediate step of transfer into Chinese Hamster Ovary cells. 
 
Title Construction of a new Dys-HAC-13 
Description We have developed a new Dys-HAC, named 13, containing a inducible Caspase9, followed by an inducible human MyoD-ER and codon-optimised human dystrophin cDNA, driven by SPC5-12 (a synthetic potent muscle specific promoter), and multi integrate and mutated p-lox sites. 
Type Of Material Cell line 
Year Produced 2014 
Provided To Others? Yes  
Impact This new construct will allow the derived human cell line, to undergo robust muscle differentiation at our will (MyoD-ER), to be susceptible to induced apoptosis (as a safety devise through i-Caspase 9) and to over produce dystropin. 
 
Title Construction of a new Dys-HAC-14 and 15 
Description We have constructed a new Dys-HAC containing Blasticidin gene, EGFP and an immortalising cassette (Bmi-1, Telomerase and HSTK suicide gene) and finally an inducible Cre-ERT2 gene that may allow to remove this flexed cassette from the HAC upon Ganciclovir treatment. The two constructs (14 and 15) differ for the location and method of integration into the HAC. 
Type Of Material Cell line 
Year Produced 2014 
Provided To Others? Yes  
Impact These novel constructs, ready to be transferred into human cells will allow the induction of unlimited proliferation (thanks to Bmi-1 and Telomerase) and at the same time, excision of the function when differentiation or transplantation will be required. Since Cre mediated excision is never 100% efficient we also have an inducible suicide gene (HSTK) which allows to selectively kill those cells (10% in preliminary experiments) where the cassette is not excised. Remarkably, human cells, with the same active cassette (though in a different molecular context) are not tumorigenic in immune deficient mice. 
 
Title Human dystrophic (DMD) reversibly immortalized mesoangioblasts containing DYS-HAC2 
Description We have succeeded in obtaining 8 different clones of lentiviral reversibly immortalised mesoangioblasts from Duchenne patients containing a novel DYS-HAC (DYS-HAC2) with reduced immunogenic potential. 
Type Of Material Cell line 
Year Produced 2014 
Provided To Others? Yes  
Impact This is the first evidence of HAC transfer in clinically relevant cell types. Transplantation studies showed engraftment in dystrophic mice and the same method is currently being applied to transfer DYS-HAC2 also to human myoblasts. 
 
Title Novel DYS-HAC sequence (and human MyoD-ER plasmid) 
Description We have almost completed the designing of the final sequence of a novel multifunctional DYS-HAC. This includes a number of genes regulating proliferation, differentiation and safety of the construct. Those genes are assembled into two large DNA vectors and we will be sending them very shortly for gene-synthesis. Afterwards we will modify our current HAC backbone by inserting those sequences by chromosome engineering. As a proof of principle for our approach, we have successfully synthesized and 
Type Of Material Biological samples 
Year Produced 2014 
Provided To Others? Yes  
Impact The development of this material is critical for the successful development of the project since it constitutes one of the main parts to engineer the novel HAC. Notably, this will be the first evindece of gene-synthesis mediated chromosomal engineering. If successful this technique will significantly impact on the chromosomal engineering community providing a way to facilitate what is usually a time consuming and technically challenging process. 
 
Description Artificial Chromosomes for cell therapy of muscular dystrophy 
Organisation Tottori University
Department Department of Biomedical Science
Country Japan 
Sector Academic/University 
PI Contribution We have strengthen our collaboration with the group of Prof. Mitsuo Oshimura (Tottori University). We are mainly engineering a new HAC and developing the platforms (in vitro and in vivo) to test its safety and efficacy once developed. Additionally, the group of Dr Elvassore is collaborating with us in order to test gene expression and protein production of DYS-HAC-corrected cells in vitro in combination with biomaterials to improve cell differentiation.
Collaborator Contribution Prof. Oshimura's group is contributing by reviewing the critical steps of cloning for the engineering of the new HAC. Dr Elvassore's group is testing by western blot dystrophin production in vitro in HAC-corrected cells.
Impact Three papers have been already published with this collaboration (Hoshiya H et al, Mol Ther 2009; Tedesco FS et al., Sci Transl Med 2011; Tedesco FS et al., Sci Transl Med 2012). The team is multidisciplinary: London: muscle regeneration, cell biology and HAC engineering. Tottori: chromosome engineering. Pavia: bioengineering.
Start Year 2008
 
Description Artificial Chromosomes for cell therapy of muscular dystrophy 
Organisation University of Pavia
Department Faculty of Engineering
Country Italy 
Sector Academic/University 
PI Contribution We have strengthen our collaboration with the group of Prof. Mitsuo Oshimura (Tottori University). We are mainly engineering a new HAC and developing the platforms (in vitro and in vivo) to test its safety and efficacy once developed. Additionally, the group of Dr Elvassore is collaborating with us in order to test gene expression and protein production of DYS-HAC-corrected cells in vitro in combination with biomaterials to improve cell differentiation.
Collaborator Contribution Prof. Oshimura's group is contributing by reviewing the critical steps of cloning for the engineering of the new HAC. Dr Elvassore's group is testing by western blot dystrophin production in vitro in HAC-corrected cells.
Impact Three papers have been already published with this collaboration (Hoshiya H et al, Mol Ther 2009; Tedesco FS et al., Sci Transl Med 2011; Tedesco FS et al., Sci Transl Med 2012). The team is multidisciplinary: London: muscle regeneration, cell biology and HAC engineering. Tottori: chromosome engineering. Pavia: bioengineering.
Start Year 2008
 
Description Artificial Chromosomes for the therapy of muscular dystrophies 
Organisation Tottori University
Department Department of Biomedical Science
Country Japan 
Sector Academic/University 
PI Contribution We previously used first generation Human artificial Chromosome encoding the the dystrohin gene in the first pre-clinical protocol of cell therapy for Duchenne Musucular Dystrophy (Tedesco et al. Sci. Transl. Med. 2011).
Collaborator Contribution Collaborators helped us to familiarize with chromosomal engineering and transfer.
Impact Tedesco et al. Science Translational Medicine 4:140ra89, 2012. Tedesco et al. Science Translational Medicine 3:96ra78, 2011 Hoshiya et al. Mol Ther. 17:309-17, 2009.
Start Year 2008
 
Description EBiSC Consortium 
Organisation European Commission
Department Innovative Medicines Initiative (IMI)
Country European Union (EU) 
Sector Multiple 
PI Contribution The role of my team is to generate cell lines and tool to study muscle diseases and develop therapies. Specific role: • WP 2.1 - Standardisation of protocols for recruitment of tissue and clinical data - Commissioned iPSC production • WP 6.2 Selection of disease types for the PoC trials - PoC differentiation - Scalable production of specific progenitor cells (FM & FST)
Collaborator Contribution From EBISC website: "The EBiSC Consortium represents all relevant stakeholders from tissue donors to clinical and academic iPSC researchers and industrial users and thus provides the scientific expertise, facilities, networks and experience to achieve the project goals and respond appropriately to advances in science and society. Led by Pfizer Ltd and managed by Roslin Cells Sciences Ltd., the Consortium comprises 8 active participant iPSC Centres with clinical and patient networks, a global leader in industrial iPSC supply, international experts in iPSC science, biobanking, bioengineering, regenerative medicine and data management, and scholars in law and ethics." Full list of partners below. EFPIA companies: - Pfizer Ltd, United Kingdom - Novo Nordisk A/S, Denmark - AstraZeneca AB, Sweden - H. Lundbeck A/S, Denmark - Janssen Pharmaceutica NV a pharmaceutical company of Johnson & Johnson, Belgium - UCB Biopharma SPRL, Belgium - Bayer Pharma AG, Germany - Eli Lilly & Co. Ltd, United Kingdom SME's: - Roslin Cell Sciences, United Kingdom - ARTTIC, France - DefiniGEN Ltd, United Kingdom - Douglas Connect GmbH (working communities), Switzerland - Bioneer A/S, Denmark Universities, research organisations, public bodies, non-profit groups - University of Edinburgh, United Kingdom - Fraunhofer-Institut für Biomedizinische Technik (IBMT), Germany - Fraunhofer-Institut für Molekularbiologie und Angewandte Oekologie IME ScreeningPort (IME-SP), Germany - Genome Research Limited (Wellcome Trust Sanger Institute), United Kingdom - European Molecular Biology Laboratory (European Bioinformatics Institute), United Kingdom - Charité Universitätsmedizin Berlin, Germany - University of Newcastle Upon Tyne, United Kingdom - Klinikum der Universität zu Köln (University of Cologne), Germany - Gottfried Wilhelm Leibniz Universität Hannover (University of Hannover) Centre for Ethics and Law in the Life Sciences, Germany - Koninklijke Nederlandse Academie van Wetenschappen (The Hubrecht Institute), Netherlands - University College London, United Kingdom - Culture Collections of Public Health England, United Kingdom - National Institute for Biological Standards and Control, United Kingdom - Universitätsklinikum Bonn (University of Bonn), Germany - Instituto de Salud Carlos III. (Spanish Stem Cell Bank), Spain - Fundacion Publica Andaluza Progreso y Salud, Spain - Center of Regenerative Medicine in Barcelona (CMR[B]), Spain - Inbiomed, Spain
Impact This collaboration is highly multi-disciplinary (please see list of collaborators above) and has already resulted in an agreement to deposit and bank two iPS cell lines from patients with rare muscle diseases.
Start Year 2016
 
Description EBiSC Consortium 
Organisation European Federation of Pharmaceutical Industries and Associations (EFPIA)
Country Belgium 
Sector Private 
PI Contribution The role of my team is to generate cell lines and tool to study muscle diseases and develop therapies. Specific role: • WP 2.1 - Standardisation of protocols for recruitment of tissue and clinical data - Commissioned iPSC production • WP 6.2 Selection of disease types for the PoC trials - PoC differentiation - Scalable production of specific progenitor cells (FM & FST)
Collaborator Contribution From EBISC website: "The EBiSC Consortium represents all relevant stakeholders from tissue donors to clinical and academic iPSC researchers and industrial users and thus provides the scientific expertise, facilities, networks and experience to achieve the project goals and respond appropriately to advances in science and society. Led by Pfizer Ltd and managed by Roslin Cells Sciences Ltd., the Consortium comprises 8 active participant iPSC Centres with clinical and patient networks, a global leader in industrial iPSC supply, international experts in iPSC science, biobanking, bioengineering, regenerative medicine and data management, and scholars in law and ethics." Full list of partners below. EFPIA companies: - Pfizer Ltd, United Kingdom - Novo Nordisk A/S, Denmark - AstraZeneca AB, Sweden - H. Lundbeck A/S, Denmark - Janssen Pharmaceutica NV a pharmaceutical company of Johnson & Johnson, Belgium - UCB Biopharma SPRL, Belgium - Bayer Pharma AG, Germany - Eli Lilly & Co. Ltd, United Kingdom SME's: - Roslin Cell Sciences, United Kingdom - ARTTIC, France - DefiniGEN Ltd, United Kingdom - Douglas Connect GmbH (working communities), Switzerland - Bioneer A/S, Denmark Universities, research organisations, public bodies, non-profit groups - University of Edinburgh, United Kingdom - Fraunhofer-Institut für Biomedizinische Technik (IBMT), Germany - Fraunhofer-Institut für Molekularbiologie und Angewandte Oekologie IME ScreeningPort (IME-SP), Germany - Genome Research Limited (Wellcome Trust Sanger Institute), United Kingdom - European Molecular Biology Laboratory (European Bioinformatics Institute), United Kingdom - Charité Universitätsmedizin Berlin, Germany - University of Newcastle Upon Tyne, United Kingdom - Klinikum der Universität zu Köln (University of Cologne), Germany - Gottfried Wilhelm Leibniz Universität Hannover (University of Hannover) Centre for Ethics and Law in the Life Sciences, Germany - Koninklijke Nederlandse Academie van Wetenschappen (The Hubrecht Institute), Netherlands - University College London, United Kingdom - Culture Collections of Public Health England, United Kingdom - National Institute for Biological Standards and Control, United Kingdom - Universitätsklinikum Bonn (University of Bonn), Germany - Instituto de Salud Carlos III. (Spanish Stem Cell Bank), Spain - Fundacion Publica Andaluza Progreso y Salud, Spain - Center of Regenerative Medicine in Barcelona (CMR[B]), Spain - Inbiomed, Spain
Impact This collaboration is highly multi-disciplinary (please see list of collaborators above) and has already resulted in an agreement to deposit and bank two iPS cell lines from patients with rare muscle diseases.
Start Year 2016
 
Description Engineering and next generation HACs and their transfer to myogenic stem cells 
Organisation Tottori University
Country Japan 
Sector Academic/University 
PI Contribution Our research team was leading this collaboration. Dr Hoshiya was the main researcher interacting with the two collaborators and succeeded in obtaining the new construct leading to the next generation HAC.
Collaborator Contribution Professor Oshimura and Dr Uno spent a sabbatical (3 months; June-September 2014) in the UCL Dept of Cell and Developmental Biology. They provided expert assistance and knowledge for the engineering and cloning of the two main plasmids containing the multi-genes that ultimately go inside the new HAC. Dr Kazuki and Dr Hiratshuka from Tottori University also assisted in this collaboration.
Impact Outputs: - New multigene plasmid containing MyoDER, dystrophin cDNAs and iCaspase9 successfully amplified - New HAC containing the above plasmid - Scientific interaction leading to a review manuscript just accepted for publication and to charity-funded PhD studentship starting in January 2015 Multidisciplinary collaboration: molecular biology, genetic engineering, cell biology, regenerative medicine.
Start Year 2009
 
Description Engineering and next generation HACs and their transfer to myogenic stem cells 
Organisation University of Manchester
Country United Kingdom 
Sector Academic/University 
PI Contribution Our research team was leading this collaboration. Dr Hoshiya was the main researcher interacting with the two collaborators and succeeded in obtaining the new construct leading to the next generation HAC.
Collaborator Contribution Professor Oshimura and Dr Uno spent a sabbatical (3 months; June-September 2014) in the UCL Dept of Cell and Developmental Biology. They provided expert assistance and knowledge for the engineering and cloning of the two main plasmids containing the multi-genes that ultimately go inside the new HAC. Dr Kazuki and Dr Hiratshuka from Tottori University also assisted in this collaboration.
Impact Outputs: - New multigene plasmid containing MyoDER, dystrophin cDNAs and iCaspase9 successfully amplified - New HAC containing the above plasmid - Scientific interaction leading to a review manuscript just accepted for publication and to charity-funded PhD studentship starting in January 2015 Multidisciplinary collaboration: molecular biology, genetic engineering, cell biology, regenerative medicine.
Start Year 2009
 
Description HACs and iPS cells for DMD 
Organisation University College London
Department Institute of Child Health
Country United Kingdom 
Sector Academic/University 
PI Contribution Generation of preliminary data to apply and secure further funding from the Muscular Dystrophy Campaign
Collaborator Contribution Professor Muntoni, a world leading paediatric neurologist, is the second supervisor of a PhD studentship secured by Dr Tedesco
Impact Muscular Dystrophy Campaign-funded 4-year PhD studentship grant to Dr Tedesco from Jan 2015 and NIHR ACF. Please see specific section for additional details.
Start Year 2013
 
Description New stem cell therapy for Duchenne muscular dystrophy 
Organisation Pompeu Fabra University
Country Spain 
Sector Academic/University 
PI Contribution My team focuses on studying the self-renewal capacity of transplantable muscle stem cells and the possibility to correct them with human artificial chromosomes.
Collaborator Contribution Dr Serrano (Pompeu Fabra University, Barcelona) is focusing on developing anti-fibrotic treatments that could be coupled with myogenic cell transplantation. Prof Cossu (Manchester University) is focusing on developing genetic treatments tat could be coupled with myogenic cell transplantation (via exon skipping).
Impact Publications (e.g. Tedesco FS et al., Methods in Molecular Bioogy. 2017;1556:129-147) and presentations at international conferences (e.g. FASEB conference "Skeletal Muscle Stem Cells and Regeneration", Keystone (CO), Jul 2016).
Start Year 2015
 
Description New stem cell therapy for Duchenne muscular dystrophy 
Organisation University of Manchester
Country United Kingdom 
Sector Academic/University 
PI Contribution My team focuses on studying the self-renewal capacity of transplantable muscle stem cells and the possibility to correct them with human artificial chromosomes.
Collaborator Contribution Dr Serrano (Pompeu Fabra University, Barcelona) is focusing on developing anti-fibrotic treatments that could be coupled with myogenic cell transplantation. Prof Cossu (Manchester University) is focusing on developing genetic treatments tat could be coupled with myogenic cell transplantation (via exon skipping).
Impact Publications (e.g. Tedesco FS et al., Methods in Molecular Bioogy. 2017;1556:129-147) and presentations at international conferences (e.g. FASEB conference "Skeletal Muscle Stem Cells and Regeneration", Keystone (CO), Jul 2016).
Start Year 2015
 
Description Plurimes Consortium 
Organisation Leiden University
Country Netherlands 
Sector Academic/University 
PI Contribution From the Plurimes project website: "The PluriMes Project: Pluripotent stem cell resources for mesodermal medicine. A consortium of 12 European partners have been awarded €6million by the European Commission for a research and development project focused on directing stem cells to become bone and muscle. Pluripotent stem cells have the potential to generate any type of cell found in the body. They are generated and multiplied in the laboratory. By harnessing the capacity of pluripotent stem cells to produce functional cell types with precision and at scale, researchers hope to enable new treatment modalities for degenerative diseases. The PluriMes project is specifically targeted at therapies for muscle, bone and cartilage. The project combines the expertise of ten academic and two industrial partners to bring together stem cell experts, genetic engineers, developmental biologists, cell therapy pioneers, bioengineers and specialist SMEs in a cross-disciplinary collaborative effort. PluriMes is supported through the European Commission's Framework 7 HEALTH research programme and Coordinated by Professor Austin Smith from the Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge." My tem and I are key members of this consortium of top European stem cell researchers and our role focuses on the development of pluripotent stem cell based technologies for skeletal muscle disease modelling, drug development, cell therapies and tissue engineering.
Collaborator Contribution Please refer to the project's website for details on specific expertise and contributions: https://www.plurimes.eu
Impact The collaboration is multidisciplinary, including also SMEs/Biotech companies. Several outputs have resulted from this partnership, including several publications (most of which listed here in ResearchFish associated to the specific awards), successful outreach activities (e.g. Pint of Science Festival 2015) and promising scientific collaborations (e.g. development of the first human pluripotent stem cell-derived artificial muscle tissue - confidential).
Start Year 2014
 
Description Plurimes Consortium 
Organisation Pluriomics BV
Country Netherlands 
Sector Private 
PI Contribution From the Plurimes project website: "The PluriMes Project: Pluripotent stem cell resources for mesodermal medicine. A consortium of 12 European partners have been awarded €6million by the European Commission for a research and development project focused on directing stem cells to become bone and muscle. Pluripotent stem cells have the potential to generate any type of cell found in the body. They are generated and multiplied in the laboratory. By harnessing the capacity of pluripotent stem cells to produce functional cell types with precision and at scale, researchers hope to enable new treatment modalities for degenerative diseases. The PluriMes project is specifically targeted at therapies for muscle, bone and cartilage. The project combines the expertise of ten academic and two industrial partners to bring together stem cell experts, genetic engineers, developmental biologists, cell therapy pioneers, bioengineers and specialist SMEs in a cross-disciplinary collaborative effort. PluriMes is supported through the European Commission's Framework 7 HEALTH research programme and Coordinated by Professor Austin Smith from the Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge." My tem and I are key members of this consortium of top European stem cell researchers and our role focuses on the development of pluripotent stem cell based technologies for skeletal muscle disease modelling, drug development, cell therapies and tissue engineering.
Collaborator Contribution Please refer to the project's website for details on specific expertise and contributions: https://www.plurimes.eu
Impact The collaboration is multidisciplinary, including also SMEs/Biotech companies. Several outputs have resulted from this partnership, including several publications (most of which listed here in ResearchFish associated to the specific awards), successful outreach activities (e.g. Pint of Science Festival 2015) and promising scientific collaborations (e.g. development of the first human pluripotent stem cell-derived artificial muscle tissue - confidential).
Start Year 2014
 
Description Plurimes Consortium 
Organisation Sapienza University of Rome
Country Italy 
Sector Academic/University 
PI Contribution From the Plurimes project website: "The PluriMes Project: Pluripotent stem cell resources for mesodermal medicine. A consortium of 12 European partners have been awarded €6million by the European Commission for a research and development project focused on directing stem cells to become bone and muscle. Pluripotent stem cells have the potential to generate any type of cell found in the body. They are generated and multiplied in the laboratory. By harnessing the capacity of pluripotent stem cells to produce functional cell types with precision and at scale, researchers hope to enable new treatment modalities for degenerative diseases. The PluriMes project is specifically targeted at therapies for muscle, bone and cartilage. The project combines the expertise of ten academic and two industrial partners to bring together stem cell experts, genetic engineers, developmental biologists, cell therapy pioneers, bioengineers and specialist SMEs in a cross-disciplinary collaborative effort. PluriMes is supported through the European Commission's Framework 7 HEALTH research programme and Coordinated by Professor Austin Smith from the Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge." My tem and I are key members of this consortium of top European stem cell researchers and our role focuses on the development of pluripotent stem cell based technologies for skeletal muscle disease modelling, drug development, cell therapies and tissue engineering.
Collaborator Contribution Please refer to the project's website for details on specific expertise and contributions: https://www.plurimes.eu
Impact The collaboration is multidisciplinary, including also SMEs/Biotech companies. Several outputs have resulted from this partnership, including several publications (most of which listed here in ResearchFish associated to the specific awards), successful outreach activities (e.g. Pint of Science Festival 2015) and promising scientific collaborations (e.g. development of the first human pluripotent stem cell-derived artificial muscle tissue - confidential).
Start Year 2014
 
Description Plurimes Consortium 
Organisation Stem Cell Technologies
Country Canada 
Sector Private 
PI Contribution From the Plurimes project website: "The PluriMes Project: Pluripotent stem cell resources for mesodermal medicine. A consortium of 12 European partners have been awarded €6million by the European Commission for a research and development project focused on directing stem cells to become bone and muscle. Pluripotent stem cells have the potential to generate any type of cell found in the body. They are generated and multiplied in the laboratory. By harnessing the capacity of pluripotent stem cells to produce functional cell types with precision and at scale, researchers hope to enable new treatment modalities for degenerative diseases. The PluriMes project is specifically targeted at therapies for muscle, bone and cartilage. The project combines the expertise of ten academic and two industrial partners to bring together stem cell experts, genetic engineers, developmental biologists, cell therapy pioneers, bioengineers and specialist SMEs in a cross-disciplinary collaborative effort. PluriMes is supported through the European Commission's Framework 7 HEALTH research programme and Coordinated by Professor Austin Smith from the Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge." My tem and I are key members of this consortium of top European stem cell researchers and our role focuses on the development of pluripotent stem cell based technologies for skeletal muscle disease modelling, drug development, cell therapies and tissue engineering.
Collaborator Contribution Please refer to the project's website for details on specific expertise and contributions: https://www.plurimes.eu
Impact The collaboration is multidisciplinary, including also SMEs/Biotech companies. Several outputs have resulted from this partnership, including several publications (most of which listed here in ResearchFish associated to the specific awards), successful outreach activities (e.g. Pint of Science Festival 2015) and promising scientific collaborations (e.g. development of the first human pluripotent stem cell-derived artificial muscle tissue - confidential).
Start Year 2014
 
Description Plurimes Consortium 
Organisation Swiss Federal Institute of Technology in Lausanne (EPFL)
Country Switzerland 
Sector Public 
PI Contribution From the Plurimes project website: "The PluriMes Project: Pluripotent stem cell resources for mesodermal medicine. A consortium of 12 European partners have been awarded €6million by the European Commission for a research and development project focused on directing stem cells to become bone and muscle. Pluripotent stem cells have the potential to generate any type of cell found in the body. They are generated and multiplied in the laboratory. By harnessing the capacity of pluripotent stem cells to produce functional cell types with precision and at scale, researchers hope to enable new treatment modalities for degenerative diseases. The PluriMes project is specifically targeted at therapies for muscle, bone and cartilage. The project combines the expertise of ten academic and two industrial partners to bring together stem cell experts, genetic engineers, developmental biologists, cell therapy pioneers, bioengineers and specialist SMEs in a cross-disciplinary collaborative effort. PluriMes is supported through the European Commission's Framework 7 HEALTH research programme and Coordinated by Professor Austin Smith from the Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge." My tem and I are key members of this consortium of top European stem cell researchers and our role focuses on the development of pluripotent stem cell based technologies for skeletal muscle disease modelling, drug development, cell therapies and tissue engineering.
Collaborator Contribution Please refer to the project's website for details on specific expertise and contributions: https://www.plurimes.eu
Impact The collaboration is multidisciplinary, including also SMEs/Biotech companies. Several outputs have resulted from this partnership, including several publications (most of which listed here in ResearchFish associated to the specific awards), successful outreach activities (e.g. Pint of Science Festival 2015) and promising scientific collaborations (e.g. development of the first human pluripotent stem cell-derived artificial muscle tissue - confidential).
Start Year 2014
 
Description Plurimes Consortium 
Organisation University College London
Department UCL Cancer Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution From the Plurimes project website: "The PluriMes Project: Pluripotent stem cell resources for mesodermal medicine. A consortium of 12 European partners have been awarded €6million by the European Commission for a research and development project focused on directing stem cells to become bone and muscle. Pluripotent stem cells have the potential to generate any type of cell found in the body. They are generated and multiplied in the laboratory. By harnessing the capacity of pluripotent stem cells to produce functional cell types with precision and at scale, researchers hope to enable new treatment modalities for degenerative diseases. The PluriMes project is specifically targeted at therapies for muscle, bone and cartilage. The project combines the expertise of ten academic and two industrial partners to bring together stem cell experts, genetic engineers, developmental biologists, cell therapy pioneers, bioengineers and specialist SMEs in a cross-disciplinary collaborative effort. PluriMes is supported through the European Commission's Framework 7 HEALTH research programme and Coordinated by Professor Austin Smith from the Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge." My tem and I are key members of this consortium of top European stem cell researchers and our role focuses on the development of pluripotent stem cell based technologies for skeletal muscle disease modelling, drug development, cell therapies and tissue engineering.
Collaborator Contribution Please refer to the project's website for details on specific expertise and contributions: https://www.plurimes.eu
Impact The collaboration is multidisciplinary, including also SMEs/Biotech companies. Several outputs have resulted from this partnership, including several publications (most of which listed here in ResearchFish associated to the specific awards), successful outreach activities (e.g. Pint of Science Festival 2015) and promising scientific collaborations (e.g. development of the first human pluripotent stem cell-derived artificial muscle tissue - confidential).
Start Year 2014
 
Description Plurimes Consortium 
Organisation University of Cambridge
Department Cavendish Laboratory
Country United Kingdom 
Sector Academic/University 
PI Contribution From the Plurimes project website: "The PluriMes Project: Pluripotent stem cell resources for mesodermal medicine. A consortium of 12 European partners have been awarded €6million by the European Commission for a research and development project focused on directing stem cells to become bone and muscle. Pluripotent stem cells have the potential to generate any type of cell found in the body. They are generated and multiplied in the laboratory. By harnessing the capacity of pluripotent stem cells to produce functional cell types with precision and at scale, researchers hope to enable new treatment modalities for degenerative diseases. The PluriMes project is specifically targeted at therapies for muscle, bone and cartilage. The project combines the expertise of ten academic and two industrial partners to bring together stem cell experts, genetic engineers, developmental biologists, cell therapy pioneers, bioengineers and specialist SMEs in a cross-disciplinary collaborative effort. PluriMes is supported through the European Commission's Framework 7 HEALTH research programme and Coordinated by Professor Austin Smith from the Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge." My tem and I are key members of this consortium of top European stem cell researchers and our role focuses on the development of pluripotent stem cell based technologies for skeletal muscle disease modelling, drug development, cell therapies and tissue engineering.
Collaborator Contribution Please refer to the project's website for details on specific expertise and contributions: https://www.plurimes.eu
Impact The collaboration is multidisciplinary, including also SMEs/Biotech companies. Several outputs have resulted from this partnership, including several publications (most of which listed here in ResearchFish associated to the specific awards), successful outreach activities (e.g. Pint of Science Festival 2015) and promising scientific collaborations (e.g. development of the first human pluripotent stem cell-derived artificial muscle tissue - confidential).
Start Year 2014
 
Description Plurimes Consortium 
Organisation University of Dresden
Department Biotechnology Center
Country Germany 
Sector Academic/University 
PI Contribution From the Plurimes project website: "The PluriMes Project: Pluripotent stem cell resources for mesodermal medicine. A consortium of 12 European partners have been awarded €6million by the European Commission for a research and development project focused on directing stem cells to become bone and muscle. Pluripotent stem cells have the potential to generate any type of cell found in the body. They are generated and multiplied in the laboratory. By harnessing the capacity of pluripotent stem cells to produce functional cell types with precision and at scale, researchers hope to enable new treatment modalities for degenerative diseases. The PluriMes project is specifically targeted at therapies for muscle, bone and cartilage. The project combines the expertise of ten academic and two industrial partners to bring together stem cell experts, genetic engineers, developmental biologists, cell therapy pioneers, bioengineers and specialist SMEs in a cross-disciplinary collaborative effort. PluriMes is supported through the European Commission's Framework 7 HEALTH research programme and Coordinated by Professor Austin Smith from the Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge." My tem and I are key members of this consortium of top European stem cell researchers and our role focuses on the development of pluripotent stem cell based technologies for skeletal muscle disease modelling, drug development, cell therapies and tissue engineering.
Collaborator Contribution Please refer to the project's website for details on specific expertise and contributions: https://www.plurimes.eu
Impact The collaboration is multidisciplinary, including also SMEs/Biotech companies. Several outputs have resulted from this partnership, including several publications (most of which listed here in ResearchFish associated to the specific awards), successful outreach activities (e.g. Pint of Science Festival 2015) and promising scientific collaborations (e.g. development of the first human pluripotent stem cell-derived artificial muscle tissue - confidential).
Start Year 2014
 
Description Plurimes Consortium 
Organisation University of Manchester
Country United Kingdom 
Sector Academic/University 
PI Contribution From the Plurimes project website: "The PluriMes Project: Pluripotent stem cell resources for mesodermal medicine. A consortium of 12 European partners have been awarded €6million by the European Commission for a research and development project focused on directing stem cells to become bone and muscle. Pluripotent stem cells have the potential to generate any type of cell found in the body. They are generated and multiplied in the laboratory. By harnessing the capacity of pluripotent stem cells to produce functional cell types with precision and at scale, researchers hope to enable new treatment modalities for degenerative diseases. The PluriMes project is specifically targeted at therapies for muscle, bone and cartilage. The project combines the expertise of ten academic and two industrial partners to bring together stem cell experts, genetic engineers, developmental biologists, cell therapy pioneers, bioengineers and specialist SMEs in a cross-disciplinary collaborative effort. PluriMes is supported through the European Commission's Framework 7 HEALTH research programme and Coordinated by Professor Austin Smith from the Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge." My tem and I are key members of this consortium of top European stem cell researchers and our role focuses on the development of pluripotent stem cell based technologies for skeletal muscle disease modelling, drug development, cell therapies and tissue engineering.
Collaborator Contribution Please refer to the project's website for details on specific expertise and contributions: https://www.plurimes.eu
Impact The collaboration is multidisciplinary, including also SMEs/Biotech companies. Several outputs have resulted from this partnership, including several publications (most of which listed here in ResearchFish associated to the specific awards), successful outreach activities (e.g. Pint of Science Festival 2015) and promising scientific collaborations (e.g. development of the first human pluripotent stem cell-derived artificial muscle tissue - confidential).
Start Year 2014
 
Description Plurimes Consortium 
Organisation University of Sheffield
Country United Kingdom 
Sector Academic/University 
PI Contribution From the Plurimes project website: "The PluriMes Project: Pluripotent stem cell resources for mesodermal medicine. A consortium of 12 European partners have been awarded €6million by the European Commission for a research and development project focused on directing stem cells to become bone and muscle. Pluripotent stem cells have the potential to generate any type of cell found in the body. They are generated and multiplied in the laboratory. By harnessing the capacity of pluripotent stem cells to produce functional cell types with precision and at scale, researchers hope to enable new treatment modalities for degenerative diseases. The PluriMes project is specifically targeted at therapies for muscle, bone and cartilage. The project combines the expertise of ten academic and two industrial partners to bring together stem cell experts, genetic engineers, developmental biologists, cell therapy pioneers, bioengineers and specialist SMEs in a cross-disciplinary collaborative effort. PluriMes is supported through the European Commission's Framework 7 HEALTH research programme and Coordinated by Professor Austin Smith from the Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge." My tem and I are key members of this consortium of top European stem cell researchers and our role focuses on the development of pluripotent stem cell based technologies for skeletal muscle disease modelling, drug development, cell therapies and tissue engineering.
Collaborator Contribution Please refer to the project's website for details on specific expertise and contributions: https://www.plurimes.eu
Impact The collaboration is multidisciplinary, including also SMEs/Biotech companies. Several outputs have resulted from this partnership, including several publications (most of which listed here in ResearchFish associated to the specific awards), successful outreach activities (e.g. Pint of Science Festival 2015) and promising scientific collaborations (e.g. development of the first human pluripotent stem cell-derived artificial muscle tissue - confidential).
Start Year 2014
 
Description Plurimes Consortium 
Organisation University of Strasbourg
Country France 
Sector Academic/University 
PI Contribution From the Plurimes project website: "The PluriMes Project: Pluripotent stem cell resources for mesodermal medicine. A consortium of 12 European partners have been awarded €6million by the European Commission for a research and development project focused on directing stem cells to become bone and muscle. Pluripotent stem cells have the potential to generate any type of cell found in the body. They are generated and multiplied in the laboratory. By harnessing the capacity of pluripotent stem cells to produce functional cell types with precision and at scale, researchers hope to enable new treatment modalities for degenerative diseases. The PluriMes project is specifically targeted at therapies for muscle, bone and cartilage. The project combines the expertise of ten academic and two industrial partners to bring together stem cell experts, genetic engineers, developmental biologists, cell therapy pioneers, bioengineers and specialist SMEs in a cross-disciplinary collaborative effort. PluriMes is supported through the European Commission's Framework 7 HEALTH research programme and Coordinated by Professor Austin Smith from the Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute at the University of Cambridge." My tem and I are key members of this consortium of top European stem cell researchers and our role focuses on the development of pluripotent stem cell based technologies for skeletal muscle disease modelling, drug development, cell therapies and tissue engineering.
Collaborator Contribution Please refer to the project's website for details on specific expertise and contributions: https://www.plurimes.eu
Impact The collaboration is multidisciplinary, including also SMEs/Biotech companies. Several outputs have resulted from this partnership, including several publications (most of which listed here in ResearchFish associated to the specific awards), successful outreach activities (e.g. Pint of Science Festival 2015) and promising scientific collaborations (e.g. development of the first human pluripotent stem cell-derived artificial muscle tissue - confidential).
Start Year 2014
 
Title Method for obtaining MAB-like cells and uses thereof 
Description The invention relates to the derivation of mesoangioblast-like (MAB-like) cells from pluripotent cells such as induced pluripotent (IPS) and embryonic stem (ES) cells, to cells obtained thereby and to medical uses of such cells, in particular in the treatment of muscular dystrophies. 
IP Reference WO2013108039 A1 
Protection Patent application published
Year Protection Granted 2013
Licensed No
Impact Industry-Academia collaboration with Takeda Pharmaceuticals funded via their New Frontier Science scheme.
 
Title Co-Investigaor in first-in-human cell therapy clinical trial 
Description Allogeneic cell therapy for Duchenne muscular dystrophy 
Type Therapeutic Intervention - Cellular and gene therapies
Current Stage Of Development Early clinical assessment
Year Development Stage Completed 2015
Development Status Under active development/distribution
Clinical Trial? Yes
Impact Further pre-clinical studies are ongoing to move towards a similar autologous strategy, including development of novel gene correction approaches (HACs, lentiviral mediated exon skipping) both at UCL and University of Manchester, where a second clinical trial is also currently being designed (funded by the Wellcome Trust to Prof Giulio Cossu). 
URL http://embomolmed.embopress.org/content/7/12/1513.long
 
Title Mesoangioblast-mediated exon skipping for genetic correction of exon 51 mutation, based upon a single injection in individual skeletal muscles of five non ambulant patients affected by Duchenne Muscular Dystrophy: a non randomized, open label, phase I/IIa study. 
Description Mesoangioblast-based gene therapy for Duchenne Muscular Dystrophy: a phase I/IIa study / DMD04 (DMD05) Background We pioneered cell therapy for muscular dystrophy with mesoangioblasts (MABS), vessel-associated myogenic progenitors, able to cross the endothelium when delivered intra-arterially and thus distribute evenly to dystrophic muscle. After satisfactorily testing the protocol in three mouse and one dog model of muscular dystrophy and toxicity studies, five DMD (age 8-12, two wheelchair-bound) were selected for a first-in-human, exploratory, prospective, non-randomized open-label phase I-IIa clinical trial of intra-arterial HLA-matched donor MABS transplantation (Eudract Number: 2011-000176-33). The trial proved safety but not efficacy, at variance with results obtained in pre-clinical models (Cossu et al. EMBO Mol. Med. 2015). Among relevant factors were the advanced age of patients and consequent low engraftment that should improve in younger patients. Even so, the level of engraftment will probably remain below the threshold of clinical efficacy. In order to approach and eventually reach clinical efficacy, several steps are being taken to implement the transplantation protocol. Among these, we moved to autologous cells, genetically corrected with a lentiviral vector expressing a small nuclear RNA (U7) engineered to recognize and skip exon 51. In other words this would be a cell-mediated exon skipping. Importantly, the snRNA from genetically corrected autologous cells should correct also neighboring nuclei, inside multinucleated fibers, and thus multiply the therapeutic effect. This novel protocol needs to be first tested locally for safety reasons and this will be done in this study. Study design The study will be a single centre, prospective, sequential, non-randomised, open-label clinical phase I-II study of cell-mediated gene therapy for DMD with autologous, genetically corrected human mesoangioblasts (auto-MABS). 1. Mesoangioblasts will be isolated from a small foot muscle (EBD) of one non-ambulant patient, with a skippable mutation in exon 51, as a separate protocol (DMD05), necessary to validate the SOPs and prepare an IMPD to be submitted to MHRA. The cells will be transduced with a GMP lentiviral vector expressing a small nuclear RNA engineered to skip exon 51, thus restoring the dystrophin reading frame. Transduced cells will be tested for proliferation, differentiation, chromosome abnormalities, pro-vector insertion sites and tumour formation in SCID/bg mice. 2. This patient will be offered the possibility of participating in the subsequent trial (DMD04). 3. Four patients (+ the one of DMD05) will be recruited and undergo a biopsy (except Patient #1 who will have already done it). As in DMD05, cells will be transduced and characterised as in DMD05. 4. After successful completion of the pre-clinical work, cells will be injected into the contra-lateral EBD. After three months a biopsy of the injected muscle will analyse engraftment of transplanted cells and dystrophin production. 5. If dystrophin accumulation is = 10% of a healthy muscle, cells will be subsequently injected in the muscles of the thumb, whose force of contraction will be measured. While safety will represent the primary outcome, increased force of the thumb might improve the quality of the patient's life. Study Objectives: 1. Safety: To assess the incidence of adverse events (any grade) in DMD patients treated with intra-muscular (foot and hand) injections of auto-MABS after genetic correction with a lentiviral vector expressing a small nuclear RNA engineered to skip exon 51 of the dystrophin gene. 2. Efficacy: A) To determine the effect of a single intra-muscular injection into the foot EBD of genetically corrected auto-MABS in inducing cell engraftment and dystrophin synthesis. B) To determine the ability of genetically corrected auto-MABS to increase the force of contraction of the thumb after single intra-muscular injections in the Adductor pollicis, Opponens pollicis, Abductor pollicis brevis and Flexor pollicis brevis of the dominant hand. Contributors: 1. University of Manchester, Institute of Inflammation and Repair: study coordination. G. Cossu (PI), pre-clinical work and analysis of biopsies (QT-PCR). Urmaas Rostalu, Francesco Galli, Bashar Aldeiri, Laricia Braggs, Alessandra Albertini (team). 2. Royal Manchester Children Hospital and NIHR / Wellcome Trust Manchester Clinical Research: Clinical work. Patient selection, treatment and follow up. Analysis of clinical endpoints. Imelda Hughes (Clinical Chief Investigaror) and Gary McCullagh. 3. Institute of Child Health, UCL: Study design, Analysis of biopsies (IF and WB) expertise in muscle cell biology. Francesco Muntoni, Silvia Torelli, Jennifer Morgan 4. University of Manchester, Cleanrooms, Core Technology Facility: isolation, expansion and characterization of MABS as Investigational Medicinal Product. Sue Kimber and Joan Benson. 5. GMP facility at the Denmark Hill campus, KCL. GMP Lentiviral production. Farzin Farzaneh, and Rebecca Prue. 6. The Cell Therapy Catapult. Regulatory aspects. Jacqueline Barry. 7. UMIP (University of Manchester Intellectual Property), Commercialization. Arnaud Garçon. 
Type Therapeutic Intervention - Cellular and gene therapies
Current Stage Of Development Initial development
Year Development Stage Completed 2016
Development Status Under active development/distribution
Clinical Trial? Yes
Impact The trial will have as first end point dystrophin production. If after cell injection, we will be able to measure an amount of dystrophin which is 10% or more of a healthy muscle, we will inject other cells, similarly corrected in the thumb of the same patient. An increase of force of contraction would greatly benefit the quality of every day life in patients. Moreover this outcome would set the basis for a subsequent trial, this time in very young patients with systemic distribution and a defined "intent to cure" 
 
Description - Muscular Dystrophy UK research strategy reviewing panel, Jan 2018 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Policymakers/politicians
Results and Impact Muscular Dystrophy UK research strategy reviewing panel
Year(s) Of Engagement Activity 2018
 
Description 19th London Myology Forum, Imperial College London (UK), Jun 2017 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Oral presentation at London Myology Forum
Year(s) Of Engagement Activity 2017
 
Description AFM - Action Duchenne - Muscular Dystrophy UK expert group on cell therapy, Jun 2017 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Policymakers/politicians
Results and Impact AFM - Action Duchenne - Muscular Dystrophy UK expert group on muscle cell therapy
Year(s) Of Engagement Activity 2017
 
Description Apollo Society London, launching event 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact Apollo Society London (launching event). Please see: http://london.apollosociety.eu
Year(s) Of Engagement Activity 2018
URL http://london.apollosociety.eu
 
Description Conference of patients affected by Duchenne Muscular Dystrophy and their families 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Patients, carers and/or patient groups
Results and Impact XIII INTERNATIONAL CONFERENCE ON DUCHENNE AND BECKER MUSCULAR DYSTROPHY
21 - 22 February 2015, The Church Palace, Rome - Italy
Year(s) Of Engagement Activity 2015
 
Description EMBO Conference "Stem Cells & Regenerative Medicine", Heidelberg (DE), May 2017 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Oral presentation at one of Europe's most prestigious conferences on stem cells
Year(s) Of Engagement Activity 2017
 
Description European Society of Gene and Cell Therapy, 2017 Congress, Berlin (DE), Oct 2017 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Three invited oral presentations from the Tedesco lab to this prestigious gene and cell therapy conference
Year(s) Of Engagement Activity 2017
 
Description Invited presentation at Institute Mondor, Creteil, France 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Paris Est - Creteil University Medical School, Paris (FR), Sep 2018
Year(s) Of Engagement Activity 2018
 
Description Invited presentation at Institute Pasteur, paris 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Institut Pasteur, Paris (FR), Sep 2018
Year(s) Of Engagement Activity 2018
 
Description Invited presentation at Kyoto University 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Center for iPS Cell Research and Application (CiRA), Kyoto University (JP), Sep 2018
Year(s) Of Engagement Activity 2018
 
Description Invited presentation at international chromosome conference 
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 Chromosome Dynamics: Emerging Concepts in Chromosome Biology, Vienna (AS), Mar 2019
Year(s) Of Engagement Activity 2019
 
Description Invited seminar at university of Barcelona 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Invited talk within the Master Programme in Gene and Cell therapy of the University of Barcelona
Year(s) Of Engagement Activity 2017
 
Description Invited talk at Friedrich-Baur-Institut, University of Munich (LUM), Munich (DE), Feb 2018 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Invited talk at Friedrich-Baur-Institut, University of Munich (LUM), Munich (DE), Feb 2018 on experimental therapies for muscle disorders. Audience of approximately 40 participants.
Year(s) Of Engagement Activity 2018
 
Description Invited talk to undergraduate and graduate students of different UCL programmes 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Invited talk to undergraduate and graduate students of different UCL programmes: "Stem Cells and Regenerative Medicine" (CELL3001), 2014 onwards; "Stem Cells and Tissue Repair" (CHLDGG04), MSc in Gene & Cell Therapy, 2013 onwards; "Tissue Engineering" (MECHGB07), MSc in Tissue Engineering, 2016 onwards.
Year(s) Of Engagement Activity 2014,2015,2016,2017,2018
 
Description Muscular dystrophy conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Patients, carers and/or patient groups
Results and Impact Action Duchenne annual conference, London, UK, Nov 2015. Parents of a child with muscular dystrophy approached for further information and also visited my laboratory a few weeks afterwards.
Year(s) Of Engagement Activity 2015
 
Description Nature Conference on Regeneration, Milan (IT), Nov 2017 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited oral presentation at very prestigious conference on stem cells and regeneration
Year(s) Of Engagement Activity 2017
URL http://www.nature.com/natureconferences/regen17/index.html
 
Description Oral presentation to UCL graduate and undergraduate students 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact Annual seminars on muscle stem cells and regeneration in three separate UCL modules: 1) "Stem Cells and Regenerative Medicine" (CELL3001), 2014 onwards; 2) "Stem Cells and Tissue Repair" (CHLDGG04), MSc in Gene & Cell Therapy, 2013 onwards; 3) "Tissue Engineering" (MECHGB07), MSc in Tissue Engineering, 2016 onwards.
Year(s) Of Engagement Activity 2013,2014,2015,2016,2017
 
Description Oxford Global 4th Annual Stem Cell Conference, London (UK), Nov 2017 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Talk at stem cell conference
Year(s) Of Engagement Activity 2017
 
Description Presentation and discussion with neuromuscular patient advocacy group (Spain) 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? Yes
Geographic Reach Regional
Primary Audience Patients, carers and/or patient groups
Results and Impact Talk sparked questions and discussion afterwards.


Patients were very happy and appeared to have understood the difficulties related to the development of new therapies for genetic diseases.
Year(s) Of Engagement Activity 2013
 
Description Presentation to Duchenne patient advocacy group 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact Improved understanding of the status of gene and cell therapy for muscular dystrophy

Request for further information and also request for job placements in my laboratory from students present in the audience
Year(s) Of Engagement Activity 2013
URL http://www.actionduchenne.org
 
Description Presentations to patient advocacy group 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact Improved understanding of the status of the research field from the Duchenne patient community.
Talk stimulated lively discussion afterwards.


After my talk a video was generated by the charity for their website.
I was contacted by several patients via email afterwards for additional information.
Year(s) Of Engagement Activity 2012,2013,2014
URL http://www.parentproject.it/wp-content/uploads/2014/01/ProgrammaProvvisorio_14-02.pdf
 
Description STEMCELL Technologies Webinar, iPSC series (streamed live worldwide), Apr 2017 
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 Media (as a channel to the public)
Results and Impact Webinar on iPSC series, streamed live worldwide, Apr 2017
Year(s) Of Engagement Activity 2017
 
Description Science Festival (London) 
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 "Pint of Science" festival (www.pintofscience.com), London, UK, May 2015. My presentation on stem cells in a local pub was also streamed by an internate radio and posted as a podcast.
Several questions where asked by members of the general public.
Year(s) Of Engagement Activity 2015
URL http://www.pintofscience.com
 
Description Seminar to general paediatricians 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact ? Weekly paediatric teaching session, University College Hospital, London, UK, Nov 2016. The clinicians reported increased interest in the subject.
Year(s) Of Engagement Activity 2016
 
Description Seminar to general paediatricians 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Patients, carers and/or patient groups
Results and Impact ? Weekly paediatric & neonatal seminar, Whittington Hospital, London, UK, Feb 2016. The clinicians reported increased interest in the subject.
Year(s) Of Engagement Activity 2016
 
Description Talk to patient association (Italy) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Patients, carers and/or patient groups
Results and Impact Int. Conferences on Duchenne and Becker Muscular Dystrophy, Rome, Italy. Organised by Duchenne Parent Project Onlus. Several questions where asked by patients on novel therapies for muscular dystrophy.
Year(s) Of Engagement Activity 2012,2013,2014
 
Description University College Hospital Neonatal Unit, London, UK, Aug 2017 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact University College Hospital Neonatal Unit, London, UK, Aug 2017
Year(s) Of Engagement Activity 2017
 
Description Visit to town hall and seminar on HACs for muscular dystrophy and on the topic of "brain drain" 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Talk sparked questions and discussion afterwards

After my talk several persons of the audience approached me to congratulate, since they had improved their knowledge in gene and cell therapy
Year(s) Of Engagement Activity 2014
 
Description World Muscle Society 2017 annual conference, Saint Malo (FR), Oct 2017 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Two oral presentations at one of the most prestigious international muscle conferences
Year(s) Of Engagement Activity 2017