Chondrocytes from Clinical Grade Embryonic Stem Cells

Lead Research Organisation: University of Manchester
Department Name: School of Biological Sciences

Abstract

Osteoarthritis is a debilitating condition of joints affecting 25% of adults over 65 yrs (WHO estimate). It causes pain, disability and loss of independence, which results in a poor quality of life. In the EU it is ranked 12th for disease burden with 35-40 million sufferers and this is set to rise with increased ageing of the population. Current clinical treatment is limited to pain relief with nothing able to delay or reverse the condition. Irreversibly damaged joints are eventually replaced by synthetic prostheses, which are successful in older patients (>65), but less so in younger patients whose life expectancy is much greater than that of the prosthesis and revision surgery is less successful. There is therefore a large unmet clinical need for improved treatment for OA. Current strategies include cartilage repair (ACI) using the patient's own chondrocytes, harvested from intact areas of tissue. However, the outcome of ACI is little better than non-cellular orthopaedic treatments and it requires 2 operations and damage to otherwise intact tissue. Patients' stem cells are also being tested from bone marrow, but this also requires 2 operations and patient specific cell culture. To overcome the problem of the variable quality of the patients' own cells and the need for 2 operations, we have developed a protocol to generate chondrocytes from human embryonic stem cells (hESCs). The protocol is entirely serum free, chemically defined and yields up to 97% chondrogenic cells. We now need to adapt this procedure to deliver cells suitable for clinical applications. This requires the use of entirely validated clinical grade reagents, scale up to generate larger numbers of cells and methods to store and deliver cells to the clinic. We will validate this protocol with a new generation of very high quality (clinical grade) stem cell lines derived in Manchester and by other Centres across the UK. The advantage of using hESCs is that they can be expanded to produce enough chondrogenic cells to treat large numbers of patients, making such a therapy cost-effective on the NHS. Follow-up tests in vivo will grade lines for success in repairing focal defects in rat knee joints (already shown for research grade cells) to allow us to assess the quality of in vivo cartilage repair and assess any incidence of tumour formation, or other adverse effects. The plan will show if hESC derived chondrocytes can be produced, delivered and are able to complete hyaline cartilage repair. This award will allow us to establish the means and methods to move forward towards phase 1 clinical trials for patients with focal cartilage defects.

Technical Summary

Current clinical treatment for osteoarthritis, which affects 25% of adults over 65 yrs (WHO estimate), is limited to pain relief and there is no treatment able to delay or reverse the condition. Irreversibly damaged joints are eventually replaced by synthetic prostheses, which are successful in older patients (>65), but less so in younger patients whose life expectancy is much greater than that of the prosthesis and revision surgery is less successful. There is therefore a large unmet clinical need for improved treatment for OA. To address this need we developed a protocol to differentiate pluripotent stem cells to chondrogenic cells at high efficiency and these cells which can repair cartilage defects in a rodent model. The protocol is entirely serum free, chemically defined and yields up to 97% chondrogenic cells. We will now adapt this procedure to deliver cells suitable for clinical applications, meeting current Good Manufacturing Practice (cGMP ) standards. We will employ strategies leading to greater cell expansion; apply our refined protocol to a range of clinical grade hESC lines derived in Manchester and in other Centres, evaluating lines for chondrogenic bias. The differentiated cells will be tested for their ability to repair a surgical cartilage defect, without evidence of adverse effects including the generation of ectopic growths or tumours in i) a rat defect model and ii) in a pilot study for a larger animal, the sheep. We will develop and test methods for the clinical delivery and storage of the cells using fibrin gels, evaluating viability and shelf life.

Planned Impact

We intend this preclinical application to lead to the development of phase 1 clinical trials which will provide direct patient benefit and new treatment options for joint trauma and OA sufferers. This will be for focal cartilage defects in the first instance but with potential for repairing larger OA lesions as clinical experience is gained. We will generate hESC-chondrocytes which are suitable for use for patients as part of planning for subsequent clinical trials with our clinical collaborators (Sanjay Anand and David Johnson, Stockport NHS Trust). The development of our protocol will lead the way in providing proof of principle for cell therapies from pluripotent cells for both academics and clinicians.

Academics will benefit from our development and refinement of reproducible protocols, increased understanding of cartilage development and homeostasis. Our work will provide in vitro and in vivo model system for understanding responses to inflammatory mediators and extracellular matrix targeted drugs. We will interact with members of the UK Regenerative Medicine platform hubs to share our findings of translation and learn from their experiences, exploiting their findings where possible e.g. in imaging and safety.

NHS: Our work will start to open up a pathway for this and similar therapies to be incorporated into NHS treatments.
Regulators: Our engagement with the regulators will also help to inform regulatory procedures for cell therapy especially those from pluripotent cells.

Patient Groups and General public: Firstly our projected therapy will provide a cell based treatment and so make a real impact on the quality of life to patients with sports injury and osteoarthritis. Secondly, we will engage with patient groups and member of the public to inform them of our activities and learn further from them regarding patient perception of need, in a continuation of our previous public engagement activities.

Biopharma: We will interact further with companies interested in cell therapy or who may wish to use our findings to generate new in vitro models for testing drugs active in modulating extracellular matrix repair. We will seek advice from biopharma and the Catapult with a view to their active engagement as we move towards the clinic.
 
Description Cochrane guideline on embryo culture temperature
Geographic Reach Europe 
Policy Influence Type Membership of a guideline committee
 
Description ESHRE working group on the impact of embryo culture medium on long term health
Geographic Reach Europe 
Policy Influence Type Influenced training of practitioners or researchers
 
Description HFEA Information for Quality programme
Geographic Reach National 
Policy Influence Type Participation in advisory committee
Impact acting as advisor to Human Fertilisation and Embryology Authority Information for quality programme which aims to improve regulatory oversight of the field including stem cell derivation, and improve information to patients and for delivery of clinical IVF services.
 
Description Influencing policy towards patenting of discoveries using hESC lines
Geographic Reach Europe 
Policy Influence Type Membership of a guideline committee
 
Description Member of TERMIS Scientific Advisory Board
Geographic Reach National 
Policy Influence Type Participation in a advisory committee
 
Description Membership of HFEA's Scientific and Clinical Advances committee
Geographic Reach National 
Policy Influence Type Participation in a advisory committee
Impact Advising the HFEA on changes in regulation on ART practice, embryology, embryonic stem cell biology
 
Description National Clinical human Embryonic Stem Cell Forum
Geographic Reach National 
Policy Influence Type Membership of a guideline committee
Impact Standardisation of requirements for clinical grade embryonic stem cell lines. Prof Brison is vice-chair of this group
 
Description Setting standards for procurement of embryonic stem cells at clinical grade
Geographic Reach National 
Policy Influence Type Influenced training of practitioners or researchers
 
Description Stem Cells for Safer Medicine Panel Member
Geographic Reach National 
Policy Influence Type Participation in a advisory committee
 
Description TCES Travel awards
Amount £300 (GBP)
Organisation Tissue and Cell Engineering Society 
Sector Charity/Non Profit
Country United Kingdom
Start 03/2019 
End 04/2019
 
Title GFP labelled human ESCs 
Description Human ESCs were labelled with GFP using lentiviral vector. 
Type Of Material Cell line 
Provided To Others? No  
Impact These cells could be used for studies requiring tracing cell migration or destination. 
 
Title Repair of cartilage defect in sheep 
Description Development of method to repair osteochondral defect in a large animal 
Type Of Material Physiological assessment or outcome measure 
Provided To Others? No  
Impact No impact yet but paper will be written shortly 
 
Title immortalisation of GMP grade fibroblasts under clean room conditions 
Description immortalisation of GMP grade fibroblasts under clean room conditions 
Type Of Material Cell line 
Year Produced 2013 
Provided To Others? Yes  
Impact Research advance and help to UKSCB 
 
Description Collaboration with Tocris 
Organisation Bio-Techne Ltd
Department Tocris
Country United Kingdom 
Sector Private 
PI Contribution Planned collaboration to develop high though put screening for drugs enhancing chondrogenesis
Collaborator Contribution Targeted drug libraries, screening support and advice; Modified compounds
Impact Seeking joint funding
Start Year 2019
 
Description Erasmus MC Rotterdam 
Organisation Erasmus MC
Country Netherlands 
Sector Academic/University 
PI Contribution Collaboration including a month's research time at host institute, Consultancy for iPSC generation and differentiation.
Collaborator Contribution Collaboration including a month's research time at host institute
Impact Shared research skills and model exchange; collaboration on research programme
Start Year 2015
 
Description GAG analysis 1 
Organisation Ben-Gurion University of the Negev
Country Israel 
Sector Academic/University 
PI Contribution We generate the cellular model , they may analyse GAGS
Collaborator Contribution Initiating
Impact initiating only
Start Year 2016
 
Description GAG analysis with Keele/Oswestry 
Organisation Keele University
Department School of Law
Country United Kingdom 
Sector Academic/University 
PI Contribution We have a protocol for chondrogenesis . Nicky Kuipper at Keele Oswestry will analyse GAGS in medium and cells for preliminary data for a grant application
Collaborator Contribution FACE and other analyses
Impact analyses of samples in progress
Start Year 2016
 
Description Miltenyi 
Organisation Miltenyi Biotec GmBH
Country Germany 
Sector Private 
PI Contribution We have submitted a large grant application to Arthritis Research Uk with Miltenyi. If successful they will contribute in kind support with cell sorting and GMP translationof our protocol for clinical therapy
Collaborator Contribution Cell sorting expertise and kits/antibodies/plates for FACs based cell sorting for enrichment of chondroprogenitors
Impact Too early
Start Year 2017
 
Description Understanding regulation of chondrogenesis by Sirt1 
Organisation Hebrew University of Jerusalem
Country Israel 
Sector Academic/University 
PI Contribution Collaboration to understand epigenetic regulation of hESC chondrogenesis through regulation by histone deacetylases
Collaborator Contribution Training in CHIP technology for Post doc in host lab. Expertise in CHIP,CHIP-CHIP, epigentic modifificaiton and analysis,cartilage biology Exchange of reagents and knowledge.
Impact ARUK grant obtained
Start Year 2013
 
Description CMFT Xmas lecture 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Xmas Royal Society-style lecture for Central Manchester NHS Foundation Trust "The Stem Cell Revolution".

School visit requests
Year(s) Of Engagement Activity 2013
 
Description Faraday Lecture Cambridge 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Faraday Public Lecture
Year(s) Of Engagement Activity 2019
 
Description Lancashire Humanists 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact 70 people attenced a public lecture on stem cell biology

Requests for other talks to Humanist groups.
Year(s) Of Engagement Activity 2013
 
Description Liverpool Medical Institue Symaposium on Stem Cell Regeneration and Osteoarthritic Tissue 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact Talk on use of stem cells in orthopedics by Dr Chris Smith (Pdra)
Year(s) Of Engagement Activity 2019
 
Description Patient information talk Manchester 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Patients, carers and/or patient groups
Results and Impact Informationa dn exchange talk with teh RUG group associated with a osteo and rheumatoid arthritis in Central Manchester Trust
Year(s) Of Engagement Activity 2019
 
Description Radio 4 interview re 'Promising' stem cell trials for blindness 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Daniel Brison interviewed and commented on news story re stem cell therapies

Raising public awareness
Year(s) Of Engagement Activity 2012
 
Description Radio 4 interview with Mark Porter 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Sue Kimber interviewed on Radio 4 Case Notes programme re stem cell therapies

Raised public awareness, led to doubled patient and GP interest in NWESCC
Year(s) Of Engagement Activity 2012
 
Description STEM Cell Day 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Talk triggered questions and interest

Students suggested they would like to apply for bio/biomed in Manchester
Year(s) Of Engagement Activity 2014
 
Description School 6th form talk 2019 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Atalk given to 5, 6th form groups about stem cells and medicine
Year(s) Of Engagement Activity 2019
 
Description School 6th form talk 2019 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Talk about Stem Cells and regenerative medicine to 5 6th form groups at local school
Year(s) Of Engagement Activity 2019
 
Description Science Media Centre press conference 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Press conference organised by SMC to discuss press release concerning submission of clinical grade embryonic stem cell lines to the UK Stem Cell Bank

Interest from media, followup stories. Followup contacts from industry resulting directly from press release.
Year(s) Of Engagement Activity 2011
 
Description Stem Cells and Regenerative medicine from chondrocytes to kidneys 
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 Talk on Regenerative medicine to U3A members
Year(s) Of Engagement Activity 2018
 
Description Stem cells magic for future medicine U3A 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact U3A and other organisations in area Public engagement and discussion about future medicine
Year(s) Of Engagement Activity 2019
 
Description What is Regenerative Medicine? 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Undergraduate students
Results and Impact Lay type presentation explaining what the opportunities are in the sector with exemplar talks form PhD students postdoctoral researchers and research fellows
Year(s) Of Engagement Activity 2017
 
Description What is Regenerative Medicine? 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Undergraduate students
Results and Impact A workshop to inform Undergraduates about what regenerative medicine is and how t can expand the possibiites for Medicine ( based on our successful workshop in 2017)
Year(s) Of Engagement Activity 2018
 
Description Women in Science Becoming the Best 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact Dissemaination of information advice and support for women's role and leadership in scince
Year(s) Of Engagement Activity 2016