Generation of human embryonic stem cell lines with clinically relevant mutations

Lead Research Organisation: King's College London
Department Name: Reproduction and Endocrinology

Abstract

Human embryonic stem (hES) cells can be isolated from early (4-8 day old) developing embryos which have been grown in the laboratory as part of, or surplus to some kinds of IVF treatment. Under certain conditions, these can be made to develop into different kinds of cells, such as nerve, or pancreatic cells which it is hoped in the future, may be useful in treating degenerative diseases such as Parkinson s disease and diabetes, or in spinal injuries by replacing the diseased or damaged cells with healthy hES cells which have been grown in the laboratory. However in addition to this potential use in therapy, the study of stem cells in the laboratory which contain clinically relevant genetic mutations - ?diseases in a dish? - may give us insights in to the early developmental mechanisms in certain genetic disorders.
With the Assisted Conception Unit (ACU) at Guy s we have been using surplus embryos donated by patients undergoing preimplantation genetic diagnosis treatment (PGD). PGD is offered to couples who carry a serious genetic disorder where the embryos are tested for the disorder prior to implantation in order to prevent transmission of diseases such as Huntington s disease, Duchenne muscular dystrophy, and spinal muscular atrophy. Embryos that are found to be free of disorder can be transferrred back to the patient s womb, or frozen for their later use. Those which will develop the genetic disorder or are of uncertain status and should not be placed back into the woman?s womb have been used to generate stem cell lines which contain the specific genetic mutation, and will be useful in understanding more about these diseases, and developing new treatments for them. All stem cell derivation from embryos is strictly monitored in the UK and requires a licence from the Human Fertilisation and Embryology Authority. It is a condition of licence that samples of any suitable stem cell lines developed must sent to the UK Stem Cell Bank from where they are made freely available to research scientists in the UK and abroad through the MRC Stem Cell Steering Committee.

Technical Summary

The development and support of the UK stem cell bank at NIBSC is a clear signal of the UK?s commitment to support both local and international research on stem cells, and to the development of lines suitable for research and clinical use. Besides the need to generate human embryonic stem (hES) cell lines for therapeutic use, there is the opportunity to use hES cell lines as an in vitro alternative to animal experiments for studying cell-cell or developing organ interactions, and the effect of pharmacological agents. Such an approach is likely to be especially useful in lines generated from embryos shown to contain specific clinically relevant genetic mutations following PGD. This strategy is predicated on the assumption that disease-carrying hESCs will differentiate faithfully and reliably into relevant differentiated cells that are functionally competent. This application seeks to test the hypothesis that disease-carrying hESCs reliably express the disease phenotype in neuroprogenitors and muscle cells after hESC differentiation in vitro. Being the largest and most successful PGD unit in the UK, and the first internationally to establish and characterise a line carrying the common deltaF508 mutation, as well as two lines containing HD expansions, we are in a leadership position to both establish and characterise additional disease carrying hESCs. It is our intention, using morphological, immunological, and molecular markers, to explore the development of neurones in stem cells with HD expansions, cardiomyocyte development and physiology in Duchenne muscular dystrophy (DMD) and neurones and myocytes in spinal muscular atrophy (SMA), which are among our most common diagnostic requests. In addition to these, we are likely to be able to generate lines with other clinically relevant mutations, which will be an invaluable research resource to others investigators which we will make available internationally through the UK Stem Cell Bank (UKSCB).

Publications

10 25 50
 
Description Advisory Committee on Safety of Cells Tissues and Organs
Geographic Reach Multiple continents/international 
Policy Influence Type Participation in advisory committee
Impact Member advising on testing and use of reproductive and stem cell tissues.
 
Guideline Title ESHRE guidelines on practice of PGD
Description ESHRE PGD Guideline
Geographic Reach Europe 
Policy Influence Type Citation in clinical guidelines
Impact Revision of Guideline to influence standards and practice of preimplantation genetic diagnosis and preimplantation genetic screening which will form the basis of other documents and compliance with EU directives
 
Description European human embryonic stem cell registry - hESCreg
Geographic Reach Europe 
Policy Influence Type Participation in advisory committee
Impact Provides information about what lines are available for distribution in Europe, their quality stage and particular characteristics which may be useful in developing lines for therapy
 
Description Joint committee on the Human Tissue and Embryos (Draft) Bill
Geographic Reach National 
Policy Influence Type Gave evidence to a government review
Impact Contributed to the demise of joint HFEA/HTA ALB to be called RATE. Remained as separate bodies and stem cell derivation remained under HFEA
 
Description Progress Educational Trust Advisory Board
Geographic Reach National 
Policy Influence Type Participation in advisory committee
 
Description Route Map for Stem Cell Regulation
Geographic Reach Multiple continents/international 
Policy Influence Type Participation in a national consultation
Impact Joint meeting of regulators to simplify processing of stem cell applications through HFEA, HTA, MHRA for clinical application
 
Description MRC Research Grant (G0701172)
Amount £600,000 (GBP)
Funding ID G0701172 
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 02/2009 
End 01/2012
 
Description MRC Research Grant (G0801061)
Amount £484,420 (GBP)
Funding ID G0801061 
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 04/2009 
End 03/2012
 
Description Refurbishment of IVF lab and stem cell laboratories
Amount £1,500,000 (GBP)
Organisation King's College London 
Sector Academic/University
Country United Kingdom
Start 01/2007 
End 12/2008
 
Description Welcome biomedical ethics strategic award
Amount £800,000 (GBP)
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 03/2009 
End 04/2014
 
Title Clinical grade hES cell lines 
Description We have derived 8 clincal grade hES cell lines from normal embryos cryopreserved in-house or from one of the ten collaborating IVF units (HFEA licence R0133). 
Type Of Material Cell line 
Provided To Others? No  
Impact We are in process of depositing lines in UKSCB. When process is completed, the lines will be available to researchers worldwide. 
 
Title Human embryonic stem cell lines with clinically relevant mutations 
Description The sixteen lines of stem cells on the NIH Registry carry genes for various hereditary disorders including Duchenne Muscular Dystrophy, Huntington's disease, cystic fibrosis, and rarer conditions such as Von Hippel-Lindau Syndrome, Wiskott-Aldrich syndrome, spinal muscular atrophy, myotonic dystrophy and neurofibromatosis. 
Type Of Material Cell line 
Provided To Others? No  
Impact Sixteen of human embryonic stem (hES) cell lines derived at King's have been approved by the US National Institutes of Health (NIH) and placed on their Stem Cell Registry, making them freely available for federally-funded research in the USA. King's is now one of the five biggest providers of disease-specific human embryonic stem cells lines on the NIH Registry, and the largest from the UK. The development is a significant milestone for King's and keeps the university at the forefront of global research into regenerative medicine. 
URL http://grants.nih.gov/stem_cells/registry/current.htm
 
Title Human embryonic stem cell lines with clinically relevant mutations 
Description We have so far derived 22 lines carrying clinically relevant genetic mutations derived from affected embryos following preimplantation genetic diagnosis (PGD) (HFEA licence R0075) which includes cystic fibrosis (4 lines), Huntington's disease (7 lines), myotonic dystrophy type 1 (1 line), neurofibromatosis type 1 (2 lines), spinal muscular atrophy type 1 (1 line), Von Hippel-Lindau cancer syndrome (3 lines), ß-thalassemia (2 line), Wiskott-Aldrich syndrome and cystic fibrosis carrer (1 line), and Turner syndrome (1 line). 
Type Of Material Cell line 
Year Produced 2013 
Provided To Others? Yes  
Impact The cells are or will be deposited in the UK Stem Cell Bank. The deposition process is ongoing. The cell lines will be then be available to other researchers. The cell lines are also sumnitted to NIH Registry 
URL http://www.kcl.ac.uk/medicine/research/divisions/wh/groups/medicine/hescell.aspx
 
Title Reseearch grade normal hES cel lines 
Description We have derived 9 lines from normal embryos cryopreserved in-house or from one of the ten collaborating IVF units (HFEA licence R0133). 
Type Of Material Cell line 
Year Produced 2007 
Provided To Others? Yes  
Impact The cell lines are either submitted or in process of submission to UKSCB. They will be available to researchers worldwide. 
 
Description hESCCO (Human Embryonic Stem Cell Coordinators) 
Organisation Saint Mary's Hospital, Manchester
Department Department of Obstetrics and Gynaecology
Country United Kingdom 
Sector Hospitals 
PI Contribution The hESCCO group was formed before this grant was approved. Therefore any collaborations made had already been established. With regard to this, we organised group meetings and established lines of communication between parties.
Collaborator Contribution Knowledge transfer and experience.Knowledge transfer and experience
Impact A group patient information and consent form has been produced in the past to give patients uniform information. The group has established a website in which to communicate. We have also produced a joint publication - Ref 18154466.
 
Description hESCCO (Human Embryonic Stem Cell Coordinators) 
Organisation Sheffield Teaching Hospitals NHS Foundation Trust
Department Assisted Conception Unit
Country United Kingdom 
Sector Hospitals 
PI Contribution The hESCCO group was formed before this grant was approved. Therefore any collaborations made had already been established. With regard to this, we organised group meetings and established lines of communication between parties.
Collaborator Contribution Knowledge transfer and experience.Knowledge transfer and experience
Impact A group patient information and consent form has been produced in the past to give patients uniform information. The group has established a website in which to communicate. We have also produced a joint publication - Ref 18154466.
 
Description hESReg European Stem Cell Registry 
Organisation European Commission
Department EC FP6 Collaborative Projects
Country European Union (EU) 
Sector Academic/University 
PI Contribution Member of Scientific Advisory board and contributor to Bank registry
Collaborator Contribution Contribute to data in registry Braude is member of scientific advisory committee
Impact First European wide stem cell registry of Embryonic and iPS stem cells http://www.hescreg.eu/
Start Year 2007
 
Description Bioethics study day ~London 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Primary Audience Policymakers/politicians
Results and Impact Glenda Cornwell & Emma Stephenson Churches Together in England Bioethics Discussion Day: Stem Cells - What are they and why might they be useful?

Advising and discussing with church ideas of stem cell research and need for use of human embryos
Year(s) Of Engagement Activity 2007
 
Description Clinical embryologist meeting 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Primary Audience Public/other audiences
Results and Impact ACE meeting Edinburgh (Jan 09). "Implications of donating embryos for stem cell research"

None
Year(s) Of Engagement Activity 2009
 
Description DKF Research Conference, Bern, Switzerland 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact More than 100 academic researchers attanded the lecture

Intense Q&A session lasting more than 30 minutes
Year(s) Of Engagement Activity 2012
 
Description Embryologist national meeting 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Primary Audience Health professionals
Results and Impact Jan 09 - "Implications of donating embryos for stem cell research, ACE meeting, Edinburgh

Information for UK embryologists about stem cell research; need, and practical and ethical implications
Year(s) Of Engagement Activity 2009
 
Description Infertility nurses meeting 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Primary Audience Health professionals
Results and Impact Glenda Cornwell Sept 07 - "Embryonic Stem Cell Research: The nurses role" National Fertility Nurse annual conference

Informing nurses involved in assisted conception units about need and use of surplus embryos for stem cell research
Year(s) Of Engagement Activity 2007
 
Description Oxford May Music 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Primary Audience Public/other audiences
Results and Impact Presentation on 'Embryonic stem cells - strings attached' - part of science and music programme held in oxford for lay audience booking ahead and attendiing for oncerts

Difficult to assess other than requests for other similar talks for schools.
Year(s) Of Engagement Activity 2009
 
Description Public Lecture Oxford 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Primary Audience Public/other audiences
Results and Impact May 2009 - Oxford May Music series- Human embryonic stem cells; Strings attached. General audience in science and music series.

Increased awareness of achievements of stem cell research and need for use of eggs and embryos
Year(s) Of Engagement Activity 2008
 
Description Seminar - London 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Primary Audience Health professionals
Results and Impact Presented update of research results to embryologists and research nurses from collaborating units

Continuing acquisition of research embryos from collaborating units
Year(s) Of Engagement Activity 2009,2010,2011
 
Description Stem cells and regenerative medicine seminars, Ljubljana, Slovenia 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Undergraduate students
Results and Impact About 30 undergraduate students and 10 faculties attanded a series of 6 lectures on stem cells and regeneratiev medicine

University of Ljubljana, Faculty of Pharmacy is currently applying for a Public Tender for cofinancing activities in years 2012-15, which stimulate openness of Slovenian Higher education into international environment released by Ministry of Education, Science, Culture and Sport - financed by the European Union and the Republic of Slovenia.

D Ilic has been invited to participate as a guest expert and leader of international Workshop in Stem cells and Regenerative Mediince within the proposed project if being selected at the public tender.
Year(s) Of Engagement Activity 2012
 
Description Workshop on stem cell & cancer research 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact CONFERENCE: King's College London and national Yang-Ming University, Taiwan, workshop on stem cell & cancer research
DATE: 17-19 Sep 2012
ORGANIZER: www.kcl.ac.uk
TITLE: Pluripotent stem cells at King's
AUTHORS: Dusko Ilic
LOCATION: King's College London, London, UK

networking, promotion of our research and resources
Year(s) Of Engagement Activity 2012
 
Description video clip on BBC website 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact SERIES: Health Explained
TITLE: How to make stem cells
DATE OF PUBLICATION: 21 Jun 2012
URL: www.bbc.co.uk/news/health-18496355

promoting our research
Year(s) Of Engagement Activity 2012