Investigation of factors that prevent endogenous regeneration of the retina by Müller stem cells
Lead Research Organisation:
University College London
Department Name: Institute of Ophthalmology
Abstract
Adult human retina harbours a population of Müller cells with stem cell characteristics. Although these cells have the ability to grow and neurally differentiate in vitro, there is no evidence that they regenerate retina in vivo. It has been thought that factors released during reactive gliosis, a glial proliferation observed in all retinal degenerative conditions, are responsible for the inhibition of retinal regeneration by endogenous stem cells. The proposed research aims to identifiy factors that may promote neural regeneration of diseased retina by stimulating the endogenous proliferation and neural differentiation of Müller stem cells. Using proteomic approaches, we will investigate the total protein and cytokine expression profile of fragments of human gliotic retina, surgically removed as part of the treatment for complicated retinal detachment, and will compare this profile with that of normal cadaveric donor retina. Factors found to be selectively increased or decreased in degenerated retina will then be examined for their ability to inhibit or promote Müller stem cell growth and differentiation in vitro. Once candidate factors have been found to be active on the inhibition or promotion of these cell functions, we will confirm their roles by blocking or promoting their activities using agonist or antagonist molecules for these factors. The work will be undertaken in collaboration with Prof. Sun Xinghai and Dr Lei Yuan from the Eye Hospital at Fudan University, Shanghai. The proteomic analysis will be performed at the UCL Institute of Ophthalmology and is expected to identify candidate factors which can be examined by both the UK and the Fudan group on Müller stem cell growth and differentiation. The project will form part of a PhD training program for a Chinese student who will visit the UCL Institute of Ophthalmology for a year. Funding for her visit and that of Prof Sun and Dr Lei, her PhD supervisors, is being sought by the Fudan University group from the Natural Science Foundation of China.
Technical Summary
Degeneration of retinal neurons is the ultimate cause of blindness resulting from prevalent conditions, such as age related macular degeneration (AMD), diabetic retinopathy (DR), end stage glaucoma, retinal detachment and retinitis pigmentosa. Although extensive research is being undertaken to replace damaged retinal neurons by neural stem cell transplantation, stimulation of endogenous proliferation and neural differentiation of retinal stem cells would be a preferred alternative to the transplantation approaches currently being investigated.
The research therefore aims to identify factors that may inhibit regeneration of diseased retina, and whose expression may be potentially altered to promote the endogenous proliferation and neural differentiation of Müller stem cells in situ. Using proteomic approaches, we will investigate the total protein and cytokine expression profile of fragments of human gliotic retina, surgically removed as part of the treatment for complicated retinal detachment, and will compare this profile with that of normal cadaveric retina. Factors selectively upregulated or downregulated in gliotic retina will be examined for their ability to inhibit or promote Müller stem cell growth and differentiation in vitro.
Candidate factors found to be active on the inhibition or promotion of these cell functions, will be further examined for their ability to block or promote stem cell activities using agonist or antagonist molecules for these factors. The work will be undertaken in collaboration with Prof. Sun Xinghai and Dr Lei Yuan from the Eye Hospital at Fudan University, Shanghai. The proteomic analysis will be performed at the UCL Institute of Ophthalmology and is expected to identify candidate factors which can be examined by both the UK and the Fudan group on Müller stem cell growth and differentiation. The project will form part of a PhD training program for a Chinese student who will visit the UCL Institute of Ophthalmology for a year.
The research therefore aims to identify factors that may inhibit regeneration of diseased retina, and whose expression may be potentially altered to promote the endogenous proliferation and neural differentiation of Müller stem cells in situ. Using proteomic approaches, we will investigate the total protein and cytokine expression profile of fragments of human gliotic retina, surgically removed as part of the treatment for complicated retinal detachment, and will compare this profile with that of normal cadaveric retina. Factors selectively upregulated or downregulated in gliotic retina will be examined for their ability to inhibit or promote Müller stem cell growth and differentiation in vitro.
Candidate factors found to be active on the inhibition or promotion of these cell functions, will be further examined for their ability to block or promote stem cell activities using agonist or antagonist molecules for these factors. The work will be undertaken in collaboration with Prof. Sun Xinghai and Dr Lei Yuan from the Eye Hospital at Fudan University, Shanghai. The proteomic analysis will be performed at the UCL Institute of Ophthalmology and is expected to identify candidate factors which can be examined by both the UK and the Fudan group on Müller stem cell growth and differentiation. The project will form part of a PhD training program for a Chinese student who will visit the UCL Institute of Ophthalmology for a year.
Planned Impact
The proposal has direct relevance to the exploration of stem cell based therapies for human application. The research aims to develop treatments for ocular conditions affecting a very large number of individuals worldwide. Degeneration of retinal neurons is the ultimate cause of blindness resulting from prevalent ocular diseases, such as age related macular degeneration (AMD), diabetic retinopathy (DR), end stage glaucoma, retinal detachment and retinitis pigmentosa, amongst others. According to Vision 2020 (www.vision2020.org), a global initiative for the prevention of blindness, AMD affects approximately 25 to 30 million people over the age of 55 worldwide, whilst glaucoma affects 60 million individuals worldwide. With recent advances in laser and surgical treatments, it is possible to prevent substantial visual loss in many patients, but in a long term, it is impossible to prevent blindness finally occurring in many individuals. The proposed research will be focused on the identification of factors that may inhibit neural regeneration of diseased retina, whose expression may be potentially altered to promote the endogenous proliferation and neural differentiation of Müller stem cells within the retina. Stimulation of endogenous proliferation and neural differentiation to replace damaged neurons would be a preferred alternative to the transplantation procedures currently being investigated. If this approach could be implemented -an injection of a given factor will be preferred to retinal transplantation surgery- it will avoid the trauma of surgical procedures and could potentially be performed with minimal resources.
People |
ORCID iD |
Gloria Limb (Principal Investigator) |
Publications
Angbohang A
(2016)
Downregulation of the Canonical WNT Signaling Pathway by TGFß1 Inhibits Photoreceptor Differentiation of Adult Human Müller Glia with Stem Cell Characteristics.
in Stem cells and development
Becker S
(2016)
Allogeneic Transplantation of Müller-Derived Retinal Ganglion Cells Improves Retinal Function in a Feline Model of Ganglion Cell Depletion.
in Stem cells translational medicine
Dellett M
(2014)
Genetic background and light-dependent progression of photoreceptor cell degeneration in Prominin-1 knockout mice.
in Investigative ophthalmology & visual science
Eastlake K
(2020)
Potential of Müller Glia for Retina Neuroprotection.
in Current eye research
Eastlake K
(2021)
Prospects for the application of Müller glia and their derivatives in retinal regenerative therapies.
in Progress in retinal and eye research
Eastlake K
(2017)
Comparison of proteomic profiles in the zebrafish retina during experimental degeneration and regeneration.
in Scientific reports
Eastlake K
(2018)
Comparative proteomic analysis of normal and gliotic PVR retina and contribution of Müller glia to this profile
in Experimental Eye Research
Jayaram H
(2014)
Optimized feline vitrectomy technique for therapeutic stem cell delivery to the inner retina.
in Veterinary ophthalmology
Limb G
(2016)
Biomaterials and Regenerative Medicine in Ophthalmology
Liu X
(2015)
IL-1ß induces IL-6 production in retinal Müller cells predominantly through the activation of p38 MAPK/NF-?B signaling pathway.
in Experimental cell research
Mansoor S
(2015)
Effects of light on retinal pigment epithelial cells, neurosensory retinal cells and Müller cells treated with Brilliant Blue G.
in Clinical & experimental ophthalmology
Moustafa MT
(2017)
Protective Effects of Memantine on Hydroquinone-Treated Human Retinal Pigment Epithelium Cells and Human Retinal Müller Cells.
in Journal of ocular pharmacology and therapeutics : the official journal of the Association for Ocular Pharmacology and Therapeutics
Ramírez C
(2013)
Hydroquinone induces oxidative and mitochondrial damage to human retinal Müller cells (MIO-M1).
in Neurotoxicology
Ramírez C
(2016)
Brimonidine Can Prevent In Vitro Hydroquinone Damage on Retinal Pigment Epithelium Cells and Retinal Müller Cells.
in Journal of ocular pharmacology and therapeutics : the official journal of the Association for Ocular Pharmacology and Therapeutics
Wakx A
(2016)
Amyloid ß Peptide Induces Apoptosis Through P2X7 Cell Death Receptor in Retinal Cells: Modulation by Marine Omega-3 Fatty Acid DHA and EPA.
in Applied biochemistry and biotechnology
Description | BIRAX |
Geographic Reach | Europe |
Policy Influence Type | Participation in a guidance/advisory committee |
Description | PhD Studentship |
Amount | £210,000 (GBP) |
Organisation | Santen Pharmaceutical Co., Ltd |
Sector | Private |
Country | Japan |
Start | 08/2016 |
End | 03/2020 |
Description | Project grant |
Amount | £75,000 (GBP) |
Organisation | Moorfields Eye Charity |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 03/2015 |
End | 11/2016 |
Description | Project grant |
Amount | £69,704 (GBP) |
Organisation | Moorfields Eye Hospital NHS Foundation Trust |
Department | The Special Trustees of Moorfields Eye Hospital General Fund |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 01/2014 |
End | 03/2015 |
Description | Studentship |
Amount | £100,000 (GBP) |
Funding ID | R180016A |
Organisation | Moorfields Eye Charity |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 02/2019 |
End | 01/2022 |
Title | Brn3b |
Description | Brn3b reporter to identify expression of Brn3b in human stem cells differentianting into retinal ganglion cells |
Type Of Material | Technology assay or reagent |
Year Produced | 2012 |
Provided To Others? | Yes |
Impact | Scentists in the USA at present using this construct to investigate embryonic stem cell differentiation into retinal ganglion cells |
Title | Development of a method to isolate Muller glial cells from pluripotent stem cells |
Description | We developed a method for isolation of Muller glia from retinal organoids derived from ESC and iPSC. We were the first to isolate these cells for use in retinal transplantation. |
Type Of Material | Technology assay or reagent |
Year Produced | 2017 |
Provided To Others? | No |
Impact | The method will facilitate the sourcing of Muller glia for the development of cell therapies to treat retinal diseases. |
Title | Muller stem cell lines |
Description | Muller stem cells derived from cadaveric human eyes |
Type Of Material | Cell line |
Year Produced | 2006 |
Provided To Others? | Yes |
Impact | Our Muller cell lines are the only cell lines available for retina research in this field. Many groups worldwide are using these cells to investigate the role of Muller glia in the pathogenesis of retinal diseases such as diabetic retinopathy and macular degeneration. So far cells have been given to approximately 100 research groups. |
Title | Refinement of an experimental model of glaucoma |
Description | We examined the efect of injecting different size and quantities of microbeads into the anterior chamber of the rat eye to develop a well charcaterized model of glaucoma in the rat. |
Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
Year Produced | 2017 |
Provided To Others? | No |
Impact | We have refined and characterized a model of experimental glaucoma for in vivo studies of this disease. |
Title | Sox2 |
Description | shRNA constructs to silence SOX2 gene in human progenitor/stem cells |
Type Of Material | Technology assay or reagent |
Year Produced | 2012 |
Provided To Others? | Yes |
Impact | Scientists are at present using the material for stem cell research |
Title | Proteomics data from zebrafish retina |
Description | Our data has been added to the ProteomeXchange to contribute to the full proteomics mapping of the zebrafish retina which is not yet complete. |
Type Of Material | Database/Collection of data |
Year Produced | 2016 |
Provided To Others? | Yes |
Impact | The mass spectrometry proteomics data have been deposited in the ProteomeXchange Consortium database via the PRIDE partner repository 47 with the dataset identifier |
URL | http://proteomecentral.proteomexchange.org |
Description | Investigation of endogenous regeneration of the retina |
Organisation | Fudan University |
Country | China |
Sector | Academic/University |
PI Contribution | Supervision of PhD student to undertake research on the project |
Collaborator Contribution | Sent student to undertake research on the project in my laboratory |
Impact | Paper submitted for publication. |
Start Year | 2012 |
Description | Proteomics assays |
Organisation | University College London |
Department | Institute of Child Health |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We performed proteomics analyses of human and zebrafish retina |
Collaborator Contribution | Provided equipment and knowledge of know how |
Impact | Paper in preparation for publication |
Start Year | 2013 |
Description | Proteomics in zebrafish retina |
Organisation | University College London |
Department | Institute of Ophthalmology UCL |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Investigated proteomics profile in the zebrafish retina |
Collaborator Contribution | Provided methodology and analysis of tissue used for proteomics |
Impact | A paper was published |
Start Year | 2013 |
Description | Sourcing human retina tissue |
Organisation | Moorfields Eye Hospital NHS Foundation Trust |
Country | United Kingdom |
Sector | Public |
PI Contribution | Investigated the proteomic profile of gliotic tissue |
Collaborator Contribution | Collaborator provided tissue from patients for research |
Impact | Paper publication |
Start Year | 2013 |
Description | ARVO meeting 2014b |
Form Of Engagement Activity | Scientific meeting (conference/symposium etc.) |
Part Of Official Scheme? | No |
Type Of Presentation | poster presentation |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Association for Research in Vision and Ophthalmology- Annual meeting. Orlando, Fl, USA, May 3-8 2014 Title of presentation: Up-regulation of the Notch and Wnt signalling pathways by HB-EGF in adult human Müller stem cells in vitro. Authors: Angshumonik Angbohang, Karen Eastlake, Na Wu, Silke Becker, G. Astrid Limb. Scientific discussions with other researchers in the field |
Year(s) Of Engagement Activity | 2014 |
Description | ARVO meeting 2014c |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Promoted interesting scientific discussions Have received expressions of interest from the industry |
Year(s) Of Engagement Activity | 2014 |
Description | ARVO meeting 2014d |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | N/A Discussions have helped forward the research |
Year(s) Of Engagement Activity | 2014 |
Description | ARVO meeting 2015a (USA) |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Poster presentation and scientific discussions with other researchers |
Year(s) Of Engagement Activity | 2015 |
Description | ARVO meeting 2015d (USA) |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Poster presentation and scientific discussions with other researchers |
Year(s) Of Engagement Activity | 2015 |
Description | Eye Research conference |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Scientific presentation and discussions with other researchers |
Year(s) Of Engagement Activity | 2016 |
Description | Eye Research conference (Poster presentation) |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Poster presentation and scientific discussions with other researchers |
Year(s) Of Engagement Activity | 2016 |
Description | Feature on visual disability |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | Yes |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | People becoming more aware of visual disability and prospects for future therapies Have received emails from the public for information on my work |
Year(s) Of Engagement Activity | 2013 |
Description | International Society for Ocular Cell Biology |
Form Of Engagement Activity | Scientific meeting (conference/symposium etc.) |
Part Of Official Scheme? | No |
Type Of Presentation | Poster Presentation |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Title: Investigation of factors that prevent endogenous regeneration of the adult human retina by Müller stem cells: proteomic comparison between normal and gliotic retina. Authors: Karen Eastlake, Wendy Heywood, Kevin Mills, Peng Khaw, Philip Banerjee, David G Charteris, G. Astrid Limb. Scientific discussion of findings with other researchers |
Year(s) Of Engagement Activity | 2013 |
Description | Invitation to speak at retina conference |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | I was invited to give a talk at a symposium organized by the German Foundation PRO RETINA. The meeting was attended by circa 400 Scientists, Young Research Fellows and Eye Charity members |
Year(s) Of Engagement Activity | 2016 |
Description | Invited talk |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Industry/Business |
Results and Impact | Circa 60 people attended the workshop which generated much discussion and informed Industry of new research developments on treatments for retinal diseases. |
Year(s) Of Engagement Activity | 2018 |
Description | Invited talk |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Postgraduate students |
Results and Impact | Circa 70 people attended talk, which generated much discussion with various individual groups on the day. |
Year(s) Of Engagement Activity | 2018 |
Description | Invited talk at Eye research Conference |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Over 100 people attended talk at Workshop organized as part of an Eye conference. Talk generated important Scientific discussions. |
Year(s) Of Engagement Activity | 2018 |
Description | Meeting of the Scientific and Medical Advisory Board of Retina International |
Form Of Engagement Activity | A formal working group, expert panel or dialogue |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Presentation generated important discussions with retina specialists N/A |
Year(s) Of Engagement Activity | 2014 |
Description | Patient symposium- Moorfields Eye Hospital |
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 | Patient- Clinical discussions to understand impact of eye disease |
Year(s) Of Engagement Activity | 2015 |
Description | Scientific session moderator |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Invited to moderate Scientific session at International Eye conference. Moderated Scientific discussions amongst attendees. |
Year(s) Of Engagement Activity | 2018 |
Description | Stem cell conference |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Discussions with Scientists an Clinicians on new developments on the application of stem cells to therapies. |
Year(s) Of Engagement Activity | 2016 |
Description | Talk |
Form Of Engagement Activity | A formal working group, expert panel or dialogue |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Policymakers/politicians |
Results and Impact | I was invited to a workshop organized by a charity to storm ideas and acquire current knowledge on new retinal therapies in order to define policy on their research funding scheme. |
Year(s) Of Engagement Activity | 2018 |
Description | Talks with patient group |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Patients attending Glaucoma clinic at Moorefield's Eye Hospital were informed of current and prospective therapies available for their treatment. Have received emails from patients asking further questions. |
Year(s) Of Engagement Activity | 2014 |
Description | Worshop invitation |
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 | Invitation to close workshop organized by the National Eye Institute of the United States in May 2016, as part of their Audacious Goal Initiative to cure blindness by 2020. The main purpose of the meeting was to discuss potential approaches for replacement of retinal ganglion cells from endogenous cell sources.. |
Year(s) Of Engagement Activity | 2016 |
URL | https://www.youtube.com/watch?v=U5pEcFPsOWU&feature=youtu.be |