Manufacturing solutions for high value induced pluripotent stem cell products (MS-iPS)
Lead Research Organisation:
University of Cambridge
Department Name: Surgery
Abstract
The project aims to develop the standards of practice and testing systems that will allow research on stem cells to be translated into the clinic. The primary focus is the use of induced pluripotent cells (iPS), but the project outputs are expected to be applicable to other types of cells, including embryonic stem cells (ES). The team will have the following roles within the project:1. To provide the cellular materials to enable assay development and cell testing2. To test the 'quality' of different cell populations i.e. their stability and differentiation potential3. To analyse the profile of gene expression of the cell populations4. To analyse the genetically imprinted regions in the cell populationsThe above approaches will allow the project members to correlate different cell characteristics with the regulation of specific genes, and/or genetic imprinting. With further development, these discoveries will form the basis of new assays and tests that are used in the production of therapeutically competent cells.
Publications
Butcher LM
(2016)
Non-CG DNA methylation is a biomarker for assessing endodermal differentiation capacity in pluripotent stem cells.
in Nature communications
Carobbio S
(2021)
Unraveling the Developmental Roadmap toward Human Brown Adipose Tissue.
in Stem cell reports
Cuomo ASE
(2020)
Single-cell RNA-sequencing of differentiating iPS cells reveals dynamic genetic effects on gene expression.
in Nature communications
Eldridge CB
(2020)
A p53-Dependent Checkpoint Induced upon DNA Damage Alters Cell Fate during hiPSC Differentiation.
in Stem cell reports
Elmentaite R
(2020)
Single-Cell Sequencing of Developing Human Gut Reveals Transcriptional Links to Childhood Crohn's Disease.
in Developmental cell
Kilpinen H
(2017)
Common genetic variation drives molecular heterogeneity in human iPSCs.
in Nature
Linker SM
(2019)
Combined single-cell profiling of expression and DNA methylation reveals splicing regulation and heterogeneity.
in Genome biology
Ortmann D
(2020)
Naive Pluripotent Stem Cells Exhibit Phenotypic Variability that Is Driven by Genetic Variation.
in Cell stem cell
Rimland CA
(2021)
Regional Differences in Human Biliary Tissues and Corresponding In Vitro-Derived Organoids.
in Hepatology (Baltimore, Md.)
| Description | The aim of my group was to derive a panel of hIPSC lines from healthy individuals of different age and sex and then to characterise the capacity of the resulting lines to differentiate into the three germ layers endoderm, mesoderm and neuroectoderm. Accordingly, 45 hIPSCs lines were derived from 16 individuals 1 to 75 years old and with an equal representation of both Sex. In addition, we used two reprogramming methods (Sendai virus and Retrovirus) and different somatic cells (Skin fibroblast and vascular cells) to increase the complexity and deepness of our studies. 35 hIPSC lines were differentiated into the three germ layers and characterised by FACS, Q-PCR and immunostaining. These analyses revealed that hIPSC lines vary in their capacity of differentiation and that this variability is likely to be provoked by genetic background. We then provide large quantity of cells for epigenetic analyses to the different members of the consortium and then help them to interpret their results. We used these data to uncover new markers characterising hIPSC lines with a high capacity of differentiation toward specific germ layers. The importance of these markers was functionally validated by performing gain and loss of function experiments. These experiments revealed new mechanisms associating cell fate decisions and genetic. |
| Exploitation Route | Explotation plan for Manufacturing solutions for high value induced pluripotent stem cell (iPSC) products (MS-iPS). The aim is to produce a minimum viable population-signature which can distinguish good from bad iPSCs in terms of differentiation capacity. The analysis will group induced pluripotent cells derived from patient tissue samples in terms of their ability to differentiate into the three cell lineages (endoderm, mesoderm and ectoderm). It will determine the key epigenetic marks which are the most important distinguishing feature to be able to divide them into "good" (those which can fully differentiate) versus "bad" (those which can't) cell lines. Initial epigenetic analysis has shown that non-CpG methylation appears to be a good discriminator, whereas CpG methylation appears not to be a good discriminator. Thus, the project will select and validate a key signature gene set in the existing cell lines. Further validation work will confirm the initial findings with the end result being the identification of a signature array which can be used as a tool. Examples of the application of this signature could be: (i) Clonal analysis of revertants i.e bad cell lines which have been rescued to good ones. (ii) Predictive capacity for cells from a completely different source/protocol but same differentiated lineages. The route to market will be via the licensing of patented gene sets for arrays. The types of products based on these innovations are well established and will initially be focussed on the non-clinical research market in the near term while pursuing clinical applications in the longer term. |
| Sectors | Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Description | International consortium large scale hIPSCs project |
| Geographic Reach | Multiple continents/international |
| Policy Influence Type | Contribution to a national consultation/review |
| Description | Manufacturing Solution for high value Induced pluripotent stem cells product |
| Amount | £656,986 (GBP) |
| Funding ID | 100757 |
| Organisation | Innovate UK |
| Sector | Public |
| Country | United Kingdom |
| Start | 09/2010 |
| End | 08/2013 |
| Title | Foregut Stem Cells |
| Description | We have identified a new type of endodermal stem cells. |
| Type Of Material | Cell line |
| Year Produced | 2014 |
| Provided To Others? | Yes |
| Impact | These cells could enable the production of homogenous population of pancreatic, lung, and hepatic cells from a broad number of hPSCs. |
| Description | Collaboration with Epigenetic Experts |
| Organisation | University College London |
| Department | UCL Cancer Institute |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | This grant has brought together experts in stem cells and epigenetics allowing the development of novel projects and innovative science. To investigate new mechanisms of regulation, to access unique expertise and to develop more competitive project. |
| Collaborator Contribution | provide expertise for data analyses |
| Impact | 1 publication |
| Start Year | 2010 |
| Description | hIPSCs and Epigenetic |
| Organisation | University of Cambridge |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | The main objective of this colaboration is to define epigenetic variability among human induced pluripotent stem cells lines. |
| Collaborator Contribution | Develop new approach to characterise hIPSCs |
| Impact | Two publications are curentrly in preparation and we obatined a grant from the TSB to continue this work. |
| Start Year | 2008 |
| Description | 4L Trophy Rally supporting Enfants du Desert |
| Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Industry/Business |
| Results and Impact | 4L Trophy Rally supporting Enfants du Desert |
| Year(s) Of Engagement Activity | 2019 |
| Description | European Researchers Night: LifeLab |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | European Researchers Night: LifeLab |
| Year(s) Of Engagement Activity | 2019 |
| Description | House of lord enquiry on regenerative medicine |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | Yes |
| Type Of Presentation | Workshop Facilitator |
| Geographic Reach | National |
| Primary Audience | Policymakers/politicians |
| Results and Impact | NA / too soon NA/ too soon |
| Year(s) Of Engagement Activity | 2012 |
| Description | Talk science festival 2019 |
| 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 at Cambridge science festival |
| Year(s) Of Engagement Activity | 2019 |