Developing biomimetic matrices for enhanced cellular reprogramming

The Nobel Prize winning discovery of induced pluripotent stem (iPS) cells by Takashi and Yamanaka in 2006 represented a groundbreaking advance in stem cells and regenerative medicine. Pluripotency is a functional state that implies the ability to form all tissues in the organism; it simulates the very beginnings of the founder tissue of a foetus. The ability to create this pluripotent state from adult cells in a laboratory liberated researchers from having to rely on embryos to produce these cells, and furthermore allowed them to have cells that were potentially competent to produce any type of cell for tissue regeneration. Given that iPS cells come from adult cells from a prospective patient, the likelihood of rejection from tissues is also significantly reduced. iPS cells therefore represent an attractive source of patient-specific cells for drug discovery, as well as more directly for genetic correction and treatment of numerous human diseases.
However, current reprogramming strategies typically take weeks and the efficiency of this process is extremely low. Furthermore, there is much work to be done to optimize the induction of pluripotency in human cells. Therefore, there is significant scope for advancing the process by which pluripotency is induced. In particular, almost nothing is known about how physical cues such as shape, topography and stiffness might regulate the establishment of pluripotency.
The lack of insight into how physical cues drive pluripotent reprogramming is particularly interesting considering that pluripotency is originally established in a highly physical environment - the developing embryo. The inspiration for the proposed research is to take what we know about the developing embryo - its spherical shape, its softness, the chemical contacts of pluripotent cells in the embryo - and attempt to create it in laboratory conditions. This is in contrast to the way most iPS research is done, in which the cells are plated on a flat, hard dish made from plastic. We are synthesizing biomimetics - meaning materials that mimic biomaterials - to simulate the embryonic environment as closely as possible to optimise the induction of pluripotency in cells. We propose that this will make the process of reprogramming more efficient, and also that these cells will be highly amenable to being guided into specific tissue cells. We can use the same biomimetic ideas to guide the cells into specific lineages. The ultimate goal of this type of research is to receive cells from a patient and create - using biomimetics - tissue competent cells for regenerating organs. This is a highly cross-disciplinary proposal that will also lend greater insight into pluripotent cell function, and how these cells interact with their environment.
The research will impact biotechnology, regenerative medicine and stem cell biology. It will bring to bear new insight into how stem cells work, and how we can investigate them. Using our connections in stem cells, biophysics, and biotechnology, we will widely circulate our results, generating impact in several academic disciplines. Given its high potential for impact in regenerative medicine and its highly cross-disciplinary nature, the proposed research is highly suited for the portfolio of the MRC.

iPS cells represent an attractive source of patient-specific cells for drug discovery and tissue regeneration, as well as more directly for genetic correction and treatment of numerous human diseases. However, current reprogramming strategies typically take weeks and the efficiency of this process is extremely low. Furthermore, there is much work to be done to optimize the induction of pluripotency in human cells. Therefore, there is significant scope for advancing the process by which pluripotency is induced. In particular, almost nothing is known about how physical cues such as shape, topography and stiffness might regulate the establishment of pluripotency. We propose to synthesise biomimetic substrate protocols to optimise the process of reprogramming. In developing these protocols, we will tune several parameters on both natural and synthetic matrices, including their stiffness, the distribution and concentration of bioactive materials on their surface, their topography, and whether they are 2D or 3D. When we have maximised the efficacy of cellular reprogramming, we will characterise the transition from a mesenchymal to epithelial phenotype these cells undergo during the process of reprogramming. We will also characterise their mechanical phenotypes to better understand this transition and what it means for the cells' physical interactions with their substrates and environment. Ultimately, we will use animal models to show that these cells possess optimal lineage plasticity and that we can use our biomimetic platforms to guide the iPS cells efficiently towards specific lineages. We will also demonstrate their utility in generating human iPS cell lines that can be used for regenerative medicine.
We will ultimately develop commercialisable biomimetic technology optimised for iPS and ESC culture. In doing so, we will improve the field of cellular reprogramming and bring significant impact to bear on the fields of regenerative medicine and bioengineering.

The proposed research will impact human health by providing novel and effective approaches for directing stem cell fate as the basis for regenerative medicine. Beyond the academic realm, we have identified the primary beneficiaries of the proposed research.
1. This research will impact commercial technologies in stem cell research and regenerative medicine by providing new insight on harnessing pluripotency. We are proposing to develop culture systems that are catered to receive a patient's cells, reprogram them in an optimised state of lineage plasticity, and specify their lineage using biomimetics. This development will be a key nucleus for future tissue engineering and healthcare technologies in the UK. The research can also benefit the biotechnology industry, in that we are proposing new methods to synthesise biomimetic substrates for cell culture. The protocols we develop in the course of this research will be highly commercialisable and will leverage new means for the biotech industry to synthesise new biomaterials.
2. General public: Our research will help identify the role of stem cells for regenerative medicine by using a physical science based approach to gain fuller insight into the process of reprogramming. The anticipated availability of new biotechnology promises to have impact on medical practice. Ultimately the efficacy of health care will be improved and the related treatment costs be reduced. This will enhance the quality of life on a national level.
Our envisioned pathway to the eventual impact described above will adopt the following route.
Personal contacts with industrial partners: The investigators have existing industrial contacts, from imaging and biotechnology (KC) to pharmaceutical (CCA) to stem cell technology (JS). These industrial connections will be used for commercialisation of intellectual property that may arise from these novel biomimetic platforms for iPS cell culture and any other results from the proposed research.
Stem Cell Institute: The WT/MRC Stem Cell Institute will provide significant exposure of our work to a large network of clinician scientists working within the Institute and also at the Addenbrooke's site in Cambridge. This will primarily occur through Institute events including the Stem Cell Club.
Conferences: We will work with the Physics of Living Matter initiative at Cambridge to organise a conference on physical biology of development and stem cells. We will also travel to numerous international conferences and seminars to disseminate our results.
Public engagement: Our public talks have resulted in tremendous feedback: the public is excited to think about biology from this unique perspective. KC and JS recently gave public lectures for the Cambridge Science Festival, both of which generated significant interest, and KC has given public lectures for the Royal Society on physics and engineering principles in stem cells. JS also speaks widely to lay audiences on reprogramming and regenerative medicine. We will amplify these efforts, and expand them to include other avenues of public engagement.
Collaboration. We will devise an appropriate series of collaboration agreements with partners to explore with suitable specialists the right way to deliver these advances into the innovation landscape. We are particularly interested in building further collaborations to enable new biotechnology for new stem cell technology. We have extensive collaborations in stem cells and biotechnology which we can rely on to further extend our network.
Exploitation. It is expected that patentable IP will arise from this project. We are already working with Cambridge Enterprise, the University's IP commercialisation subsidiary, on filing patents for the technology and at the point where proof-of-premise is established. We will continue to communicate with existing industrial links outlined above to gauge value of seeking patent protection for any developments.