Stem Cell Epigenetics and Biomolecular Imaging
- Principal Investigator:
- Professor Udo Oppermann
- Grant support:
- NIHR Oxford Biomedical Research Unit
- Oxford Stem Cell Institute (OSCI)
- Group Members:
- Clarence Yapp (DPhil student)
- Anneke Kramm (DPhil student)
- Sharif Halai (DPhil student)
Research vision and objectives
The landmark discovery that lineage-restricted cells can be reprogrammed directly to a state of pluripotency through ectopic expression of defined transcription factors (e.g. Oct4, SOX2, KLF4, and c-Myc) has opened a new frontier in the field of regenerative medicine. In this project, we wish to assess the potential of induced pluripotent stem (iPS) cells in regenerative medicine of musculoskeletal diseases. Stem cell therapy based on iPS cells obtained from patients and retransplanted after differentiation into functional tissue, avoids immunological rejection and offers great potential as possible therapeutic option in musculoskeletal diseases. However, before advancement of patient-derived iPS cells and differentiation into desired tissue can proceed into clinical, regenerative applications, several important aspects have to be addressed, and strategies overcoming these hurdles have to be developed. These include (i) improvement of reprogramming efficiency by utilizing our in-house expertise and availability of small molecule and siRNA libraries (focused kinase, signalling pathways and epigenetic ie methylation and acetylation inhibitors), (ii) strict avoidance of viral methods or animal material to prevent adverse genomic modifications and immunological complications, (iii) methods to assess iPS and differentiation states of iPS derived differentiated target cells, (iv) methods to monitor and manipulate possible teratogenic potentials of iPS derived cells using CHIP-seq and large-scale sequencing to detect chromatin/epigenetic modifications, and (v) methods to monitor and promote integration of differentiated cells into functional tissue.
Our vision is to successfully implement the novel strategies into a platform where we will generate iPS cells from patient-derived somatic cells, and generate from these differentiated tenocytes and chondrocytes on synthetic scaffolds ready for autologous transplantion into the patient. To begin with, we will develop alternative, non-viral transfection methods based on femtosecond laser technology (optoporation), will investigate optimal vectors for obtaining iPS cells and will systematically manipulate key pathways (senescence, inflammation, epigenetics ie DNA/protein methylations, acetylation) to obtain high numbers of stable and well-characterized iPS cells. We will develop novel non-invasive monitoring and assessment strategies for iPS and somatic cells based on near-infrared multiphoton and Raman spectroscopy; and will devise strategies (dynamic mechanical analysis, Fourier transform infrared spectroscopy) to assess iPS derived cells and matrix ex vivo in scaffold materials which would be used for transplantation.
Recent Key Publications
- Tirlapur UK, Yapp C Near Infrared Three-Dimensional Nonlinear Optical Monitoring of Stem Cell Differentiation Chapter 13.In A. Diaspro (ed.) Optical Fluorescence Microscopy. Springer-Verlag Berlin Heidelberg (2011)
- Xu X, Urban JP, Tirlapur UK, Cui Z Osmolarity effects on bovine articular chondrocytes during three-dimensional culture in alginate beads. Osteoarthritis Cartilage. 2010 Mar;18(3):433-9. Epub 2009 Oct 12.
- Mansfield J, Yu J, Attenburrow D, Moger J, Tirlapur U, Urban J, Cui Z, Winlove P The elastin network: its relationship with collagen and cells in articular cartilage as visualized by multiphoton microscopy. J Anat. 2009 Dec;215(6):682-91. Epub 2009 Oct 1.
- Guehring T, Wilde G, Sumner M, Grünhagen T, Karney GB, Tirlapur UK, Urban JP Notochordal intervertebral disc cells: sensitivity to nutrient deprivation. Arthritis Rheum. 2009 Apr;60(4):1026-34.