A developmental bioengineering approach to dissecting the cartilaginous mechanostat

Project No: KTPS-NC-15
Intake: 2021 KTPS-NC

PROJECT OUTLINE

During development, health and in disease cells are bombarded by a plethora of signals to both integrate and elicit a response to. One set of machinery that cells use is collectively associated with the primary cilium, a singular organelle that only in the last 15 years has established to be a nexus for signalling. It is both associated with modulating a range of cell behaviours, including those downstream to biological cues, but also regulating the response to mechanical forces. However, its role in mechanotransduction is still putative (R et al. 2019).

The Wann lab has been exploring the role of ciliary genes and proteins in MSK cells and tissues. In vitro the cilium is important to the response of cells cultured in 3D to compressive forces and mechanically regulated matrix remodelling pertinent to osteoarthritis. We have previously explored how the cilium regulates and integrates extracellular cues in isolated cells but more recently we have recently built a body of evidence from tissue, to support the concept that the cilium acts as part of a tissue ‘mechanostat’ in vivo. For example in the context of epiphyseal fusion, a ciliary protein acts as a mechano-dampener, critical to instructing the cessation of growth.

We are now addressing key questions in tissue mechanotransduction, attempting to define the signals and machinery that comprise a system capable of ‘tuning’ the appropriate tissue remodelling response to physical forces, against a landscape of complex biological cues. This is relevant to MSK health, including arthritis, but also the human ciliopathies.

We now aim to create and exploit organoid limb assembly (Fernando et al. 2018), use engineered tissue gradient approaches (Li et al. 2019) and organ-on-a-chip technologies, combined with multiple imaging modalities, molecular tagging (McLeod and Mauck 2016), and omics to explore the integration of bio-physico cues in 3D models to complement in vivo studies. This project will aim to use human iPSC, including those derived from ciliopathy patients.

Aims

Validate a hydrogel/matrix/cell organoid model system for studying MSK mechanobiology.
Exploit non-canonical amino acid tagging and pulsed-SILAC to visualise and measure matrix remodelling in vitro.
‘Humanise’ system with hiPSC and develop organ-on-a chip approaches.
Define the signals and machinery that comprise the cartilaginous ‘mechanostat’.

KEYWORDS

Developmental Bioengineering
Mechanobiology
Stem cell
Tissue remodelling
Imaging

TRAINING OPPORTUNITIES

This is a timely, multidisciplinary project seeking to address biological questions within the Wann lab, using established and exciting emerging technologies, supported by the interdisciplinary supervisory team and the wider Versus Arthritis centre for OA pathogenesis. Engineering approaches will be supported by Professor E.Stride and Professor M Knight, molecular biochemistry (SILAC, Professor T.Vincent KIR) and Imaging (Professor M.Dustin, KIR). There will be an opportunity to collaborate and ultimately forge a joint effort with the centre for predictive models (Professor M.Knight, Emulate-QMUL). The project will entail training in cell and tissue culture, scaffolds, matrix biology, mechanobiology and organ-on-a-chip approaches combined with imaging and molecular biology.

KEY PUBLICATIONS

Fernando, W. A., I. Papantoniou, L. F. Mendes, G. N. Hall, K. Bosmans, W. L. Tam, L. M. Teixeira, M. Moos, Jr., L. Geris, and F. P. Luyten. 2018. 'Limb derived cells as a paradigm for engineering self-assembling skeletal tissues', J Tissue Eng Regen Med, 12: 794-807.
Li, C., L. Ouyang, I. J. Pence, A. C. Moore, Y. Lin, C. W. Winter, J. P. K. Armstrong, and M. M. Stevens. 2019. 'Buoyancy-Driven Gradients for Biomaterial Fabrication and Tissue Engineering', Adv Mater, 31: e1900291.
McLeod, C. M., and R. L. Mauck. 2016. 'High fidelity visualization of cell-to-cell variation and temporal dynamics in nascent extracellular matrix formation', Sci Rep, 6: 38852.
R, R. Ferreira, H. Fukui, R. Chow, A. Vilfan, and J. Vermot. 2019. 'The cilium as a force sensor-myth versus reality', J Cell Sci, 132.
Fernando, W. A., I. Papantoniou, L. F. Mendes, G. N. Hall, K. Bosmans, W. L. Tam, L. M. Teixeira, M. Moos, Jr., L. Geris, and F. P. Luyten. 2018. 'Limb derived cells as a paradigm for engineering self-assembling skeletal tissues', J Tissue Eng Regen Med, 12: 794-807.
Li, C., L. Ouyang, I. J. Pence, A. C. Moore, Y. Lin, C. W. Winter, J. P. K. Armstrong, and M. M. Stevens. 2019. 'Buoyancy-Driven Gradients for Biomaterial Fabrication and Tissue Engineering', Adv Mater, 31: e1900291.
McLeod, C. M., and R. L. Mauck. 2016. 'High fidelity visualization of cell-to-cell variation and temporal dynamics in nascent extracellular matrix formation', Sci Rep, 6: 38852.
R, R. Ferreira, H. Fukui, R. Chow, A. Vilfan, and J. Vermot. 2019. 'The cilium as a force sensor-myth versus reality', J Cell Sci, 132.