Advanced engineering of the human neuromuscular and myotendinous junctions for next generation in vitro 3D modelling of severe muscle disorders

Project background and description

Current in vitro models of human skeletal muscle are transforming our understanding of neuromuscular biology, yet important limitations remain. In particular, their inability to faithfully recapitulate interactions with key supporting tissues constrains both mechanistic insight and therapeutic development for neuromuscular diseases. Over the past years, our laboratories at the Francis Crick Institute (www.tedescolab.org; www.seriolab.org) have developed complementary expertise in stem cell biology, organoid engineering, and neuromuscular modelling. This collaboration has successfully contributed to large-scale initiatives such as the MAGIC consortium (www.magic-horizon.eu) to harness muscle models for gene therapy development. Despite these advances, modelling the critical interfaces between skeletal muscle and its two major regulatory systems - motor neurons (MNs) and tendons - remains a major unmet challenge.

Here, we propose to engineer next-generation human 3D models that integrate both the neuromuscular junction (NMJ) and the myotendinous junction (MTJ), enabling a more physiologically relevant representation of muscle function and disease. Our central hypothesis is that the combined incorporation and optimisation of these key input/output interfaces within muscle organoids will allow unprecedented dissection of disease mechanisms affecting currently incurable neuromuscular conditions primarily affecting those two compartments. We will leverage proprietary technologies from both laboratories, including iPSC-based differentiation, organoid assembly, advanced imaging, and functional interrogation platforms, alongside bioengineering approaches developed in collaboration with the Crick Making Lab. We will focus on disease-relevant paradigms affecting each interface: collagen VI- and Laminin ?2-related muscular dystrophies (impacting the MTJ), and congenital myasthenic syndromes (affecting the NMJ).

Aim 1 will generate and characterise isogenic iPSC-derived cellular components, including myogenic cells, motor neurons, fibro-adipogenic progenitors (FAPs), and tenocytes.

Aim 2 will establish a bioengineered 3D scaffold to support the assembly, maturation, and functional connectivity of neuromuscular and myotendinous interfaces.

Aim 3 will define disease-specific phenotypes using structural, molecular, and functional readouts across both junctions.

Aim 4 will provide proof-of-principle therapeutic validation by testing candidate gene therapies and repurposed small molecules interventions emerging from our MAGIC and DREAMS (www.dreamshorizon.eu) consortia, respectively.

This interdisciplinary project integrates stem cell biology, bioengineering, and computational analysis, positioning the Crick as the central hub while benefiting from strong links with UCL, King's College London, UK Dementia Research Institute and Great Ormond Street Hospital for Children. By reconstructing functional neuromuscular and myotendinous interfaces in vitro, this project will redefine how human muscle diseases are modelled, providing a transformative platform for mechanistic discovery and therapy development.

Candidate background

This project would suit candidates with a background in bioengineering, stem cell biology and computation biology, and an interest in disease modelling, OMICS- and organoid-based approaches.

Lab-specific question

Imagine that you want to differentiate human iPSCs into a specialised cell type for which no published differentiation protocol exists. How would you go about developing a differentiation strategy from first principles? Please explain your reasoning and experimental approach.

Funding Notes

Successful applicants will be awarded a non-taxable annual stipend of £27,715 plus payment of university tuition fees. Students of all nationalities are eligible to apply.

References

1. Choi, S., Ferrari, G., Moyle, L.A., Mackinlay, K., Naouar, N., Jalal, S., . . . Tedesco, F.S. (2022) Assessing and enhancing migration of human myogenic progenitors using directed iPS cell differentiation and advanced tissue modelling. EMBO Molecular Medicine 14: e14526. PubMed abstract
2. Hagemann, C., Bailey, M.C.D., Carraro, E., Stankevich, K.S., Lionello, V.M., Khokhar, N., . . . Serio, A. (2024) Low-cost, versatile, and highly reproducible microfabrication pipeline to generate 3D-printed customised cell culture devices with complex designs. PLOS Biology 22: e3002503. PubMed abstract
3. Joshi, S., Moreno-Gonzalez, C., Suklai, P., Carraro, E., Ratcliffe, C.D.H., Boezio, G.L.M., . . . Serio, A. (2025) Preprint: VISIBLE: An imaging-driven system for sampling, biofabrication, and manipulation of complex biological models. Available at: bioRxiv https://www.biorxiv.org/content/10.1101/2025.06.12.659321v2.full.pdf
4. Moore, D., Steele-Stallard, H., Pinton, L., Lionello, V.M., Rossi, L., Aghaeipour, A., . . . Tedesco, F.S. (2025) Preprint: Advanced human iPSC-based modelling of LMNA-related congenital muscular dystrophy enables development of targeted genetic therapies for muscle laminopathies. Available at: bioRxiv https://www.biorxiv.org/content/10.1101/2025.06.22.660928v2.full.pdf
5. Pinton, L., Khedr, M., Lionello, V.M., Sarcar, S., Maffioletti, S.M., Dastidar, S., . . . Tedesco, F.S. (2023) 3D human induced pluripotent stem cell-derived bioengineered skeletal muscles for tissue, disease and therapy modeling. Nature Protocols 18: 1337–1376. PubMed abstract