Introduction: Type 1 neurofibromatosis (NF1) is an autosomal dominant multisystemic disorder caused by aberrations in the neurofibromin gene. The population incidence is approximately 1 per 3000. Typically, patients develop multiple cutaneous tumours that grown from axon of peripheral nerve, called neurofibromas. These benign tumours are generally composed of Schwann cells (SC) and fibroblasts, but others cells type can also be found. Highly variable clinical manifestations between NF1 patients are observed. Actually, there is no specific treatment for this stigmatizing disease. The purpose of this study is to develop a tissue engineered human skin model derived from NF1 patients to characterized and understand the formation of neurofibromas.
Methods: The auto-assembly model was used to generate tissue-engineered skin (TES) in vitro with fibroblasts and keratinocytes isolated from NF1 patients (n=3). We used spheroid suspension culture to generated neurofibroma-like tumours; one composed of immortalized SC and another with an equal number of SC and fibroblasts. Then, spheroids were added on the dermis 10 days before keratinocytes seeding.
Results: We first determined the best conditions for the formation of spheroids. Densification area was significantly increased already at day 3 and continued until day 10. Immunofluorescence revealed that spheroids/neurofibromas-like, seeded with NF1-TES, are in a proliferative state. Furthermore, non-apparent activation of apoptosis within spheroid is detected. Seeding of keratinocytes suggests an upregulation of cells proliferation, but more investigation need to be done. Finally, results were significantly similar between NF1 cell lineages.
Conclusions: Our NF1 skin model could become a unique tool to better characterize the mechanism of action of a new drug on NF1 tumor shape and growth as well as to assess tumorigenic properties of each of the tested NF1 gene mutation, and ultimately provide better tools to develop new therapies for patients through development of precision/personalized medicine strategies.
Patient Care: This could provide a tool on which therapeutic modalities targeted toward neurofibromas may be tested in vitro
Learning Objectives: By the conclusion of this session, participants should be able to describe the methods used to replicate a skin model from NF1 patients