Spatial transcriptomic analysis of a pre-clinical NF1 pseudarthrosis fracture model

Patients with Neurofibromatosis Type 1 (NF1) present with fracture pseudarthroses, though the exact mechanism underlying this abnormal healing remains unknown. Here, we performed spatial transcriptomics to spatially-define the molecular signatures across endochondral healing following fracture. Integrating single-cell sequencing of patient fracture-derived primary cells, our results provide a dynamic cellular context to the molecular dysregulation associated with somatic fracture healing defects in NF1. Overall design: 8-week old control and Postn-cre;Nf1[flox/-] mice underwent distal tibia fracture surgery. Fractures were stabilized with an intramedullary pin. Ten days post-fracture, calluses were harvested, fixed in formallin for 24h, decalcified in 14% EDTA for 3 days prior to embedding in paraffin. Both fracture calluses were embedded together, and two sections through the fracture callus were analyzed using the 10X Genomics Visium spatial transcriptome platform following manufacturer's recommendations. Following sequencing, bioinformatic analyses were conduced using Partek Flow.