TrkA^WT;NOD-Scid and TrkA^F592A;NOD-Scid xenograft osteosarcoma (orthotopic) implants at 12 d post-143B OS cell innoculation for scRNA-Seq.

Bone pain is a presenting feature of osteosarcoma (OS) mediated by skeletal-innervating peripheral afferent neurons. Potential regulatory roles of skeletal-innervating neurons in OS beyond pain sensation are unknown. Here, a chemical-genetic approach was utilized to perturb innervation during osteosarcoma growth and disease progression. TrkA inhibition in transgenic mice led to a significant reduction in sarcoma-associated innervation and vascularization, tumor growth, and metastasis, and prolonged overall survival. Single-cell transcriptomics showed that chemical denervation led to prominent dysregulations in neurotrophin and BMP signaling among sarcoma cells, as well as VEGF signaling within tumor-associated endothelial cells. The importance of sarcoma innervation and neural-sarcoma crosstalk was further confirmed using human OS samples and sequencing datasets. In a translational effort, chemical inhibition of peripheral nerve growth via FDA-approved bupivacaine liposomes both alleviated tumor-associated bone and significantly delayed tumor growth. In sum, TrkA-expressing peripheral neurons positively regulate OS progression and neural inhibition appears to disrupt neurovascular coupling within the sarcoma leading to significantly reduced tumor growth and improved disease progression. These data suggest that interventions to prevent pathological innervation of osteosarcoma may represent a novel adjunctive therapy to improve clinical management. Tumor xenograft implants were dissected from TrkA^WT;NOD-Scid and TrkA^F592A; NOD-Scid animals at 12 d post-tumor cell innoculation for scRNA-Seq. The two sequenced samples were pooled triplicates of TrkA^WT and TrkA^F592A animals.