The functional interdependence of nerves and blood vessels is a well-established concept during tissue morphogenesis, yet the role of neurovascular coupling in proper and aberrant tissue repair is an emerging field of interest. Here, we sought to define the regulatory relationship of peripheral nerves on vasculature in a severe extremity trauma model in mice, which results in aberrant cell fate and heterotopic ossification (HO). First, a high spatial degree of neurovascular congruency was observed to exist within extremity injury associated heterotopic ossification. Vascular and perivascular cells demonstrate characteristic responses to injury, as assessed by single cell RNA sequencing. This vascular response to injury was blunted in neurectomized mice, including a decrease in endothelial proliferation and type H vessel formation, and a downregulation of key transcriptional networks associated with angiogenesis. Independent mechanisms to chemically or genetically inhibit axonal ingrowth led to similar deficits in HO site angiogenesis, a reduction in type H vessels, and heterotopic bone formation. Finally, a combination of single cell transcriptomic approaches within the dorsal root ganglia identified key neural-derived angiogenic paracrine factors that may mediate neuron-to-vascular signaling in HO. These data provide further understanding of nerve-to-vessel crosstalk in traumatized soft tissues, which may reflect a key determinant of mesenchymal progenitor cell fate after injury.

神经和血管的功能相互依赖性是组织形态发生过程中一个公认的概念，但神经血管耦合在正确和异常组织修复中的作用是一个新兴的研究领域。在这里，我们试图在小鼠严重四肢创伤模型中定义周围神经对脉管系统的调节关系，这会导致异常的细胞命运和异位骨化（HO）。首先，观察到在四肢损伤相关的异位骨化中存在高度的神经血管一致性。通过单细胞 RNA 测序评估，血管和血管周围细胞表现出对损伤的特征性反应。这种血管对损伤的反应在神经切除小鼠中减弱，包括内皮增殖和H型血管形成的减少，以及与血管生成相关的关键转录网络的下调。化学或遗传抑制轴突向内生长的独立机制导致 HO 位点血管生成的类似缺陷、H 型血管的减少和异位骨形成。最后，背根神经节内单细胞转录组学方法的组合确定了可能介导 HO 中神经元至血管信号传导的关键神经源性血管生成旁分泌因子。这些数据提供了对受伤软组织中神经与血管串扰的进一步了解，这可能反映了损伤后间充质祖细胞命运的关键决定因素。