Abstract The neurocutaneous tumor predisposition disorder neurofibromatosis Type I (NF1) results from deregulation of RAS signaling in the MAPK pathway that leads to a wide spectrum of clinical presentations. Dermal or cutaneous neurofibromas (cNF), a Schwann cell tumor in the skin, affect more than 95% of individuals with NF1 and are a major source of emotional, physical, and social distress as NF1 patients can have thousands of these tumors covering most of their skin. Thus, patients with NF1 often identify these tumors as their greatest burden. To date, there is no available medical treatment for cNF and no known way to prevent them from developing. The major barriers that impede progress in this field are the lack of accurate models of these common cNF tumors for drug evaluation and a limited understanding of their pathogenesis as well as the identity of specific cell of origin that directly gives rise to cNF and signaling pathways essential for tumor development. We take advantage of genetic labeling for cell lineage tracing to identify mouse neural crest Cre lines that are expressed in the subpopulation of Schwann cell lineage that give rise to cNF when NF1 is deleted. We discovered that the Homeobox B7 transcription factor serves as the lineage marker to trace the developmental origin of cNF neoplastic cells that completely recapitulates human neurofibromatosis, generating a novel mouse model that spontaneously develops both cutaneous and plexiform neurofibroma. In addition, we discovered that modulation of the Hippo pathway acts as a modifier that enhances the MAPK pathway activation by NF1 loss to promote neurofibromagenesis, suggesting that dampening the Hippo pathway may serve as part of the comprehensive treatment approach for neurofibroma. This novel mouse model has begun to yield vital clues to neurofibroma pathogenesis and now opens the doors for deciphering the evolution of cNF to identify effective therapies, where none exist today. Citation Format: Zhiguo Chen, Juan Mo, Jean-Philippe Brosseau, Tracey Shipman, Yong Wang, Chung-Ping Liao, Jonathan M. Cooper, Robert J. Allaway, Sara J.C. Gosline, Justin Guinney, Thomas J. Carroll, Lu Q. Le. A novel model of neurofibroma that deciphers its developmental origin and susceptibility to modification by the Hippo pathway [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr B23.