Abstract 4128: Loss of p53 and Rb enhances ligand-dependent hedgehog signaling in development and cancer
Authors: Catherine R. Cochrane, Anette Szczepny, Samantha W. Jayasekara, Vijesh Vaghjiani, D. Watkins, Jason E. Cain
Journal: Cancer Research (2018)

Abstract The evolutionary conserved Hedgehog (Hh) signaling pathway plays fundamental morphogenic and mitogenic roles in tissue development and homeostasis, as well as in the initiation and progression of various cancers. Signaling is driven by the binding of Sonic hedgehog (Shh) ligand to the receptor Patched (Ptch1), to initiate primary cilia dependent Smoothened (Smo)-Gli activation. Although mutations in Hh pathway components are described in some cancers, the majority of Hh tumors exhibit ligand-dependent Hh signaling, where Hh ligand is produced by the stroma and/or tumor cells to maintain pathway activation and self-renewal. Here, we have utilized various Hh-dependent developmental models to better understand the tumorigenic mechanisms of Hh ligand-dependent signaling in cancer. During cerebellar development, Shh is secreted from cerebellar purkinje cells for rapid expansion of the cerebellar granule cell precursor (GCP) population in the external granule layer (EGL). Inactivation of Shh in purkinje cells (Pcp2Cre;Shhfl/fl) leads to reduced GCP proliferation, premature eradication of the EGL and reduced cerebellar size. In contrast, Shh overexpression by purkinje cells (Pcp2Cre;ShhTg) results in increased GCP expansion, a sustained EGL and increased cerebellar size. Indeed, signs of remnant EGL persist into adulthood, similar to the pre-neoplastic lesions observed in Ptch1+/- mice, believed to be the origin of SHH medulloblastoma. During skeletal development, Indian hedgehog secreted from pre-hypertrophic chondroblasts is required for osteoblast progenitor specification from multipotent mesenchymal stem cells. Sustained Hh signaling in osteoblast precursors (OsxCre;SmoM2 or OsxCre;ShhTg) results in reduced ossification of the calvaria, maxilla and mandible, reduced long bone width, and reduced expression of mature osteoblast markers. Together, these models implicate sustained Hh expression in the maintenance of an immature cell state at the expense of differentiation. Moreover, in a mouse model of osteosarcoma, inactivation of Smo (OsxCre;p53fl/fl;Rbfl/fl;Smofl/fl) abolished the malignant phenotype of OsxCre;p53fl/fl;Rbfl/fl mice, leading to benign osteoid osteoma consistent with osteoblast differentiation. Increased Hh signaling in cancers with prevalent p53 and Rb mutations led us to investigate a role for p53 and Rb in Hh ligand-dependent signaling. Genetic inactivation of p53 and/or Rb in the developing mouse neural tube resulted in expansion of the Hh-dependent Nkx2.2 ventral domain, consistent with a Hh gain of function phenotype. Similarly, siRNA knockdown of p53 and/or Rb in mouse mesenchymal stem cell line C3H10T1/2 markedly increased Hh-dependent osteoblast specification in response to Hh ligand. Taken together, we propose that p53 and Rb mutations enhance ligand-dependent Hh pathway activation that maintains cells in a primitive state and promotes tumorigenesis. Citation Format: Catherine R. Cochrane, Anette Szczepny, Samantha W. Jayasekara, Vijesh Vaghjiani, D. Watkins, Jason E. Cain. Loss of p53 and Rb enhances ligand-dependent hedgehog signaling in development and cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4128.