title: Slit-Robo GTPase-Activating Protein 2 as a metastasis suppressor in osteosarcoma.
abstract: Osteosarcoma is the most common primary bone tumor, with metastatic disease responsible for most treatment failure and patient death. A forward genetic screen utilizing Sleeping Beauty mutagenesis in mice previously identified potential genetic drivers of osteosarcoma metastasis, including Slit-Robo GTPase-Activating Protein 2 (Srgap2). This study evaluates the potential role of SRGAP2 in metastases-associated properties of osteosarcoma cell lines through Srgap2 knockout via the CRISPR/Cas9 nuclease system and conditional overexpression in the murine osteosarcoma cell lines K12 and K7M2. Proliferation, migration, and anchorage independent growth were evaluated. RNA sequencing and immunohistochemistry of human osteosarcoma tissue samples were used to further evaluate the potential role of the Slit-Robo pathway in osteosarcoma. The effects of Srgap2 expression modulation in the murine OS cell lines support the hypothesis that SRGAP2 may have a role as a suppressor of metastases in osteosarcoma. Additionally, SRGAP2 and other genes in the Slit-Robo pathway have altered transcript levels in a subset of mouse and human osteosarcoma, and SRGAP2 protein expression is reduced or absent in a subset of primary tumor samples. SRGAP2 and other axon guidance proteins likely play a role in osteosarcoma metastasis, with loss of SRGAP2 potentially contributing to a more aggressive phenotype.
authors.affiliation: University of Minnesota, Masonic Cancer Center Minneapolis, MN, USA.
authors.forename: Tracy A
authors.initials: TA
authors.lastname: Marko
authors.affiliation: Department of Biomedical Engineering University of Minnesota, Minneapolis, MN, USA.
authors.forename: Ghaidan A
authors.initials: GA
authors.lastname: Shamsan
authors.affiliation: University of Minnesota, Masonic Cancer Center Minneapolis, MN, USA.
authors.forename: Elizabeth N
authors.initials: EN
authors.lastname: Edwards
authors.affiliation: University of Minnesota, Masonic Cancer Center Minneapolis, MN, USA.
authors.forename: Paige E
authors.initials: PE
authors.lastname: Hazelton
authors.affiliation: University of Minnesota, Masonic Cancer Center Minneapolis, MN, USA.
authors.forename: Susan K
authors.initials: SK
authors.lastname: Rathe
authors.affiliation: University of Minnesota, Masonic Cancer Center Minneapolis, MN, USA.|Comparative Pathology Shared Resource, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Cornax
authors.affiliation: University of Minnesota, Masonic Cancer Center Minneapolis, MN, USA.|Comparative Pathology Shared Resource, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Paula R
authors.initials: PR
authors.lastname: Overn
authors.affiliation: University of Minnesota, Masonic Cancer Center Minneapolis, MN, USA.
authors.forename: Jyotika
authors.initials: J
authors.lastname: Varshney
authors.affiliation: Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Brandon J
authors.initials: BJ
authors.lastname: Diessner
authors.affiliation: University of Minnesota, Masonic Cancer Center Minneapolis, MN, USA.|Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.|Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Branden S
authors.initials: BS
authors.lastname: Moriarity
authors.affiliation: University of Minnesota, Masonic Cancer Center Minneapolis, MN, USA.|Comparative Pathology Shared Resource, University of Minnesota, Minneapolis, MN, USA.|College of Veterinary Medicine, Department of Veterinary Population Medicine, University of Minnesota, Minneapolis, MN, USA.
authors.forename: M Gerard
authors.initials: MG
authors.lastname: O'Sullivan
authors.affiliation: Department of Biomedical Engineering University of Minnesota, Minneapolis, MN, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Odde
authors.affiliation: University of Minnesota, Masonic Cancer Center Minneapolis, MN, USA.|Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Largaespada
mesh_terms: D000818:Animals; D001859:Bone Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D020690:GTPase-Activating Proteins / Q000235:genetics* / Q000378:metabolism*; D055786:Gene Knockout Techniques; D016147:Genes, Tumor Suppressor; D005820:Genetic Testing; D006801:Humans; D051379:Mice; D060787:Neoplasm Grading; D009362:Neoplasm Metastasis; D012516:Osteosarcoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D017423:Sequence Analysis, RNA
chemical_list: D020690:GTPase-Activating Proteins; C501995:SRGAP2 protein, human; C491402:Srgap2 protein, mouse