{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T06:41:41Z","timestamp":1784616101402,"version":"3.55.0"},"reference-count":47,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2024,8,9]],"date-time":"2024-08-09T00:00:00Z","timestamp":1723161600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Berner-Garde Foundation and the Bernese Mountain Dog Club of America"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Histiocytic sarcoma (HS) is a rare and highly aggressive cancer in humans and dogs. In dogs, it has a high prevalence in certain breeds, such as Bernese mountain dogs (BMDs) and flat-coated retrievers. Hemophagocytic histiocytic sarcoma (HHS) is a unique form of HS that presents with erythrophagocytosis. Due to its rareness, the study of HHS is very limited, and mutations in canine HHS patients have not been studied to date. In previous work, our research group identified two major PTPN11\/SHP2 driver mutations, E76K and G503V, in HS in dogs. Here, we report additional mutations located in exon 3 of PTPN11\/SHP2 in both HS and HHS cases, further supporting that this area is a mutational hotspot in dogs and that mutations in tumors and liquid biopsies should be evaluated utilizing comprehensive methods such as Sanger and NextGen sequencing. The overall prevalence of PTPN11\/SHP2 mutations was 55.8% in HS and 46.2% in HHS. In addition, we identified mutations in KRAS, in about 3% of HS and 4% of HHS cases. These findings point to the shared molecular pathology of activation of the MAPK pathway in HS and HHS cases. We evaluated the efficacy of the highly specific MEK inhibitor, cobimetinib, in canine HS and HHS cell lines. We found that the IC50 values ranged from 74 to 372 nM, which are within the achievable and tolerable ranges for cobimetinib. This finding positions cobimetinib as a promising potential candidate for future canine clinical trials and enhances our understanding of the molecular defects in these challenging cancers.<\/jats:p>","DOI":"10.3390\/genes15081050","type":"journal-article","created":{"date-parts":[[2024,8,12]],"date-time":"2024-08-12T05:19:38Z","timestamp":1723439978000},"page":"1050","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Canine Histiocytic and Hemophagocytic Histiocytic Sarcomas Display KRAS and Extensive PTPN11\/SHP2 Mutations and Respond In Vitro to MEK Inhibition by Cobimetinib"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8512-5336","authenticated-orcid":false,"given":"Ya-Ting","family":"Yang","sequence":"first","affiliation":[{"name":"Department of Microbiology, Genetics and Immunology, Michigan State University, East Lansing, MI 48824, USA"},{"name":"Department of Small Animal Clinical Sciences, Michigan State University, East Lansing, MI 48824, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2057-4306","authenticated-orcid":false,"given":"Alexander I.","family":"Engleberg","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics and Immunology, Michigan State University, East Lansing, MI 48824, USA"},{"name":"Department of Small Animal Clinical Sciences, Michigan State University, East Lansing, MI 48824, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ishana","family":"Kapoor","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics and Immunology, Michigan State University, East Lansing, MI 48824, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7484-9793","authenticated-orcid":false,"given":"Keita","family":"Kitagawa","sequence":"additional","affiliation":[{"name":"Comparative Medicine and Integrative Biology Program, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara A.","family":"Hilburger","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics and Immunology, Michigan State University, East Lansing, MI 48824, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tuddow","family":"Thaiwong-Nebelung","sequence":"additional","affiliation":[{"name":"Veterinary Diagnostic Laboratory, College of Veterinary Medicine, Michigan State University, Lansing, MI 48910, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9720-1279","authenticated-orcid":false,"given":"Vilma","family":"Yuzbasiyan-Gurkan","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics and Immunology, Michigan State University, East Lansing, MI 48824, USA"},{"name":"Department of Small Animal Clinical Sciences, Michigan State University, East Lansing, MI 48824, USA"},{"name":"Comparative Medicine and Integrative Biology Program, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Boerkamp, K.M., van der Kooij, M., van Steenbeek, F.G., van Wolferen, M.E., Groot Koerkamp, M.J., van Leenen, D., Grinwis, G.C.M., Penning, L.C., Wiemer, E.A.C., and Rutteman, G.R. (2013). Gene expression profiling of histiocytic sarcomas in a canine model: The predisposed flatcoated retriever dog. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0071094"},{"key":"ref_2","first-page":"115","article-title":"Mortality in a cohort of flat-coated retrievers in the UK","volume":"7","author":"Dobson","year":"2009","journal-title":"Vet. Comp. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"S19","DOI":"10.1093\/jhered\/esp039","article-title":"Epidemiology, pathology, and genetics of histiocytic sarcoma in the Bernese mountain dog breed","volume":"100","author":"Abadie","year":"2009","journal-title":"J. Hered."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1177\/0300985813510413","article-title":"A review of histiocytic diseases of dogs and cats","volume":"51","author":"Moore","year":"2014","journal-title":"Vet. Pathol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1158\/1055-9965.EPI-12-0190-T","article-title":"The MTAP-CDKN2A locus confers susceptibility to a naturally occurring canine cancer","volume":"21","author":"Shearin","year":"2012","journal-title":"Cancer Epidemiol. Biomark. Prev."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Evans, J.M., Parker, H.G., Rutteman, G.R., Plassais, J., Grinwis, G.C.M., Harris, A.C., Lana, S.E., and Ostrander, E.A. (2021). Multi-omics approach identifies germline regulatory variants associated with hematopoietic malignancies in retriever dog breeds. PLoS Genet., 17.","DOI":"10.1101\/2021.04.05.438235"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"830","DOI":"10.1038\/s41379-018-0200-x","article-title":"Identification of diverse activating mutations of the RAS-MAPK pathway in histiocytic sarcoma","volume":"32","author":"Shanmugam","year":"2019","journal-title":"Mod. Pathol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1038\/s41392-023-01705-z","article-title":"Targeting the RAS\/RAF\/MAPK pathway for cancer therapy: From mechanism to clinical studies","volume":"8","author":"Bahar","year":"2023","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1111\/vco.12357","article-title":"Gain-of-function mutation in PTPN11 in histiocytic sarcomas of Bernese Mountain Dogs","volume":"16","author":"Thaiwong","year":"2018","journal-title":"Vet. Comp. Oncol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Takada, M., Smyth, L.A., Thaiwong, T., Richter, M., Corner, S.M., Schall, P.Z., Kiupel, M., and Yuzbasiyan-Gurkan, V. (2019). Activating Mutations in PTPN11 and KRAS in Canine Histiocytic Sarcomas. Genes, 10.","DOI":"10.3390\/genes10070505"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3984","DOI":"10.1182\/bloodadvances.2022009013","article-title":"Successful treatment of non-Langerhans cell histiocytosis with the MEK inhibitor trametinib: A multicenter analysis","volume":"7","author":"Aaroe","year":"2023","journal-title":"Blood Adv."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2439","DOI":"10.1158\/1535-7163.MCT-17-1273","article-title":"Targeting MEK in a Translational Model of Histiocytic Sarcoma","volume":"17","author":"Takada","year":"2018","journal-title":"Mol. Cancer Ther."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"22","DOI":"10.30802\/AALAS-CM-18-000065","article-title":"Development of an Orthotopic Intrasplenic Xenograft Mouse Model of Canine Histiocytic Sarcoma and Its Use in Evaluating the Efficacy of Treatment with Dasatinib","volume":"69","author":"Takada","year":"2019","journal-title":"Comp. Med."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Takada, M., Kitagawa, K., Zhang, Y., Bulitta, J.B., Moirano, S., Jones, A., Borgen, J., Onsager, A., Thaiwong, T., and Vail, D.M. (2024). Population Pharmacokinetics, Pharmacodynamics and Safety Properties of Trametinib in Dogs With Cancer: A Phase I Dose Escalating Clinical Trial. Vet. Comp. Oncol., ahead of print.","DOI":"10.1111\/vco.12989"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1657","DOI":"10.1002\/ijc.32991","article-title":"PTPN11 mutations in canine and human disseminated histiocytic sarcoma","volume":"147","author":"Hedan","year":"2020","journal-title":"Int. J. Cancer"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1111\/vco.12524","article-title":"Canine histiocytic sarcoma cell lines with SHP2 p.Glu76Gln or p.Glu76Ala mutations are sensitive to allosteric SHP2 inhibitor SHP099","volume":"18","author":"Tani","year":"2020","journal-title":"Vet. Comp. Oncol."},{"key":"ref_17","unstructured":"U.S. Food and Drug Administration (FDA) (2024, July 01). Available online: https:\/\/www.accessdata.fda.gov\/drugsatfda_docs\/nda\/2015\/206192Orig1s000PharmR.pdf."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1007\/s10637-016-0374-3","article-title":"A first-in-human phase I study to evaluate the MEK1\/2 inhibitor, cobimetinib, administered daily in patients with advanced solid tumors","volume":"34","author":"Rosen","year":"2016","journal-title":"Investig. New Drugs"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1837","DOI":"10.1039\/D3MD00145H","article-title":"MEK inhibitors in cancer treatment: Structural insights, regulation, recent advances and future perspectives","volume":"14","author":"Ram","year":"2023","journal-title":"RSC Med. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1177\/1060028016672037","article-title":"Cobimetinib","volume":"51","author":"Signorelli","year":"2017","journal-title":"Ann. Pharmacother."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Takada, M., Parys, M., Gregory-Bryson, E., Vilar Saavedra, P., Kiupel, M., and Yuzbasiyan-Gurkan, V. (2018). A novel canine histiocytic sarcoma cell line: Initial characterization and utilization for drug screening studies. BMC Cancer, 18.","DOI":"10.1186\/s12885-018-4132-0"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"eadg7492","DOI":"10.1126\/science.adg7492","article-title":"Accurate proteome-wide missense variant effect prediction with AlphaMissense","volume":"381","author":"Cheng","year":"2023","journal-title":"Science"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1038\/nmeth0410-248","article-title":"A method and server for predicting damaging missense mutations","volume":"7","author":"Adzhubei","year":"2010","journal-title":"Nat. Methods"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1038\/nbt.1754","article-title":"Integrative genomics viewer","volume":"29","author":"Robinson","year":"2011","journal-title":"Nat. Biotechnol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1340726","DOI":"10.3389\/fimmu.2024.1340726","article-title":"Tyrosine phosphatase PTPN11\/SHP2 in solid tumors\u2014Bull\u2019s eye for targeted therapy?","volume":"15","author":"Chen","year":"2024","journal-title":"Front. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1038\/s41586-021-03819-2","article-title":"Highly accurate protein structure prediction with AlphaFold","volume":"596","author":"Jumper","year":"2021","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"D368","DOI":"10.1093\/nar\/gkad1011","article-title":"AlphaFold Protein Structure Database in 2024: Providing structure coverage for over 214 million protein sequences","volume":"52","author":"Varadi","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1002\/pro.3943","article-title":"UCSF ChimeraX: Structure visualization for researchers, educators, and developers","volume":"30","author":"Pettersen","year":"2021","journal-title":"Protein Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"50","DOI":"10.3109\/00498254.2016.1157645","article-title":"Absorption, metabolism and excretion of cobimetinib, an oral MEK inhibitor, in rats and dogs","volume":"47","author":"Takahashi","year":"2017","journal-title":"Xenobiotica"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1016\/S0092-8674(00)80938-1","article-title":"Crystal structure of the tyrosine phosphatase SHP-2","volume":"92","author":"Hof","year":"1998","journal-title":"Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"8816","DOI":"10.1158\/0008-5472.CAN-04-1923","article-title":"Activating mutations of the noonan syndrome-associated SHP2\/PTPN11 gene in human solid tumors and adult acute myelogenous leukemia","volume":"64","author":"Paez","year":"2004","journal-title":"Cancer Res."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Prouteau, A., Denis, J.A., De Fornel, P., Cadieu, E., Derrien, T., Kergal, C., Botherel, N., Ulv\u00e9, R., Rault, M., and Bouzidi, A. (2021). Circulating tumor DNA is detectable in canine histiocytic sarcoma, oral malignant melanoma, and multicentric lymphoma. Sci. Rep., 11.","DOI":"10.1038\/s41598-020-80332-y"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1038\/s41586-019-1012-y","article-title":"Efficacy of MEK inhibition in patients with histiocytic neoplasms","volume":"567","author":"Diamond","year":"2019","journal-title":"Nature"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2398","DOI":"10.1056\/NEJMc1713676","article-title":"Rosai-Dorfman Disease with Activating KRAS Mutation\u2014Response to Cobimetinib","volume":"377","author":"Jacobsen","year":"2017","journal-title":"N. Engl. J. Med."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1136\/vr.158.24.825","article-title":"Histiocytic sarcoma of the spleen in flat-coated retrievers with regenerative anaemia and hypoproteinaemia","volume":"158","author":"Dobson","year":"2006","journal-title":"Vet. Rec."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1354\/vp.43-5-632","article-title":"Canine hemophagocytic histiocytic sarcoma: A proliferative disorder of CD11d+ macrophages","volume":"43","author":"Moore","year":"2006","journal-title":"Vet. Pathol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.jcpa.2011.05.006","article-title":"Paraneoplastic syndrome in haemophagocytic histiocytic sarcoma in a dog","volume":"146","author":"Soare","year":"2012","journal-title":"J. Comp. Pathol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1111\/avj.12767","article-title":"Lomustine chemotherapy for the treatment of presumptive haemophagocytic histiocytic sarcoma in Flat-coated Retrievers","volume":"96","author":"Elliott","year":"2018","journal-title":"Aust. Vet. J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1154","DOI":"10.1002\/vms3.480","article-title":"Case report: Haemophagocytic histiocytic sarcoma in an english setter","volume":"7","author":"Kerboeuf","year":"2021","journal-title":"Vet. Med. Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1177\/1040638717701521","article-title":"Atypical cytologic presentation of a histiocytic sarcoma in a Cavalier King Charles Spaniel dog","volume":"29","author":"Clarke","year":"2017","journal-title":"J. Vet. Diagn. Investig."},{"key":"ref_41","first-page":"1103","article-title":"Doxorubicin and zoledronate treatment in a dog with hemophagocytic histiocytic sarcoma","volume":"64","author":"Soileau","year":"2023","journal-title":"Can. Vet. J."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e1496","DOI":"10.1002\/cnr2.1496","article-title":"Primary splenic histiocytic sarcoma associated with hemophagocytic lymphohistiocytosis: A case report and review of literature of next-generation sequencing involving FLT3, NOTCH2, and KMT2A mutations","volume":"5","author":"Montalvo","year":"2022","journal-title":"Cancer Rep. (Hoboken)"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1078\/0344-0338-00362","article-title":"Primary histiocytic sarcoma of the spleen associated with erythrophagocytic histiocytosis","volume":"199","author":"Audouin","year":"2003","journal-title":"Pathol. Res. Pract."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.vetimm.2014.11.006","article-title":"Passage-dependent morphological and phenotypical changes of a canine histiocytic sarcoma cell line (DH82 cells)","volume":"163","author":"Heinrich","year":"2015","journal-title":"Vet. Immunol. Immunopathol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1007\/BF02623551","article-title":"A macrophage-monocyte cell line from a dog with malignant histiocytosis","volume":"24","author":"Wellman","year":"1988","journal-title":"In Vitro Cell Dev. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1111\/bjh.18964","article-title":"Outcomes after interruption of targeted therapy in patients with histiocytic neoplasms","volume":"203","author":"Reiner","year":"2023","journal-title":"Br. J. Haematol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1812","DOI":"10.1182\/blood-2023-187508","article-title":"Phase 2 Trial of Single-Agent Cobimetinib for Adults with Histiocytic Neoplasms","volume":"142","author":"Diamond","year":"2023","journal-title":"Blood"}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/15\/8\/1050\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:34:11Z","timestamp":1760110451000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/15\/8\/1050"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,8,9]]},"references-count":47,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2024,8]]}},"alternative-id":["genes15081050"],"URL":"https:\/\/doi.org\/10.3390\/genes15081050","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,8,9]]}}}