title: Multiepitope tissue analysis reveals SPPL3-mediated ADAM10 activation as a key step in the transformation of melanocytes.
abstract: The evolution of cancer is characterized by the appearance of specific mutations, but these mutations are translated into proteins that must cooperate to induce malignant transformation. Using a systemic approach with the multiepitope ligand cartography (MELC) technology, we analyzed protein expression profiles (PEPs) in nevi and BRAFV600E-positive superficial spreading melanomas (SSMs) from patient tissues to identify key transformation events. The PEPs in nevi and SSMs differed predominantly in the abundance of specific antigens, but the PEPs of nevi- and melanoma-associated keratinocytes gradually changed during the transformation process. A stepwise change in PEP with similar properties occurred in keratinocytes cocultured with melanoma cells. Analysis of the individual steps indicated that activation of the metalloproteinase ADAM10 by signal peptide peptidase-like 3 (SPPL3) triggered by mutant BRAFV600Ewas a critical transformation event. SPPL3-mediated ADAM10 activation involved the translocation of SPPL3 and ADAM10 into Rab4- or Rab27-positive endosomal compartments. This endosomal translocation, and hence ADAM10 activation, was inhibited by the presence of the tumor suppressor PTEN. Our findings suggest that systematic tissue antigen analysis could complement whole-genome approaches to provide more insight into cancer development.
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Translational Research Center, Schwabachanlage 12, 91054 Erlangen, Germany.
authors.forename: Christian
authors.identifier: http://orcid.org/0000-0002-6676-0497
authors.initials: C
authors.lastname: Ostalecki
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Translational Research Center, Schwabachanlage 12, 91054 Erlangen, Germany.
authors.forename: Jung-Hyun
authors.identifier: http://orcid.org/0000-0001-8677-8961
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Translational Research Center, Schwabachanlage 12, 91054 Erlangen, Germany.
authors.forename: Jochen
authors.identifier: http://orcid.org/0000-0003-2365-8502
authors.initials: J
authors.lastname: Dindorf
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Translational Research Center, Schwabachanlage 12, 91054 Erlangen, Germany.
authors.forename: Lena
authors.initials: L
authors.lastname: Collenburg
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Translational Research Center, Schwabachanlage 12, 91054 Erlangen, Germany.
authors.forename: Stephan
authors.identifier: http://orcid.org/0000-0001-8869-1798
authors.initials: S
authors.lastname: Schierer
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Translational Research Center, Schwabachanlage 12, 91054 Erlangen, Germany.
authors.forename: Beate
authors.initials: B
authors.lastname: Simon
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Translational Research Center, Schwabachanlage 12, 91054 Erlangen, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Schliep
authors.affiliation: Institute of Molecular Immunology, Helmholtz-Zentrum München, Marchioninistraße 25, D-81377 Munich, Germany.
authors.forename: Elisabeth
authors.identifier: http://orcid.org/0000-0002-8133-8374
authors.initials: E
authors.lastname: Kremmer
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Translational Research Center, Schwabachanlage 12, 91054 Erlangen, Germany.
authors.forename: Gerold
authors.identifier: http://orcid.org/0000-0002-5031-7562
authors.initials: G
authors.lastname: Schuler
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Translational Research Center, Schwabachanlage 12, 91054 Erlangen, Germany. andreas.baur@uk-erlangen.de.
authors.forename: Andreas S
authors.identifier: http://orcid.org/0000-0003-1783-3853
authors.initials: AS
authors.lastname: Baur
mesh_terms: D000072197:ADAM10 Protein / Q000378:metabolism*; D016282:Aspartic Acid Endopeptidases / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000378:metabolism*; D018920:Coculture Techniques; D018604:Epitope Mapping / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D008544:Melanocytes / Q000378:metabolism*; D008545:Melanoma / Q000378:metabolism / Q000473:pathology; D009154:Mutation; D051059:PTEN Phosphohydrolase / Q000378:metabolism; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics / Q000378:metabolism
chemical_list: D048493:Proto-Oncogene Proteins B-raf; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human; D016282:Aspartic Acid Endopeptidases; C513914:signal peptide peptidase like 3, human; D000072197:ADAM10 Protein