title: Targeted killing of rhabdomyosarcoma cells by a MAP-based human cytolytic fusion protein.
abstract: The treatment of rhabdomyosarcoma (RMS) is challenging, and the prognosis remains especially poor for high-grade RMS with metastasis. The conventional treatment of RMS is based on multi-agent chemotherapy combined with resection and radiotherapy, which are often marked by low success rate. Alternative therapeutic options include the combination of standard treatments with immunotherapy. We generated a microtubule-associated protein (MAP)-based fully human cytolytic fusion protein (hCFP) targeting the fetal acetylcholine receptor, which is expressed on RMS cells. We were able to express and purify functional scFv35-MAP from Escherichia coli cells. Moreover, we found that scFv35-MAP is rapidly internalized by target cells after binding its receptor, and exhibits specific cytotoxicity toward FL-OH1 and RD cells in vitro. We also confirmed that scFv35-MAP induces apoptosis in FL-OH1 and RD cells. The in vivo potential of scFv35-MAP will need to be considered in further studies.
authors.affiliation: Department of Experimental Medicine and Immunotherapy, Institute of Applied Medical Engineering, University Hospital RWTH Aachen, Aachen, Germany.
authors.forename: Hannes
authors.initials: H
authors.lastname: Brehm
authors.affiliation: Department of Experimental Medicine and Immunotherapy, Institute of Applied Medical Engineering, University Hospital RWTH Aachen, Aachen, Germany.
authors.forename: Dmitrij
authors.initials: D
authors.lastname: Hristodorov
authors.affiliation: Department of Experimental Medicine and Immunotherapy, Institute of Applied Medical Engineering, University Hospital RWTH Aachen, Aachen, Germany.
authors.forename: Alessa
authors.initials: A
authors.lastname: Pardo
authors.affiliation: Department of Experimental Medicine and Immunotherapy, Institute of Applied Medical Engineering, University Hospital RWTH Aachen, Aachen, Germany.
authors.forename: Radoslav
authors.initials: R
authors.lastname: Mladenov
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.
authors.forename: Judith
authors.initials: J
authors.lastname: Niesen
authors.affiliation: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany; Institute of Molecular Biotechnology (Biology VII), RWTH Aachen University, Aachen, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Fischer
authors.affiliation: Institute for Pathology, Experimental Pathology and Immunotherapy, UKGM Giessen, Germany.
authors.forename: Mehmet K
authors.initials: MK
authors.lastname: Tur
authors.affiliation: Department of Experimental Medicine and Immunotherapy, Institute of Applied Medical Engineering, University Hospital RWTH Aachen, Aachen, Germany; Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany. Electronic address: stefan.barth@uct.ac.za.
authors.forename: Stefan
authors.initials: S
authors.lastname: Barth
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D002166:Camptothecin / Q000494:pharmacology; D045744:Cell Line, Tumor; D018680:Cholinergic Antagonists / Q000494:pharmacology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007167:Immunotherapy; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D011485:Protein Binding; D011950:Receptors, Cholinergic / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology*; D012208:Rhabdomyosarcoma / Q000188:drug therapy*; D020298:U937 Cells
chemical_list: D018680:Cholinergic Antagonists; D008869:Microtubule-Associated Proteins; D011950:Receptors, Cholinergic; D011993:Recombinant Fusion Proteins; D002166:Camptothecin
keywords: Fetal acetylcholine receptor; Human cytolytic fusion protein; Immunotherapy; Microtubule-associated protein; Rhabdomyosarcoma