title: Establishment of a rhabdomyosarcoma xenograft model in human-adapted mice.
abstract: The outcome of patients with advanced stage rhabdomyosarcoma (RMS) is still sobering. This outcome has not improved through conservative treatments. Therefore, novel treatment approaches such as immunotherapy need to be evaluated in human-adapted animal models. The aim of this study was to develop a humanized mouse model of childhood RMS as a basis for the study of immunotherapeutic approaches. Therefore, NOD/LtSz-scid IL2rgammanull-mice were used for all the experiments (n=19). The animals underwent sublethal irradiation on days 1 and 2 (1 x 300 cGy). After irradiation, the transplantation of human CD34+-cells (1,000,000 cells per animal i.v.) was carried out. Five animals served as the control and did not undergo stem cell transplantation. The engraftment of human cells was assessed in peripheral blood on days 21 and 55 by FACS analysis. Eight weeks after transplantation, the subcutaneous xenotransplantation of human alveolar and embryonal RMS cell lines was carried out. Tumor growth was monitored and tumors were resected 93 days after CD34+-transplantation. The tumor specimens were evaluated histologically. The successful engraftment of human cells with the establishment of a human immune system was observed in 12 out of 14 animals. B and T cells were mostly detected in the peripheral blood. There were only a few monocytes and almost no natural killer cells. The xenotransplantation of alveolar RMS resulting in subcutaneous tumor growth was feasible in 7 animals. The xenotransplantation of embryonal RMS was performed in 5 animals and led to tumor growth in 1 animal. A histological work up showed either alveolar or embryonal RMS cells with central necrosis. This is the first time a xenotransplantation model of human RMS has been developed in a humanized mouse model. The establishment of subcutaneous tumor xenografts was more effective in the alveolar subtype. This model offers a basic tool for further analyzing novel immunotherapeutic approaches in RMS, and could possibly be used in other solid pediatric tumors.
authors.affiliation: Department of Pediatric Surgery, University Children's Hospital, 72076 Tübingen, Germany. guido.seitz@med.uni-tuebingen.de
authors.forename: Guido
authors.initials: G
authors.lastname: Seitz
authors.forename: Matthias
authors.initials: M
authors.lastname: Pfeiffer
authors.forename: Jörg
authors.initials: J
authors.lastname: Fuchs
authors.forename: Steven W
authors.initials: SW
authors.lastname: Warmann
authors.forename: Ivo
authors.initials: I
authors.lastname: Leuschner
authors.forename: Christian
authors.initials: C
authors.lastname: Vokuhl
authors.forename: Peter
authors.initials: P
authors.lastname: Lang
authors.forename: Rupert
authors.initials: R
authors.lastname: Handgretinger
authors.forename: Sorin
authors.initials: S
authors.lastname: Armeanu-Ebinger
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002469:Cell Separation; D002648:Child; D004195:Disease Models, Animal; D005434:Flow Cytometry; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D008817:Mice, Mutant Strains; D016513:Mice, SCID; D012208:Rhabdomyosarcoma; D012983:Soft Tissue Neoplasms; D014183:Transplantation, Heterologous / Q000379:methods*