Insertional mutagenesis by replication-deficient retroviral vectors encoding the large T oncogene.
Insertion sites of replication-deficient retroviral vectors may trigger clonal dominance of hematopoietic cells in vivo. Here, we tested whether this would also be the case when using vectors that express powerful oncogenes, such as the large tumor antigen (TAg) of simian virus 40. TAg inactivates the tumor-suppressor proteins p53 and Rb by virtue of a chaperone-like activity. Primary hematopoietic stem/progenitor cells transduced with retroviral vectors encoding TAg-induced histiocytic sarcoma (HS) or myeloid leukemia (ML) in transplanted mice (average survival of 21 weeks). Retrovirally introducing TAg into pretransformed 32D cells generated a monocytic leukemia, with faster kinetics ( approximately 8 weeks). Leukemic clones showed retroviral insertions in genes contributing to all known TAg cooperation pathways, acting mitogenic and/or modulating apoptosis (such as BclX, Crk, Pim2, Csfr1/Pdgfrb, Osm/Lif, Axl, Fli, Sema4b, Sox4). 32D-derived monocytic leukemias showed hits in Pim2 and Max proto-oncogenes, or the chaperone Hspa4, plus additional signaling genes. Vector-mediated insertional mutagenesis thus revealed a broad spectrum of potential TAg complementation genes. These findings have important implications for the use of retroviral transgenesis in cancer research, and the expression of signaling genes in somatic gene therapy.
Li
Zhixiong
Z
Department of Experimental Hematology, OE6960, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.
Kustikova
Olga S
OS
Kamino
Kenji
K
Neumann
Thomas
T
Rhein
Mathias
M
Grassman
Elke
E
Fehse
Boris
B
Baum
Christopher
C
D000818:Animals; D000957:Antigens, Viral, Tumor / Q000378:metabolism / Q000502:physiology*; D001854:Bone Marrow Cells / Q000166:cytology; D016026:Bone Marrow Transplantation; D016158:Genes, p53; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000166:cytology; D016403:Lymphoma, Large B-Cell, Diffuse / Q000235:genetics; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D016296:Mutagenesis; D012190:Retroviridae / Q000235:genetics* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology
D000957:Antigens, Viral, Tumor