A Rapid Embryonic Stem Cell-Based Mouse Model for B-cell Lymphomas Driven by Epstein-Barr Virus Protein LMP1.
The Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) contributes to oncogenic human B-cell transformation. Mouse B cells conditionally expressing LMP1 are not predisposed to B-cell malignancies, as LMP1-expressing B cells are eliminated by T cells. However, mice with conditional B-cell LMP1 expression and genetic elimination of α/β and γ/δ T cells ("CLT" mice) die early in association with B-cell lymphoproliferation and lymphomagenesis. Generation of CLT mice involves in-breeding multiple independently segregating alleles. Thus, although introduction of additional activating or knockout mutations into the CLT model is desirable for further B-cell expansion and immunosurveillance studies, doing such experiments by germline breeding is time-consuming, expensive, and sometimes unfeasible. To generate a more tractable model, we generated clonal CLT embryonic stem (ES) cells from CLT embryos and injected them into RAG2-deficient blastocysts to generate chimeric mice, which, like germline CLT mice, harbor splenic CLT B cells and lack T cells. CLT chimeric mice generated by this RAG2-deficient blastocyst complementation ("RDBC") approach die rapidly in association with B-cell lymphoproliferation and lymphoma. Because CLT lymphomas routinely express the activation-induced cytidine deaminase (AID) antibody diversifier, we tested potential AID roles by eliminating the AID gene in CLT ES cells and testing them via RDBC. We found that CLT and AID-deficient CLT ES chimeras had indistinguishable phenotypes, showing that AID is not essential for LMP1-induced lymphomagenesis. Beyond expanding accessibility and utility of CLT mice as a cancer immunotherapy model, our studies provide a new approach for facilitating generation of genetically complex mouse cancer models.
Howard Hughes Medical Institute, Program in Cellular and Molecular Medicine, Children's Hospital Boston, Department of Genetics, Harvard Medical School, Boston, Massachusetts.
Zhaoqing
Z
Ba
Howard Hughes Medical Institute, Program in Cellular and Molecular Medicine, Children's Hospital Boston, Department of Genetics, Harvard Medical School, Boston, Massachusetts.
Fei-Long
FL
Meng
Howard Hughes Medical Institute, Program in Cellular and Molecular Medicine, Children's Hospital Boston, Department of Genetics, Harvard Medical School, Boston, Massachusetts.
Monica
M
Gostissa
Howard Hughes Medical Institute, Program in Cellular and Molecular Medicine, Children's Hospital Boston, Department of Genetics, Harvard Medical School, Boston, Massachusetts.
Pei-Yi
PY
Huang
Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts. Department of Diagnostics, School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
Qiang
Q
Ke
Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
Zhe
Z
Wang
Howard Hughes Medical Institute, Program in Cellular and Molecular Medicine, Children's Hospital Boston, Department of Genetics, Harvard Medical School, Boston, Massachusetts.
Mai N
MN
Dao
Howard Hughes Medical Institute, Program in Cellular and Molecular Medicine, Children's Hospital Boston, Department of Genetics, Harvard Medical School, Boston, Massachusetts.
Yuko
Y
Fujiwara
Max-Delbrueck-Center for Molecular Medicine, Berlin, Germany.
Klaus
K
Rajewsky
Howard Hughes Medical Institute, Program in Cellular and Molecular Medicine, Children's Hospital Boston, Department of Genetics, Harvard Medical School, Boston, Massachusetts. Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts. Alt@enders.tch.harvard.edu Baochun_Zhang@dfci.harvard.edu.
Baochun
B
Zhang
Howard Hughes Medical Institute, Program in Cellular and Molecular Medicine, Children's Hospital Boston, Department of Genetics, Harvard Medical School, Boston, Massachusetts. Alt@enders.tch.harvard.edu Baochun_Zhang@dfci.harvard.edu.
Frederick W
FW
Alt
D000818:Animals; D002460:Cell Line; D002472:Cell Transformation, Viral; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics; D004195:Disease Models, Animal; D053595:Embryonic Stem Cells / Q000378:metabolism*; D055786:Gene Knockout Techniques; D018390:Gene Targeting; D056426:Genetic Loci; D005838:Genotype; D006801:Humans; D007157:Immunologic Surveillance / Q000235:genetics; D016130:Immunophenotyping; D016393:Lymphoma, B-Cell / Q000209:etiology* / Q000401:mortality / Q000473:pathology; D051379:Mice; D018345:Mice, Knockout; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D014763:Viral Matrix Proteins / Q000235:genetics*
D004268:DNA-Binding Proteins; C037662:EBV-associated membrane antigen, Epstein-Barr virus; C499308:Rag2 protein, mouse; D011948:Receptors, Antigen, T-Cell; D014763:Viral Matrix Proteins