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Induction of Chronic Myelogenous Leukemia in Mice by the P210 bcr/abl Gene of the Philadelphia Chromosome

Daley, George Q., Van Etten, Richard A., Baltimore, David·1990·Science
DOI 10.1126/science.2406902
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Abstract

In tumor cells from virtually all patients with chronic myelogenous leukemia, the Philadelphia chromosome, a fusion of chromosomes 9 and 22, directs the synthesis of the P210 bcr/abl protein. The protein-tyrosine kinase activity and hybrid structure of P210 bcr/abl are similar to the oncogene product of the Abelson murine leukemia virus, P160 gag /v- abl , which induces acute lymphomas. To determine whether P210 bcr/abl can induce chronic myelogenous leukemia, murine bone marrow was infected with a retrovirus encoding P210 bcr/abl and transplanted into irradiated syngeneic recipients. Transplant recipients developed several hematologic malignancies; prominent among them was a myeloproliferative syndrome closely resembling the chronic phase of human chronic myelogenous leukemia. Tumor tissue from diseased mice harbored the provirus encoding P210 bcr/abl . These results demonstrate that P210 bcr/abl expression can induce chronic myelogenous leukemia. Retrovirus-mediated expression of the protein provides a murine model system for further analysis of the disease.

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Fig. 1 The retroviral construct used to transduce the P210berlabl gene. The construct pGD210 was made from a derivative of pZIPNeoSV(X) that had been modified to include the 3′ long terminal repeat (LTR) of the myeloproliferative sarcoma virus (16) and the B2 mutation (38) shown to enhance retroviral expression in undifferentiated embryonal carcinoma cell types [pSV(X)B2-MPSV] (39); the neo gene of pSV(X)B2-MPSV was deleted, and replaced by the 172/215 bcr/abl cDNA (19, 40), such that the bcr/abl gene sequences would be expressed from the spliced subgenomic proviral mRNA; the promoter element and neo gene sequences of the pMCINeo plasmid (Stratagene) were inserted at the Cla I site. The pGD210 plasmid construct was used to make a helper-free retroviral producer cell line. Énzyme abbreviations: B, Bam HI; C, Cla I; E, Eco RI; Xb, Xba I; Xh, Xho I.
Fig. 1 The retroviral construct used to transduce the P210berlabl gene. The construct pGD210 was made from a derivative of pZIPNeoSV(X) that had been modified to include the 3′ long terminal repeat (LTR) of the myeloproliferative sarcoma virus (16) and the B2 mutation (38) shown to enhance retroviral expression in undifferentiated embryonal carcinoma cell types [pSV(X)B2-MPSV] (39); the neo gene of pSV(X)B2-MPSV was deleted, and replaced by the 172/215 bcr/abl cDNA (19, 40), such that the bcr/abl gene sequences would be expressed from the spliced subgenomic proviral mRNA; the promoter element and neo gene sequences of the pMCINeo plasmid (Stratagene) were inserted at the Cla I site. The pGD210 plasmid construct was used to make a helper-free retroviral producer cell line. Énzyme abbreviations: B, Bam HI; C, Cla I; E, Eco RI; Xb, Xba I; Xh, Xho I.CC-BY · From the PDF
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Fig. 5 Morphology of acute lymphoid leukemia cells. (A) Peripheral blood smear of mouse FN, showing predominance of immature blast cells. (B) Cytospin preparation of tumor cells from large paraspinal mass of mouse GR-1. This sâme immature blast cell type was detected in cytospin preparations from liver and spleen as well.
Fig. 5 Morphology of acute lymphoid leukemia cells. (A) Peripheral blood smear of mouse FN, showing predominance of immature blast cells. (B) Cytospin preparation of tumor cells from large paraspinal mass of mouse GR-1. This sâme immature blast cell type was detected in cytospin preparations from liver and spleen as well.CC-BY · From the PDF
Fig. 6 Pathologic analysis of mice with tumors of macrophage cells. (A) Histologic section of the liver of mouse DR-2, demonstrating the palestaining nodular infiltrates of tumor cells (×40). (B) High-power image of (A) showing the border area between normal hepatic tissue and the pale-staining tumor infiltrate, marked by arrow (×400). (C) Cytospin preparation of cells in the ascites of mouse CL-2, showing the highly vacuolated cytoplasm. (D) Histochemical stain of ascites cells for α-naphthyl acetate esterase, showing the red-brown rêaction product characteristic of the monocyte-macrophage lineage.
Fig. 6 Pathologic analysis of mice with tumors of macrophage cells. (A) Histologic section of the liver of mouse DR-2, demonstrating the palestaining nodular infiltrates of tumor cells (×40). (B) High-power image of (A) showing the border area between normal hepatic tissue and the pale-staining tumor infiltrate, marked by arrow (×400). (C) Cytospin preparation of cells in the ascites of mouse CL-2, showing the highly vacuolated cytoplasm. (D) Histochemical stain of ascites cells for α-naphthyl acetate esterase, showing the red-brown rêaction product characteristic of the monocyte-macrophage lineage.CC-BY · From the PDF
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Research works printed but not linked: 33

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Metadata

Authors
Daley, G. Q., Van Etten, R. A., & Baltimore, D.
Affiliations
Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge 02142.
Venue
Science
Publisher
American Association for the Advancement of Science (AAAS)
Published
1990-02-16
Volume
247
Issue
4944
Pages
824-830
ISSN
  • 0036-8075
  • 1095-9203
DOI
10.1126/science.2406902
Cited by
1,756
References
1
Language
en

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