The low grade lymphomas of man (follicular (FL) and small lymphocytic (SLL)) occasionally undergo transformation to more aggressive diffuse large cell lymphomas (DLCL). There exists a clonal relationship between these tumors arising in a single individual as evidenced by constancy of immunoglobulin gene rearrangements and translocation breakpoint (t(11;14) and t(14;18) in SLL and of FL respectively). The molecular basis of this transformation in unknown. These studies propose to isolate and characterize sequences that become activated as a consequence of this transformation event. Isolation of sequences specific to the DLCL will be accomplished by preparing a cell-specific probe by subtraction hybridization thereby eliminating sequences commonly expressed in the follicular and diffuse lymphomas. Unfortunately, the follicular lymphoma tissue cannot be grown in vitro, thus mRNA from the follicular lymphomas is limited. To circumvent this limitation, a novel approach to cloning mRNA as sense-strand single-stranded DNA is proposed that is generally applicable to tissue specimens that cannot be expanded in vitro. cDNA will be directionally cloned into lambda ZAP from which the cloned insert can be expanded as a defective filamentous bacteriophage; the ssDNA genome can then be used in substraction hybridizations in place of RNA as it represents the sense strand. The cell-specific probe will be used to screen cDNA libraries prepared from the DLCL. The clones isolated by this strategy will be characterized as to their expression in follicular and large cell lymphomas as well as normal and stimulated B- lymphocytes. In addition, the transforming potential of these genes will be determined by electroporation of normal B- lymphocytes and FL cells with these gene subcloned into a mammalian expression vector, pRSY-gpt. Further investigations will explore the inability to propagate FL cells in vitro. Utilizing somatic cell hybrids between transformed lymphoma cell lines of EBY-transformed lymphocytes and FL cells, we will determine if the transformed phenotype (assayed as growth in semisolid media) can be transferred to the FL cells.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08CA001396-02
Application #
3079877
Study Section
Cancer Institutional Fellowship Review Committee (CT)
Project Start
1988-07-01
Project End
1991-06-30
Budget Start
1989-07-01
Budget End
1990-06-30
Support Year
2
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Stanford University
Department
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
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Zhang, Yujing; Shen, Jing; Ming-Whei et al. (2007) Telomere length in hepatocellular carcinoma and paired adjacent non-tumor tissues by quantitative PCR. Cancer Invest 25:668-77
Zelenetz, A D; Cleary, M L; Levy, R (1993) A submicroscopic interstitial deletion of chromosome 14 frequently occurs adjacent to the t(14;18) translocation breakpoint in human follicular lymphoma. Genes Chromosomes Cancer 6:140-50
Zelenetz, A D (1992) Construction of complex directional complementary DNA libraries in SfiI. Methods Enzymol 216:517-30
Zelenetz, A D; Chen, T T; Levy, R (1992) Clonal expansion in follicular lymphoma occurs subsequent to antigenic selection. J Exp Med 176:1137-48
Campbell, M J; Zelenetz, A D; Levy, S et al. (1992) Use of family specific leader region primers for PCR amplification of the human heavy chain variable region gene repertoire. Mol Immunol 29:193-203
Zelenetz, A D; Campbell, M J; Bahler, D W et al. (1991) Follicular lymphoma: a model of lymphoid tumor progression in man. Ann Oncol 2 Suppl 2:115-22
Zelenetz, A D; Chen, T T; Levy, R (1991) Histologic transformation of follicular lymphoma to diffuse lymphoma represents tumor progression by a single malignant B cell. J Exp Med 173:197-207
Zelenetz, A D; Chu, G; Galili, N et al. (1991) Enhanced detection of the t(14;18) translocation in malignant lymphoma using pulsed-field gel electrophoresis. Blood 78:1552-60
Zelenetz, A D; Levy, R (1990) Directional cloning of cDNA using a selectable SfiI cassette. Gene 89:123-7

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