This project is a continuation of a portion of the project entitled The Molecular Pathogenesis of Multiple Myeloma, Z01 SC 006581. Progress during the past year has resulted in several publications, and added to previous accomplishments. First, comprehensive FISH analyses on metaphase chromosomes in 47 multiple myeloma cell lines and 48 advanced multiple myeloma tumors showed the following: 1) primary IgH translocations, mostly balanced, involving 5 recurrent partners were present in 68% of cell lines, 70% of non-hyperdiploid tumors, but only 12% of hyperdiploid tumors;2) secondary translocations (IgH translocations not involving the five recurrent loci, IgL and IgK translocations, and MYC translocations) usually are unbalanced, with complex structures, and occur with a similar prevalence in hyperdiploid and non-hyperdiploid tumors;3) two of the 5 recurrent translocations rarely occur within the same tumor cell;and 4) the 5 recurrent translocations sometimes can be secondary events. Second, we summarized FISH and array comparative genomic hybridization analyses on 47 multiple myeloma cells lines that enabled us to detect and characterize MYC rearrangements in 43 (>90%) of the cell lines. Together with our previous results, this gave us further support for our hypothesis that MYC rearrangements represent a late progression event that provides a general paradigm for secondary translocations in multiple myeloma. Third, we published two papers in which we cloned and/or characterized by FISH translocation breakpoints that are found in complex translocations that involve MYC. Fourth, we have cloned from two MM cell MYC translocations in which no Ig sequences are involved, but one of which involves BLIMP, a master regulator of normal plasma cell differentiation. Fifth, we have compared array CGH data on 50 MM cell lines with array CGH data on 155 MM tumors that was made publically available by the MMRC. These results show that MYC rearrangements are much more frequent in primary MM tumors than found by others with FISH analyses, i.e., about 32% of newly diagnosed MM tumors and 46% of previously treated tumors. Sixth, we analyzed the MMRC data set that included gene expression profiling and CGH arrays for 155 tumors to show that MYC expression is significantly increased in tumors that have rearrangements in the MYC locus. Finally, were were able to identify polymorphisms in MYC in two of the four MM cell lines that have no detectable rearrangement by either FISH or array CGH analyses;both informative lines express both MYC alleles to a similar extent. This contrasts to previous results on thirteen MM cell lines that have MYC rearrangements and MYC polymorphisms, for which we showed selective expression of only one MYC allele. So, for the first time we can concludes that MYC expression in MM cell lines need not be associated with rearrangements or other structural changes in the MYC locus that dysregulate the expression of only one MYC gene. Results for MYC and also other loci suggest different mechanisms for primary vs secondary genomic rearrangements. One new finding is that secondary breakpoints often are associated with duplicated sequences present on both sides of the breakpoint. The duplicated sequences can give rise to tandem duplications, insertions of other DNA fragments up to 100 kb or more between the duplicated sequences, and reciprocal translocations with other chromosomes so that the duplicated sequences are present on both derivative chromosomes. It remains to be determined if the duplications are also found with chromosome inversions.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC010910-05
Application #
8552887
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
2012
Total Cost
$446,555
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Onozawa, Masahiro; Zhang, Zhenhua; Kim, Yoo Jung et al. (2014) Repair of DNA double-strand breaks by templated nucleotide sequence insertions derived from distant regions of the genome. Proc Natl Acad Sci U S A 111:7729-34
Affer, M; Chesi, M; Chen, W D et al. (2014) Promiscuous MYC locus rearrangements hijack enhancers but mostly super-enhancers to dysregulate MYC expression in multiple myeloma. Leukemia 28:1725-35
Bergsagel, P L; Kuehl, W M (2013) Degree of focal immunoglobulin heavy chain locus deletion as a measure of B-cell tumor purity. Leukemia 27:2067-8
Kuehl, W Michael; Bergsagel, P Leif (2012) Molecular pathogenesis of multiple myeloma and its premalignant precursor. J Clin Invest 122:3456-63
Kuehl, W Michael; Bergsagel, P Leif (2012) MYC addiction: a potential therapeutic target in MM. Blood 120:2351-2
Zingone, Adriana; Kuehl, W Michael (2011) Pathogenesis of monoclonal gammopathy of undetermined significance and progression to multiple myeloma. Semin Hematol 48:4-12
Zingone, A; Cultraro, C M; Shin, D-M et al. (2010) Ectopic expression of wild-type FGFR3 cooperates with MYC to accelerate development of B-cell lineage neoplasms. Leukemia 24:1171-8
Dib, Amel; Glebov, Oleg K; Shou, Yaping et al. (2009) A der(8)t(8;11) chromosome in the Karpas-620 myeloma cell line expresses only cyclin D1: yet both cyclin D1 and MYC are repositioned in close proximity to the 3'IGH enhancer. DNA Repair (Amst) 8:330-5
Vatsveen, Thea Kristin; Tian, Erming; Kresse, Stine H et al. (2009) OH-2, a hyperdiploid myeloma cell line without an IGH translocation, has a complex translocation juxtaposing MYC near MAFB and the IGK locus. Leuk Res 33:1670-7
Dib, Amel; Gabrea, Ana; Glebov, Oleg K et al. (2008) Characterization of MYC translocations in multiple myeloma cell lines. J Natl Cancer Inst Monogr :25-31

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