和癌症（regulationof microrna processing and its implications for development and cancer）

1,Regulation of microRNA processing and its implications for development and cancer J. Michael Thomson The Hammond Lab,2,Thomson, J. M.,MicroRNA expression profiles,Thomson and Hammond, unpublished data,4,Evolutionary Conserved miRNA Cluster,5,miR17-92 is Conserved in Vertebrates,6,miR-17-92 is Amplified in Lymphoma,Ota, et. al., Cancer Res. 2004 64:3087-95,7,The mir17-92 cluster resides in a region with known amplifications in cases of DLBCL, follicular and mantle cell lymphoma, and several lung and head/neck carcinomas Lymphoma cell lines with known amplification for C13orf25 exhibit high level expression for miR17-92 compared to cell lines lacking amplification. All cell lines exhibit reduced expression of let7 family members compared to cord blood B-cells,miR-17-92 is Amplified in Lymphoma,He, et al Nature 435(7043):828-33 2005,8,Overexpression of miR-17-19b in a Mouse Model of Human Lymphoma,9,Expression of miR17-19b Results in B-cell Lymphoma,He, et al Nature 435(7043):828-33 2005,10,miR-17-92 is regulated by E2F transcription factors,MicroRNA expression profiles,Thomson and Hammond, unpublished data,OncomiR-1 and Let-7 family are inversely expressed in development and cancer,Tumorigenesis,Development,Thomson and Hammond, unpublished data,14,The microRNA hypothesis of cancer,Tumors down-regulate many microRNAs to promote a “stem cell” like state Selected microRNAs (oncomiRs) are expressed at a high level to promote proliferation and suppress apoptosis What is the mechanism for widespread alterations in gene Expression? Transcription, processing, turnover?,15,Transcriptional reactivation with methylase and HDAC Inhibitors results in the activation of 17/313 microRNAs A similar study of 15 microRNAs often down-regulated in Cancer from 91 cancer cell lines revealed lesions that often disrupted the secondary structure of the pri-miRNA, but have little or no effect on expression Genetic deletions can explain some cases of microRNA loss, but these are often limited to specific individual cases. Another possibility could be regulated processing, as many microRNAs reside in the introns of common housekeeping mRNAs.,Saito, Y., et al Cancer Cell 2006; Diederichs Calin, GA., et al PNAS 2002, 2004,Exonic Non-coding,MicroRNAs are transcribed by RNA Polymerase II,Intronic Non-coding and coding,Lee, et al., EMBO J. 23:4051-60 2004,MicroRNAs are transcibed by RNA Polymerase II,Exonic Non-coding,Intronic Non-coding and coding,-OH,/-OH,Profiling the mature and primary microRNA transcripts,Pri-miRNA,Drosha Cleavage site,Mature,19,ES cells P19 10.5d 14.5d 18.5d,Let-7 family is post-transcriptionally regulated,Thomson and Hammond, unpublished data,ES cells P19 10day 12 day 14 day 18 day,20,MicroRNA are Post-transcriptionally Regulated,Thomson, et al Genes Dev. 20(16):2202-7 2006,21,Thomson, et al Genes Dev. 20(16):2202-7 2006,MicroRNA are Post-transcriptionally Regulated,22,23,Thomson, et al Genes Dev. 20(16):22202-7 2006,Differentiation of mouse P19 cells9 cells:,24,P19 embryonic carcinoma differentiation,Thomson, et al Genes Dev. 20(16):2202-7 2006,25,Drosha,DGCR8,Oct-4,Primary processing machinery is unchanged in P19 differentiatation,Thomson, et al Genes Dev. 20(16):22202-7 2006,Oct4,Differentiated Neuroblastoma cell line N1E-115,Differentiated,Undifferentiated,Serum starve 4 days,27,Global Reduction in miRNAs in the Context of Cancer,Red = Abundant,Blue= Depleted,Lu, J., et al Nature 435:834 2005,28,Thomson, et al Genes Dev. 20(16):22202-7 2006,29,P19 embryonic carcinoma differentiation,KS test, p 0.01,Thomson, et al Genes Dev. 20(16):22202-7 2006,30,Acknowledgements,UNC HAMMOND LAB Scott Hammond Keith Woods Summer Goodson Marty Newman Handan Kaygun Andy Baraniak Vidya Mani Tricia Wright LIEB LAB Jason Lieb Cheol Koo Lee MILGRAM LAB Beth Morin-Kensicki Paul Barnes OBRIEN LAB Jim Tsuruta,Memorial Sloan-Kettering Cancer Center CORDON-CARDO LAB Carlos Cordon-Cardo Eva Hernando-Monge,CSHL HANNON LAB Greg Hannon Lin He Jidong Liu Michelle Carmell LOWE LAB Scott Lowe David Mu Scott Powers,Constella Group Joel Parker,Stratagene Becky Mullinax,31,32,33,MicroRNA qPCR,Shi and Chiang Biotechniques 39(4):519-25 2005,Hsa-miR-18,Hsa-miR-1