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Mirna regulated genes and pathways as targets for therapeutic intervention

A pathway and gene technology, applied in the fields of molecular biology and medicine, can solve the problem of unknown details of regulatory pathways and networks

Inactive Publication Date: 2010-01-13
ASURAGEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current limitation of this approach is that, as noted above, the details of the regulatory pathways and networks affected by any given miRNA remain largely unknown

Method used

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  • Mirna regulated genes and pathways as targets for therapeutic intervention

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0277] Analysis of gene expression after transfection with HSA-MIR-16

[0278] miRNAs are believed to affect gene expression mainly at the translation level. Translational regulation that leads to up- or down-regulation of protein expression can lead to changes in the activity and expression of downstream gene products and genes, which in turn are regulated by these proteins. These regulatory effects will be shown as changes in the overall mRNA expression profile. In addition, it has recently been reported that in some cases, miRNAs can reduce the mRNA content of their direct targets (Bagga et al., 2005; Lim et al., 2005), and the changes can be observed after microarray gene expression analysis. Microarray gene expression analysis was performed to identify genes misregulated by hsa-miR-16.

[0279] The synthetic precursor miR-16 (Ambion) was reverse-transfected into quadruplicate samples of A549 cells, and each sample was performed at three time points. According to the manufa...

Embodiment 2

[0290] Cellular pathways affected by HSA-MIR-16

[0291] Misregulation of gene expression by hsa-miR-16 (Table 1) affects many cellular pathways that represent potential therapeutic targets for the control of cancer and other diseases and disorders. The inventors determined the identity and nature of the cytogenetic pathways affected by the regulatory cascade induced by hsa-miR-16 expression. Use Ingenuity Pathways Analysis ( Systems, Redwood City, CA) performs cell path analysis. The most affected pathways after hsa-miR-16 overexpression in A549 cells are shown in Table 2.

[0292] Table 2. Functional cellular pathways that are significantly affected after hsa-miR-16 is overexpressed in human cancer cells

[0293]

[0294] These data prove that hsa-miR-16 directly or indirectly affects the expression of numerous metabolism, cell proliferation, cell development and cell cycle related genes, and therefore mainly affects the functional pathways related to cell growth, development a...

Embodiment 3

[0296] Predicted gene target of HSA-MIR-16

[0297] Use proprietary algorithm miRNATarget TM (Asuragen) predicted gene targets for binding hsa-miR-16-1 and regulated by hsa-miR-16-1 and are shown in Table 3.

[0298] Table 3. Predicted target genes of hsa-miR-16

[0299]

[0300] ACACB

[0301]

[0302] AGTR2

[0303] ANAPC11

[0304]

[0305] ARHGEF4

[0306] ASPH

[0307] ATXN2

[0308] BCAS1

[0309] BNIP1

[0310] C10orf46

[0311] C14orf129

[0312] C18orf1

[0313] C1orf62

[0314] C21orf25

[0315] C6orf47

[0316] C9orf48

[0317] CAMSAP1L1

[0318] CCDC28A

[0319] CD22

[0320] CDC7

[0321] CEP350

[0322] CHST10

[0323] CLTB

[0324] COPS7A

[0325] CRSP9

[0326] CX3CL1

[0327] DCAMKL1

[0328]

[0329] DKFZp666G0

[0330] DNAJC5G

[0331] DUSP3

[0332] EFNB2

[0333] ELOVL6

[0334] ETS2

[0335] FAM3A

[0336] FBXO21

[0337]

[0338] FLJ14816

[0...

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Abstract

The present invention concerns methods and compositions for identifying genes or genetic pathways modulated by miR-16, using miR-16 to modulate a gene or gene pathway, using this profile in assessing the condition of a patient and / or treating the patient with an appropriate miRNA.

Description

[0001] This application relates to U.S. Patent Application No. 11 / 141,707 filed on May 31, 2005 and U.S. Patent Application No. 11 / 273,640 filed on November 14, 2005, each of which is incorporated herein by reference in its entirety in. Technical field [0002] The invention relates to the fields of molecular biology and medicine. More specifically, the present invention relates to methods and compositions for treating diseases or conditions affected by miR-16 microRNA, microRNA expression, and genes and cell pathways directly and indirectly regulated by them. Background technique [0003] In 2001, several organizations used cloning methods to isolate and identify a large group of "microRNA" (miRNA) from C.elegans, Drosophila and humans (Lagos-Quintana et al., 2001; Lau et al., 2001; Lee and Ambros, 2001). Hundreds of miRNAs have been identified in plants and animals (including humans) that do not seem to have endogenous siRNA. Therefore, although similar to siRNA, miRNA is dif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11
CPCC12Q2600/178C12Q2600/112C12Q1/6886C12Q1/6809C12Q2600/118C12Q2525/207
Inventor 麦克·拜罗姆查尔斯·D·约翰逊大卫·布朗安德里斯·G·巴德
Owner ASURAGEN
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