1. miRNAs and their Applications Laura Martí Sánchez Genomics Advanced Genetic Master December, 2015

2. INDEX  Introduction  History  Biogenesis  Applications: Cancer Rossetta Genomics kit  Bibliography

3. IntroductionHistoryBiogenesisApplicationsBibliography  non-coding RNA sequences (average length of 22 nucleotides)  founded in eukaryotes (animals, plants and viruses)  Regulation of gene expression thereby controlling many cellular processes  Human genome may encode 28645 miRNAs miRNAs Todd Smith. May, 2009 FinchTalk. Small RNAs get smaller

4. IntroductionHistoryBiogenesisApplicationsBibliography First miRNA: lin-4 (1993)  By efforts of Victor Ambros (1987) and Gary Ruvkun (1988) in lin-4  Study of the gene lin-14 en C. Elegans development  Negative regulation of gene lin-14  They showed that this RNA doesn’t encode a protein  In December 1993, in the same issue of Cell, Ambros and Ruvkun independently reported that the small and non-protein coding transcript lin 4 regulates the gene lin-14 Lin-4 Transcripts and Complementarity between lin-4 and lin-14

5. IntroductionHistoryBiogenesisApplicationsBibliography Processing of miRNA Transcription Nuclear Processing Nuclear Export Cytoplasmatic Processing

6. IntroductionHistoryBiogenesisApplicationsBibliography miRNA inhibition pathways Interaction with protein translation process: M1: Preventing assembling of the initiation process M2: Recluting 60S ribosomic subunit M3: on the translation process M6,7: Degradation

7. IntroductionHistoryBiogenesisApplicationsBibliography  The development of miRNA microarrays, RT-PCR platforms and deep sequencing is a good advantage for the knowledge of these RNAs.  During the past decade, our knowledge about role of miRNA in human diseases has grown exponentially, specially in cancer

8. IntroductionHistoryBiogenesisApplicationsBibliography miRNAs and Cancer Use of miRNA based classifier to identify tissue of origin for cancers of unknown primaries Utilizing a signature of altered miRNA expression to differentiate cancer tissue from normal tissue Study SNPs in the miRNA genes, miRNA binding sites in the target mRNA genes or in the miRNA processing/machinery pathway genes to predict cancer predisposition Distinguishing tumor subtypes using a panel of miRNAs that show differential expression with one cancer type Profiling circular blood or tumor derived exosomal miRNAs, surprassing the invasive procedures to aid in early detection of cancers miRNAs in Cancer Diagnosis

9. IntroductionHistoryBiogenesisApplicationsBibliography mi-Lung TM Test (Rosetta Genomics) Types of lung Cancer Non- squamos Carcinoid Small cell Squamous cell Carcinoma miRNAFunctionUses Hsa-miR-106a regulates the tumor suppressor retinoblastoma High levels in retinoblastoma Hsa-miR-125a-5p involved in the invasion and migration of NSCLC cells High levels in NSCLC Hsa-miR-129-3p has not yet been demonstrated underexpressed in lung adenocarcinoma Hsa-miR-205 regulates epithelial to mesenchymal transition be used to distinguish squamous from nonsquamous NSCLC Hsa-miR-21 Epidermal growth factor receptor at higher levels in NSCLC versus SCLC Hsa-miR-29b acts as a tumor suppressor via the targeting of methyltransferases at higher levels in NSCLC versus SCLC Hsa-miR-375regulates cancer growth up-regulated in lung cancers with neuroendocrine features Hsa-miR-7regulate epidermal growth factor receptor High levels in NSCLC 451 samples Agilent microarray (675 probes) RT-qPCR

10. IntroductionHistoryBiogenesisApplicationsBibliography mi-Lung TM Test (Rosetta Genomics)

11. IntroductionHistoryBiogenesisApplicationsBibliography Other tests from Rosetta Genomics

12. IntroductionHistoryBiogenesisApplicationsBibliography Farazi TA, Spitzer JI, Morozov P, Tuschl T(2011): miRNAs in human cancer. J Pathol 223:102–115 Gilad S, Lithwick-Yanai G, Barshack I, Benjamin S, Krivitsky I, Edmonston TB, Bibbo M, Thurm C, Horowitz L, Huang Y, Feinmesser M, Hou JS, St Cyr B,Burnstein I, Gibori H, Dromi N, Sanden M, Kushnir M, Aharonov R (2012) Classification of the four main types of lung cancer using a microRNA-based diagnostic assay. J Mol Diagn 14(5):510-7 Lee RC, Feinbaum RL, Ambros V. (1993) The C. elegans heterochronic gene lin- 4 encodes small RNAs with antisense complementarity to lin-14. Cell. 75(5):843-54 Moura RS, Vaz-Cunha P, Silva-Gonçalves C, Correia-Pinto J. (2015) Characterization of miRNA processing machinery in the embryonic chick lung. Cell Tissue Res 362(3):569- 575 Leonov G, Shah K, Yee D, Timmis J, Sharp TV, Lagos D. (2015) Suppression of AGO2 by miR-132 as a determinant of miRNA-mediated silencing in human primary endothelial cells. Int J Biochem Cell Biol. 69:75-84 Jacek Krol, Inga Loedige & Witold Filipowicz. (2010) The widespread regulation of microRNA biogenesis, function and decay. Nature Reviews Genetics 11, 597-610

13. Thanks for your attention! Laura Martí Sánchez