GeneExpression Systems of USA and University of Cambridge of UK
Appasani Research Conferences (ARCEI.ORG)
Jointly Presents
Fifth International

MicroRNAs Europe 2010 Meeting

on
MicroRNAs: Biology to Development and Disease

November 1 - 2, 2010
Venue: Peterhouse, University of Cambridge, Cambridge, UK

A Unique Theme Conference & Exhibition in the MicroRNomics!

Meeting Place:

The Venue Peterhouse (middle) is the oldest College of the University of Cambridge, founded in 1284 and an institution dedicated to education and research. Throughout its history, Petreans have been at the heart of the political, social and religious controversies that have shaped today’s society. Some of the influential Petreans include: Henry Cavendish, Lord Kelvin, Sir Frank Whittle and Christopher Cockerill, and Nobel Laureates – Sir John Kendrew, Sir Aaron Klug, Archer Martin and Max Perutz , who gave a twentieth century lead in the field of Molecular Biology. Since many centuries it remained as a hub for innovation and successive generations of the brightest young people around the world.

Why do you wait to join for an intellectual gathering in the ‘microRNomics’ arena at the prestigious college Peterhouse in the Premier University campus!

(click here for meeting poster)

Click Here for Agenda

Monday, November 1, 2010
8:00 A.M: Registration Open
9:00 - 6:00 P.M: Scientific Sessions

Tuesday, November 2, 2010
8:00 A.M: Registration Open
9:00 - 3:30 P.M: Scientific Sessions

Organizer:

	Krishnarao Appasani, PhD., MBA Founder & CEO GeneExpression Systems, Inc. Waltham, MA USA

Keynote Speakers:

	Chris Boshoff, MD, PhD, MRCP            Professor & Director of Cancer Institute University College of London London, United Kingdom Title: Kaposi sarcoma: the role of viral and cellular miRNAs
	Arthur A. Levin, PhD. Industry Keynote Speaker Vice President & Chief Development Officer Sataris Pharma, US Operations San Diego, CA, USA Title: Development of miravirsen an LNA-modified oligonucleotide inhibitor of miR-122

Other Speakers:

	Pavel Sumazin, Ph.D. Research Scientist Sulzberger Columbia Genome Center Columbia University, New York, NY, USA Title: Glioblastoma-related microRNA regulators and targets
	Thomas Brefort                                            Director Application Development Febit Biomed GmbH & Heidelberg, Germany Title: miRNA biomarker profiling from blood – A promising approach to non-invasive diagnostic testing
	Laura Pazzaglia, PhD.                                  Laboratory of Experimental Oncology                        Itituto Ortopedico Rizzoli Bologna, Italy Title: miRNAs in Osteosarcoma
	Christina Roberts, PhD. student Evolutionary Biology Centre Uppsala University Uppsala, Sweden Title: Cell signaling by miR-165/166 in plant pattern and organ development
	Cindy Horwedel                                            Graduate Student in the Lab. Of Dr. Ulrich Tschulena Division of Molecular Genome Analysis Deutsches Krebsforschungszentrum (DKFZ) Heidelberg, Germany Title:A novel large-scale screen to identify modulators of oncomir miR-21
	Matthew J.A. Wood, BM BCh., DPhil.         Lecturer in Biomedical Science University of Oxford, Oxford, UK Title: Non coding RNAs in neurodegenerative diseases
	Alain Sewer, PhD.                                         Scientist - Disease Modeling & Simulation                 Phillip Morris International R&D Neuchatel, Switzerland Title:Evaluation of a novel miRNA-specific normalization method and comparison between Affymetrix and Exiqon platforms
	Beate Niesler, PhD.                                     Scientist & Group Leader Institute of Human Genetics, University of Heidelberg Heidelberg, Germany Title: A functional variant in the microRNA-510 target site of the serotonin receptor type 3E gene is associated with diarrhea predominant irritable bowel syndrome
	Kyriacos N. Felekkis, PhD.                          Scientist in the Biological Sciences University of Cyprus Nicosia, Cyprus Title: The role of microRNAs (miRNA) in the development of Polycystic Kidney Disease
	Michael J. Kerin, MCh., FRCSI.                  Professor and Head of Surgery National University of Ireland, School of Medicine & University Hospital, Galway, Ireland Title: microRNAs in breast cancer
	Pieter Mestdagh                                           Ph.D Student in the Center for Medical Genetics Ghent University Hospital, Ghent, Belgium Title: The microRNA body map: dissecting microRNA function through integrative genomics
	Maria Roubelakis, DPhil.                             Cell and Gene Therapy Laboratory                             Biomedical Research Foundation, Academy of Athens, Athens, Greece Title: Identification and functional analysis of miRNAs expressed in human mesenchymal stem cells
	Francesca Ruberti, PhD.                             CNR-Institute of Neurobiology and Molecular Medicine Roma, Italy Title: microRNAs in neurodegenerative diseases
	Erik Vassella, PhD.                           Group Leader in the Inst. of Pathology, University of Bern Bern, Switzerland Title: miR-34a and miR-15a/16 are co-regulated in non-small cell lung cancer and control cell cycle progression in a synergistic and Rb-dependent manner
	Paolo Ceppi, PhD Student University of Turin at San Luigi Hospital Department of Clinical and Biological Sciences Orbassano, Italy Title: Loss of mir-200c promotes an aggressive, invasive and chemoresistant phenotype in NSCLC
	Iris Lavon PhD.                                             Professor & Head of Molecular Neuro-Oncology Hadassah Hebrew University Medical Center Jerusalem, Israel Title: Do microRNA clusters that share reminiscent expression profile between glioma and neural precursor cells have a role in tumorigenesis
	Muhammad I. Aslam, PhD.               Scientist in the Molecular Medicine University of Leicester & Leicester Royal Infirmary Leicester, UK Title: MicroRNAs are novel biomarkers of colorectal cancer
	Nagy Habib, MB Ch., PhD., FRCS.             Professor of Surgery & Director of HPB Services Imperial College London & Hammersmith Hospital, London, UK Title: Characterization of short RNAs that activate KLF4 expression
	Stephanie Urschel, PhD. Senior Field Scientist Europe Thermo Fisher Scientific, Germany Title:Sustained Over-expression of microRNA Mimics using a Lentiviral Delivery System
	Hazel Pinheiro, PhD. Application Specialist Exiqon A/S Vedbaek, Denmark Title: Early detection of colorectal cancer from patient blood plasma using microRNA-based RT-qPCR
	Bassam Badran, PhD.                                  Professor of immunology Dept of Biochemistry                        Lebanese University, Faculty of Sciences Hadath Beirut, Lebanon Title: Valproate treatment of CD4+CD25- T cells transiently confers on them a Treg microRNA profile

Key Sessions:

The most-up-to-date developments will be addressed:
MicroRNAs Discovery and Biogenesis
MicroRNAs in Development
Bioinformatics of miRNAs
MicroRNAs in Virology & Diagnostics
MicroRNAs in Stem Cell Biology
MicroRNAs in Disease Biology

Exhibitors are welcome to reserve their booth space early!

Please contact if you are interested in speaking in the scientific or Technology workshops of this meeting.

GeneExpression Systems, Inc.
P.O. Box 540170
Waltham, MA 02454 USA
Tel: (781) 891-8181
Fax: (781) 891-8234
E-mail: Genexpsys@expressgenes.com
www.expressgenes.com

Poster Abstract Submission by October 1, 2010

Abstracts

MiRNAs   in Osteosarcoma
Laura Pazzaglia1, Chiara Novello1, Amalia Conti1, Irene Quattrini1, Luisa Montanini2, Piero Picci1 and Maria Serena Benassi1
1Laboratory of Experimental Oncology,  Istituti Ortopedici Rizzoli, Bologna – Italy
2Center for Molecular and Translational Oncology, University of Parma - Italy

Abstract: Osteosarcoma (OS) is the most common primary malignant bone tumor in children and young adults. Conventional therapy for OS has reached a plateau of 60-70%, a 5-year survival rate that has changed little in two decades, highlighting the need for new therapeutic approaches by the research  of new targets. We performed different  high-throughput analysis on OS cell lines and samples and we have identified  several differentially expressed  miRNA (in particular mir-196a, mir-1, mir-133b).
In vitro functional studies are on-going  to better investigate  the role of these miRNAs as potential prognostic marker and drug target.

 The microRNA body map: dissecting microRNA function through integrative genomics
Pieter Mestdagh1*, Steve Lefever1*, Filip Pattyn1,2, Dana Ridzon3,  Annelies Fieuw1, Joëlle Vermeulen1, Anne De Paepe1, Linda Wong3,  Frank Speleman1, Caifu Chen3, Jo Vandesompele1
1Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium. 2German Cancer Research Center (DKFZ), Heidelberg, Germany. 3Life Technologies, Foster City, California, USA. *Equally contributing authors

Abstract: MicroRNAs are tiny regulators of coding gene expression. While a growing body of evidence implicates deregulated microRNA expression in various aspects of human disease, including cancer, insights in global microRNA function remain limited. Here, we present the microRNA body map, an interactive online compendium and mining tool of high-dimensional newly generated and published microRNA expression profiles along with functional annotation inferred through integrative transcriptomics. The microRNA body map enables prioritization of candidate microRNAs based on their expression profile across tissue or disease subgroup and based on a new functional annotation. The microRNA body map project has great potential to become a community resource.

Anovellarge-scale screen to indentify modulators of oncomir miR-21
Horwedel C, Keklikoglou I, Zhang J, Diederichs S, Wiemann S, Tschulena U

Division Molecular Genome Analysis, DKFZ (German Cancer Research Center), Im Neuenheimer Feld 580, 69120 Heidelberg, Germany

Abstract: MicroRNA-21 is upregulated in different cancers and modulates cancer-relevant processes. To identify genes involved in the regulation of miR-21, we have screened a large-scale siRNA-library. We have identified 100 genes that significantly regulate miR-21 activity. These candidates were followed up in different assays to analyze whether these genes function at the transcriptional or post-transcriptional level. Moreover, these genes were also analyzed for their ability to modulate other microRNAs to identify general regulators of miRNA biogenesis. Individual candidates are currently analyzed for their specific mode of miR-21-activity modulation. Thus, by these screens we identified miR-21 specific and general regulators of miR-processing.

Do microRNA Clusters That Share Reminiscent Expression Profile Between Glioma and Neural Precursor Cells Have a Role in Tumorigenesis?
Iris Lavon1,2, Daniel Zrihan1,2, Avital Granit1,2, Ofira Einstein2, Yoav Smith3,
Tamir Ben-Hur2 and Tali Siegal1.
1Gaffin Center for Neuro-Oncology, 2Department of Neurology, 3Genomic Data Analysis Unit, Hadassah Hebrew University Medical Center

Abstract: We showed that gliomas exhibit a miRNA expression profile reminiscent of neural precursor cells. About half of the miRNAs expressed in the shared profile, cluster in seven genomic regions susceptible to genetic/epigenetic alterations in cancers. Our study provided the first evidence for association between these clusters and gliomas. Some of these clusters such as mir17-92 were reported to have a pro-oncogenic role, while the potential tumorigenic role of clusters, such as the 53 miRNAs cluster on chromosome 14q32.31, remains unknown. Our current study which aims to reveal the tumorigenic role of the cluster on chromosome 14q32.31 and previous results will be discussed.

Identification and functional analysis of miRNAs expressed in human mesenchymal stem cells
Maria Roubelakis, DPhil., Cell and Gene Therapy Laboratory, Foundation for Biomedical Research of the Academy of Athens, Athens, Greece;

M.G. Roubelakis1,2 , O. Trohatou1,2,  D. Zagoura1,2,  P. Zotos1,3, V. Bitsika1,2, S. Kossida3, Κ.Ι. Pappa1,2,4 , A. Antsaklis4 and N.P. Anagnou1,2

1Cell and Gene Therapy Laboratory, Foundation for Biomedical Research of the Academy of Athens, Athens, Greece; 2Laboratory of Biology, University of Athens School of Medicine, Athens, Greece; 3Bioinformatics Department, Foundation for Biomedical Research of the Academy of Athens, Athens, Greece; 4Department of Obstetrics and Gynecology, University of Athens School of Medicine, Athens, Greece.

Abstract: Human mesenchymal stem cells (hMSCs) represent a population of multipotent stem cells, easily expanded in culture and able to differentiate into many lineages. Our group has isolated MSCs the amniotic fluid (AF). To further decipher the molecular mechanisms as they relate to the MSCs from bone marrow (BM) and umbilical cord blood (UCB), we investigated the comparative post-transcriptional regulation mechanisms of MSCs from the three sources at the miRNA level. More specifically, the objectives of the study were i) the detection of miRNA populations in AF, BM and UCB-MSCs, ii) the validation of their expression levels using Real Time PCR, iii) the generation of a new algorithm for the in silico detection of miRNA target-genes, iv) the validation of miRNA binding on specific targets predicted by the algorithm application and v) the determination of specific miRNA funcrional role.

Valproate treatment of CD4+CD25- T cells transiently confers on them a Treg microRNA profile
Badran Bassam, PhD., Professor of Immunology, Department of Biochemistry, Laboratory of Immunology, Faculty of Sciences; EDST-PRASE, Lebanese University, Hadath Beirut, Lebanon

Fayyad-Kazan Hussein1†, Rouas Redouane1†, Zeineddine Ibrahim2, Makhour Yolla2, Hamade Eva2, Badran Rabih1, Fayyad-Kazan Mohammad1, Lewalle Philippe1, Jebbawi Fadi1, Romero Pedro3, Burny Arsène1, Martiat Philippe*1 and Badran Bassam*2. 

1. Laboratory of Experimental Hematology, Institut Jules Bordet, Université Libre de
Bruxelles, 121, Boulevard de Waterloo, 1000, Bruxelles, Belgium
2. Department of Biochemistry, Laboratory of Immunology, Faculty of Sciences; EDST-
PRASE, Lebanese University, Hadath Beirut, Lebanon.
3. Division of Clinical Onco-Immunology, Ludwig Institute for Cancer Research, Lausanne
branch, 4 Av. Pierre-Decker, 1005 Lausanne, Switzerland

Abstract: Regulatory T-cells (Tregs) play a key role in immune system homeostasis and tolerance to antigens, thereby preventing autoimmunity, and may be partly responsible for the lack of an appropriate immune response against tumor cells. Although not sufficient, a high expression of FOXP3 is necessary for their suppressive function. Recent reports have shown that HDAC inhibitors (SAHA) modulated microRNA expression profiles in various cell types. We therefore decided to investigate the effect of valproate on microRNA expression profile in CD4+CD25- T cells purified from cord blood. We found that, valproate treatment induced the acquisition of the miRs Tregs signature. To elucidate whether the changes in the miRs expression could be due to the increased FOXP3 expression, we transduced these non-Tregs with a FOXP3 lentiviral expression vector, and found no changes in miRs expression. Therefore the modification in their miRs expression profile is not due to an increased expression of FOXP3, but directly results from HDAC inhibition.
We conclude that valproate treatment of human non-Tregs confers them a microRNA profile similar to that of their regulatory counterpart.

Prognostic Impact of miRNAs in NSCLC Evaluated by High Throughput in Situ Hybridization and LNA Microarrays
 Kenneth Lonvik, PhD Student, Dept of Clinical Pathology, The University Hospital of
Northern Norway, Tromsoe, Norway

Abstract: For the past two years, our research group has investigated the prognostic impact of different miRNAs in Non-Small Cell Lung Cancer (NSCLC), by in situ hybridization (ISH) and LNA microRNA microarray analysis. We have previously published several papers on expression of proteins and their receptors in NSCLC. We have a biobank containing tumor specimens from 335 patients diagnosed with NSCLC stage I – IIIA. From these specimens we have made TMAs (Tissue MicroArrays), making it possible to analyse many patients simultaneously by in situ hybridization. For LNA microarrays we selected ten patients with high survival, ten with low survival and ten normal lung tissue samples. By LNA microarray we found more than 60 microRNAs to be differently expressed in lung tumors compared to normal lung tissue (p < 0.012, FDR < 0.1). We also found significant differences between patients with long and short postoperative survival. More than 30 microRNAs were differently expressed between these two groups (p < 0.02, FDR < 0.33). We have so far performed ISH on two different miRNAs; has-miR-155 and has-miR-126. We found miR-155 to have some prognostic impact in NSCLC, depending on histological subtype and nodal status. miR-126 expression was shown to have a significant impact on disease-specific survival, where high expression was linked to a poorer prognosis than low expression. The results have been verified by qRT-PCR

Characterization of short RNAs that activate KLF4 expression
Nagy Habib, MB Ch., PhD., FRCS., Professor of Surgery & Director of HPB Services, Faculty of Medicine-Imperial College London & Hammersmith Hospital, London, UK

Pål Sætrom1,2,3, John J. Rossi4, Noriyuki Kasahara5, Jessica Alluin4, Paul Mintz6,  Nagy A. Habib6
1Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, NO-7489 Trondheim, Norway
2Department of Computer and Information Science, Norwegian University of Science and Technology, NO-7491 Trondheim, Norway
3Interagon AS, Laboratoriesenteret, NO-7489 Trondheim, Norway
4Division of Molecular Biology, Beckman Research Institute of City of Hope, Duarte, CA 91010, USA
5Department of Medicine, UCLA School of Medicine, 675 Charles E. Young Drive South, MRL-1551, Los Angeles, CA 90095-7019
6Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, London, UK

It is well recognized that short interfering RNA (siRNA) sequences play an important role in gene expression. These are a class of short double-stranded RNA molecules involved in the RNA interference (RNAi) pathway, which regulates the expression of specific genes. In recent years, siRNA sequences have been exploited to study the molecular and biological properties of stem cells primarily by down-regulating specific genes. We have taken a novel genomic-bioinformatic approach to design short activating RNA (saRNA) molecules that can up-regulate specific genes. We show proof-of-principle that saRNAs could be designed to up-regulate specific genes such as Kruppel-like factor 4 (KLF4). The KLF4 gene is a transcription factor necessary for maintaining embryonic and somatic stem cells that control the expression of pluripotency genes including POU5F1, Sox2, c-Myc, and Nanog. We have designed and tested several saRNAs to up-regulate the KLF4 gene. We demonstrate that some of the saRNAs can up-regulate KLF4 in addition to Sox2, c-Myc, and Nanog expressions in hematopoietic and mesenchymal stem cells. The genomic-bioinformatic method of identifying saRNAs for the study of stem cell properties could have broad implications in stem cell research.

Glioblastoma-related microRNA regulators and targets
Pavel Sumazin, Ph.D., Research Scientist & Associate Director for Bioinformatics
Sulzberger Columbia Genome Center, Columbia University, New York, NY, USA

Pavel Sumazin, Wei-Jen Chung, Hua-Sheng Chiu, Xuerui Yang, Mukesh Bansal, Andrea Califano

Recent evidence suggests that MicroRNAs (miRs), including MiR-26a, miR-221/2 and a battery of differentially expressed miRs across tumors and prognosis may be key regulators of Glioblastoma multiforme (GBM) tumorigenesis. To characterize their regulatory role, we constructed an integrated gene-miR regulatory network that is being used to identify GBM master regulators. We will present our work on identifying miR regulators and targets in GBM tumors. We will present models, algorithms, and results from validation experiments, including validated predictions of post transcriptional regulators of miR biogenesis, modulators of miR regulation, and synergistic miR targeting.

MicroRNAs in neurodegenerative diseases
Francesca Ruberti, PhD., CNR-Institute of Neurobiology and Molecular Medicine
European Brain Research Institute, Rome-Italy

Francesca Ruberti 1, Elisa Vilardo1, Christian Barbato1 Maria Teresa Ciotti1, Carlo Cogoni2
INMM-Istituto di Neurobiologia e Medicina Molecolare, CNR, Rome-Italy
EBRI-European Brain Research Institute, Rome-Italy

In the brain, amyloid precursor protein (APP) and Aβ are critical in the pathogenesis of  Alzheimer’s Disease (AD), which is the most common form of dementia in the elderly. We recently demonstrated by gain and loss of function experiments, that miR-101 is a negative regulator of APP expression and Aβ load in primary rat hippocampal neurons. We are planning to investigate the role of miR-101 in the regulation of amyyloid precursor protein and in the onset and progression of AD by using established primary cell cultures and AD murine models.

The role of microRNAs (miRNA) in the development of Polycystic Kidney Disease
Kyriacos N. Felekkis, PhD., Member of Executive Committee, Center for Research in Molecular Medicine, Department of Biological Sciences, University of Cyprus, Nicosia, Cyprus

Kyriacos N. Felekkis1, Carsten Sticht2, Gregoris Papagregoriou1, Bettina Kranzlin2, Norbert Gretz2 and Constantinos Deltas1
1 Department of Biological Sciences, University of Cyprus
2 Medical Research Center, Klinikum Manheim, University of Heidelberg
MicroRNAs (miRNAs) are recently discovered small non-coding RNA species that post-transcriptionally regulate gene expression. MicroRNAs have the ability to simultaneously regulate the expression of multiple genes by interfering with the cell’s translational machinery. As a result, they are implicated in the pathology of various diseases, which are characterized by deregulation in gene expression.

Sustained Over-expression of microRNA Mimics using a Lentiviral Delivery System
Stephanie Urschel, PhD., Senior Field Scientist Europe, Thermo Scientific Genomics
Erembodegem, Belgium

microRNA (miRNA) mimics and inhibitors are frequently employed to better understand the contributions of non-coding RNAi to cell physiology. Currently, the predominant method for over-expressing mature miRNAs involves delivery of synthetic mimics into target cells that can be readily transfected. To further expand the collection of tools available to researchers, and expand into cell lines that are difficult to transfect, we have developed a new lentiviral-based miRNA expression platform. This uses a unique miRNA scaffold for efficient processing and improved functionality. Strategies used to optimize robust miRNA expression will be presented along with data describing their overall performance in mammalian cell culture models.

Cell signaling by miR-165/166 in plant pattern and organ development
Christina Roberts, PhD. student, Uppsala University, Evolutionary Biology Centre
Uppsala, Sweden

We are using the Arabidopsis thaliana root as a model system to analyze pattern organization and cell fate. Class III HOMEODOMAIN LEUCINE ZIPPER (HD-ZIP III) proteins are transcription factors, involved in shoot dorsoventral specification that are post-transcriptionally regulated by miR165/166. We have determined that they also dose-dependently regulate the water conducting xylem cell type and patterning in roots. This regulation occurs through a pathway of bidirectional non-cell autonomous signaling by the miRNA and another transcription factor, restricting the HD-ZIP III domain to the root centre. The mobility of these factors relies on plasmodesmatal pores for their non-cell autonomous action.

MicroRNAs are novel biomarkers of colorectal cancer
Muhammad I. Aslam, MBBS, PhD Student, University of Leicester, Department of Cancer Studies and Molecular Medicine, Leicester Royal Infirmary, Leicester, UK

Colorectal Cancer (CRC) screening with the faecal occult blood testing, although effective, is generally considered to lack convenience, sensitivity and specificity for use as a general screening test. This explains why the search for an improved, non invasive and more accurate screening test has continued despite a recent commencement of National Bowel Screening Programme in the UK. In this search the presence of cancer related microRNAs in the circulating blood at levels sufficient to be measurable for the detection of tumours have been identified. Our feasibility study investigated the potential use of circulating microRNAs for the early detection of colorectal cancers.

miRNA biomarker profiling from blood – A promising approach to non-invasive diagnostics
Thomas Brefort, PhD. 1, 2 Director Application Development, 1febit biomed gmbh, Heidelberg; 2Biomarker Discovery Center Heidelberg, Germany

MicroRNAs are specifically involved in the regulation of distinct biological processes. Accumulating evidence for deregulated miRNA expression in several diseases, qualify non-coding RNAs as ideal candidates for novel diagnostic markers. Here, we present miRNA expression profiling studies on various human diseases, including different cancers. We analysed blood-derived miRNAs from respective patient and control groups on the automated Geniom® microarray platform and identified signatures of significantly deregulated miRNAs that allowed the specific, sensitive and accurate discrimination of diseased from healthy samples. Our findings emphasize the potential of blood-derived miRNA signatures for non-invasive diagnostics, improved surveillance of disease progression and response to treatment.

Development of miravirsen an LNA-modified oligonucleotide inhibitor of miR-122
Arthur A. Levin, PhD. Vice President & Chief Development Officer, Division of Drug Development, Santaris Pharma, Hørsholm DK and San Diego, CA, USA

Locked Nucleic Acid modified oligonucleotides that bind to and sequester specific microRNAs (miRs) have been widely used to characterize miR functions. Miravirsen (SPC3649) is an LNA modified oligonucleotide complementary to miR-122, a critical factor in hepatitis C viral replication.  Phase 1 studies in healthy volunteers produced results consistent with those predicted by laboratory studies and miravirsen has now entered Phase 2 clinical trials in patients infected with HCV.  Results of non-clinical safety studies and initial clinical results demonstrate that targeting miRs for therapeutic use in humans is possible and as the first miR-targeting therapeutic in clinical trials, miravirsen is leading the way for a new field of miR-targeting therapeutics.

Loss of mir-200c promotes an aggressive, invasive and chemoresistant phenotype in NSCLC
Paolo Ceppi, PhD Student, University of Turin at San Luigi Hospital, Department of Clinical and Biological Sciences, Orbassano,  Italy

Paolo Ceppi1,2, Giridhar Mudduluru2, Regalla Kumarswamy2, Ida Rapa1, Giorgio V Scagliotti1, Mauro Papotti1, Heike Allgayer2.
1Department of Clinical and Biological Sciences, University of Turin at San Luigi Hospital, Orbassano, Italy. 2Department of Experimental Surgery and Molecular Oncology of Solid Tumors, Medical Faculty Mannheim, University of Heidelberg, and German Cancer Research Center (DKFZ)- Heidelberg, Germany.

We have investigated the role of miR-200c, previously shown to control epithelial-to-mesenchymal transition (EMT), a process by which cancer cells acquire dedifferentiated/invasive characteristics, in non–small cell lung cancer (NSCLC). MiR-200c forced overexpression resulted in a reversal of EMT, reduced invasion and metastasis formation, and in enhanced sensitivity to chemotherapeutic agents. The hypermethylation of the promoter region was found responsible for the loss of miR-200c in invasive cells. Moreover, lower miR-200c levels were associated with higher lymph-nodal invasion, poor grade of differentiation, and lower E-cadherin expression in NSCLC patients. These results highlight a pivotal role of miR-200c in NSCLC progression.

miR-34a and miR-15a/16 are co-regulated in non-small cell lung cancer and control cell cycle progression in a synergistic and Rb-dependent manner
Erik Vassella, PhD., Group Leader in the Inst. of Pathology, University of Bern, Bern, Switzerland

MicroRNAs are small non-coding RNAs that regulate important cellular processes including proliferation and apoptosis. Although they often target common genes, little information is available how they interact with each other to control biological functions. MiR-34a and miR-15a/16 are functionally related. They share common targets and control similar processes including cell cycle progression and apoptosis. Here we show that miR-34a and miR-15a/16 act synergistically in inducing cell cycle arrest in a Rb-dependent manner. The concerted action of these miRNAs on cell cycle arrest is due to down-regulation of targets unique to either one of these miRNAs. This is based on the finding that down-regulation of cyclin E1, a unique target of miR-15a/16, by RNA interference abrogated the synergistic effect. In contrast, both miRNAs act on common targets in an additive rather than synergistic manner. Consistent with a functional relatedness, miR-34a and miR-15a/16 significantly correlate in their expression in adenocarcinomas of the lung. Co-regulation of these miRNAs is not directly linked nor is it due to defects in miRNA processing. To our knowledge this is the first example of miRNAs in human acting in a synergistic manner. These results may contribute to the understanding of the integrated network of miRNAs involved in gene expression and may also have therapeutic implications.

A functional variant in the microRNA-510 target site of the serotonin receptor type 3E gene is associated with diarrhea predominant irritable bowel syndrome
Beate Niesler, PhD., Scientist & Group Leader, University of Heidelberg, Institute of Human Genetics, Dept. of Human Molecular Genetics, Heidelberg, Germany

We found the HTR3E SNP c.*76G>A (rs62625044) to be associated with diarrhea predominant irritable bowel syndrome (IBS-D) in women (P = 0.0002, OR = 5.39). The SNP affected binding of miR-510 to the HTR3E 3´UTR and caused elevated expression. HTR3E and miR-510 co-localize in gastrointestinal tissue. This is the first example indicating microRNA related expression regulation of a serotonin receptor gene with a cis-regulatory variant affecting this regulation and appearing to be associated with female IBS-D.

Evaluation of a novel miRNA-specific normalization method and comparison between Affymetrix and Exiqon platforms
 Alain Sewer, PhD., Scientist - Disease Modeling & Simulation, R & D, PMI, Neuchatel, Switzerland

Abstract:
Obtaining reliable miRNA expression profiles from high-throughput array data is an essential concern in many studies. One crucial step is the raw data normalization that allows unbiased between-array comparisons in the downstream analyses. However, miRNA data differs slightly from "standard" gene expression profiles. In this talk we present a novel miRNA-specific normalization method based on controlled assumptions. This method is evaluated based on quality control criteria and on its ability to detect the biology stimulated in the underlying experiment. These two metrics allow classifying other normalization methods and are applied to an Affymetrix/Exiqon platform comparison, complemented by qRT-PCR validations.