4th RNAi Research & Therapeutics Conference

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4th RNAi Research & Therapeutics Conference

  1. 1. 3rd RNAi Research & Therapeutics – May 30-31, 2012 – Boston, MA 9:15 Novel Delivery Reagents for in Vivo Delivery of Wednesday, May 30, 2012 Low Dose of siRNA8:00 Welcome & Opening Remarks Xavier de Mollerat du Jeu, Ph.D., Senior Staff Scientist, Research and Development, Life Technologies KEYNOTE PRESENTATION siRNA is poised to be the next therapeutic drug. However, delivery of these molecules to the appropriate tissue8:05 Functional Analyses of RNAi Triggers remains the major bottleneck. The goal of this study was to develop new in vivo delivery reagents to deliver siRNA inJohn J. Rossi, Ph.D., Professor & Chair, Division of vivo by screening a library of lipid formulations complexedMolecular Biology, Beckman Research Institute, City of with siRNA. An siRNA targeting FactorVII (FVII) wasHope complexed with each formulation and injected intravenously at a 1mg/kg to 0.0125 mg/kg doses. The formulationsSmall interfering RNAs trigger post-transcriptional gene resulting in initial FVII knockdown were further optimized bysilencing by serving as guides for Argonaute proteins which mixture DOE and evaluated for their ability to deliver otherin combination with other proteins in the RNA induced siRNA as well as microRNA mimics and inhibitors. After asilencing complex (RISC) are the effectors of gene single intravenous injection of FVII siRNAs (0.050mg/kg)silencing. Having guide sequences that are efficiently complexed with this new reagent, we observed more thanincorporated into the Argonaute effectors critically impacts 90% mRNA and protein level reduction in liver cells forof the efficacy of RNAi. We have been studying the more than 2 weeks and this silencing was dose dependentfunctional differences and similarities between 19+2 siRNAs with an ED50 of 0.0125mg/kg. We also observed aand asymmetric 25/27mer Dicer substrate siRNAs as well reduction of Choleserol, LDL and triglycerides when weas Pol II and Pol III promoter expressed short hairpin RNAs silenced APOB using this reagent. In addition, we were able(shRNAs). Our studies have investigated incorporation of to knockdown 4 genes at the same time after a singlethese RNAs by the argonaute proteins via deep sequence injection of a mix of different siRNA complexed with thisanalyses as well as more standard Northern blotting reagent. This reagent can also deliver other payloads suchapproaches. The presentation will summarize the results of as antimirs and microRNA mimics. Finally, these newthese studies and provide insight into the factors that reagents are safe and did not trigger any sustained IFNenhance incorporation into RISC and subsequent target response or liver toxicity.knockdown . 9:40 Combinatorial Development of Synthetic siRNA Delivery Systems RNAi Delivery Methods Daniel G. Anderson, Associate Professor, Chemical Moderator: Devin Leake, Global Director of Research & Engineering,Harvard-MIT Division of Health Sciences & Development, Genomics, Thermo Fisher Scientific Technology, David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology FEATURED SPEAKER High throughput, combinatorial approaches have revolutionized small molecule drug discovery. Here we8:40 Recent Advances in Lipid Nanoparticle (LNP) describe our work on high throughput methods forMediated siRNA Delivery developing and characterizing siRNA delivery systems. Libraries of degradable polymers and lipid-like materialsMatthew Stanton, Ph.D., Head, RNA Optimization, Merck have been synthesized, formulated and screened for theirResearch Laboratories ability to delivery siRNA, both in vitro and in vivo. A number of siRNA delivery formulations have been developed with inLipid nanoparticles (LNP) represent the most advanced vivo efficacy, and show potential therapeutic application fordelivery platform for siRNA therapeutics. Our efforts in LNP the treatment of genetic disease, viral infection, and cancer.mediated siRNA delivery have focused upon optimization ofthe individual lipid components within the LNP (chemical 10:05 Networking & Refreshment BreakSAR), optimization of the formulation parameters(compositional and process SAR) and mechanistic 10:30 The Development of Dynamic Poly Conjugatesunderstanding of LNP mediated delivery. This presentationwill focus on the discovery of low MW cationic lipids as a Dave Rozema, Ph.D., Vice President, Chemistry,novel class of siRNA delivery agents with improved in vivo Arrowhead Researchtolerability and expanded therapeutic margins. Safe and efficient in vivo delivery of siRNA to the appropriate target cell would be a major advance in RNAi therapeutics. Hepatocytes, the key parenchymal cells of the liver, are a particularly attractive target cell type for siRNA delivery not only for their central role in infectious diseases Updated 4/27/12
  2. 2. 3rd RNAi Research & Therapeutics – May 30-31, 2012 – Boston, MAand metabolic disorders, but also their accessibility through Recent clinical trials of small interfering RNAs (siRNAs)liver fenestrae. We have developed a vehicle for thehave highlighted the need for robust delivery and detectiondelivery of siRNA to hepatocytes in vivo, which we have techniques that will enable the application of thesenamed Dynamic PolyConjugates. Key features of the therapeutics to increasingly complex disease and organDynamic PolyConjugate technology include: a new class of systems. Recent evidence points to synthetic RNA ligandsmembrane-active polymers, the ability to reversibly mask (aptamers) as an emerging class of pharmaceuticals withthe activity of these polymers until they reach the acidic great potential for targeted diagnostics and therapy. Whileenvironment of endosomes, and the ability to target these encouraging, the extended use of RNA aptamers as amodified polymers and their siRNA cargo specifically to delivery tool for siRNAs awaits the identification of RNAhepatocytes in vivo after intravenous injection. aptamer sequences capable of targeting and entering the cytoplasm of many different cell types. Here, we describeAttendees will learn latest developments in DPC novel selection methodologies for the rapid identificationtechnology and characterization of RNA aptamers capable of delivering- size of therapeutic window siRNAs into the cytoplasm of target cells. We coupled these- duration of gene knockdown from single and repeat methodologies with state-of-the-art RNA chemistries and injections fluorescent technologies resulting in effective, targeted- mechanism of targeting and intracellular delivery image-guided RNA reagents (TIGRs) that can be easily tracked in vivo. Importantly, these TIGRs may represent a10:55 Design and Production of Lipid Nanoparticle crucial first step in the transition of siRNAs from the bench-Systems for the Systemic Delivery of siRNA side into the clinic.Pieter R. Cullis, Ph.D., FRSC Director, NanoMedicines 11:45 Novel siRNA Delivery Technology TargetingResearch Group, Professor, Biochemistry and Molecular STAT3 in Tumor and Tumor Microenvironment forBiology, University of British Columbia Treatment of CancerRNAi-based drugs such as siRNA require sophisticated Hua Yu, Ph.D., Associate Chair and Professor, Cancerdelivery systems in order to achieve therapeutic benefits. Immunotherapeutics & Tumor Immunology, BeckmanThese delivery systems must protect encapsulated siRNA Research Institute at City of Hope Comprehensivefrom degradation in the circulation, promote accumulation in Cancer Centertarget tissue and facilitate intracellular delivery into targetcells. In order to be suitable for clinical use, these delivery STAT3 is persistently activated in many types of tumors. Itsystems must also be relatively non-toxic and must plays a crucial role in tumor invasion and resistance toencapsulate siRNA efficiently into well-defined, reproducible therapy, enabling tumor growth while turning off the body’snanomedicines using a scalable manufacturing process. anti-tumor immune response. Silencing STAT3 couldLipid nanoparticles (LNP) are currently the leading delivery therefore have significant implications for improved cancersystems for satisfying these demands. Efficient loading into therapy. However, as a transcription factor lacking its ownLNP can be achieved using ionizable cationic lipids that are enzymatic activity, STAT3 is a challenging target forrelatively non-toxic and can be optimized to achieve conventional small molecule drugs. While siRNA has beenmaximum intracellular delivery of siRNA following uptake effectively used to silence a host of genes, delivering it tointo target cells. With regard to manufacture of LNP siRNA specific tumor or tumor stromal cells while avoidingsystems, formulation processes require rapid mixing of an unrelated cells, and thus unwanted side effects, has provenaqueous stream, containing siRNA, with an enthanolic difficult. Dr Yu and colleagues have recently developed ansolution containing cationic lipid and PEG-lipid. We have in vivo targeted siRNA delivery technology - by covalentlydevised scalable microfluidic mixing technology that results linking siRNA to the CpG moiety, they are able to deliverin the formation of LNP siRNA systems over the size range siRNA to those cells expressing the endosomal Toll-like20-100 nm with siRNA encapsulation efficiencies receptor 9 (TLR9), which recognizes CpG, TLR9’s ligand.approaching 100%. It is shown that LNP siRNA systems These include immune cells such as dendritic cells,containing optimized ionizable cationic lipids are highly macrophages and B cells, and a variety of tumor cells,potent and relatively non-toxic agents for silencing including lymphoma and glioma. In addition to serving as ahepatocyte target genes following i.v. injection, achieving delivery vehicle for the siRNA, CpG’s binding and activation50% or greater target gene silencing of 10 ?g siRNA/kg of the TLR9 receptor has its own anti-tumorbody weight with therapeutic indices of 1000 or higher. This immunostimulatory effects. The CpG-Stat3 siRNA servesis currently the world-leading “gold standard” for the as an ideal weapon against many cancers, delivering a dualpotency of siRNA-based therapeutics in vivo. blow by simultaneously activating immune cells in the microenvironment and promoting tumor self-destruction. Dr11:20 TIGRs: Targeted Image-Guided RNA Therapies Yu and colleagues at City of Hope Comprehensive Cancer Center are moving this approach towards clinical trials.Paloma H. Giangrande, Ph.D., Assistant Professor,Hematology, Oncology and Blood & Marrow 12:10 Lunch On Your OwnTransplantation Faculty, University of Iowa Updated 4/27/12
  3. 3. 3rd RNAi Research & Therapeutics – May 30-31, 2012 – Boston, MA Therapeutic, Diagnostics and 2:55 Asymmetric, Hydrophobically-Modified RNAi Clinical Applications Compounds: From Mechanism of Uptake to Clinical Development Moderator: Ralph A. Tripp, Professor & GRA Chair, Infectious Diseases, University of Georgia James Cardia, Ph.D., Scientist II, Technology Development, RXi Pharmaceuticals RNAi-based therapeutics offer the potential to efficiently FEATURED PRESENTATION and specifically inhibit expression of any target gene in the human genome. To realize RNAi’s full potential as a viable1:40 RNAi Therapeutics: from Discovery to Clinical therapeutic, challenges in the delivery of RNAi compoundsDevelopment to target tissues and individual cells must be overcome. This presentation will provide an overview of our recentRachel Meyers, Ph.D., Vice President, Research and RLD, work using a medicinal chemistry approach to generateAlnylam improved RNAi compounds that are readily taken up by human cells without any delivery vehicle or formulation.2:05 Advances in Orally Administered RNAi This novel type of RNAi compound, termed self-deliveringTherapeutics rxRNAs (sd-rxRNA™), are extensively chemically-modified, asymmetric RNA duplexes, with a short duplex region ofRichard T. Ho, M.D., Ph.D., Executive Vice President, 11-15 bp and a fully phosphorothioated single stranded tail.Research and Development, Marina Biotech As demonstrated by total internal reflection fluorescence (TIRF) microscopy, efficient cellular internalization of sd-2:30 siRNA Therapeutics for Eye Diseases rxRNA compounds occurs within minutes of exposure to cells. Uptake is driven by the RAB5/EEA1 associatedElena Feinstein, Chief Scientific Officer, Quark branch of the endocytotic pathway. Preclinical dataPharmaceuticals supporting RXI-109, the first clinical candidate based on the sd-rxRNA platform will also be presented. RXI-109 isTwo of our three siRNA drug candidates that are currently designed to reduce the expression of CTGF (connectivein clinical development are being tested for ophthalmic tissue growth factor), a critical regulator of several biologicalindications. One of these siRNAs, PF-655, targets our pathways involved in fibrosis, including scar formation inproprietary gene RTP801, a stress-induced mTOR inhibitor. human skin. Pending FDA review, a clinical trial to evaluateThis drug has finished Phase IIa clinical trials in diabetic RXI-109 for safety, tolerability and initial efficacy will bemacular edema where dose dependent and substantial initiated in 2012.improvement of visual acuity has been observed. Relativelyslow kinetics of the development of the observed - Describe a novel class of small asymmetrictherapeutic effects as well as data from animal models hydrophobically-modified RNAi compounds, which areindicate that intravitral administration of siRNA targeting active in vitro and in vivo in the absence of a deliveryRTP801 causes both neuroprotection and stimulation of vehicleaxonal outgrowth. Another siRNA, QPI-1007, targeting - Data demonstrating mechanism of sd-rxRNA cellularcaspase 2 is in Phase I clinical trials in non-arteritic uptakeischemic optic neuropathy (NAION). Intravitreal injections of - Describe RXI-109 - an anti-CTGF sd-rxRNA and newthis siRNA showed significant and substantial anti-fibrotic compound for the treatment of dermal anti-neuroprtection in five different animal models of retinal scarringganglion cell injury including the glaucoma. The - Data characterizing RXI-109 efficacy in animal modelsintermediate analysis of human data points to the inhibitionof further visual deterioration in NAION patients treated with 3:20 Networking & Refreshment Breaksingle intravitreal injection of QPI-1007 within 2 weeks afterthe symptoms occur. In animal studies, both these siRNAs 3:50 Pre-Clinical and Clinical Development of Atu027displayed a quick distribution throughout retinal layersfollowing intravitreal injection, did not cause any type of Klaus Giese, Ph.D., Chief Scientific Officer, Research andlocal innate immune response including interferon response Development, Silence Therapeuticsand elicited RNAi-dependent cleavage of target mRNAs. Atu027, a novel RNAi therapeutic composed is currentlyThe presented data: being tested in a Phase 1 clinical trial in oncology. This(1) Confirms safety and RNAi activity of intravitreally investigational drug targets the expression of PKN3 in the delivered siRNA drugs; vascular endothelium and shows strong anti-tumor and anti-(2) Confirms predictability of animal efficacy results with metastatic activity in various pre-clinical models. Latest siRNA therapeutics in human trials; developments on Atu027 will be discussed.(3) Demonstrates clinical efficacy of siRNA therapeutics in clinical trials with efficacy rather than with - RNA interference pharmacodynamic endpoints - Delivery systeme - Oncology - Clinical trial Updated 4/27/12
  4. 4. 3rd RNAi Research & Therapeutics – May 30-31, 2012 – Boston, MA4:15 siGENOME Discovery and Validation of Novel opportunities and challenges of using RNAi technology toAnti-influenza Drugs help advancing oncology pipeline - with a focus on lessons learnt from our experience.S. Mark Tompkins, Associate Professor, InfectiousDiseases, University of Georgia 5:30 Networking ReceptionInfluenza A virus causes widespread infection in humansoften with severe clinical manifestations, thus there is a Thursday, May 31, 2012need for vaccines and therapeutic drugs. As influenza virusrelies on host cell proteins and their associated pathways tocomplete its life cycle, identifying the host moleculesrequired for virus replication provides valuable new targets Plenary Keynote Sessionfor antiviral therapy. In this study, we performed a genome- Moderator: Shidong Jia, Scientist, Genentechwide RNA interference (RNAi) screen in a humanrespiratory epithelial cell line infected with H1N1 virusesA/WSN/33, A/New Caledonia/20/99, or A/California/04/09 to KEYNOTE PRESENTATIONidentify host genes important for influenza virus replication.From the RNAi screen numerous host genes were found to 8:00 Steven Burrill, Chief Executive Officer, Burrill &be critical for influenza replication or act as host resistance Companygenes using three assay endpoints that included influenzaNP localization, viral genome replication, and infectious Pharmaceutical R&D spending is falling, the demand forvirus production. The key signaling pathways linked to innovation is increasing, and the traditional business modelthese genes were identified and validated by reporter for the industry is failing. What does innovation look likesystems. MicorRNAs that regulate these pathways were today in the new austerity and what will it take to bealso identified and shown to modulate viral replication. successful? What’s required is creativity in raising money,Pathway analysis showed that several genes functioned as making deals, and forging new business models. Burrillapex genes in host cell pathway, and targeting these genes brings his 45 years of successful dealmaking, companywith re-purposed small molecule drugs phenotyped RNAi- building, and investing to this discussion.based gene silencing and inhibition influenza virusreplication. These novel druggable targets were sensitive tonanomolar levels of drugs in vitro and in mouse models of KEYNOTE PRESENTATIONinfection suggesting new disease intervention strategieswith new classes of antiviral drugs for chemoprophylaxis 8:45 The Personal Genome Project - Open Access toand treatment. Genome Sequences + Trait data.BENEFITS George Church, Ph.D., Professor, Genetics; Director,1. Use of RNAi for novel drug target discovery Center for Computational Genetics, Harvard Medical2. Targeting host processes mitigates potential for viral School escape mutants3. The potential for viral use of common pathways may The PGP enables open observation and critique of a large enable broad anti-viral activity cohort "test-driving" comprehensive participatory4. Pathway analysis identifies microRNAs that may serve personalized medicine. Since 2004, we have helped push as global regulators of infection the cost of reading and writing DNA (and biological5. Potential for rapid development by repurposing existingsystems) down by a million-fold (5-fold faster exponential compounds. than Moores law) and enabled fully open-access human Genome+Environment=Trait (GET) data, stem cells, and4:40 DsiRNA/EnCore Hepatocellular Carcinoma clinical community curation/interpretation toolsProgram: Preclinical Delivery,Efficacy and Tolerability (Evidence.PersonalGenomes.org). This involves inherited genomes plus day-to-day genomic variation -- cancers,Sujit K. Basu, Ph.D.,Vice President, Formulation, Dicerna microbes, allergens, vaccines, & subcellular-resolutionPharmaceuticals epigenomics. We are also sequencing centenarians and long-lived mammals. Benefits include human genome5:05 RNAi for Cancer Target Discovery and Cancer engineering technologies for personalized diagnostics asTherapy: Opportunities and Challenges well as stem cell, synthetic organ, microbiome andYu Shen, Senior Group Leader, Cancer Research, Abbott immunome transplantation therapies.RNAi is not only an invaluable research tool for studyinggene functions in cell and animals but also holds promiseas a novel therapeutic modality. Over the last 10 years, we KEYNOTE PRESENTATIONhave been applying the RNAi technology for cancer targetidentification/validation and for the development of siRNAtherapy. In this presentation, we will highlight the Updated 4/27/12
  5. 5. 3rd RNAi Research & Therapeutics – May 30-31, 2012 – Boston, MA9:30 RNAi and Immortality: Recognition of Self/non- the FAS-induced apoptosis, in vitro screening data also Self Nucleic Acids enabled us to select targets that protected mice from FAS- induced hepatic failure. These results demonstrate thatCraig C Mello, Ph.D., Nobel Laureate, Blais Professor in complex pooled shRNA libraries provide a highly efficient,Molecular Medicine, University of Massachusetts flexible, and cost-effective alternative to array-based RNAiMedical School; Howard Hughes Investigator, HHMI screening methods for identifying genes regulating biological responses and possible new therapeutic targets.Organisms exhibit a fascinating array of gene-silencingpathways, which have evolved, in part, to confront invasive 11:10 Improving Reproducibility of Pooled Viralnucleic acids such as transposons and viruses. Not Screens - No shRNA Left Behindsurprisingly, these pathways are highly active in thegermline and can be elicited upon the introduction of Devin Leake, Global Director of Research& Development,transgenes. A key question raised by the existence of these Genomics, Thermo Fisher Scientificpathways is how do they distinguish self- from non-selfnucleic acids? Evidence exists for a number of cues that RNAi screens are an essential tool for gene discovery andmight facilitate the recognition of foreign sequences validation. Two predominant RNAi screening strategiesincluding, copy-number sensing, sensing of unpaired DNA, employ arrayed siRNAs or pooled shRNAs. In the formeror the sensing of aberrant RNA (e.g. dsRNA). Here we strategy, a large collection of siRNAs are arrayed acrossreport on a remarkable silencing pathway that can thousands of wells so that each well targets one gene. Inpermanently silence even single-copy transgenes. We the latter strategy, a large collection of lentivirus-expressedshow that the initiation of silencing depends on the piwi shRNAs target thousands of genes in a single well. CellsArgonaute PRG-1 and its genomically encoded piRNA with shRNAs targeting certain genes respond differently tocofactors. Our findings support a model in which PRG-1 phenotypic selection and become enriched or depletedscans for foreign sequences while two other Argonaute during the screen. Identifying the exact shRNA producingpathways serve as epigenetic memories of "self" and "non- the phenotype is then determined using next generationself" RNAs. These findings suggest that organisms utilize sequencing of genomic DNA from the cell(s). However,RNAi-related mechanisms to keep inventory of all genes many variables affect the reproducibility of this powerfulexpressed in the germ-line, and to recognize and silence screening strategy. For example, the size and compositionforeign genes. of the shRNA collection impacts shRNAs representation in the pool (and in transduced cells). Further, the amount of10:15 Networking & Refreshment Break genomic DNA needed for analysis is influenced by the amount of shRNA represented. Our results show that optimizing the level of shRNA by controlling the number of Target Discovery and Validation viral-transduced cells as well as the amount of genomic Moderator: Benjamin Haley, Scientist, Genentech DNA dramatically increase the screens reproducibility.10:45 RNAi Genetic Screening for Drug Target • Value of conducting pooled RNAi discovery screeningDiscovery without automation • Benefits of using lentiviral shRNA expression for targetPaul Diehl, Ph.D., Director, Business Development, validationCellecta • Techniques for increasing precision of shRNA discovery screeningPhenotypic loss-of-function RNAi screens with complex • Experimental design that will improve discovery rate oflentiviral-based shRNA expression libraries that target and pooled lentiviral shRNA collectionssilence several thousand genes provide a realistic andworkable approach to identify genes that functionally 11:35 A Bioinformatics Method to Identify Off-targetmodulate a cellular response such as viability of cancer Effects in RNAi Screenscells or apoptosis. As long as the shRNA libraries areproperly constructed so that hairpin representation is well Randall W. King, M.D., Ph.D., Associate Professor, Cellcharacterized and reasonably constrained, and changes in Biology, Harvard Medical SchoolshRNA representation in selected vs. control cellpopulations can be efficiently measured by HT sequencing, Because off-target effects hamper interpretation andpooled RNAi screens produce robust and reproducible validation of RNAi screens, we developed a bioinformaticsresults in a range of cell models. method, Genome-wide Enrichment of Seed Sequence matches (GESS), to identify candidate off-targetedWe will present results from two analyses: one “drop-out” transcripts from direct analysis of primary screening data.screen to identify genes essential for viability in a panel of GESS identified a prominent off-targeted transcript inleukemic cells, and a second “rescue” screen to identify several screens, including MAD2 in a screen forgenes required for FAS induced apoptosis. Both screens components of the spindle assembly checkpoint. I willfound a combination of known and novel signaling pathway discuss how incorporation of the results of GESS analysisand regulatory genes whose functions were confirmed to be can enhance the validation rate in RNAi screens.required to produce the biological responses. In the case of Updated 4/27/12
  6. 6. 3rd RNAi Research & Therapeutics – May 30-31, 2012 – Boston, MA12:00 Discovering Novel Regulators of Tumor 2:45 Application of genome-wide RNAi screens for theAngiogenesis through Integrated in vivo and in vitro identification of novel drug targets, combinationGenetic Analyses therapies, and development of potential patient selection biomarkersBenjamin Haley, Scientist, Genentech Attila Seyhan, Sr. Biomarker Discovery and DevelopmentThe survival benefits of antiangiogenic cancer therapies Leader, Biotherapeutics Clinical R&D, Pfizerhave been proven over the last decade. To betterunderstand the gene networks that promote or maintain Neratinib (HKI-272) is a small molecule tyrosine kinasetumor angiogenesis, and potentially enhance clinical inhibitor of the ErbB receptor family currently in Phase IIIoutcomes, we have developed a focused RNAi-based clinical trials. Despite its efficacy, the mechanism ofscreen in cultured primary endothelial cells. Our approach potential cellular resistance to neratinib and genes involvedmakes use of an in vivo-validated genetic signature with it remains unknown.combined with real-time phenotypic analyses in vitro. I willdiscuss the results and significance of the screen, along We undertook a genome-wide pooled lentiviral RNAi screenwith the unique challenges presented upon interpretation of to identify synthetic lethal or enhancer genes that interactreal-time datasets. with neratinib in a human breast cancer cell line (SKBR-3).12:25 Lunch Provided by GTC We discovered a diverse set of genes and pathways whose inhibition selectively impaired or enhanced the viability of cancer cells in the presence of subeffective concentrations MicroRNA and Diseases of neratinib. Examining the changes of these genes and Enal Razvi, Biotechnology Analyst, Nova Bioreports their protein products also led to a rationale for clinically relevant drug combination treatments. Treatment of cells1:30 [Oral Presentation from Exemplary Submitted with either paclitaxel or cytarabine in combination withAbstracts] neratinib resulted in a strong antiproliferative effect.To be considered for an oral presentation, please submit an Notably, our findings support a paclitaxel and neratinibabstract here by April 30. phase III clinical trial in breast cancer patients.1:55 Respiratory Syncytial Virus (RSV) Modifies In addition, we performed an RNAi screen to identify genesMicroRNA Gene Regulation during Infection Affecting involved in resistance to lethal concentrations of neratinib,RSV Replication using a long term survival study with pooled RNAi libraries and identified genes involved in chemoresistance to lethalRalph A.Tripp, Professor & GRA Chair, Infectious Diseases, concentrations of neratinib.University of Georgia The identification of novel mediators of cellular resistance toRespiratory syncytial virus (RSV) causes substantial neratinib could lead to the identification of new ormorbidity and life-threatening lower respiratory tract disease neoadjuvant drug targets and their use as patient orin infants, young children, and the elderly. Understanding treatment selection genetic biomarkers could make thethe host response to RSV infection is critical for developing application of anti-ErbB therapeutics more clinicallydisease intervention approaches. The role of microRNAs effective.(miRNAs) in post-transcriptional regulation of host genesresponding to RSV infection is not well understood. In this - The study identified novel chemosensitizer andstudy, we show that RSV infection of a type II lung epithelial chemoresistance targets for an experimental cancer(A549) cell line induces five miRNAs (let-7f, miR-24, miR- drug neratinib.337-3p, miR-26b and miR-520a-5p) and represses two - The identification of novel targets and pathways tomiRNAs (miR-198 and miR-595), and show that RSV G neratinib could lead to the development of novelprotein is a major inducer of let-7f. Luciferase-UTR selective therapies and combination therapies thatreporters and miRNA mimics and inhibitors validated a overcome therapeutic resistance to neratinib and othersubset of predicted target genes for let-7f, specifically ErbB2-targeted therapies.showing let-7f regulates cell cycle genes (CCND1, DYRK2 - The screen also identified a set of novel genes whoseand ELF4), a chemokine gene (CCL7), and a suppressor of silencing by RNAi caused long term chemoresistance tocytokine signaling 3 (SOCS3) gene. Together, these results lethal concentrations of neratinib.show that RSV G protein affects let-7f regulation of host - The identification of novel mediators of cellulargene networks, a feature that affects RSV replication. resistance to neratinib has profound implications for both basic and translational research.A benefit of this talk will be a better understanding of the - These genes can provide a rationale to stratify patientsmechanisms that contribute to RSV disease and new who may most likely benefit from neratinib therapy.pathways for RSV disease intervention. 3:15 Conference Concludes2:20 TBA Updated 4/27/12

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