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GTC, Hyatt at Fisherman's Wharf, San Francisco, CA
Jul 07-08, 2011
| 7:00 | Registration & Continental Breakfast | | | | | 7:55 | Welcome & Opening Remarks | | | | | | | | | KEYNOTE PRESENTATION | | 8:00 | Gymnotic Delivery and Activity of Antisense Oligonucleotides | | | | | |  | Cy Stein, M.D., Ph.D.
Director of Medical Genitourinary Oncology
Einstein-Montefiore Medical Center | | | | | | The delivery problem is the single most important current problem that requires resolution for the field of oligonucleotide therapeutics to progress. Two years ago, we reported that LNA-phosphorothioate gap-mers, under the correct culture conditions and concentrations, do not require any transfection reagents for either delivery or highly efficient gene silencing. We now extend the general, robust phenomenon of gymnotic delivery to FANA-phosphorothioate gap-mer oligos, using the androgen receptor as model target. We will present evidence that the gymnotic process can be significantly modified by agents that affect cell membrane function, and additional data on the mechanism of gene silencing after gymnotic delivery. | | | | | | | | Session I: RNAi Delivery Methods | | | | | | | 8:45 | Steven F. Dowdy, Ph.D., Investigator, Howard Hughes Medical Institute; Professor, Cellular and Molecular Medicine,UC San Diego | | | | | 9:10 | Biomaterials for siRNA Delivery | | | Daniel Anderson, Ph.D., Associate Professor, Chemical Engineering, Massachusetts Institute of Technology | | | | | 9:35 | Non-invasive Administration of LNA Antisense to the Liver and Kidneys | | | Sterghios Moschos, Ph.D., Senior Scientist, Pfizer | | | | | 10:00 | Networking & Refreshment Break | | | | | 10:30 | Peter Nolan, Ph.D., Executive Director and Senior Vice President, Oxford BioMedica | | | | | Session II: Target Validation | | | | | | | 10:55 | Impact of Promoter Activity on Lentiviral microRNA-based Gene Silencing in Specific Cell Contexts | | | Devin Leake, Ph.D., Global Director of Research & Development, Genomics, Thermo Fisher Scientific | | | | | | Viral systems provide an effective approach for delivering genetic information into cells enabling researchers to modulate gene expression for target identification and validation studies in pre-clinical development. Those based on pseudotyped lentiviral vectors provide 1) delivery of large genomic payloads, 2) broad tropism, and 3) long-term transgene expression by integration into the host genome. To successfully utilize lentiviral delivery in RNAi-mediated gene silencing, careful attention must be paid to the design of the short hairpin RNA (shRNA), the vector backbone, and the choice of the promoter to drive expression. Depending on the cell type, promoter activity may be weak or inactivated by epigenetic mechanisms. We have previously described a wholly unique lentiviral shRNA expression platform based on an optimized microRNA expression cassette (SMARTvector 2.0). Here we present data linking the potency of shRNA-induced knockdown with promoter activity in the context of different cellular environments. We will show that promoter performance varies significantly among cell lines which, in turn, influences potency of gene knockdown and introduce a unique approach for the selection of appropriate promoters for successful vector-based RNAi.
Benefits of this talk:| • Information regarding potential promoter expression issues when using viral delivery to integrate into a host genome | | • Effective method for silencing genes in relevant cell types | | • Information about alternative promoters for expression experiments | | • Introduction of strategy for identifying optimal promoter in experimental system |
| | | | | 11:20 | Targeting EVI-1 in Solid Tumors | | | Thomas Primiano, Ph.D., Founder and Chief Executive Officer, NanoOncology | | | | | 11:45 | siGENOME Discovery and Validation of Novel Anti-influenza Therapeutic Drugs | | | S. Mark Thompkins, Ph.D., Associate Professor, Infectious Diseases, University of Georgia Medical School | | | | | | Influenza A virus causes widespread infection in humans often with severe clinical manifestations, thus there is a need for vaccines and therapeutic drugs. As influenza virus relies on host cell proteins and their associated pathways to complete its life cycle, identifying the host molecules required for virus replication provides valuable new targets for antiviral therapy. In this study, we performed a genome-wide RNA interference (RNAi) screen in a human respiratory epithelial cell line infected with H1N1 viruses A/WSN/33, A/New Caledonia/20/99, or A/California/04/09 to identify host genes important for influenza virus replication. From the RNAi screen numerous host genes were found to be critical for influenza replication or act as host resistance genes using three assay endpoints that included influenza NP localization, viral genome replication, and infectious virus production. The key signaling pathways linked to these genes were identified and validated by reporter systems. The results showed that several genes functioned as apex genes in host cell pathway, and targeting these genes with re-purposed small molecule drugs phenotyped RNAi-based gene silencing and inhibition influenza virus replication. These novel druggable targets were sensitive to nanomolar levels of drugs in vitro and in mouse models of infection suggesting new disease intervention strategies with new classes of antiviral drugs for chemoprophylaxis and treatment.
Benefits of this talk:| • Use of RNAi for novel drug target discovery | | • Targeting host processes mitigates potential for viral escape mutants | | • The potential for viral use of common pathways may enable broad anti-viral activity | | • Potential for rapid development by repurposing existing compounds |
| | | | | 12:10 | Lunch On Your Own | | | | | 1:40 | [Oral Presentations from Exemplary Submitted Abstracts] | | | To be considered for an oral presentation, please submit an abstract here. | | | | | Session III: Gene Activation and Silencing by Small RNAs | | | | | | | 2:05 | Target Knockdown in Tumor with Nanoparticle Dicer Substrate RNA (DsiRNA) Delivery System | | | Sujit Basu, Ph.D., Senior Director, Formulation, Dicerna | | | | | | RNA interference (RNAi) is a naturally occurring cellular process able to silence the expression of specific genes. The ability to trigger RNAi in mammalian cells using double-stranded RNA has stimulated great interest in therapeutic applications of RNAi to silence disease causing genes. This presentation will describe novel Dicer Substrate RNA-based therapeutics (DsiRNA) that utilize an earlier entry step in the gene silencing process, namely the engagement of the enzyme Dicer, which is a natural initiation point of the RNAi cascade. Introduction of double-stranded, sequence-specific RNA oligonucleotides that are substrates for Dicer action allows reduction of expression of a target gene in a highly selective and specific fashion. Compared to other RNAi approaches, this next generation RNAi strategy demonstrates greater potency and longer duration of action. We have formulated DsiRNA payload in lipid nanoparticle delivery systems to achieve >80% knockdown in mouse liver and >50% knockdown in orthotopic Hep3B liver tumor model. | | | | | 2:30 | W. Michael Flanagan, Ph.D., Senior Director, RNA Sciences, Sirna Therapeutics of Merck | | | | | 2:55 | Networking & Refreshment Break | | | | | 3:30 | Non-coding RNAs; The Art of Epigenetically Modulating Gene Transcription in Human Cells | | | Kevin Morris, Ph.D., Assistant Professor, Molecular Medicine, The Scripps Research Institute | | | | | | Observations over the last few years have demonstrated that exogenously introduced small non-coding antisense RNAs can transcriptionally modulate gene expression in human cells. The mechanism involved in small antisense RNA directed transcriptional gene silencing (TGS) appears to involve Argonaute 1, Histone Deactylase 1, and DNA methyltransferase 3a. However, an endogenous RNA trigger directing these or other epigenetic regulatory proteins to targeted genomic loci in human cells had until recently remained unknown. We present evidence here suggesting that long antisense non-coding RNAs function in human cells as effector molecules driving TGS. These long antisense non-coding RNAs function to guide epigenetic remodeling complexes consisting of Enhancer of Zeste and G9a to target loci. When these regulatory antisense non-coding RNAs are degraded using small non-coding RNAs the result can be a concomitant activation of their protein coding counter part. Importantly, this RNA based transcriptional regulatory mechanism can be taken advantage of to either transcriptionally silence a genes expression in a long-term manner or activate a genes transcription by the targeted degradation of the regulatory long antisense non-coding RNAs.
Benefits of this talk:| • Define endogenous mechanism of ncRNA regulation in human cells | | • Provide examples of utilizing the endogenous mechanism to silence and activate several different genes involved in human disease. | | • Provide details of new algorithm that builds on mechanistic data, show examples of function. | | • Provide information on targeted delivery approaches that employ the transcriptional activating and silencing small RNAs in human cells to target human diseases such as Cancer and HIV-1. |
| | | | | 3:55 | Peter A. Beal, Ph.D., Principal Investigator, Chemical Biology, UC Davis | | | | | | | | | FEATURED PRESENTATION | | 4:20 | | | | | | |  | John Rossi, Ph.D.
Dean, Irell and Manella Graduate School of Biological Sciences
Beckman Research Institute, City of Hope | | | | | | | | | | | | | | 5:00 | Networking Reception & Poster Session | | | |
| | | | | Day 2 - Friday, July 8, 2011 | | | | | 7:30 | Continental Breakfast | | | | | | | | | KEYNOTE PRESENTATION | | 8:00 | RNAi Therapy and the Immune System – A Good Thing? | | | | | |  | Nigel McMillan, M.D.
Principal Research Fellow and Deputy Director
University of Queensland Diamantina Institute | | | | | | RNA Interference holds great promise as a therapy for cancers an viral infections via its ability to directly kill cells via down-regulation of critical target genes. We now have siRNAs that work in the pM range and much of the current clinical trial efforts are targeting towards diseases that benefit from direct introduction of siRNA eg AMD via intraocular injection. We need to extend our delivery thinking beyond these sites and consider therapies that evoke beneficial immune responses. This is because all current cancer therapy merely reduces disease burden - cure or clearance relies on the immune system. I will describe our technologies to delivery to alternative sites such as tumours (stealth lipoplexes), the ano-genital mucosa (the PLAS system) and skin (nanoneedles). We have also investigated ways to improve the efficacy of RNAi and have developed siRNAs and shRNAs that are able silencing target genes and evoke innate and adaptive immune responses against targeted cancer cells. This new class of “bifunctional” and “trifunctional” siRNAs should be much more effective at treating cancers and viral infections.
Making RNAi better.| • Can we deliver to sites of clinical relevance? | | • Can we improve RNAi by evoking immune responses? | | • Which immune responses are desired? |
| | | | | | | | Session IV: An Examination of Therapeutic and Clinical Applications | | | | | | | 8:45 | Wayne Jiang, M.D. Ph.D., Senior Research Scientist, Idera Pharmaceuticals | | | | | 9:10 | David V. Schaffer, Ph.D., Principal Investigator, Chemical and Biomolecular Engineering, UC Berkeley | | | | | 9:35 | TBA | | | | | 10:00 | Networking & Refreshment Break | | | | | 10:30 | Harnessing the Power of RNAi – Target-Specific Approaches through Multiple Delivery Systems | | | Michael V. Templin, Ph.D., Senior Vice President, Preclinical Development, Marina Biotech | | | | | | Nucleic acid-based therapeutics provide an opportunity for targeting disease-causing proteins or protein pathways that are not appropriate for small molecule or monoclonal antibody approaches. Down-regulation of an individual protein, by targeting a specific mRNA, can be achieved with double-stranded (e.g., siRNA) or single-stranded oligonucleotides. MicroRNA (miRNA) pathways can be targeted with single-stranded inhibitors or double-stranded miRNA mimetics can increase the level of an essential miRNA lost in the disease process. Unique constructs and chemistry, e.g., UsiRNA and conformationally restricted nucleotides (CRN), increase the specificity of this class of drugs. Delivery systems ranging from simple “saline-like” formulations to dynamic formulations based on liposomes or engineered bacteria add further power. When required, a discovery process can utilize these diverse platforms to identify the most appropriate modality. However, focused efforts to understand the intended target, the biology required for efficacy, and the disease indication, can indicate the modality with the highest probability for success in development.
Benefits of this talk:| • A broad nucleic acid-based platform provides a superior drug discovery engine; key considerations included, | | o Diverse delivery systems | | ? Saline-like, liposome, bacterial | | o Multiple routes of administration | | ? Systemic, local, oral | | o Diverse therapeutic approaches | | ? Double-stranded RNAi-based therapeutics | | ? Single-stranded oligonucleotide therapeutics | | ? MicroRNA-based therapeutics (single- and double-stranded) | | • Balance of platform diversity and focused efforts provides the power to bridge from discovery into development |
| | | | | 10:55 | Preclinical Development of an sd-rxRNATM Based Therapy for Treatment of Fibrosis and Retinal Disorders | | | James Cardia, Ph.D., Scientist II, Chemistry, RXi Pharmaceuticals | | | | | 11:20 | [Oral Presentations from Exemplary Submitted Abstracts] | | | To be considered for an oral presentation, please submit an abstract here. | | | | | 11:50 | Lunch | | | | | Session V: The Role of MicroRNAs | | | | | | | 1:30 | microRNAs with Broad Antiviral Activity | | | Amy Buck, Ph.D., Assistant Professor, Pathway Medicine, University of Edinburgh Medical School | | | | | 1:55 | William S. Marshall, Ph.D., Co-Founder, President and Chief Executive Officer, miRagen Therapeutics | | | | | 2:20 | Regulation of Liver Growth by miRNAs in Liver Transplantation | | | Jian Wu, M.D. Ph.D., Professor, Gastroenterology & Hepatology, UC Davis | | | | | | Liver transplantation is the only established treatment for end-stage liver disease and fulminant liver failure. Due to severe shortage of donor livers, living donor liver transplantation (LDLT) emerged as an alternative approach to shorten the waiting duration. However, the molecular mechanisms underlying the growth of small size grafts and the remaining livers after living donation are poorly understood. miRNAs negatively modulate expression of genes that are involved in cellular function and metabolism. The present study aims to identify critical miRNA species that modulate the growth of small grafts and the remaining livers after partial hepatectomy (PH). Small size graft liver transplantation (SSGLT) in rats w as employed as a model of LDLT. We found that miR_122a, Let_7b and miR_26a were reduced by more than 90% in 45% volume grafts. In the remaining livers after 50% PH, 30 miRNAs were down-regulated by more than 50%. A negative correlation existed between down-regulated miRNAs and highly up-regulated genes involved in cell cycle and proliferation in the remaining livers. Moreover, overexpression of miR_26a markedly down-regulated cyclin E2 protein levels, and significantly decreased proliferation of HepG2 cells.
In conclusion: Down-regulated miRNAs play a pivotal role in promoting the growth of small size grafts and the remaining livers. The negative correlation between down-regulated miRNAs and up-regulated genes suggests that these specific miRNAs participate in the modulation of a growth response in both living donors and small size graft recipients. Cyclin E2 is the target gene of miR_26a, which was markedly reduced during liver growth. | | | | | 2:50 | Respiratory Syncytial Virus (RSV) Proteins Dysregulate Host MicroRNA Governance of Cellular Pathways | | | Ralph A. Tripp, Ph.D., Georgia Research Alliance Chair & Professor, Infectious Diseases, University of Georgia | | | | | | Respiratory syncytial virus (RSV) causes substantial morbidity and life-threatening lower respiratory tract disease in infants, young children, and the elderly. Understanding the host response to RSV infection is critical for developing disease intervention approaches and fundamental to this is understanding microRNA (miRNA) governance of host genes. In this presentation, we will show that RSV infection in human respiratory epithelial cells induces (let-7f, miR-24, miR-337-3p, miR-26b and miR-520a-5p) and represses (miR-198 and miR-595) host cell miRNAs important in governing the response to infection. Notably, we will show that specific RSV proteins induce specific miRNAs, a feature that affects cell cycle and chemokine genes, and suppressor of cytokine signaling genes, whose expression controls anti-viral cytokines and responses. The benefit from these findings are that understanding which RSV proteins affect miRNA expression during infection allows for development of novel disease intervention approaches, and understanding miRNA governance of the host response offers new strategies to control or manipulate the regulation of multiple gene networks, a feature that may reduce or eliminate disease pathogenesis. | | | | | 3:45 | Conference Concludes | | | | |
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Organized by:
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GTC |
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Invited Speakers:
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| KEYNOTE SPEAKER | KEYNOTE SPEAKER | | | Cy Stein, M.D., Ph.D.
Director
Medical Genitourinary Oncology
Einstein-Montefiore Medical Center | Nigel McMillan, M.D.
Principal Research Fellow
University of Queensland
Deputy Director, Diamantina Institute | | | | FEATURED SPEAKER | | John Rossi, Ph.D.
Dean, Irell and Manella Graduate School of Biological Sciences
Beckman Research Institute, City of Hope | | |
| | | | DISTINGUISHED SPEAKERS | | | Daniel G. Anderson, Ph.D.
Associate Professor, Chemical Engineering
Massachusetts Institute of Technology
| | | Sujit Basu, Ph.D.
Senior Director, Formulation
Dicerna Pharmaceuticals
| | | Amy Buck, Ph.D.
Assistant Professor, Pathway Medicine
University of Edinburgh Medical School
| | | James Cardia, Ph.D.
Scientist II, Chemistry
RXi Pharmaceuticals
| | | Steven F. Dowdy, Ph.D.
Investigator, Howard Hughes Medical Institute
Professor, Cellular and Molecular Medicine, UC San Diego
| | | W. Michael Flanagan, Ph.D.
Senior Director, RNA Sciences
Sirna Therapeutics, Merck
| | | Wayne Jiang, Ph.D.
Senior Research Scientist
Idera Pharmaceuticals
| | | Devin Leake, Ph.D.
Director, Research & Development
Thermo Fisher Scientific
| | | William S. Marshall, Ph.D.
Co-Founder, President and Chief Executive Officer
miRagen Therapeutics
| | | Kevin Morris, Ph.D.
Assistant Professor, Molecular Medicine
The Scripps Research Institute
| | | Sterghios Moschos, Ph.D.
Senior Scientist
Pfizer
| | | Peter Nolan, Ph.D.
Executive Director and Senior Vice President
Oxford BioMedica
| | | Thomas Primiano, Ph.D.
Founder and Chief Executive Officer
NanoOncology
| | | David V. Schaffer, Ph.D.
Principal Investigator, Chemical and Biomolecular Engineering
UC Berkeley
| | | Michael Templin, Ph.D.
Senior Vice President, Preclinical Development
Marina Biotech
| | | S. Mark Thompkins, Ph.D.
Associate Professor, Infectious Diseases
University of Georgia Medical School
| | | Ralph A. Tripp, Ph.D.
Professor and Georgia Research Alliance Chair, Vaccine Studies
University of Georgia
| | | Jian Wu, Ph.D.
Professor, Gastroenterology & Hepatology
UC Davis |
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Deadline for Abstracts:
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June 7, 2011
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Registration:
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https://www.gtcbio.com/index.php?option=com_register&cn=2nd+RNAi+Research&cid=32
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