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WEDNESDAY, NOVEMBER 3

8:30 am – 11:30 pm Pre-Conference Short Course* Course 5 Best Practices for Setting Up Effective RNAi Screens  The course is designed to provide in-depth information on how to go about setting up RNAi screening experiments and how to design assays for getting optimal results. The challenges working with siRNAs and shRNAs and the delivery reagents needed to get them into the appropriate cells and tissues will be discussed. The instructors will also provide their input on best practices for the execution of experiments and interpretation of results when dealing with complex biology and informatics. Course Instructors: Hakim Djaballah, Ph.D., Director, HTS Core Facility, Memorial Sloan Kettering Cancer Center Namjin Chung, Ph.D., Senior Research Investigator, Applied Genomics, Bristol-Myers Squibb Co. Serena Silver, Ph.D., RNAi Screening Projects Group Leader, Broad Institute at MIT Marc Ferrer, Ph.D., Team Leader, NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health * Separate registration required.

 

Seeking Novel Drug Targets

1:30 Chairperson’s Remarks

Albert B. Seymour, Ph.D., Director of Biological Profiling, Target Generation Unit, Biotherapeutics and Bioinnovation Center, Pfizer Inc.

1:40 A Lentivirus-mediated RNAi Screen Identifies Novel Regulators of Wnt/b-Catenin Signaling

Michele Cleary, Ph.D., Senior Director, Automated Biotechnology, Merck & Co., Inc.

To identify novel regulators of canonical Wnt signaling, we performed a lentivirus-mediated RNAi screen with a 14,000 vector shRNA library targeting 5,000 human druggable genes. Screen hits included multiple known regulators of Wnt/beta-catenin signaling as well as novel pathway regulators. Our follow up studies suggest potential for therapeutic manipulation of Wnt/beta-Catenin signaling through modulation of a well-established druggable target.

2:10 An Image-based siRNA Screen for Kinase Genes That are Important for Cancer Cells to Adapt to Hypoxia

Steven Haney, Ph.D., Associate Fellow, Applied Quantitative Genotherapeutics, Pfizer BioTherapeutics and Bioinnovation Center, Pfizer Inc.

The talk will describe the challenge in developing a morphology-based screen that identifies genes promoting the growth of cancer cells under hypoxia, selecting hits from primary screening data, and establishing the relevance of hits to cancer cell survival mechanisms. Assay development, statistical criteria for hit analysis and biological validation of hits will be discussed.

2:40 RNAi Screen With a High-Throughput qRT-PCR Assay Readout for Novel Target Identification

Namjin Chung, Ph.D., Senior Research Investigator, Applied Genomics, Bristol Myers Squibb Co.

3:10 Networking Refreshment Break in the Exhibit Hall

3:45 Synthetic Lethal siRNA Screens Using Preclinical Compounds

Alex Gaither Ph.D., Research Investigator II, Developmental and Molecular Pathways, Novartis Institutes for Biomedical Research

We are using siRNA screens to look for new synthetic lethal targets in drug resistant menlanoma cell lines. One of the challenges facing targeted low molecular weight inhibitors is the rapid emergence of resistance to drug. We have drug resistant cell lines in genetically defined backgrounds where we are trying to identify new targets and/or pathways that could sensitize to targeted therapies. The goal of our work is to better understand what combinations could be taken to the clinic to help prevent drug-induced resistance and improve the efficacy of our compounds in man.

4:15 Strategies for Improving RNAi Screening Success: Using a Ubiquitin-EGFP Assay to Identify Druggable Genes Required for Proteasome FunctionSponsored by
 

James Goldmeyer, Ph.D., Field Application Scientist , Thermo Scientific Genomics, Dharmacon RNAi Technologies
RNA interference (RNAi) is a powerful tool to perform gene knockdown and phenotype correlation studies.  When performed on a whole-genome scale, RNAi screens represent an effective strategy to identify novel therapeutic targets and to elucidate mechanisms of action or sensitivity to drugs.  In spite of its immense potential, RNAi library screening presents unique challenges that must be addressed to ensure success.  The authors will describe their RNAi screening facility and the strategies employed to ensure meaningful screening results.  As a case study, a cell-based assay using a ubiquitin-EGFP cell line to screen for drug target genes that when silenced inhibit proteasome function and/or affect cell viability will be discussed.  In particular, the authors will describe the approaches used to ensure effective delivery of highly potent siRNA molecules and to identify appropriate siRNAs that function as robust positive and negative controls.  The strategies used to validate compatibility of the RNAi automation platform with the phenotype being analyzed will also be provided.  Finally, the authors will discuss the data analysis and hit identification methods commonly used along with approaches for follow-up analysis of potential hits from primary screening, and methods to stratify these potential hits.

4:45 Translating Human Genetic Discoveries to Novel Targets and Therapeutics

Albert B. Seymour, Ph.D., Director of Biological Profiling, Target Generation Unit, Biotherapeutics and Bioinnovation Center, Pfizer Inc.

 

5:15 PANEL DISCUSSION: The Current Fate of Drug Targets Being Uncovered by RNAi Screens Moderator: Albert B. Seymour, Ph.D., Director of Biological Profiling, Target Generation Unit, Biotherapeutics and Bioinnovation Center, Pfizer Inc.

5:45 End of Day

 

6:00 pm – 9:00 pm Conference Short Course* DINNER COURSE Course 6 In vivo Discovery and Validation Using RNAi  In vivo experiments with RNAi are proving to be more complex and intricate than those done in vitro. This course is designed to provide experimental and technical details involving the design and use of such RNAi-based in vivo screens. Questions surrounding the choice and use of appropriate reagents and animal models, criteria for experimental design and quality control, immunogenic and off-target effects, and other important issues will be discussed in extensive detail. The course will also go over the challenges surrounding efficient delivery of siRNA or shRNA molecules into the cell, tissue uptake and retention, success of systemic and local delivery, and instructors will offer guidance and alternatives based on their expertise and experiences. These in vivo-based RNAi screens cover a diverse range of applications such as, screens for functional analysis of cellular pathways and moieties, for understanding the biology of a disease, for validation of drug targets, and many others. Scientists involved in these areas of research or those looking to learn more about how these screens are effectively designed and used are encouraged to participate. Course Instructors: Annie Conery, Ph.D., Director, C. elegans Screening Core, Department of Molecular Biology, Massachusetts General Hospital Siew Ho, Ph.D., Senior Principal Scientist, Applied Biotechnology, Bristol-Myers Squibb Co. * Separate registration required.