Thursday, February 2

7:30-8:15 Technology Workshop 

Sponsorship Available. Contact: Carol Dinerstein at 781-972-5471 or dinerstein@healthtech.com.

GENOME-WIDE RNAI SCREENING FOR TARGET IDENTIFICATION

8:30 Chairperson’s Opening Remarks

8:30-9:00 Sensitized RNAi Screen of Human Kinases and Phosphatases Identifies Novel Regulators of Apoptosis and Chemoresistance
Dr. John Blenis, Professor of Cell Biology, Harvard Medical School
Given the adaptability of tumor cells, drug resistance is a major cause of failure in conventional chemotherapy. Tilting the cellular balance towards apoptosis by activating cell death signals with conventional chemotherapeutic agents, combined with inhibiting tumor-specific survival signals, may be crucial in determining the fate of cancer cells. The combination of targeted and conventional therapies has the potential to maximize efficacy, while minimizing toxicity. Using large-scale RNAi-based screens, we report the identification of known and novel human kinases and phosphatases that promote cell survival, and a new group of phosphatases with tumor-suppressor-like activity. In addition, functional screens in the presence of low dose apoptosis-inducing chemotherapeutic agents has identified a group of kinases whose loss of function sensitizes cells to undergo cell death, highlighting their importance as potential drug targets.

9:00-9:30 Fast siRNA Transfection Method for Automated High-Throughput Screen of Genome-Scale siRNA Libraries
Dr. Namjin Chung, Senior Research Scientist, Automated Biotechnology, Merck Research Laboratories
By understanding the efficacy, stability, and toxicity of lipid-based transfection reagents, we developed a fast and efficient method for transfecting siRNA into cultured mammalian cells. Transfection throughput is only limited by the speed of liquid and microplate handling on the robotic platform, and triplicate transfections of a genome-scale siRNA library of more than 20,000 genes can be completed in 16 hours. Transfection efficiency, behaviors of various control siRNAs, and other quality metrics were comparable to or better than conventional, low-throughput methods, while the cost of screening was significantly lower. The current method provides an efficient means for investigating gene functions in the large scale.

9:30-10:00 A Systematic RNAi Screen for C. Elegans Longevity Genes
Dr. Siu Sylvia Lee, Assistant Professor, Department of Molecular Biology and Genetics, Cornell University
We are using genome-wide RNAi screenings to identify C. elegans longevity genes. We have screened over 16,000 RNAi clones, and identified about 90 RNAi inactivations that reproducibly extended C. elegans lifespan. These RNAi clones correspond to 90 distinct C. elegans genes. The candidate longevity genes we identified participate in a wide variety of cellular processes, indicating that diverse biological functions can influence longevity in C. elegans.

10:00-10:30 HT RNAi Screen and Analysis of Cholesterol Synthesis Pathway in Human Hepatoma Cells
Dr. Christophe J. Echeverri, CEO/CSO, Cenix BioScience GmbH
The talk will describe a recently-completed RNAi screen of over 5,000 therapeutically-relevant genes for new atherosclerosis targets, conducted in collaboration with Bayer Healthcare. The screening assay was based on the identification of genes whose inhibition caused an increase in LDL uptake, as a direct consequence of inhibiting cholesterol synthesis. Genes that were thereby newly implicated in this complex pathway were further analyzed by a new validation approach known as Pathway Titration™. I will particularly focus on the range of scientific and strategic decisions that led us through the many challenges of this large study.

10:30-11:30 Coffee Break with Exhibit and Poster Viewing

INTEGRATING SIRNA- AND COMPOUND-LIBRARY SCREENING

11:30-12:00 Targeting the HIF-1 Pathway: An Experience with siRNA and Small Molecule Screens
Dr. Yu Shen, Group Leader, Cancer Exploratory Biology, Abbott Laboratories
HIF-1 emerges as an attractive target for cancer therapy. However, the lack of apparent “druggable” targets in the pathway hinders the development of small molecule inhibitors targeting HIF-1. In order to identify “druggable” targets in the HIF-1 pathway, we carried out an HIF-1 reporter screen using both a siRNA library against the “druggable genome” and a compound library consisting of 800,000 small molecule compounds. The siRNA library screen resulted in overwhelming “off-target hits.” An analysis of some of the off-target hits revealed that siRNA-mediated off-target gene silencing could be triggered by as low as 7-nt complementation between a siRNA and an mRNA. In contrast, screening of the compound library using the HIF-1 reporter assay resulted in the identification of a class of HIF-1 inhibitors with sub-nanomolar cellular activity. Further analysis of this class of compounds highlights the importance of an often-overlooked mechanism in HIF-1 regulation.

12:00-12:30 Advanced Analytical Approaches to Small Molecule and RNAi Studies Using High-Content Screening and Informatics
Dr. Steven Haney, Senior Scientist, Biological Technologies, Wyeth Research
High-Content Screening is a powerful technology for studying effects of small molecule compounds and RNAi on cells. In fact, it is a challenge to integrate all of the data that is available. We have been interested in leveraging these data, and have begun to develop methods for extracting and analyzing these complex data sets. Our goal is to be able to identify responses of cells in an unbiased approach that will maximize our understanding of how gene and target perturbations affect signaling pathways.

12:30-1:00 High-Content Functional Genomics Screening to Enhance Target Confidence in Safety and Mechanism
To Be Announced
A limited understanding of genes’ mechanistic properties is a major factors contributing to the unacceptably high attrition rate observed for many drug targets. Current cell-based screening assays typically measure a discrete event and not the cause, as such a negative result does not distinguish between a target or compound driven response. Here we describe a molecular genetics screening approach incorporating siRNA duplexes as a tool to modulate the expression of molecules within “druggable” target space. The Cellomics ArrayScan platform is subsequently utilized to monitor and evaluate multiple functional endpoints. The integration of these phenotypic measurements with existing chemical and informatics data will enhance the understanding of target mechanism and secondary pharmacology enabling better decision making within a therapeutic program. 

1:00-2:30 Lunch with Exhibit and Poster Viewing (last chance to view)

2:30 Close of Main Conference

For more information please contact: 
Julia Boguslavsky, Conference Director, Cambridge Healthtech Institute
Phone: 781-972-5482 or E-mail: juliab@healthtech.com

For exhibit and sponsorship information, please contact:
Carol Dinerstein, Manager, Business Development
Phone: 781-972-5471 • E-mail: dinerstein@healthtech.com