SLAS2012 Live Streaming Brings Worlds Together
SLAS is pleased to offer the exclusive opportunity to participate in SLAS2012 via live streaming of two select sessions — completely free. These two sessions allow the global scientific community to come together in person and online through live video webcasts and real-time networking. In addition, for those participating via live streaming the sessions deliver all the benefits of a physical presence no matter your location. You will be able to ask questions and get direct answers from the presenters and panelists who are live at SLAS2012 in San Diego.
To participate via live streaming, simply go to SLAS2012.org to log-in at the times noted below for each session. There is no need to pre-register. Live streaming is delivered in an Adobe Flash format and is not compatible with iPhone or iPad devices. Both sessions will be available on-demand for a limited time following SLAS2012.
Live Streaming Now Available On-Demand
Monday, February 6, 2012
10:30 am - 12:30 pm U.S. Pacific Standard Time
Bridging the Valley of Death; How Can Academia and Pharma Best Work Together?
Moderator: Derek Lowe, In the Pipeline
Panelists: John Luk, National University of Singapore; Rudy Juliano, University of North Carolina; Mao Mao, Pfizer; Alan Palkowitz, Eli Lilly and Company; John Reed, Sanford Burnham Medical Research Institute
This session brings together opinion leaders from academic drug discovery and pharmaceutical companies to examine how the recent increase in academic drug discovery impacts drug discovery and models for these interactions are discussed. The discussion is a moderated panel style with questions gathered from the SLAS membership through social media sites, expert opinions and audience participation.
Monday, February 6, 2012
3 - 5 pm U.S. Pacific Standard Time
Advances in High-Throughput Screening Technologies
Moving HCS Into HTS... The Journey of Moving From 384 to 1536-Well Automated High Content Assays
Debra Nickischer, Bristol-Myers Squibb Company
Co-Authors: Lisa Elkin, Gerald Duke, Dan Meyers, Parker Rex, Andrea Westin, Bristol-Myers Squibb Company
Within the Drug Discovery industry, and certainly within BMS, there has been a growing recognition of the value of high-content screening, as researchers aim to screen compounds and identify hits using assays that more closely mimic the relevant physiological environment. While HCS has been in routine use by many for a while now, a number of hurdles have historically prohibited very large high-throughput screening efforts with this platform. The need for suitable cell washing capabilities and HCS readers that are ammenable to 1536-well assays, a reliable informatics framework to accommodate the scale of HCS data, and novel approaches to data analysis represent only a few of the many considerations for implementing high-throughput, high-content screening. Despite these hurdles, within BMS, there has been a growing interest in screening large compound inventories using this platform, often against targets have been prosecuted using more traditional HTS approaches in the past with no tractable leads. Focusing on a target that has been illusive and challenging for the cardiovascular program at BMS, the relative challenges and strengths of high-content screening observed at BMS will be outlined, as the design and screening of a phenotypic assay for this target is described alongside the hit assessment strategy to identify and prioritize hits.
High Content Mass Cytometry Screening to Study Single Cell Signaling Networks in Biology and Disease
Bernd Bodenmiller, Stanford University School of Medicine
Co-Authors: Eli Zunder, Rachel Finck, Robert Bruggner, Erica Savig, Erin Simonds, Sean Bendall, Tiffany Chen, Karen Sachs, Garry Nolan, Peter Krutzik, Stanford University
Cancer cells and their signaling networks are highly heterogeneous, thereby driving and maintaining the disease. The structure of these networks and their change during disease or therapy is often unknown. Therefore the analyses of the signaling networks between cell subsets and states and how these respond to drug treatment and other perturbations at the single-cell is highly desirable.
Here a novel high throughput workflow based on a next-generation single cell mass cytometry instrument is presented. Mass cytometry allows quantifying up to 100 molecules on the single-cell level, thereby the signaling and cellular state can be measured within accurately defining cell types and subpopulations. To add high-throughput capacity to mass cytometry, a cell-based multiplexing technique was developed, called mass-tag cellular barcoding (MCB). Here each cell sample is labeled with a unique combination of mass-tags, mixed with other samples before antibody staining, and then deconvoluted after measurement. This strategy now allows measuring thousands of samples per day, strongly reduces antibody costs and greatly increases the quality of the data due to the homogenous cell labeling. Potential applications for high-content MCB analysis range from drug and RNAi screens to biological discovery. To illustrate the power of the approach for screening applications, the effects of kinase inhibitors on peripheral blood mononuclear cells signaling networks was comprehensively profiled. Each inhibitor was tested at eight dilutions against 12 conditions and levels of 14 intracellular signaling molecules in 14 cell types were evaluated, resulting in 18,816 individual quantified cell populations from a single tube of multiplexed cells. These analyses revealed complex signaling network responses and correlations, enabling to classify each cell population and inhibitor with unprecedented accuracy.
Two High Content Screening Approaches for the Rare Neurological Disease Amyotrophic Lateral Sclerosis
Marcie Glicksman, Brigham and Women
Neurodegenerative diseases are challenging from a drug discovery perspective with virtually no disease modifying agents available on the market. Amyotrophic Lateral Sclerosis (ALS, Lou Gehrig's disease) is a rare neurodegenerative disease. From the time of diagnosis to death is usually 3-5 years. So far, there are six genes identified associated with ALS. The genetic causes of ALS can provide a number of clues to the mechanism of the disease as well as good potential therapeutic targets. We have done two high throughput high content screening assays to screen our compound library for modulators of two genes associated with ALS. The genes TDP-43 (TAR DNA-binding protein 43) and Fus/TLS (FUsed in Ewing's Sarcoma/ Translated in LipoSarcoma) were identified in a genome-wide screen among autosomal dominant ALS patients and are both DNA/RNA binding proteins. They are found to aggregate into stress granules that are believed to be toxic to cells. We developed an assay for TDP-43 measuring aggregate formation in cells and an assay for Fus/TLS measuring translocation of tagged protein between the nucleus and cytoplasm. Multiple stages are required for developing an HCS assay. The first stage of assay development is to quantify the phenotypic changes and develop an accurate analysis method. The middle stage of assay development is to establish robustness and reproducibility in the assay. The last stage is to establish scalability of the assay. The unique features for developing these two high content imaging assays will be discussed. Highlights will include the challenges and the benefits of this type of assay.
Can Peptide Substrates Properly Represent Physiological Substrates in Drug Discovery?
Min Liu, LDDN
Leucine-rich repeat kinase2 (LRRK2), a large and complex protein that possesses two enzymatic properties, kinase and GTPase, is one of the major genetic factors in Parkinsonâ€™s disease (PD). Over 40 mutations in LRRK2 have been found in the most common familial forms and some sporadic forms of PD, and have been associated with typical idiopathic, late-onset PD. Some LRRK2 mutations have increased kinase activity, which correlates with increased neuronal cytotoxicity. Accumulated evidence has suggested that kinase activity of LRRK2 plays a critical role in the pathogenesis of PD Therefore, identification of LRRK2 kinase inhibitors has become priority in drug discovery for the treatment of PD. Due to the unknown physiological substrates of LRRK2, a peptide substrate LRRKtide has been widely used for characterizing LRRK2 inhibitors. However, our recent enzyme kinetic studies using structurally related kinase of LRRK2, B-Raf, strongly suggest that peptide and protein substrates of B-Raf bind on distinct binding sites and inhibitors behave dramatically different toward the peptide and physiological protein substrates. B-Raf, which has 33% sequence identity compared to LRRK2, can catalyze the same LRRKtide phosphorylation as LRRK2 does, suggesting that the two kinases have very similar active site. However, the actions of B-Raf toward its physiological substrate MEK1 and LRRKtide are kinetically dramatically different — Km of ATP differs by 2000-fold (0.1 mM in MEK1 phosphorylation and 200 mM in LRRKtide phosphorylation). The kinetic difference was also reflected in inhibitor potency. The well characterized potent B-Raf inhibitors completely lost their potency toward LRRKtide phosphorylation catalyzed by B-Raf. The significant changes in ATP and inhibitor binding affinity were explained by the distinct binding site of LRRKtide on B-Raf, a site different from the MEK1 binding site, as evidenced by the alternative substrate studies. The fact that LRRK2 shares high structure similarity with B-Raf raises the questionsâ€”whether the potency of LRRK2 inhibitors determined by using peptide substrate can truly represent the potency in vivo and whether the information gathered using peptide substrate is proper to steer drug discovery efforts. It has long been believed that peptide substrates behave similarly to the physiological protein substrates. Investigators have more often than not chosen peptides as the substrate, this choice being motivated by ease of assay design and the ability to design peptide substrate with a single phosphorylation site. These results will not only change the way in which many view protein and peptide substrates in drug development, but also highly impact the field.