DAY 2

Thursday, January 27

8:30-9:00   Reports from ThinkTank Sessions

Data Interpretation

9:00-9:05   Chairperson’s Opening Remarks
Ms. Ann Hoffman, Senior Principal Scientist, Roche Discovery Technologies, Hoffmann-La Roche, Inc.

9:05-9:20    Solutions Applied to Manage, Analyze and Archive the TeraBytes of High-Content Screening Data: The End Users Perspective
Mr. Leon S. Garfinkel, Director, Informatics & Infrastructure, Preclinical Discovery Research, Hoffmann-La Roche, Inc.
The expansion of HCS as an automated, multifaceted technology continues to produce results that add value to comprehensive databases. Reliance on these proprietary databases to provide pertinent knowledge to query compound fingerprints is vital in the decision making process of turning compounds into drugs. Here we describe variations on the management of high-content screening (HCS) data that allows simple workflows and user interfaces for the scientists with cost-effective documentation and archiving that remains expandable for administration by the information technologists. Using a tiered storage solution implemented and co-developed by Roche with Cellomics and EMC Corporation, we compared our data handling abilities with the commonly used basic but large file share structure. The evaluation included assessing the work flow regarding data acquisition, analysis, and processing of HCS high throughput and secondary screening data. This presentation will highlight the advantages of both approaches which are currently being utilized. One conclusion of these experiences is that Enterprise solutions are effectively supporting the HCS initiative and in a cost effective manner as well as making the best use of the technology available.

9:20-9:35   Managing Terabytes of Data in an Open Concept
Dr. Miroslav Cik, Principal Scientist, Department of AD&HTS, Johnson & Johnson Pharmaceutical Research & Development
We combined a high-speed imaging instrument, called MIAS-2, with analysis software packages from different manufacturers. To allow such combinations, and to tackle the resulting bottlenecks of terabytes of data, the MIAS-2 reader was connected with an image database from Nugenesis. This open integration concept of image acquisition, analysis and storage allows the use of HCS at its full capacity.

9:35-9:50   Building a Systematics for Protein Subcellular Location using Automated Microscopy
Dr. Robert F. Murphy, Professor of Biological Sciences and Biomedical Engineering; and Director, Merck Computational Biology and Chemistry Program, Carnegie Mellon University
Drug screens using automated microscopy are usually based on major changes in the distribution of a reporter, and comparison of patterns between screens is not common. My group has been developing methods for building a informatics framework in which proteins can be organized by their location patterns and by how they change under different conditions. The images from a single drug screen can be seen as providing untapped, deep information on the patterns displayed by a single target, and there is a need for approaches that can combine such information with shallow information on large numbers of proteins. We have developed and characterized many features for describing subcellular patterns, with a specific focus on making these as independent as possible of cell type and microscopy details. We have shown that these can be used to build systems for classifying proteins by location and for identifying subtle changes in protein patterns. These have been incorporated into the Protein Subcellular Location Image Database to facilitate post-acquisition, cross platform, cross cell type analysis of protein patterns.

9:50-10:05   Discovery of a Cell-Active Inhibitor of Mitogen-Activated Protein Kinase Phosphatase-1 (MKP-1) by a High-Content Screen and an Unbiased Statistical Approach to Hit Selection
Dr. Andreas Vogt, Research Assistant Professor, Department of Pharmacology; and Associate Director, Fiske Drug Discovery Laboratory, University of Pittsburgh School of Medicine 
High-content screening (HCS), when performed on defined cellular systems, can be a powerful method to definitively identify agents that interact with their intended targets in the context of the whole cell. Because of the complexity and size of HCS data sets, methods to analyze HCS data need to balance the desire to fully exploit rich information content against the need to reduce the data to a manageable and meaningful size. We present here the development of a novel analysis tool to select positives from an HCS data set. The method is based on an unbiased, simultaneous, quantitative comparison of the cumulative distribution functions (CDF) of two fluorescence parameters in individual cell subpopulations within the same well of a 384 well microplate. Application of the algorithm to an HCS data set from a chemical complementation screen using a mixed cell population of genetically modified and unmodified cells increased signal-to-noise ratio by 8-fold and reduced the number of false positives by 70% compared to our previously published method (Vogt et al., Chem. Biol. 10, 733-742, 2003). The method, while simplifying the data, exploited all of the available information contained in the high-content data set. The screen identified a potent and selective inhibitor of mitogen-activated protein kinase phosphatase-1 with cellular activity. 

10:05-10:20 Visualizing Multi-Dimensional Data Associated with Imaging-Based Systems Biology
Dr. James G. Evans, CSBi Research Scientist, Whitehead MIT BioImaging Center
This talk will discuss approaches used to visualize a diverse range of image data and the multi-dimensional arrays of associated meta-data. The types of image data will range from vast numbers of multi-channel 2-D images to time-resolved 3-D image data to multi-terabyte high-resolution tissue volumes. Methods employed to visualize associations between large numbers of meta-data parameters and for visualizing trends in dynamic data sets will also be described.

Phenotypic Targets in HCS With image-based screening, function- or phenotype-based screening is now possible. Here we will explore some different types of  morphological measurements and what targets they may be useful for. CO-ORGANIZED WITH     1. Multi-Cellular Organization Multi-cellular organization is a critical component of functions such as angiogenesis, metastasis, and all types of development. Measurements of the degree of association of cells into structures such as tubes and colonies is applicable to studying cell scattering, tube formation, stem cell differentiation, and foci formation. Correlation of expression and signaling in cells with these downstream functional events can provide insights into new drug targets and understanding of structure-function relationships. 2. Proximity Among Cell Populations The association of cells in a culture and arrangement of similar and dissimilar cell populations has profound implications in processes such as cell signaling, chemotaxis, immune response, and muscle cell migration. The average and minimum distance between a cell and its neighbors can be automatically determined for cells of the same type or across types. The ability to screen for compounds involved with cell scattering, gap junction formation, and contact activation could lead to identification of novel drug targets and classes. 3. Texture Changes Proteins and other cellular components moving within a cell can often be identified by analyzing their distribution pattern, capturing more subtle changes than would be measurable by identifying discreet objects. With these types of measurements one could determine whether a target of interest is distributed evenly throughout the cytoplasm, is somewhat punctate, or has a radial distribution (concentration gradient from the center to the edge of the cell). Such measurements could be made to understand functional targets including, but not limited to, cytoskeletal rearrangement, receptor internalization, nuclear structure, and focal adhesions or podosomes. 4. Shape Changes Measurements of an object’s overall shape, presence of processes or outgrowths, perimeter smoothness, and orientation can establish a screenable phenotype. Examples of biological applications of these types of phenotypic changes include: neurite outgrowth, cytotoxicity (rounding of cells), elongation of cells prior to tube formation, and nuclear morphology changes due to normal division or apoptosis. Discussion Leader:  Dr. Sarah Burroughs Tencza, Product Manager, BioApplications, Cellomics, Inc. Panelists: Dr. James G. Evans, CSBi Research Scientist, Whitehead MIT BioImaging Center Dr. Myles Fennell, Principal Research Scientist I, Neuroscience, Wyeth Research Dr. Richik Ghosh, Director of Assay Feasibility, Cellomics, Inc. Dr. O. Joseph Trask, Jr., Associate Senior Biochemist, Eli Lilly and Co.

1:00-2:30   Luncheon in the Exhibit Hall (last chance for exhibit and poster viewing)

2:00-7:00   Concurrent User Group Meetings
Cellomics User Group Meeting	IN Cell Analyzer Platform Technology
This user group meeting will be a forum for you to interact with other scientist who are using HCS as a tool for furthering their discovery efforts. Submit an abstract to present what your laboratory is doing with HCS, or attend to learn how your colleagues are optimizing their HCS experience. We will have oral and poster presentations by leading scientists in the industry. All Cellomics’ customers are welcome to attend. For more information on this event or to register to attend, please contact us at usergroup@cellomics.com or visit our User Group Web site at http:\\usergroup.cellomics.com. IN Cell Analyzer Platform Technology Day	The IN Cell Analyzer Platform Technology Day will be useful for both current users of IN Cell Analyzer instruments & reagents, and for those interested in learning more about this developing technology. During this session current IN Cell users will provide the insights on use of IN Cell Analyzers for HCA. We will also provide information on newly launched products and will give some insights on products that are in development. You will also get an opportunity to talk to GE healthcare experts on HCA. For more information, please contact your local GE Healthcare representative. Register now for this workshop!
Don’t miss the chance to co-locate your company’s user group meeting with High-Content Analysis 2005! These opportunities are limited.  Please contact Carol Dinerstein at 781-972-5471 or dinerstein@healthtech.com.