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Thursday, February 20

Day 1 | Day 2 | Download Brochure 

7:30-8:15 am Breakfast Presentation (Sponsorship Opportunity Available) or Morning Coffee


3-D Cell Culture for Drug Discovery

8:30-8:35 Chairperson’s Opening Remarks

Thomas R. Broker, Ph.D., Professor, Biochemistry & Molecular Genetics, University of Alabama at Birmingham

8:35-9:00 The Third Dimension for High-Throughput Screening in Cancer Biology

Geoffrey A. Bartholomeusz, Ph.D., Assistant Professor and Director, siRNA Core Facility, Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center

Tumor microenvironments are complex 3-D microenvironments. 3-D multicellular tumor spheroid cell culture models demonstrate properties that show significant co-relation to tumors. The ease at which spheroid models can be applied in high-throughput screens has resulted in the realization of their importance to address relevant questions in tumor biology. We are developing 3-D spheroid cell culture models to be used in high-throughput screening. The design of one such model for target identification utilizing high-throughput RNAi screens will be discussed.

9:00-9:25 3-D Organotypic Epithelial Raft Cultures Provide an Authentic Environment for Antiviral Drug Testing

Thomas R. Broker, Ph.D., Professor, Biochemistry & Molecular Genetics, University of Alabama at Birmingham

Organotypic epithelial tissue cultures grown at the liquid medium-air interface stratify and differentiate into squamous epithelia that faithfully recapitulate the tissue source as well as possible disease state of the keratinocytes. The cells can be challenged with natural pathogens, or individual genes can be transfected and expressed in the cultures. The tissues can be harvested for microarray and molecular analyses or microscopically visualized for histology and key protein and nucleic acid biomarkers. Anti-viral drug studies in raft cultures are especially informative.

9:25-9:50 Microscale 3-D Cultures for Testing Anti-Cancer and Anti-Fibrosis Drug Candidates

Shuichi Takayama, Ph.D., Professor, Biomedical Engineering, University of Michigan

I will present technologies for cell-based assays in cancer and autoimmune disease/fibrosis. Devices include a microfluidic cell tri-culture model, a 384 hanging drop array for cancer spheroid formation, and a microscale fibroblast-collagen gel contraction assay enabled by aqueous two-phase system microdroplets. These platforms are more physiological in terms of chemokine-mediated chemotaxis, biochemical transport, and mechanical microenvironments. Initial biological validation of the platforms and drug testing results will be presented.

9:50-10:15 Q & A with the Speakers

10:15-11:10 Coffee Break in the Exhibit Hall with Poster Viewing


Stem Cell-Derived Cellular Models

11:10-11:35 Image-Based Screening Platform for Simultaneous Measurement of Calcium and Voltage Transients of Human Stem Cell-Derived Cardiomyocytes

Manuel Ruidiaz, Ph.D., Imaging and Analysis Scientist, CPCCG, Sanford-Burnham Medical Research Institute

Alex Savtchenko, Ph.D., Staff Scientist, Muscle Development and Regeneration Program, Sanford-Burnham Medical Research Institute

Stem cell research holds a promise of a break-through solution by providing “disease-in-a-dish” models of cardiac disorders. Typical cardiomyocyte-based assays have relied heavily on single-cell electrophysiology recordings, which are labor intensive and low-throughput. To enable medium-throughput screening applications, a high-content screening platform for simultaneous quantification of two biologically distinct parameters from the same cardiomyocyte, changes in cellular membrane potential and a dynamic distribution of intracellular calcium ions, has been developed. Images of fluorescent calcium and voltage dyes are simultaneously recorded on a single optical channel with 100 Hz frame rate in 384-well format using commercially available imaging equipment. Calcium and voltage signals were subsequently deconvolved into separate traces, thus eliminating the need for expensive multi-camera systems. Potential applications include cardio-toxic counter-assays and screening of cardio-active compounds on induced pluripotent stem cell derived cardiomyocytes (iPSC-CM).

11:35-12:00 pm New Insights Gained from Using Human iPSC-Derived Motor Neurons to Study Motor Neuron Diseases

Lee Rubin, Ph.D., Professor, Stem Cell & Regenerative Medicine, Harvard University; Director, Translational Medicine, HSCI

Recent advances in stem cell biology have made it possible to study human disease in a relatively convenient cell culture setting. My lab has investigated pathological aspects of two motor neuron disorders—spinal muscular atrophy and amyotrophic lateral sclerosis—by generating large numbers of motor neurons from multiple lines of individual patient-derived induced pluripotent stem cells (iPSCs). I will describe new insights that we have made using this approach.

12:00-1:30 Enjoy Lunch on Your Own

More Predictive Cellular Co-Culture Models

1:30-1:35 Chairperson’s Opening Remarks

Ray Mattingly, Ph.D., Professor, Pharmacology, Wayne State University

1:35-2:00 An Engineered Co-Culture System for NF1 (Type 1 Neurofibromatosis) Neurofibromas

Ray Mattingly, Ph.D., Professor, Pharmacology, Wayne State University

Neurofibromas include a driving, deranged Schwann cell component plus the peripheral nerve, fibroblasts, mast cells, endothelial cells, and secreted collagens and other matrix proteins. The complex mix of cell types is likely to: 1) promote the growth and phenotypic transformation of the tumors; and 2) present complications for their treatment. We are bioengineering 3-D models of neurofibromas that have an upper layer of immortalized neurofibroma cells, co-cultured with fibroblasts and endothelial cells, above a lower layer that incorporates a hyaluronic acid-based nanofiber scaffold to mimic the neuron. We propose that these engineered co-cultures will facilitate identification and screening of effective pharmaceutical approaches, alone and in combination, for the prevention, management and treatment of tumors in NF1 patients.

2:00-2:25 Screening the Micro-Colon: An HTS Platform to Identify Stromal Targets that Support Tumor Spheroid Formation

Shane Horman, Ph.D., Research Investigator, Genomics, Genomics Institute of the Novartis Research Foundation

An HTS co-culture colon mini tumor model consisting of normal colon stromal cells and colon cancer cells was created to screen for stromal genes that support tumor spheroid formation. 1,000 stromal genes were knocked down using a custom shRNA lentiviral library and their effects on colon tumor spheroid formation were quantitated using laser scanning fluorescence cytometry. Novel stromal targets that support tumor spheroid formation were revealed which may constitute attractive future drug discovery targets.

2:25-2:50 An Improved Glycosamino Glycan Clearance Assay Platform to Measure Relative Potency of Lysosomal Sulfatases

Vijay Chhajlani, Ph.D., Associate Director, Analytical Development, Shire

Glycosamino glycans (GAGs) are linear and variably sulfated oligosaccharide chains that serve as key components of the extracellular matrix and are also involved in cellular signaling in a wide range of processes. A pathogenic accumulation of GAGs in the lysosome in such cell type subsets as neurons and fibroblasts ultimately leads to impairment of cellular function and a wide range of clinical manifestations. An in vitro method for detecting intracellular GAGs takes advantage of normal proteoglycan synthesis and is used to assess replacement enzyme potency in primary patient fibroblasts. This method allows the determination of sulfatase activity on physiologically relevant substrate in a cellular model.

2:50-3:50 Refreshment Break in the Exhibit Hall with Poster Viewing


High-Content Analysis of Live Cells and Tissues

Chairperson's Opening Remarks

David Nolte, Ph.D., Professor, Physics, Purdue University; President, Animated Dynamics, Inc.

3:50-4:15 Biodynamic Imaging for Phenotypic Profiling of Three-Dimensional Tissue

David Nolte, Ph.D., Professor, Physics, Purdue University; President, Animated Dynamics, Inc.

Biodynamic imaging (BDI) delivers better drug assessment in three-dimensional (3-D) living tissue samples, revealing a comprehensive phenotypic response that is more physiologically meaningful than from conventional 2-D or monolayer screens. The technique is label-free and probes up to 1 mm deep in living tissue to study drug response in heterogeneous tissue environments. Biodynamic imaging converts all forms of functional cellular motions into contrast maps of cellular mechanisms of action. We present the first comparison of biodynamic imaging against morphological image analysis of two-dimensional cell culture, and find a strong correlation that enables a projection approach to translate between HCA and biodynamic feature spaces. There are significant 2-D versus 3-D phenotypic differences exhibited by 25% of the drugs/cell-lines which could relate to therapeutic efficacy and toxicity in 3-D that would be missed by 2-D screens.

4:15-4:40 Imaging and Quantifying the DNA Damage Response in Live 3-D Mammary Acini

Sylvain Costes, Ph.D., Principal Investigator, Cancer and DNA Damage Response, Lawrence Berkeley National Laboratory

4:40-5:05 Live-Cell Imaging of Tandem Fluorescent-LC3 to Measure Autophagic Flux

Christopher Hale, Ph.D., Scientist, Discovery Technologies, Amgen

Upregulation of autophagic flux has been proposed as a therapeutic strategy for alleviating dysfunction in neurodegenerative disease. To identify genes involved in autophagy, we carried out a live-cell, image-based, high-content siRNA screen using U-2 OS cells stably transfected with RFP-GFP-LC3, or tandem fluorescent LC3 (tfLC3). Cytosolic RFP-GFP-LC3 is lipidated and translocated to an autophagosomal isolation membrane; once an autophagosome has fused with an acidic lysosome, the GFP signal is quenched due to its instability at low pH, leaving a GFP- RFP+ autolysosome. We developed an analysis algorithm to quantitate autophagosomes and autolysosomes per cell over time to identify genes that both increased and decreased autophagic flux.

5:05-5:30 Cellular Biochemistry and Multiplexing for Live-Cell, High-Content Studies of Prostate Cancer

Fred Schaufele, Ph.D., Associate Professor, Center for Reproductive Sciences, University of California San Francisco

Screening strategies depend on the definition of the target activity, which in many instances is altered by the cellular context in which the target protein finds itself. An androgen receptor function that drives treatment-resistant prostate cancer was found to be disturbed specifically in treatment-resistant cells. We describe screens to identify newer interventions that are conducted in treatment-resistant cell lines that phenocopy the disease.

5:30 Close of Conference

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