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Human induced pluripotent stem cell derived tissue cells as a relevant model for phenotypic screening
Blake Anson, Product Manager, Cellular Dynamics International
Human induced pluripotent stem cell (hiPSC)-derived tissue cells offer limitless quantities of highly pure and functional terminal cell types for a variety of basic and applied research efforts. By providing unprecedented access to human biology in easy to handle industrial quantities, hiPSC-tissue cells uniquely enable large-scale investigations and make phenotypic screening with functionally relevant human-sourced material a reality. Cellular Dynamics International has developed a robust manufacturing process that produces pure human tissue cells at rates greater than 0.5 to 1x109/day to enable large-scale screens. This presentation will focus on the industrialization and use of iCell® Cardiomyocytes and Neurons. Their potential for use as a biologically relevant source material will be established by presenting data demonstrating the functional recapitulation of in-vivo physiology. Their utility in phenotypic screening paradigms will be will be demonstrated with data showing how these tissue cells are being engineered to express relevant reporters as well as how they are being used on a variety of higher throughput screening platforms to screen against endogenous cellular processes.
Label-free phenotypic screening using real-time cellular analysis
Jeffrey Irelan, Director, ACEA Biosciences
The xCELLigence Real-Time Cellular Analysis (RTCA) System is a label-free platform for dynamic monitoring of cellular phenotypic responses. Cells are seeded onto plates containing biosensors that allow for precise measurement of changes in electrical impedance correlating to changes in cell morphology, attachment, and proliferation. Most disease-relevant pathways couple to one or more of these phenotypes, thus allowing for evaluation of endogenously expressed targets in a therapeutically relevant cellular context. The 384-well RTCA HT System was evaluated in screens for modulators of receptor tyrosine kinase mediated signaling as well as G-coupled protein receptors. First, a small scale proof of concept “duplex” screen (for both agonists and antagonists) was performed on cells expressing, a Gi – coupled GPCR in either the endogenous or recombinant expression context, resulting in identification of both known and novel modulators of this receptor. “Oncogene addiction” is the dependence of cancer cells on a single oncogenic pathway (often driven by aberrant RTK function) for sustained proliferation and/or survival. A large scale screen using RTCA on an addicted cell line was performed, leading to identification and development of a novel inhibitor of the “driver” RTK. The benefits and challenges of phenotypic screening on the RTCA system will be discussed.
Random Homozygous Gene Perturbation (RHGP) as a Tool for Drug Target Discovery and Validation
Wu-Bo Li, Director, Functional Genetics, Inc.
Random homozygous gene perturbation (RHGP) can identify any host gene that causes a desired phenotype. The central feature of RHGP is a gene search vector (GSV) designed to interrogate the entire genome and identify target genes that cause the phenotype of interest. The GSV that contains an inducible promoter is transduced into a target cell population. Transduction results in random integrations of the GSV into the genome of the target cells. In the presence of inducer, the GSV promoter is activated and triggers the transcription of the RNA extending into the host genome sequence flanking the GSV. RHGP can sample every gene in a cell for both over expression and loss of expression and provides an efficient, robust and high-throughput means to improve target discovery using a function first approach. With the RHGP approach, Functional Genetics has identified the host target genes involved in pathogenesis of cancer metastasis and drug resistance, Alzheimer’s disease, and viral infections. The first infectious disease therapeutic developed with RHGP technology is now in clinic trial. Two additional infectious disease therapeutics will be entering preclinical development shortly.
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