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Friday, October 4 • 11:30am - 11:50am
Poster Presentations in Exhibition- #2 Automated Analysis Of Sarcomere Organization In IPSC Cardiomyocytes

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Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs), in combination with CRISPR genome editing, now offer unparalleled opportunities to study cardiac biology and disease. Mature and functional iPSC-CMs are frequently characterized by the presence of visually distinct sarcomere bands. However, few methods exist to comprehensively quantify sarcomere organization. These methods can be relatively low-throughput (requiring manual selection for regions of interest), employ Fast Fourier Transforms (requiring a minimum number of detected sarcomeres), and/or only assess well-aligned sarcomeres from mature cardiac tissues. Here we take advantage of a pixel-based image analysis method to overcome these limitations and evaluate iPSC-CM sarcomere structure. Our approach, or sarcomere organization analysis (SOA), incorporates automated cell segmentation and extracts sarcomere information from Haralick texture features. This quantification enables SOA to process any geometric shape, account for noise, and identify sarcomere persistence. These are especially important features since iPSC-CM cell shape is quite variable and sarcomere orientations and lengths can be heterogeneous. Thus, SOA allowed us to effectively distinguish levels of sarcomere organization to define CMs with highly and moderately organized sarcomeres compared to CMs with visibly disorganized sarcomeres. These differences were further apparent when comparing WT iPSC-CMs to iPSC-CMs with a genetic sarcomere mutation. This mutation resulted in more CMs with sarcomeres longer than 2 µm and a reduced organization. Our SOA method is intended to help provide a more systematic approach to examining cardiac disease biology resulting from genetic variants. Current work is aimed at incorporating deep learning for enhanced analysis and a more complete description of sarcomere organization and iPSC-CM phenotypes.

Speakers
DW

Devin Wakefield, Ph.D.

Sr. Associate Scientist, Amgen
*Sr Associate Scientist, Amgen: 2018-present*Genome Analysis Unit imaging research*Postdoctoral Fellow, Beckman Research Institute at City of Hope: 2015-2018*Super-resolution microscopy of cancer targets*Graduate Student, Cornell University: 2010-2015*IgE receptor-mediated signaling... Read More →


Friday October 4, 2019 11:30am - 11:50am
Washington Ballroom (5th Fl) - Courtyard Boston Downtown 275 Tremont Street, Boston, MA 02116