Multimodal spatiotemporal phenotyping of human retinal organoid development
Philipp Wahle, Giovanna Brancati, Christoph Harmel, Zhisong He, Gabriele Gut, Jacobo Sarabia del Castillo, Aline Xavier da Silveira dos Santos, Qianhui Yu, Pascal Noser, Jonas Simon Fleck, Bruno Gjeta, Dinko Pavlinić, Simone Picelli, M. W. Hess, Gregor W. Schmidt, Tom T. A. Lummen, Yanyan Hou, Patricia Galliker, David Goldblum, Márton Balogh, Cameron S. Cowan, Hendrik P. N. Scholl, Botond Roska, Magdalena Renner, Lucas Pelkmans, Barbara Treutlein, J. Gray Camp
Abstract
Organoids generated from human pluripotent stem cells provide experimental systems to study development and disease, but quantitative measurements across different spatial scales and molecular modalities are lacking. In this study, we generated multiplexed protein maps over a retinal organoid time course and primary adult human retinal tissue. We developed a toolkit to visualize progenitor and neuron location, the spatial arrangements of extracellular and subcellular components and global patterning in each organoid and primary tissue. In addition, we generated a single-cell transcriptome and chromatin accessibility timecourse dataset and inferred a gene regulatory network underlying organoid development. We integrated genomic data with spatially segmented nuclei into a multimodal atlas to explore organoid patterning and retinal ganglion cell (RGC) spatial neighborhoods, highlighting pathways involved in RGC cell death and showing that mosaic genetic perturbations in retinal organoids provide insight into cell fate regulation.