Droplet-based forward genetic screening of astrocyte-microglia cross-talk

• 成果类型: Article
• 署名作者: Wheeler, Michael A.; Clark, Iain C.; Lee, Hong-Gyun; Li, Zhaorong; Linnerbauer, Mathias; Rone, Joseph M.; Blain, Manon; Akl, Camilo Faust; Piester, Gavin; Giovannoni, Federico; Charabati, Marc; Lee, Joon-Hyuk; Kye, Yoon-Chul; Choi, Joshua; Sanmarco, Liliana M.; Srun, Lena; Chung, Elizabeth N.; Flausino, Lucas E.; Andersen, Brian M.; Rothhammer, Veit; Yano, Hiroshi; Illouz, Tomer; Zandee, Stephanie E. J.; Daniel, Carolin; Artis, David; Prinz, Marco; Abate, Adam R.; Kuchroo, Vijay K.; Antel, Jack P.; Prat, Alexandre; Quintana, Francisco J.
• 署名单位: Harvard University; Harvard University Medical Affiliates; Brigham & Women's Hospital; Harvard Medical School; Harvard University; Massachusetts Institute of Technology (MIT); Broad Institute; University of California System; University of California Berkeley; McGill University; University of Rochester; Harvard University; Harvard Medical School; Harvard University; Harvard University Medical Affiliates; Brigham & Women's Hospital; Harvard University; Harvard Medical School; University of Erlangen Nuremberg; Cornell University; Universite de Montreal; Helmholtz Association; Helmholtz-Center Munich - German Research Center for Environmental Health; University of Munich; Cornell University; Weill Cornell Medicine; University of Freiburg; University of Freiburg; University of Freiburg; University of Freiburg; University of California System; University of California San Francisco; Chan Zuckerberg Initiative (CZI)
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-10081
• DOI: 10.1126/science.abq4822
• 发表日期: 2023-03-10
• 页码: 1023-1030
• 关键词: regulatory t-cells; in-vivo; il-33 promotes; inflammation; SYSTEM; amphiregulin; macrophages; dna

摘要

Cell-cell interactions in the central nervous system play important roles in neurologic diseases. However, little is known about the specific molecular pathways involved, and methods for their systematic identification are limited. Here, we developed a forward genetic screening platform that combines CRISPR-Cas9 perturbations, cell coculture in picoliter droplets, and microfluidic-based fluorescence-activated droplet sorting to identify mechanisms of cell-cell communication. We used SPEAC-seq (systematic perturbation of encapsulated associated cells followed by sequencing), in combination with in vivo genetic perturbations, to identify microglia-produced amphiregulin as a suppressor of disease-promoting astrocyte responses in multiple sclerosis preclinical models and clinical samples. Thus, SPEAC-seq enables the high-throughput systematic identification of cell-cell communication mechanisms.