Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
Science. 2023 Mar 10;379(6636):1023-1030. doi: 10.1126/science.abq4822. Epub 2023 Mar 9.
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.
中枢神经系统中的细胞间相互作用在神经疾病中起着重要作用。然而，目前对于涉及的具体分子途径知之甚少，而且系统识别这些途径的方法也很有限。在这里，我们开发了一种正向遗传筛选平台，该平台结合了 CRISPR-Cas9 干扰、皮升级微滴中的细胞共培养和基于微流控的荧光激活微滴分选，以鉴定细胞间通讯的机制。我们使用 SPEAC-seq（包裹相关细胞的系统扰动，然后进行测序），结合体内遗传干扰，鉴定出小胶质细胞产生的 Amphiregulin 作为多发性硬化症临床前模型和临床样本中促进疾病的星形胶质细胞反应的抑制剂。因此，SPEAC-seq 能够高通量系统地鉴定细胞间通讯机制。
