Identification of astrocyte regulators by nucleic acid cytometry.
Identification of astrocyte regulators by nucleic acid cytometry.
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DOI：
10.1038/s41586-022-05613-0
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发表时间：
2023-03
期刊：
影响因子：
64.8
通讯作者：
Abate, Adam R. R.
中科院分区：
文献类型：
--
作者：
Clark, Iain C. C.;Wheeler, Michael A. A.;Lee, Hong-Gyun;Li, Zhaorong;Sanmarco, Liliana M. M.;Thaploo, Shravan;Polonio, Carolina M. M.;Shin, Seung Won;Scalisi, Giulia;Henry, Amy R. R.;Rone, Joseph M. M.;Giovannoni, Federico;Charabati, Marc;Akl, Camilo Faust;Aleman, Dulce M. M.;Zandee, Stephanie E. J.;Prat, Alexandre;Douek, Daniel C. C.;Boritz, Eli A. A.;Quintana, Francisco J. J.;Abate, Adam R. R.
多发性硬化症是一种慢性中枢神经系统炎症性疾病。星形胶质细胞是一种存在于中枢神经系统的异质胶质细胞，参与多发性硬化及其实验性自身免疫性脑脊髓炎的发病机制。然而，很少有独特的表面标记可用于分离星形胶质细胞亚群，阻止其分析和确定候选治疗靶点；致病星形胶质细胞的稀有性进一步放大了这些局限性。在这里，为了应对这些挑战，我们开发了通过核酸检测和测序(FIND-SEQ)对细胞进行重点询问，这是一种高通量微流控细胞术方法，它结合了液滴中细胞的包裹、基于PCR的目标核酸检测和液滴分类，从而能够在单细胞分辨率下对感兴趣的细胞进行深入的转录分析。我们应用FIND-SEQ来研究星形胶质细胞的调控，其特征是转录因子XBP1的剪接驱动激活，该转录因子促进多发性硬化症和实验性自身免疫性脑脊髓炎的疾病病理。使用Find-Seq结合条件基因敲除小鼠，体内CRISPR-Cas9驱动的遗传扰动研究，以及小鼠实验性自身免疫性脑脊髓炎和人类多发性硬化症样本的批量和单细胞RNA测序分析，我们确定了核受体NR3C2及其辅阻遏子NCOR2在限制XBP1驱动的致病星形胶质细胞反应中的新作用。综上所述，我们使用FIND-SEQ来确定限制XBP1驱动的致病星形胶质细胞反应的治疗靶向机制。Find-Seq能够研究以前无法接触到的细胞，包括由独特的基因表达特征或其他核酸标记定义的稀有细胞亚群。
Multiple sclerosis is a chronic inflammatory disease of the central nervous system. Astrocytes are heterogeneous glial cells that are resident in the central nervous system and participate in the pathogenesis of multiple sclerosis and its model experimental autoimmune encephalomyelitis. However, few unique surface markers are available for the isolation of astrocyte subsets, preventing their analysis and the identification of candidate therapeutic targets; these limitations are further amplified by the rarity of pathogenic astrocytes. Here, to address these challenges, we developed focused interrogation of cells by nucleic acid detection and sequencing (FIND-seq), a high-throughput microfluidic cytometry method that combines encapsulation of cells in droplets, PCR-based detection of target nucleic acids and droplet sorting to enable in-depth transcriptomic analyses of cells of interest at single-cell resolution. We applied FIND-seq to study the regulation of astrocytes characterized by the splicing-driven activation of the transcription factor XBP1, which promotes disease pathology in multiple sclerosis and experimental autoimmune encephalomyelitis. Using FIND-seq in combination with conditional-knockout mice, in vivo CRISPR–Cas9-driven genetic perturbation studies and bulk and single-cell RNA sequencing analyses of samples from mouse experimental autoimmune encephalomyelitis and humans with multiple sclerosis, we identified a new role for the nuclear receptor NR3C2 and its corepressor NCOR2 in limiting XBP1-driven pathogenic astrocyte responses. In summary, we used FIND-seq to identify a therapeutically targetable mechanism that limits XBP1-driven pathogenic astrocyte responses. FIND-seq enables the investigation of previously inaccessible cells, including rare cell subsets defined by unique gene expression signatures or other nucleic acid markers.
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影响因子：
25
作者：
Habib, Naomi;McCabe, Cristin;Medina, Sedi;Varshavsky, Miriam;Kitsberg, Daniel;Dvir-Szternfeld, Raz;Green, Gilad;Dionne, Danielle;Nguyen, Lan;Marshall, Jamie L.;Chen, Fei;Zhang, Feng;Kaplan, Tommy;Regev, Aviv;Schwartz, Michal
通讯作者：
Schwartz, Michal
影响因子：
25
作者：
Hasel, Philip;Rose, Indigo V. L.;Liddelow, Shane A.
通讯作者：
Liddelow, Shane A.
影响因子：
64.8
作者：
Calfon, M;Zeng, HQ;Ron, D
通讯作者：
Ron, D
影响因子：
30.5
作者：
Hiltensperger M;Beltrán E;Kant R;Tyystjärvi S;Lepennetier G;Domínguez Moreno H;Bauer IJ;Grassmann S;Jarosch S;Schober K;Buchholz VR;Kenet S;Gasperi C;Öllinger R;Rad R;Muschaweckh A;Sie C;Aly L;Knier B;Garg G;Afzali AM;Gerdes LA;Kümpfel T;Franzenburg S;Kawakami N;Hemmer B;Busch DH;Misgeld T;Dornmair K;Korn T
通讯作者：
Korn T
星形胶质细胞疤痕形成有助于中枢神经系统轴突再生。
影响因子：
64.8
作者：
Anderson MA;Burda JE;Ren Y;Ao Y;O'Shea TM;Kawaguchi R;Coppola G;Khakh BS;Deming TJ;Sofroniew MV
通讯作者：
Sofroniew MV