Efficient generation of lower induced motor neurons by coupling Ngn2 expression with developmental cues.
Efficient generation of lower induced motor neurons by coupling Ngn2 expression with developmental cues.
复制标题
DOI：
10.1016/j.celrep.2022.111896
复制
发表时间：
2023-01-31
期刊：
影响因子：
8.8
通讯作者：
中科院分区：
文献类型：
--
作者：
人类多能干细胞（hPSC）是一种强大的工具，用于难以接近的组织（如大脑）的疾病建模。目前的方案要么用小分子指导神经元分化，要么使用转录因子介导的编程。在这项研究中，我们将转录因子Neurogenin 2（Ngn 2）的过表达与小分子图案化相结合，以将hPSC分化为低诱导运动神经元（liMoNes/liMN）。该方法在超过95%的细胞中诱导包括MN特异性Hb 9/MNX 1的典型MN标志物。limMN类似于真正的hPSC衍生的MN，表现出自发电活动，表达突触标记，并且可以在体外接触肌细胞。对50个hPSC系进行的合并、多重单细胞RNA测序揭示了宫颈和肱肌MN的不同亚型的可重复群体，这些亚型类似于它们在体内的胚胎对应物。将小分子模式化与Ngn 2过表达相结合有助于高产量、可重复地产生疾病相关的MN亚型，这是推动我们了解MN生物学及其在疾病中的破坏的基础。Limone等人通过小分子模式化和TF过表达诱导hPSC神经化为脊髓MN。多重、合并的单细胞RNA测序在数十种细胞系中显示出高重现性。这些MN村类似于体内脊髓MN，并产生疾病相关的MN人群。
Human pluripotent stem cells (hPSCs) are a powerful tool for disease modeling of hard-to-access tissues (such as the brain). Current protocols either direct neuronal differentiation with small molecules or use transcription-factor-mediated programming. In this study, we couple overexpression of transcription factor Neurogenin2 (Ngn2) with small molecule patterning to differentiate hPSCs into lower induced motor neurons (liMoNes/liMNs). This approach induces canonical MN markers including MN-specific Hb9/MNX1 in more than 95% of cells. liMNs resemble bona fide hPSC-derived MN, exhibit spontaneous electrical activity, express synaptic markers, and can contact muscle cells in vitro. Pooled, multiplexed single-cell RNA sequencing on 50 hPSC lines reveals reproducible populations of distinct subtypes of cervical and brachial MNs that resemble their in vivo, embryonic counterparts. Combining small molecule patterning with Ngn2 overexpression facilitates high-yield, reproducible production of disease-relevant MN subtypes, which is fundamental in propelling our knowledge of MN biology and its disruption in disease. Limone et al. induce neuralization of hPSCs into spinal MNs by small molecule patterning and TF overexpression. Multiplexed, pooled single-cell RNA-sequencing showcases high reproducibility in dozens of cell lines. These MN villages resemble in vivo spinal MNs and produce disease-relevant MN populations.
登录
查看更多内容
影响因子：
8.8
作者：
Jones RA;Harrison C;Eaton SL;Llavero Hurtado M;Graham LC;Alkhammash L;Oladiran OA;Gale A;Lamont DJ;Simpson H;Simmen MW;Soeller C;Wishart TM;Gillingwater TH
通讯作者：
Gillingwater TH
影响因子：
64.5
作者：
Dasen, Jeremy S.;De Camilli, Alessandro;Jessell, Thomas M.
通讯作者：
Jessell, Thomas M.
影响因子：
3.7
作者：
Ashoti A;Limone F;van Kranenburg M;Alemany A;Baak M;Vivié J;Piccioni F;Dijkers PF;Creyghton M;Eggan K;Geijsen N
通讯作者：
Geijsen N
影响因子：
64.5
作者：
Dasen, JS;Tice, BC;Jessell, TM
通讯作者：
Jessell, TM
影响因子：
46.9
作者：
Chambers, Stuart M.;Fasano, Christopher A.;Papapetrou, Eirini P.;Tomishima, Mark;Sadelain, Michel;Studer, Lorenz
通讯作者：
Studer, Lorenz