Machine learning-based classification of dual fluorescence signals reveals muscle stem cell fate transitions in response to regenerative niche factors.
Machine learning-based classification of dual fluorescence signals reveals muscle stem cell fate transitions in response to regenerative niche factors.
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DOI：
10.1038/s41536-023-00277-4
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发表时间：
2023-01-14
影响因子：
7.2
通讯作者：
Blau, Helen M. M.
中科院分区：
文献类型：
--
作者：
Togninalli, Matteo;Ho, Andrew T. V.;Madl, Christopher M. M.;Holbrook, Colin A. A.;Wang, Yu Xin;Magnusson, Klas E. G.;Kirillova, Anna;Chang, Andrew;Blau, Helen M. M.
肌肉干细胞（MuSC）命运的适当调控是骨骼肌再生的关键。由于人群水平终点分析的局限性，促进再生的生态位因子如何控制MuSC命运决定的动力学仍然未知。为了解决这一知识差距，我们开发了一种双荧光成像时移（dual - flit）显微镜方法，该方法利用机器学习分类策略以高时间分辨率跟踪单细胞命运决定。使用两个读出干性维持和肌源性承诺的荧光报告，我们为单个musc及其后代构建了详细的谱系树，将每个分裂事件分类为对称自我更新，不对称或对称承诺。我们的分析显示，脂质代谢物前列腺素E2 （PGE2）随着时间的推移加速了MuSC的增殖速度，同时使分裂事件偏向对称自我更新。相比之下，IL6家族成员Oncostatin M （OSM）在第一代之后降低了增殖率，同时阻断了肌原性承诺。通过我们的Dual-FLIT方法，这些对生态位线索调控MuSC动态的见解是唯一可行的。我们预计，类似的二进制活细胞读数来源于双flit将显著扩大我们的理解如何生态位因子控制组织再生的实时。
The proper regulation of muscle stem cell (MuSC) fate by cues from the niche is essential for regeneration of skeletal muscle. How pro-regenerative niche factors control the dynamics of MuSC fate decisions remains unknown due to limitations of population-level endpoint assays. To address this knowledge gap, we developed a dual fluorescence imaging time lapse (Dual-FLIT) microscopy approach that leverages machine learning classification strategies to track single cell fate decisions with high temporal resolution. Using two fluorescent reporters that read out maintenance of stemness and myogenic commitment, we constructed detailed lineage trees for individual MuSCs and their progeny, classifying each division event as symmetric self-renewing, asymmetric, or symmetric committed. Our analysis reveals that treatment with the lipid metabolite, prostaglandin E2 (PGE2), accelerates the rate of MuSC proliferation over time, while biasing division events toward symmetric self-renewal. In contrast, the IL6 family member, Oncostatin M (OSM), decreases the proliferation rate after the first generation, while blocking myogenic commitment. These insights into the dynamics of MuSC regulation by niche cues were uniquely enabled by our Dual-FLIT approach. We anticipate that similar binary live cell readouts derived from Dual-FLIT will markedly expand our understanding of how niche factors control tissue regeneration in real time.
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DOI：
10.1093/bioinformatics/btu080
发表时间：
2014-06-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Maška M;Ulman V;Svoboda D;Matula P;Matula P;Ederra C;Urbiola A;España T;Venkatesan S;Balak DM;Karas P;Bolcková T;Streitová M;Carthel C;Coraluppi S;Harder N;Rohr K;Magnusson KE;Jaldén J;Blau HM;Dzyubachyk O;Křížek P;Hagen GM;Pastor-Escuredo D;Jimenez-Carretero D;Ledesma-Carbayo MJ;Muñoz-Barrutia A;Meijering E;Kozubek M;Ortiz-de-Solorzano C
通讯作者：
Ortiz-de-Solorzano C
影响因子：
64.8
作者：
Sacco, Alessandra;Doyonnas, Regis;Kraft, Peggy;Vitorovic, Stefan;Blau, Helen M.
通讯作者：
Blau, Helen M.
影响因子：
10.6
作者：
Magnusson KE;Jalden J;Gilbert PM;Blau HM
通讯作者：
Blau HM
影响因子：
8.8
作者：
Rodgers, Joseph T.;Schroeder, Matthew D.;Rando, Thomas A.
通讯作者：
Rando, Thomas A.
影响因子：
23.9
作者：
Bentzinger, C. Florian;Wang, Yu Xin;von Maltzahn, Julia;Soleimani, Vahab D.;Yin, Hang;Rudnicki, Michael A.;Rudnicki, A.
通讯作者：
Rudnicki, A.