Phenotypic Screening Following Transcriptomic Deconvolution to Identify Transcription Factors Mediating Axon Growth Induced by a Kinase Inhibitor.
Phenotypic Screening Following Transcriptomic Deconvolution to Identify Transcription Factors Mediating Axon Growth Induced by a Kinase Inhibitor.
复制标题
DOI：
10.1177/24725552211026270
复制
发表时间：
2021-12
期刊：
影响因子：
3.1
通讯作者：
Lemmon, Vance P.
中科院分区：
文献类型：
--
作者：
Lowell, Jeffrey A.;O'Neill, Nicholas;Danzi, Matt C.;Al-Ali, Hassan;Bixby, John L.;Lemmon, Vance P.
中枢神经系统（CNS）损伤后，神经元对再生反应的内在限制和受损CNS环境中的抑制因子都限制了再生轴突的生长。成功的轴突再生的研究提供了机会，以确定这些情况下，从正常的成人中枢神经系统的区别功能。一个这样的机会是由激酶抑制剂RO 48提供的，其显著增强体外神经元的轴突生长，并且当在单侧延髓切开术后脑室内注入时，显著增加皮质脊髓束神经元的对侧发芽。作者在这里提出了RO 48相关轴突生长的转录组学去卷积，目的是确定与CNS中轴突生长相关的转录调节因子。通过使用RNA测序（RNA-seq）和转录因子结合位点富集分析，作者确定了一系列在体外RO 48诱导的大鼠海马神经元轴突生长过程中驱动差异基因表达的转录因子puplatin。以这种方式鉴定的82个转录因子基序包括一些已知与轴突生长调节相关的基序，如Jun、Klf 4、Myc、Atf 4、Stat 3和Nfatc 2，以及许多与轴突生长无关的基序。进行了一个表型功能丧失筛选，以评估这些转录因子在神经突生长中的作用，该筛选确定了几个潜在的生长调节因子。随后的验证表明，叉头盒（Fox）家族转录因子Foxp 2限制神经突生长，而FoxO亚家族成员Foxo 1和Foxo 3a促进神经突生长。因此，作者的联合转录组-表型筛选策略允许鉴定多靶点激酶抑制剂下游神经突生长的新型转录调节因子。
After injury to the central nervous system (CNS), both neuron-intrinsic limitations on regenerative responses and inhibitory factors in the injured CNS environment restrict regenerative axon growth. Instances of successful axon regrowth offer opportunities to identify features that differentiate these situations from that of the normal adult CNS. One such opportunity is provided by the kinase inhibitor RO48, which dramatically enhances neurite outgrowth of neurons in vitro and substantially increased contralateral sprouting of corticospinal tract neurons when infused intraventricularly following unilateral pyramidotomy. The authors present here a transcriptomic deconvolution of RO48-associated axon growth, with the goal of identifying transcriptional regulators associated with axon growth in the CNS. Through the use of RNA sequencing (RNA-seq) and transcription factor binding site enrichment analysis, the authors identified a list of transcription factors putatively driving differential gene expression during RO48-induced neurite outgrowth of rat hippocampal neurons in vitro. The 82 transcription factor motifs identified in this way included some with known association to axon growth regulation, such as Jun, Klf4, Myc, Atf4, Stat3, and Nfatc2, and many with no known association to axon growth. A phenotypic loss-of-function screen was carried out to evaluate these transcription factors for their roles in neurite outgrowth; this screen identified several potential outgrowth regulators. Subsequent validation suggests that the Forkhead box (Fox) family transcription factor Foxp2 restricts neurite outgrowth, while FoxO subfamily members Foxo1 and Foxo3a promote neurite outgrowth. The authors’ combined transcriptomic-phenotypic screening strategy therefore allowed identification of novel transcriptional regulators of neurite outgrowth downstream of a multitarget kinase inhibitor.
登录
查看更多内容
影响因子：
8.8
作者：
Fink KL;López-Giráldez F;Kim IJ;Strittmatter SM;Cafferty WBJ
通讯作者：
Cafferty WBJ
影响因子：
3.5
作者：
Blackmore, Murray G.;Moore, Darcie L.;Smith, Robin P.;Goldberg, Jeffrey L.;Bixby, John L.;Lemmon, Vance P.
通讯作者：
Lemmon, Vance P.
影响因子：
5.3
作者：
Broude, E;McAtee, M;Bregman, BS
通讯作者：
Bregman, BS
影响因子：
16.2
作者：
Gao, Y;Deng, KW;Filbin, MT
通讯作者：
Filbin, MT
影响因子：
4
作者：
Al-Ali, Hassan;Schuerer, Stephan C.;Bixby, John L.
通讯作者：
Bixby, John L.