Department of Biochemistry and Molecular Biology, Medical University of Lublin, 21-093, Lublin, Poland.
Faculty of Science and Engineering, Biosciences, Åbo Akademi, Turku, Finland.
Cell Commun Signal. 2022 May 18;20(1):67. doi: 10.1186/s12964-022-00885-5.
The Notch signaling pathway is a crucial regulator of cell differentiation as well as tissue organization, whose deregulation is linked to the pathogenesis of different diseases. NOTCH1 plays a key role in breast cancer progression by increasing proliferation, maintenance of cancer stem cells, and impairment of cell death. NOTCH1 is a mechanosensitive receptor, where mechanical force is required to activate the proteolytic cleavage and release of the Notch intracellular domain (NICD). We circumvent this limitation by regulating Notch activity by light. To achieve this, we have engineered an optogenetic NOTCH1 receptor (optoNotch) to control the activation of NOTCH1 intracellular domain (N1ICD) and its downstream transcriptional activities. Using optoNotch we confirm that NOTCH1 activation increases cell proliferation in MCF7 and MDA-MB-468 breast cancer cells in 2D and spheroid 3D cultures, although causing distinct cell-type specific migratory phenotypes. Additionally, optoNotch activation induced chemoresistance on the same cell lines. OptoNotch allows the fine-tuning, ligand-independent, regulation of N1ICD activity and thus a better understanding of the spatiotemporal complexity of Notch signaling. Video Abstract.
Notch 信号通路是细胞分化和组织组织的关键调节剂，其失调与不同疾病的发病机制有关。NOTCH1 通过增加增殖、维持癌症干细胞和损害细胞死亡来促进乳腺癌的进展。NOTCH1 是一种机械敏感受体，需要机械力来激活 Notch 细胞内结构域（NICD）的蛋白水解切割和释放。我们通过光来调节 Notch 的活性来规避这一限制。为此，我们设计了一种光遗传学 NOTCH1 受体（optoNotch）来控制 NOTCH1 细胞内结构域（N1ICD）及其下游转录活性的激活。使用 optoNotch，我们证实 NOTCH1 的激活增加了 MCF7 和 MDA-MB-468 乳腺癌细胞在 2D 和球体 3D 培养物中的增殖，尽管导致了不同的细胞类型特异性迁移表型。此外，optoNotch 的激活诱导了相同细胞系的化疗耐药性。OptoNotch 允许对 N1ICD 活性进行精细调节、无需配体的调节，从而更好地理解 Notch 信号的时空复杂性。视频摘要。
