Zhang Mengqi, Jiang Wei I, Arkelius Kajsa, Swanson Raymond A, Ma Dengke K, Singhal Neel S
Department of Neurology, University of California- San Francisco, San Francisco CA, 94158, USA.
Department of Physiology, University of California- San Francisco, San Francisco CA, 94158, USA; Cardiovascular Research Institute, University of California- San Francisco, San Francisco CA, 94158, USA.
Redox Biol. 2025 May 31;85:103709. doi: 10.1016/j.redox.2025.103709.
PALS1-associated tight junction (PATJ) protein is linked to metabolic disease and stroke in human genetic studies. Despite the recognized role of PATJ in cell polarization, its specific functions in metabolic disease and ischemic stroke recovery remain largely unexplored. We explored the functions of PATJ in an in vitro model and in vivo in C. elegans and mice. Using a mouse model of stroke, we found post-ischemic stroke duration-dependent increase of PATJ abundance in endothelial cells. PATJ knock-out (KO) HEK293 cells generated by CRISPR-Cas9 suggest roles for PATJ in cell proliferation, migration, mitochondrial stress response, and interactions with the Yes-associated protein (YAP)-1 signaling pathway. Notably, PATJ deletion altered YAP1 nuclear translocation. PATJ KO cells demonstrated transcriptional reprogramming based on RNA sequencing analysis, and identified dysregulation in genes central to vascular development, stress response, and metabolism, including RUNX1, HEY1, NUPR1, and HK2. Furthermore, we found that mpz-1, the homolog of PATJ, was significantly upregulated under hypoxic conditions in C. elegans. Knockdown of mpz-1 resulted in abnormal neuronal morphology and increased mortality, both of which were exacerbated by hypoxia exposure, indicating a critical protective role of PATJ/MPZ-1 in maintaining neuronal integrity and survival, particularly during oxygen deprivation stress relevant to ischemic stroke. These insights offer a new understanding of PATJ's regulatory functions within cellular and vascular physiology and help lay the groundwork for therapeutic strategies targeting PATJ-mediated pathways for stroke rehabilitation and neurovascular repair.
在人类遗传学研究中，PALS1相关紧密连接（PATJ）蛋白与代谢性疾病和中风有关。尽管人们已经认识到PATJ在细胞极化中的作用，但其在代谢性疾病和缺血性中风恢复中的具体功能仍 largely未被探索。我们在体外模型以及线虫和小鼠体内探索了PATJ的功能。利用中风小鼠模型，我们发现在缺血性中风后，内皮细胞中PATJ丰度呈持续时间依赖性增加。通过CRISPR-Cas9技术构建的PATJ基因敲除（KO）HEK293细胞表明，PATJ在细胞增殖、迁移、线粒体应激反应以及与Yes相关蛋白（YAP）-1信号通路的相互作用中发挥作用。值得注意的是，PATJ缺失改变了YAP1的核转位。基于RNA测序分析，PATJ基因敲除细胞表现出转录重编程，并确定了血管发育、应激反应和代谢相关基因的失调，包括RUNX1、HEY1、NUPR1和HK2。此外，我们发现PATJ的同源物mpz-1在秀丽隐杆线虫缺氧条件下显著上调。敲低mpz-1会导致神经元形态异常和死亡率增加，而缺氧暴露会加剧这两种情况，表明PATJ/MPZ-1在维持神经元完整性和存活方面具有关键的保护作用，特别是在与缺血性中风相关的缺氧应激期间。这些见解为PATJ在细胞和血管生理学中的调节功能提供了新的认识，并有助于为针对PATJ介导的中风康复和神经血管修复途径的治疗策略奠定基础。
