PDF(Pubmed)
Abstract：
SDF-1/CXCR4 chemokine signaling are indispensable for cell migration, especially the Primordial Germ Cell (PGC) migration towards the gonadal ridge during early development. We earlier found that this signaling is largely conserved in the Japanese anchovy (Engraulis japonicus, EJ), and a mere treatment of CXCR4 antagonist, AMD3100, leads to germ cell depletion and thereafter gonad sterilization. However, the effect of AMD3100 was limited. So, in this research, we scouted for CXCR4 antagonist with higher potency by employing advanced artificial intelligence deep learning-based computer simulations. Three potential candidates, AMD3465, WZ811, and LY2510924, were selected and in vivo validation was conducted using Japanese anchovy embryos. We found that seven transmembrane motif of EJ CXCR4a and EJ CXCR4b were extremely similar with human homolog while the CXCR4 chemokine receptor N terminal (PF12109, essential for SDF-1 binding) was missing in EJ CXCR4b. 3D protein analysis and cavity search predicted the cavity in EJ CXCR4a to be five times larger (6,307 ų) than that in EJ CXCR4b (1,241 ų). Docking analysis demonstrated lower binding energy of AMD3100 and AMD3465 to EJ CXCR4a (Vina score -9.6) and EJ CXCR4b (Vina score -8.8), respectively. Furthermore, we observed significant PGC mismigration in microinjected AMD3465 treated groups at 10, 100 and 1 × 105 nM concentration in 48 h post fertilized embryos. The other three antagonists showed various degrees of PGC dispersion, but no significant effect compared to their solvent control at tested concentrations was observed. Cumulatively, our results suggests that AMD3645 might be a better candidate for abnormal PGC migration in Japanese anchovy and warrants further investigation.
摘要：
SDF-1/CXCR4趋化因子信号是细胞迁移不可或缺的，特别是在早期发育过程中，原始生殖细胞（PGC）向性腺脊迁移。我们早些时候发现，这种信号在日本an鱼（Engraulisjaponicus，EJ),仅仅是CXCR4拮抗剂的治疗，AMD3100导致生殖细胞耗尽，然后性腺灭菌。然而,AMD3100的作用有限。所以,在这项研究中，我们通过采用先进的基于人工智能深度学习的计算机模拟,搜寻了具有更高效力的CXCR4拮抗剂.三个潜在的候选人,选择AMD3465、WZ811和LY2510924,并使用日本an鱼胚胎进行体内验证。我们发现EJCXCR4a和EJCXCR4b的七个跨膜基序与人类同源物非常相似，而EJCXCR4b中缺少CXCR4趋化因子受体N末端（PF12109，对SDF-1结合至关重要）。3D蛋白质分析和腔搜索预测，EJCXCR4a中的腔比EJCXCR4b（1,241，λ）大五倍（6,307，λ）。对接分析表明AMD3100和AMD3465对EJCXCR4a（Vina评分-9.6）和EJCXCR4b（Vina评分-8.8）的结合能较低，分别。此外，我们在受精后48小时内观察到10、100和1×105nM浓度的显微注射AMD3465处理组中明显的PGC误移。其他三种拮抗剂均表现出不同程度的PGC分散性，但与测试浓度下的溶剂对照相比，没有观察到明显的影响。累计,我们的结果表明,AMD3645可能是日本an鱼中PGC异常迁移的更好候选者,值得进一步调查.
