Department of Critical Care Medicine, The Affiliated Cancer Hospital of Zhengzhou University & Henan Cancer Hospital, Zhengzhou, Henan, China.
Department of Cardiovascular Medicine, The Second School of Clinical Medicine, Southern Medical University, Guangzhou, Guangdong, China.
Infect Immun. 2023 Aug 16;91(8):e0010223. doi: 10.1128/iai.00102-23. Epub 2023 Jul 5.
Fusobacterium nucleatum colonization contributes to the occurrence of portal vein thrombosis in patients with gastric cancer (GC). However, the underlying mechanism by which F. nucleatum promotes thrombosis remains unclear. In this study, we recruited a total of 91 patients with GC and examined the presence of F. nucleatum in tumor and adjacent non-tumor tissues by fluorescence hybridization and quantitative PCR. Neutrophil extracellular traps (NETs) were detected by immunohistochemistry. Extracellular vesicles (EVs) were extracted from the peripheral blood and proteins in the EVs were identified by mass spectrometry (MS). HL-60 cells differentiated into neutrophils were used to package engineered EVs to imitate the EVs released from NETs. Hematopoietic progenitor cells (HPCs) and K562 cells were used for megakaryocyte (MK) differentiation and maturation to examine the function of EVs. We observed that F. nucleatum-positive patients had increased NET and platelet counts. EVs from F. nucleatum-positive patients could promote the differentiation and maturation of MKs and had upregulated 14-3-3 proteins, especially 14-3-3ε. 14-3-3ε upregulation promoted MK differentiation and maturation . HPCs and K562 cells could receive 14-3-3ε from the EVs, which interacted with GP1BA and 14-3-3ζ to trigger PI3K-Akt signaling. In conclusion, we identified for the first time that F. nucleatum infection promotes NET formation, which releases EVs containing 14-3-3ε. These EVs could deliver 14-3-3ε to HPCs and promote their differentiation into MKs via activation of PI3K-Akt signaling.
具核梭杆菌定植有助于胃癌（GC）患者发生门静脉血栓形成。然而，具核梭杆菌促进血栓形成的潜在机制尚不清楚。在这项研究中，我们共招募了 91 名胃癌患者，并通过荧光杂交和定量 PCR 检测肿瘤和相邻非肿瘤组织中具核梭杆菌的存在。通过免疫组织化学检测中性粒细胞胞外诱捕网（NETs）。从外周血中提取细胞外囊泡（EVs），并通过质谱（MS）鉴定 EVs 中的蛋白质。HL-60 细胞分化为中性粒细胞，用于包装工程化的 EVs，以模拟来自 NETs 的 EVs。使用造血祖细胞（HPCs）和 K562 细胞进行巨核细胞（MK）分化和成熟，以检查 EVs 的功能。我们观察到具核梭杆菌阳性患者的 NET 和血小板计数增加。具核梭杆菌阳性患者的 EVs 可促进 MK 的分化和成熟，并上调 14-3-3 蛋白，特别是 14-3-3ε。14-3-3ε 的上调促进了 MK 的分化和成熟。HPCs 和 K562 细胞可以从 EVs 中接收 14-3-3ε，其与 GP1BA 和 14-3-3ζ相互作用，触发 PI3K-Akt 信号。总之，我们首次发现具核梭杆菌感染可促进 NET 的形成，从而释放含有 14-3-3ε 的 EVs。这些 EVs 可以通过激活 PI3K-Akt 信号将 14-3-3ε 传递给 HPCs，并促进其分化为 MKs。
