圖1. 調(diào)控自殺性中性粒細(xì)胞溶解的機(jī)制示意圖。

不同的刺激 [如PMA（phorbol 12-myristate 13-acetate，佛波醇）、離子霉素、晶體、CXCL8、Fc（可結(jié)晶的片段）-受體和病原體] 誘導(dǎo)自殺性中性粒細(xì)胞溶解，有時(shí)稱為 "NETosis"。PMA，一個(gè)經(jīng)典的非生理性刺激物，直接激活蛋白激酶C（PKC）和Raf-MEK-ERK-MAP激酶途徑。MAP激酶的激活將啟動(dòng)NADPH氧化酶復(fù)合物的形成，導(dǎo)致快速生成活性氧（ROS）。中性粒細(xì)胞彈性酶（NE）和骨髓過氧化物酶（MPO）有助于核膜通透性和染色質(zhì)的進(jìn)一步展開。肽基精氨酸脫氨酶4（PAD4）是一種Ca2+特異性酶，主要定位在細(xì)胞核內(nèi)。PAD4通過將精氨酸變?yōu)楣习彼醽硇揎椊M蛋白，從而導(dǎo)致染色質(zhì)解聚，在核破裂時(shí)，瓜氨酸組蛋白和核DNA一起被釋放出來。釋放的DNA會(huì)進(jìn)一步被顆粒蛋白（NE，MPO）和細(xì)胞溶質(zhì)蛋白（calpain）裝飾。中性粒細(xì)胞溶解也被證明依賴于細(xì)胞周期蛋白激酶4和6（CDK4/6）。CDK4/6的激活導(dǎo)致視網(wǎng)膜母細(xì)胞瘤蛋白（Rb-P）和核纖層B（lamin B）的磷酸化，以及核包膜破裂。細(xì)胞內(nèi)ROS的增加會(huì)進(jìn)一步激活受體交互蛋白激酶3（RIPK3）和混合系激酶域樣蛋白（MLKL），促使質(zhì)膜破裂。Gasdermin D是一種成孔的細(xì)胞質(zhì)蛋白，在質(zhì)膜通透性的后期階段牽涉其中。質(zhì)膜破裂后，含有顆粒蛋白的瓜氨酸核DNA支架以云的形式釋放在細(xì)胞外空間，導(dǎo)致中性粒細(xì)胞死亡。

原文

Fig. 1. Schematic representation of mechanisms regulating suicidal neutrophil cytolysis. Different stimuli [e.g., PMA [59], ionomycin [198], crystals [301], CXCL8 [204], Fc (fragment crystallizable)-receptors [243,302] and pathogens [303]] induce suicidal neutrophil lysis, sometimes called “NETosis”. PMA, a classical non-physiological stimulus, directly activates protein kinase C (PKC) and the Raf-MEK-ERK-MAP kinase pathway [66]. MAP kinases activation would initiate NADPH oxidase complex formation leading to rapid generation of reactive oxygen species (ROS) [60, 229]. Neutrophil elastase (NE) [68] and myeloperoxidase (MPO) [200] contribute to nuclear membrane permeabilization and further unfolding of chromatin [246]. Peptidyl arginine deiminase 4 (PAD4) is a Ca2+-specific enzyme primarily localized in the nucleus. PAD4 modifies histone by changing arginine to citrulline which leads to chromatin decondensation, and upon nuclear rupture, citrullinated-histone together with nuclear DNA is released [70, 304,305]. Released DNA becomes further decorated with granular (NE, MPO) [15] and cytosolic (calpain) proteins [306,307]. Neutrophil cytolysis was also shown to be dependent on cyclin kinases 4 and 6 (CDK4/6) [308]. Activation of CDK4/6 leads to phosphorylation of the retinoblastoma protein (Rb-P) and lamin B, and nuclear envelope breakdown [308]. Increased intracellular ROS would further activate receptor-interacting protein kinase 3 (RIPK3) and mixed lineage kinase domain-like (MLKL), contributing to the plasma membrane rupture [309,310]. Gasdermin D, a pore-formin cytoplasmic protein, is implicated during later stages of plasma membrane permeabilization [207,257]. After plasma membrane rupture, the citrullinated nuclear DNA scaffold containing granule proteins are released in the extracellular space in a cloud format leading to neutrophil death [14,59].

圖 2. 調(diào)節(jié)vital中性粒細(xì)胞胞外誘捕網(wǎng) (NETs) 形成的機(jī)制示意圖。

不同的刺激物 [例如金黃色葡萄球菌、LPS、GM-CSF 和 C5a、甲狀腺癌條件培養(yǎng)基 、免疫復(fù)合物迅速（30 分鐘內(nèi) ) 誘導(dǎo) NET 的形成。中性粒細(xì)胞的刺激導(dǎo)致 NADPH 氧化酶快速激活，從而導(dǎo)致細(xì)胞內(nèi) ROS 增加。Vital NET 形成在沒有核膜破壞的情況下發(fā)生，據(jù)報(bào)道 DNA 通過囊泡離開細(xì)胞質(zhì)。DNA 被顆粒蛋白（NE、MPO、PR3）修飾。中性粒細(xì)胞仍然存活以發(fā)揮進(jìn)一步的功能，并利用增加的細(xì)胞內(nèi) ROS 動(dòng)員細(xì)胞骨架將顆粒和線粒體運(yùn)輸?shù)交罨闹行粤＜?xì)胞的前緣。線粒體可以在Vital NET 形成中扮演三個(gè)角色。首先，它們可以介導(dǎo)線粒體 ROS (mtROS) 的形成，其次，線粒體 DNA (mtDNA) 可以釋放到細(xì)胞外，再者，它們?yōu)樘墙徒?ATP 的產(chǎn)生提供支持。還有報(bào)道稱，mtROS 可以通過線粒體外膜中電壓依賴性陰離子通道 (VDAC) 寡聚體形成的孔隙釋放短的 mtDNA 片段。mtROS 和 mtDNA 可能參與 NET 形成引起了極大的興趣，因?yàn)?mtDNA 在人和小鼠中發(fā)揮免疫調(diào)節(jié)作用。
關(guān)于翻譯這里插幾句：neutrophil extracellular traps（NETs）的翻譯，有些人用的是中性粒細(xì)胞胞外陷阱，我個(gè)人傾向于中性粒細(xì)胞胞外誘捕網(wǎng)這個(gè)翻譯，因?yàn)樗目s寫剛好是NET，這樣的話有點(diǎn)形意兼?zhèn)涞奈兜?，也比較好記。NETosis有兩種，分別是suicidal NETosis（腫瘤組織中主要是suicidal型[1]）和vital NETosis，根據(jù)前面suicidal 的意思，老編覺得vital翻譯為重要的顯然是不合適的。根據(jù)文獻(xiàn)[1-2]（Vital NET formation occurs independently of cell death in the absence of membrane disruption within minutes after stimulation of neutrophils. Suicidal NETosis, in which neutrophils die after expelling the filaments; and vital NETosis, in which expulsion appears without altering the membrane.）或許活力型NETosis比較合適（中性粒細(xì)胞沒死；可能也正因如此，有些大牛用Vital NET formation來來描述這個(gè)生物學(xué)過程，而不是vital NETosis[1]），跟ChatGPT（GPT-3.5 Turbo）想的意思差不多（見下圖）。還有NETosis，這些名詞到底怎么翻譯才最好，就留給大牛們來做了。

▲截自：https://sci.justscience.cn/chatgpt 聊天記錄

原文

Fig. 2. Schematic representation of mechanisms regulating vital neutrophil extracellular traps (NET) formation. NET formation. Different stimuli [e.g., S. aureus [30,58], LPS [75,227,311], GM-CSF and C5a [16,29], conditioned media of thyroid cancer [72], immune complexes [29,290,312]] rapidly (within 30 min) induce the formation of NETs. Stimulation of neutrophils leads to rapid NADPH oxidase activation, resulting in increased intracellular ROS [16,60]. Vital NET formation occurs without nuclear membrane disruption and has been reported with DNA leaving the cytoplasm in vesicles [30,58,75]. DNA becomes decorated with granule proteins (NE, MPO, PR3) [15,16,29]. The neutrophil remains alive for further functions [46,72], and utilizes the increased intracellular ROS to mobilize the cell cytoskeleton to transport granules and mitochondria to the leading edge of activated neutrophils [73]. Mitochondria can play three roles in vital NET formation. First, they can mediate mitochondrial ROS (mtROS) formation [72, 290], and second, mitochondria DNA (mtDNA) can be released extracellularly [290,312,313], and third, they provide support for glycolytic ATP production [29]. It has also been reported that mtROS can release short mtDNA fragments via pores formed by the voltage-dependent anion channel (VDAC) oligomers in the mitochondrial outer membrane [314]. The possible involvement of mtROS and mtDNA in NET formation is of great interest because mtDNA plays an immunomodulatory role in human and mouse [54].

圖 3. A) 髓過氧化物酶 (MPO) 和瓜氨酸化組蛋白 H3 (H3Cit)，分別是中性粒細(xì)胞和 NET 的標(biāo)志物，在乳腺癌和結(jié)腸癌患者的肝轉(zhuǎn)移中含量豐富。肝臟中 NETs (NET-DNA) 的 DNA 成分通過與 CCDC25（coiled-coil domain containing protein 25；是一種在癌細(xì)胞上表達(dá)的跨膜蛋白）蛋白的高親和力相互作用，對(duì)癌細(xì)胞具有趨化作用。DNA-CCDC25 相互作用觸發(fā)細(xì)胞內(nèi)信號(hào)級(jí)聯(lián)，促進(jìn)癌細(xì)胞的定向遷移和轉(zhuǎn)移的形成。在小鼠模型中，體內(nèi)給予抗 CCDC25 中和抗體減少了 NET 介導(dǎo)的遠(yuǎn)處轉(zhuǎn)移的形成。B) NET 喚醒休眠的癌細(xì)胞。在某些癌癥（例如乳腺癌、前列腺癌）中，癌細(xì)胞在蘇醒之前可以保持休眠狀態(tài)并且在臨床上檢測(cè)不到達(dá)數(shù)年，如轉(zhuǎn)移性癌癥。有人提出炎癥在癌細(xì)胞休眠和覺醒之間的轉(zhuǎn)換中發(fā)揮作用。在乳腺癌和前列腺癌的小鼠模型中，休眠的癌細(xì)胞與細(xì)胞外基質(zhì) (ECM) 蛋白層粘連蛋白結(jié)合。反復(fù)滴注脂多糖或尼古?。▋煞N NET 誘導(dǎo)劑），促進(jìn)肺部嗜中性粒細(xì)胞炎癥和 NET 形成。兩種 NET 相關(guān)蛋白酶，中性粒細(xì)胞彈性蛋白酶 (NE) 和基質(zhì)金屬肽酶 9 (MMP9)，依次切割層粘連蛋白，暴露表位，通過整合素激活觸發(fā)休眠癌細(xì)胞的增殖。一種針對(duì) NET 重塑層粘連蛋白的阻斷抗體可減少尼古丁或 LPS 誘導(dǎo)的炎癥，這些炎癥會(huì)喚醒小鼠休眠的癌細(xì)胞。C) 不同的原發(fā)性黑色素瘤和結(jié)腸癌細(xì)胞系誘導(dǎo)人 PMN（polymorphonuclear neutrophil leukocyte，即neutrophil）和 PMN-MDSC（polymorphonucler myeloid-derived suppressor cells, 多形核髓源性抑制細(xì)胞）形成 NET。Reparixin 是一種 CXCR1/CXCR2 趨化因子受體拮抗劑，可抑制 NET 形成。NETs 包裹和包裹腫瘤細(xì)胞并保護(hù)它們免受 CD8+ T 細(xì)胞和自然殺傷 (NK) 細(xì)胞介導(dǎo)的細(xì)胞毒性。D) 人中性粒細(xì)胞胞外誘捕網(wǎng) (NETs) 調(diào)節(jié)血管生成。PMA 激活的人嗜中性粒細(xì)胞釋放 NET，從而誘導(dǎo)人肺動(dòng)脈內(nèi)皮細(xì)胞 (HPAEC) 的促血管生成反應(yīng)（例如，增加毛細(xì)管長(zhǎng)度、新芽數(shù)量和新芽面積）。這些作用被 DNase 1 或髓過氧化物酶 (MPO) 抑制劑抑制。NET 還增加了 HPAEC 的增殖活性。此外，單獨(dú)或聯(lián)合使用血管生成素 1 (ANGPT1) 和血管生成素 2 (ANGPT2) 可誘導(dǎo)人嗜中性粒細(xì)胞釋放 NET。ANGPT 誘導(dǎo)的 NET 形成被 PAD4 抑制劑 GSK484 和 NADPH 氧化酶抑制劑抑制。血管生成素介導(dǎo)的 NETs 增加了人血管內(nèi)皮細(xì)胞 (HUVEC) 管的長(zhǎng)度和環(huán)的數(shù)量。

原文

Fig. 3. A) Myeloperoxidase (MPO) and citrullinated histone H3 (H3Cit), markers for neutrophils and NET, respectively, are abundant in the liver metastases of patients with breast and colon cancers [125]. The DNA components of NETs (NET-DNA) in the liver is chemotactic for cancer cells by interacting with high affinity with the coiled-coil domain containing protein 25 (CCDC25), which is a transmembrane protein expressed on cancer cells. DNA?? CCDC25 interaction triggers an intracellular signaling cascade promoting directional migration of cancer cells and the formation of metastasis. In mouse models, the administration in vivo of a neutralizing antibody anti-CCDC25 reduced the formation of NET-mediated distant metastases [125]. B) NET awaken dormant cancer cells. In several cancers (e.g., breast, prostate) cancer cells can remain dormant and clinically undetectable for years before awakening, as metastatic cancer [127,128]. It has been suggested that inflammation plays a role in the switch between dormancy and awakening of cancer cells [129,130]. In mouse models of breast and prostate cancers, dormant cancer cells bind to the extracellular matrix (ECM) protein laminin. Repeated instillation of lipopolysaccharide [54,131] or nicotine, two inducers of NETs [247], promote neutrophil inflammation and NET formation in the lung. Two NET-associated proteases, neutrophil elastase (NE) and matrix metallopeptidase 9 (MMP9), sequentially cleave laminin exposing an epitope that triggers proliferation of dormant cancer cells through integrin activation. A blocking antibody against NET-remodeled laminin reduces nicotine- or LPS-induced inflammation from awakening dormant cancer cells in mice [131]. C) Different primary melanoma and colon carcinoma cell lines induce NET formation from human PMNs and PMN-MDSCs [116]. Reparixin, a CXCR1/ CXCR2 chemokine receptor antagonists, inhibits NET formation. NETs wrap and coat tumor cells and shields them from the cytotoxicity, mediated by CD8+ T cells and natural killer (NK) cell [116]. D) Human neutrophil extracellular traps (NETs) modulate angiogenesis. PMA-activated human neutrophils release NETs, which induce pro-angiogenic responses (e.g., increased capillary tube length, number of sprouts, and sprouting area) of human pulmonary artery endothelial cells (HPAECs). These effects are inhibited by DNase 1 or a myeloperoxidase (MPO) inhibitor. NETs also increases the proliferative activity of HPAECs [152]. In addition, angiopoietin 1 (ANGPT1) and angiopoietin 2 (ANGPT2) alone or combined induce NET release from human neutrophils. NET formation induced by ANGPTs is inhibited by the PAD4 inhibitor GSK484, and by a NADPH oxidase inhibitor. Angiopoietin-mediated NETs increase human vascular endothelial cell (HUVEC) tube length and the number of loops [155].

參考文獻(xiàn)
1. Fang Q, Stehr A M, Naschberger E, et al. No NETs no TIME: Crosstalk between neutrophil extracellular traps and the tumor immune microenvironment[J]. Frontiers in Immunology, 2022, 13.

2. Guillotin F, Fortier M, Portes M, et al. Vital NETosis vs. suicidal NETosis during normal pregnancy and preeclampsia[J]. Frontiers in Cell and Developmental Biology, 2022, 10.

主要譯自：Seminars in Cancer Biology (10.1016/j.semcancer.2021.07.011)，僅作學(xué)術(shù)分享使用；侵刪