Brefeldin A (BFA) is a fungal macrocyclic lactone and a potent, reversible inhibitor of intracellular vesicle formation and protein trafficking between the endoplasmic reticulum (ER) and the Golgi apparatus[1][2].Brefeldin A is an ATPase inhibitor with IC50 value of 0.2 µM[9].Brefeldin A and its analogs are promising inhibitors in drug development due to a number of key features such as apoptosis?inducing properties as well as antitumor, antifungal, and antiviral effects [3,6].Brefeldin A is a CRISPR/Cas9 activator. Brefeldin A inhibits HSV-1 and has anti-cancer activity[4].
Perturbation of ER-Golgi trafficking by brefeldin A (BFA) treatment attenuated nucleotide-binding oligomerization domain-like receptor family, pyrin-domain-containing 3 (NLRP3) inflammasome activation in mouse bone marrow-derived macrophages (BMDMs) [5].ADP-ribosylation of BARS is mediated by formation of a conjugate between Brefeldin A and ADPR. BARS shows BAC binding when incubated with the medium from the brefeldin A-treated CD38+ HeLa cells[3].Brefeldin A induces anchorage-independent cell death in MDA-MB-231 breast cancer cells with an EC50 of 0.016 µg/mL, inhibits the formation of MDA-MB-231 colonies in 3D and 2D cultures and inhibits the migration and MMP 9 activity of MDA-MB-231[2].
In tumor-bearing mice, M-brefeldin A can prolong blood circulation, improve tumor accumulation ability, and show effective inhibition of tumor growth, M-brefeldin A 10 mg/kg group showed effective antitumor effect and significantly delayed tumor progression, while M-brefeldin A 5 mg/kg mice did not show significant inhibitory effect[7]. Mice were treated with the Golgi blocker Brefeldin A. Since most cytokines are processed and secreted via the classical secretion pathway through the Golgi, brefeldin A blocks cytokine secretion, leading to their accumulation within immune cells, which are eventually detected by flow cytometry. Thus, treatment of mice with brefeldin A allows in situ assessment of cytokine production without the use of reporter mice [8].
References:
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[2]: Tseng CN, Hong YR, et,al. Brefeldin A reduces anchorage-independent survival, cancer stem cell potential and migration of MDA-MB-231 human breast cancer cells. Molecules. 2014 Oct 29;19(11):17464-77. doi: 10.3390/molecules191117464. PMID: 25356567; PMCID: PMC6271931.
[3]: Wang J, Fang Y, et,al. Erythroleukemia cells acquire an alternative mitophagy capability. Sci Rep. 2016 Apr 19;6:24641. doi: 10.1038/srep24641. PMID: 27091640; PMCID: PMC4835698.
[4]: Yu C, Liu Y, et,al. Small molecules enhance CRISPR genome editing in pluripotent stem cells. Cell Stem Cell. 2015 Feb 5;16(2):142-7. doi: 10.1016/j.stem.2015.01.003. PMID: 25658371; PMCID: PMC4461869.
[5]: Hong S, Hwang I, et,al. Brefeldin A-sensitive ER-Golgi vesicle trafficking contributes to NLRP3-dependent caspase-1 activation. FASEB J. 2019 Mar;33(3):4547-4558. doi: 10.1096/fj.201801585R. Epub 2018 Dec 28. PMID: 30592629.
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[7]: Zhang JM, Jiang YY, et,al. Brefeldin A delivery nanomicelles in hepatocellular carcinoma therapy: Characterization, cytotoxic evaluation in vitro, and antitumor efficiency in vivo. Pharmacol Res. 2021 Oct;172:105800. doi: 10.1016/j.phrs.2021.105800. Epub 2021 Aug 4. PMID: 34363949.
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[9]: Wierzbicki PM, Kogut-Wierzbicka M, et,al. Protein and siRNA delivery by transportan and transportan 10 into colorectal cancer cell lines. Folia Histochem Cytobiol. 2014;52(4):270-80. doi: 10.5603/FHC.a2014.0035. Epub 2014 Dec 16. PMID: 25511292.
Brefeldin A（BFA）是一种真菌大环内酯，能够有效、可逆地抑制内质网和高尔基体之间的蛋白质运输和囊泡形成[1][2]。Brefeldin A是一种ATP酶抑制剂，其IC50值为0.2微米[9]。由于具有诱导细胞凋亡、抗肿瘤、抗真菌和抗病毒等多种特性，Brefeldin A及其类似物在药物开发中被广泛应用[3,6]。此外，Brefeldin A还可以作为CRISPR/Cas9激活剂使用，并且对HSV-1具有抑制作用并具有抗癌活性[4]。
在小鼠骨髓源性巨噬细胞（BMDMs）中，使用卟啉菌素A（BFA）处理扰乱了内质网-高尔基体运输，减弱了核苷酸结合寡聚化域类受体家族、含有pyrin域的3号（NLRP3）炎症小体的激活[5]。通过形成卟啉菌素A和ADPR之间的共轭物介导对BARS进行ADP核糖化。当与经过卟啉菌素A处理的CD38+ HeLa细胞培养基一起孵育时，BARS显示出与BAC结合[3]。卟啉菌素A诱导MDA-MB-231乳腺癌细胞无需附着即可死亡，EC50为0.016 µg/mL，并抑制了MDA-MB-231在三维和二维培养中形成集落以及抑制其迁移和MMP 9活性[2]。
在携带肿瘤的小鼠中，M-布雷菲定A可以延长血液循环时间，提高肿瘤积累能力，并显示出有效抑制肿瘤生长的作用。M-布雷菲定A 10毫克/千克组表现出有效的抗肿瘤效果并显著延缓了肿瘤进展，而M-布雷菲定A 5毫克/千克组则没有显示出明显的抑制作用[7]。小鼠接受了高尔基体阻断剂Brefeldin A治疗。由于大多数细胞因子是通过经典分泌途径通过高尔基体加工和分泌的，Brefeldin A会阻止细胞因子分泌，导致它们在免疫细胞内积累，并最终被流式细胞术检测到。因此，在小鼠中使用Brefeldin A治疗可以对细胞因子产生进行原位评估，无需使用报告小鼠[8]。