González Epelboim Victoria Rebeca Dana, Lamas Diego G, Huck-Iriart Cristián, Caputo Ezequiel Nicolas, Altube Maria Julia, Jerez Horacio Emanuel, Simioni Yamila Roxana, Ghosal Kajal, Morilla Maria Jose, Higa Leticia Herminia, Romero Eder Lilia
Nanomedicine Research and Development Centre (NARD), Science and Technology Department, National University of Quilmes, Roque Saenz Peña 352, Bernal 1876, Buenos Aires, Argentina.
Laboratorio de Cristalografía Aplicada (LCA), Instituto de Tecnologías Emergentes y Ciencias Aplicadas (ITECA), Escuela de Ciencia y Tecnología (ECyT), UNSAM-CONICET, Campus Miguelete, San Martín 1650, Buenos Aires, Argentina.
Int J Mol Sci. 2025 Sep 4;26(17):8607. doi: 10.3390/ijms26178607.
The membranes of halophilic archaea are a source of novel biomaterials, mainly of isoprenoid nature, with therapeutic properties practically unraveled. Here, we explored the antitumoral activity of neutral archaeolipids (NAs, such as bacterioruberin, astaxanthin, and dihydrosqualene) present in the total archaeolipids (TAs) (a fraction from the first step of lipid extraction by the modified Blight and Dyer technique) extracted from halophilic archaea Halorubrum tebenquichense, and formulated as TA-nanoarchaeosomes (TA: polar archaeolipids (PAs): Tween 80, 5:5:4 ::, TA-nanoARC). The structure of 300.3 ± 84.2 nm TA-nanoARC of 0.59 ± 0.12 polydispersity index and -20 ± 3.7 mV ζ potential as determined by SAXS modelling, revealed that NA reduced the hydrophobic core and enlarged its hydrophilic section in comparison to TA-lacking bilayers (nanoARC), while preserving the width (50 Å) and unilamellarity. Stable to storage and nebulization, TA-nanoARC was cytotoxic on A549 cells after 48 h, with an IC50 expressed as [bacterioruberin] of 0.15 μg/mL (0.20 µM), comparable to or lower than the IC50 of docetaxel or cisplatin. Such cytotoxicity was exerted at a concentration harmless to macrophages (mTHP-1 cells). Besides, the conditioned medium from TA-nanoARC nebulized on A549 cells reduced the expression of the CD204/SRA-1, an M2 phenotype marker, and induced pro-inflammatory activity, comparable to or to a greater extent than that induced by lipopolysaccharide, including IL-6 and TNF-α, in mTHP-1 as a model of tumor-associated macrophages. The endocytosis of TA-nanoARC by A549 cells induced Lysotracker red fluorescence to fade and blur. This suggested the internalization of the highly viscous and ordered TA-nanoARC rich in NAs and subsequent lysosomal dysfunction (and not its antioxidant activity), as responsible for the selective damage on A549 cells. These are the first results showing that nebulized TA-nanoARC, lethal to A549 cells and modulating mTHP-1 cell phenotype, may act as antitumorals in the absence of cytotoxic drugs.
嗜盐古菌的膜是新型生物材料的来源，主要具有类异戊二烯性质，其治疗特性几乎尚未被揭示。在此，我们探究了从嗜盐古菌特本奎琴盐红菌中提取的总古菌脂质（TAs）（通过改良的布利特和戴尔技术进行脂质提取第一步得到的一个组分）中存在的中性古菌脂质（NAs，如细菌红素、虾青素和二氢角鲨烯）的抗肿瘤活性，并将其制成TA-纳米古菌脂质体（TA：极性古菌脂质（PAs）：吐温80，5:5:4 ::，TA-纳米ARC）。通过小角X射线散射（SAXS）建模确定，TA-纳米ARC的结构为300.3±84.2纳米，多分散指数为0.59±0.12，ζ电位为-20±3.7毫伏，结果显示与不含NA的双层膜（纳米ARC）相比，NA减小了疏水核心并扩大了亲水部分，同时保持了宽度（约50 Å）和单分子层性。TA-纳米ARC在储存和雾化过程中稳定，48小时后对A549细胞具有细胞毒性，以[细菌红素]表示的半数抑制浓度（IC50）为0.15微克/毫升（约0.20微摩尔），与多西他赛或顺铂的IC50相当或更低。这种细胞毒性在对巨噬细胞（mTHP-1细胞）无害的浓度下发挥作用。此外，在A549细胞上雾化TA-纳米ARC得到的条件培养基降低了M2表型标志物CD204/SRA-1的表达，并诱导了促炎活性，与脂多糖诱导的促炎活性相当或更强，脂多糖诱导的促炎活性包括白细胞介素-6和肿瘤坏死因子-α，这里以mTHP-1作为肿瘤相关巨噬细胞的模型。A549细胞对TA-纳米ARC的内吞作用导致溶酶体示踪剂红色荧光褪色和模糊。这表明富含NA的高粘性且有序的TA-纳米ARC被内化，随后溶酶体功能障碍（而非其抗氧化活性）是对A549细胞选择性损伤的原因。这些是首批结果，表明雾化的TA-纳米ARC对A549细胞具有致死性并能调节mTHP-1细胞表型，在没有细胞毒性药物的情况下可能具有抗肿瘤作用。
