在黑色素瘤微环境中,通过miR-150-3p共传递,工程纳米囊泡平台同时触发yap依赖性铁凋亡和重编程t细胞免疫。

IF 13.3 1区 医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL
Theranostics Pub Date : 2025-07-25 eCollection Date: 2025-01-01 DOI:10.7150/thno.115860
Jiemin Wang, Zhenguo Zhao, Haopeng Yang, Ruixuan Wang, Shu Wang, Jiale Yu, Yujia Wang, Ruihua Liu, Yani Chen, Yueshi Liu, Kesong Shi, Pengyong Han, Miao Liu, Jing Miao, Xiaoyang Li, Xiangnan Li, Haiquan Yu
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引用次数: 0

摘要

理由:黑色素瘤仍然是一种高度侵袭性的恶性肿瘤,有效的治疗方法有限,并且经常对免疫检查点封锁(ICB)产生耐药性。细胞外囊泡(EVs)是基于rna的治疗方法的一个很有前途的平台,但它们的临床转化受到低效率的货物装载和肿瘤特异性靶向不足的阻碍。为了解决这些局限性,我们开发了一种工程化的EV策略,将有效的miRNA包装与肿瘤靶向表面修饰结合起来,以提高黑色素瘤的治疗效果。方法:在HEK293T细胞中共表达miR-150-3p和Annexin A2 (ANXA2),然后用靶向肿瘤的iRGD肽对其表面进行修饰,从而产生工程化ev (ev -150)。通过RNA测序、RNA免疫沉淀(RIP)、染色质免疫沉淀(ChIP)和荧光素酶报告基因测定获得机制见解。通过脂质过氧化分析、线粒体膜电位测定和透射电子显微镜(TEM)来评估铁下垂诱导。使用皮下和转移性黑色素瘤小鼠模型评估体内治疗效果和生物分布。通过流式细胞术分析患者来源的CD8 + T细胞和用ev -150处理的黑色素瘤细胞共培养中的CD8 + T细胞活化,研究了免疫调节。结果:miR-150-3p在黑色素瘤源性ev中升高,ANXA2被鉴定为关键的rna结合蛋白,选择性地促进其装载到ev中。ev -150在体内被黑色素瘤细胞吸收增强,并改善肿瘤特异性积累。在机制上,iEV-150抑制NF2表达,破坏NF2- lats1相互作用,激活YAP信号,随后上调铁凋亡相关基因ACSL4和CHAC1,从而通过NF2- hippo -YAP轴诱导铁凋亡。除了具有直接的抗肿瘤作用外,ev -150还能促进CD8 + T细胞在肿瘤微环境内的浸润和活化,显著增强ICB在黑色素瘤模型中的治疗效果。结论:iEV-150整合了anxa2介导的miRNA装载、肿瘤特异性靶向、铁凋亡诱导和免疫微环境重编程。这种工程化的EV策略提供了一种有效的基于rna的治疗平台,可以克服ICB耐药性,提高黑色素瘤的精准治疗。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Engineered nanovesicle platform simultaneously triggers YAP-dependent ferroptosis and reprograms T-cell immunity through miR-150-3p codelivery in melanoma microenvironment.

Rationale: Melanoma remains a highly aggressive malignancy with limited effective therapies and frequent resistance to immune checkpoint blockade (ICB). Extracellular vesicles (EVs) represent a promising platform for RNA-based therapeutics, but their clinical translation is impeded by inefficient cargo loading and insufficient tumor-specific targeting. To address these limitations, we developed an engineered EV strategy integrating efficient miRNA packaging with tumor-targeting surface modifications to enhance therapeutic outcomes in melanoma. Methods: Engineered EVs (iEV-150) were generated by co-expressing miR-150-3p and Annexin A2 (ANXA2) in HEK293T cells, followed by surface modification with tumor-targeting iRGD peptides. Mechanistic insights were obtained using RNA sequencing, RNA immunoprecipitation (RIP), chromatin immunoprecipitation (ChIP), and luciferase reporter assays. Ferroptosis induction was evaluated through lipid peroxidation analysis, mitochondrial membrane potential assays, and transmission electron microscopy (TEM). Therapeutic efficacy and biodistribution were assessed in vivo using subcutaneous and metastatic melanoma mouse models. Immune modulation was examined by analyzing CD8⁺ T cell activation via flow cytometry in co-cultures of patient-derived CD8⁺ T cells and melanoma cells treated with iEV-150. Results: miR-150-3p was elevated in melanoma-derived EVs, and ANXA2 was identified as a key RNA-binding protein that selectively facilitated its loading into EVs. iEV-150 exhibited enhanced uptake by melanoma cells and improved tumor-specific accumulation in vivo. Mechanistically, iEV-150 suppressed NF2 expression, disrupted the NF2-LATS1 interaction, activated YAP signaling, and subsequently upregulated ferroptosis-related genes ACSL4 and CHAC1, thereby inducing ferroptosis through the NF2-Hippo-YAP axis. In addition to its direct anti-tumor effects, iEV-150 promoted CD8⁺ T cell infiltration and activation within the tumor microenvironment, and significantly enhanced the therapeutic efficacy of ICB in melanoma models. Conclusions: iEV-150 integrates ANXA2-mediated miRNA loading, tumor-specific targeting, ferroptosis induction, and immune microenvironment reprogramming. This engineered EV strategy provides an effective RNA-based therapeutic platform to overcome ICB resistance and enhance precision treatment in melanoma.

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来源期刊
Theranostics
Theranostics MEDICINE, RESEARCH & EXPERIMENTAL-
CiteScore
25.40
自引率
1.60%
发文量
433
审稿时长
1 months
期刊介绍: Theranostics serves as a pivotal platform for the exchange of clinical and scientific insights within the diagnostic and therapeutic molecular and nanomedicine community, along with allied professions engaged in integrating molecular imaging and therapy. As a multidisciplinary journal, Theranostics showcases innovative research articles spanning fields such as in vitro diagnostics and prognostics, in vivo molecular imaging, molecular therapeutics, image-guided therapy, biosensor technology, nanobiosensors, bioelectronics, system biology, translational medicine, point-of-care applications, and personalized medicine. Encouraging a broad spectrum of biomedical research with potential theranostic applications, the journal rigorously peer-reviews primary research, alongside publishing reviews, news, and commentary that aim to bridge the gap between the laboratory, clinic, and biotechnology industries.
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