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Article(id=1212693249131724812, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1212693246539649865, articleNumber=1001-2494(2024)19-1781-08, orderNo=null, doi=10.11669/cpj.2024.19.001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1717689600000, receivedDateStr=2024-06-07, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1767058201343, onlineDateStr=2025-12-30, pubDate=1728316800000, pubDateStr=2024-10-08, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1767058201343, onlineIssueDateStr=2025-12-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1767058201343, creator=13701087609, updateTime=1767058201343, updator=13701087609, issue=Issue{id=1212693246539649865, tenantId=1146029695717560320, journalId=1190317699101192196, year='2024', volume='59', issue='19', pageStart='1781', pageEnd='1880', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1767058200723, creator=13701087609, updateTime=1767059042003, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1212696775207600634, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1212693246539649865, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1212696775207600635, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1212693246539649865, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1781, endPage=1788, ext={EN=ArticleExt(id=1212693249542766616, articleId=1212693249131724812, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Advances of Bionic Nano-decoy System in Diseases Treatment, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

As an innovative diseases treatment strategy, bionic nano-decoy system which can neutralize a variety of pathogenic substances has attracted extensive attention in the biomedical field in recent years. Compared with traditional diseases treatment methods, bionic nano-decoy system shows the characteristics of high-efficiency pathogenic molecule clearance, excellent biocompatibility and sustainable or repeated drug delivery, which make it have great application potential in the field of diseases neutralization and treatment. In this paper, the concept, characteristics, preparation technology and application scope of bionic nano-decoy system are reviewed, aiming to provide important reference for researchers and medical professionals in the design and development of new bionic nano-decoy system, in order to promote the development of this field and ultimately achieve clinical application.

, correspAuthors=Qiang ZHANG, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Long YANG, Xueqing WANG, Qiang ZHANG), CN=ArticleExt(id=1212693251153379412, articleId=1212693249131724812, tenantId=1146029695717560320, journalId=1190317699101192196, language=CN, title=仿生纳米诱饵系统在疾病治疗中的研究进展, columnId=1190352408384471863, journalTitle=中国药学杂志, columnName=综述, runingTitle=null, highlight=null, articleAbstract=

能够中和多种致病物质的仿生纳米诱饵系统作为一种创新性的疾病治疗策略,近年来在生物医学领域引起了广泛关注。与传统疾病治疗方法相比,仿生纳米诱饵系统展现出了高效的致病分子清除能力、卓越的生物相容性以及可持续或重复给药的特点,这些特性使其在疾病的中和治疗领域具有巨大的应用潜力。笔者综述了仿生纳米诱饵系统的概念和特点、制备技术以及应用范围,旨在为科研人员和医疗专业人士提供设计和开发新型仿生纳米诱饵系统的重要参考,以期推动该领域的发展并加快实现临床应用。

, correspAuthors=张强, authorNote=null, correspAuthorsNote=
* 张强,男,博士,教授 研究方向:分子药剂学与创新制剂临床转化 Tel: (010)82802791
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杨龙,男,硕士研究生 研究方向:仿生纳米药物递送系统

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杨龙,男,硕士研究生 研究方向:仿生纳米药物递送系统

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Drug Deliv, 28(1): 1109-1119., articleTitle=Recent advances in cell membrane-camouflaged nanoparticles for inflammation therapy, refAbstract=null)], funds=[Fund(id=1212795877471736173, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, awardId=2023YFC2605000, language=CN, fundingSource=国家重点研发计划资助(2023YFC2605000), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1212795873659113674, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, xref=null, ext=[AuthorCompanyExt(id=1212795873667502283, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, companyId=1212795873659113674, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=School of Pharmaceutical Sciences, Peking University, Beijing 100191, China), AuthorCompanyExt(id=1212795873671696588, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, companyId=1212795873659113674, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=北京大学药学院, 北京 100191)])], figs=[ArticleFig(id=1212795875718517024, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=EN, label=null, caption=null, figureFileSmall=L9KlXDfkntXfgZgaa2pdpw==, figureFileBig=lK73bgf22E9vO9RQ05F6tQ==, tableContent=null), ArticleFig(id=1212795875785625895, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=CN, label=图1, caption=常见的仿生纳米诱饵系统(BNDS)类型, figureFileSmall=L9KlXDfkntXfgZgaa2pdpw==, figureFileBig=lK73bgf22E9vO9RQ05F6tQ==, tableContent=null), ArticleFig(id=1212795875903066413, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=EN, label=null, caption=null, figureFileSmall=JqkAlGtdaOs1CZpg0XNN1A==, figureFileBig=MPPuCTVVHRaxaM5HZS1Omw==, tableContent=null), ArticleFig(id=1212795875986952496, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=CN, label=图2, caption=BNDS的常见应用范围, figureFileSmall=JqkAlGtdaOs1CZpg0XNN1A==, figureFileBig=MPPuCTVVHRaxaM5HZS1Omw==, tableContent=null), ArticleFig(id=1212795876066644279, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=EN, label=null, caption=null, figureFileSmall=q2HzGKp2kjje3GUBA3dDSA==, figureFileBig=wNsNEthtX0X8NvjIMFdLmw==, tableContent=null), ArticleFig(id=1212795876158918970, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=CN, label=图3, caption=BNDS在生物医学中的应用[23], figureFileSmall=q2HzGKp2kjje3GUBA3dDSA==, figureFileBig=wNsNEthtX0X8NvjIMFdLmw==, tableContent=null), ArticleFig(id=1212795876263776574, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
方法 纯化机制 优点 缺点 参考文献
差速离心法 尺寸、密度 成本低、应用广泛 含蛋白质聚集体等杂质 [30]
密度梯度离心法 密度 纯度高 操作复杂,成本高 [31]
超滤法 大小 快速 EV会吸附到膜上而损失 [32]
切向流过滤法 大小 快速、EV吸附少 设备复杂 [33]
聚合物沉淀法 溶解度、表面电荷 高通量 可能引入聚合物杂质 [34]
尺寸排阻色谱法 大小 纯度高、所需样品量少 产量低、蛋白质污染 [35]
离子交换色谱法 表面电荷 纯度高 特异性低 [36]
相分离法 物理性质 快速廉价 EV膜完整性降低 [37]
微流控法 大小、密度 快速高效 成本高、样品少 [38]
亲和捕获法 表面生物标志物 特异性强 产量低、特异性抗体/适配体昂贵 [39]
), ArticleFig(id=1212795876385411394, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=CN, label=表1, caption=

胞外囊泡(EV)的分离纯化方法

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方法 纯化机制 优点 缺点 参考文献
差速离心法 尺寸、密度 成本低、应用广泛 含蛋白质聚集体等杂质 [30]
密度梯度离心法 密度 纯度高 操作复杂,成本高 [31]
超滤法 大小 快速 EV会吸附到膜上而损失 [32]
切向流过滤法 大小 快速、EV吸附少 设备复杂 [33]
聚合物沉淀法 溶解度、表面电荷 高通量 可能引入聚合物杂质 [34]
尺寸排阻色谱法 大小 纯度高、所需样品量少 产量低、蛋白质污染 [35]
离子交换色谱法 表面电荷 纯度高 特异性低 [36]
相分离法 物理性质 快速廉价 EV膜完整性降低 [37]
微流控法 大小、密度 快速高效 成本高、样品少 [38]
亲和捕获法 表面生物标志物 特异性强 产量低、特异性抗体/适配体昂贵 [39]
), ArticleFig(id=1212795876523823430, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
方法 原理 优点 缺点 参考文献
挤出法 机械力破碎膜、热力学自组装 操作简单 批次间差异大 [41]
冻融循环法 细胞内部形成冰晶破碎膜、热力学自组装 成本低 粒径均一性差 [42-44]
超声法 超声波破碎膜、热力学自组装 快速 不适合大规模生产 [45]
化学诱导法 化学诱导细胞膜起泡 操作简单 膜蛋白活性降低 [46]
氮气空化法 高压破碎膜、热力学自组装 操作简单 批次间差异大 [47]
微流体切割法 物理切割破碎膜、热力学自组装 高通量 装置复杂 [48]
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细胞衍生纳米囊泡的制备方法

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方法 原理 优点 缺点 参考文献
挤出法 机械力破碎膜、热力学自组装 操作简单 批次间差异大 [41]
冻融循环法 细胞内部形成冰晶破碎膜、热力学自组装 成本低 粒径均一性差 [42-44]
超声法 超声波破碎膜、热力学自组装 快速 不适合大规模生产 [45]
化学诱导法 化学诱导细胞膜起泡 操作简单 膜蛋白活性降低 [46]
氮气空化法 高压破碎膜、热力学自组装 操作简单 批次间差异大 [47]
微流体切割法 物理切割破碎膜、热力学自组装 高通量 装置复杂 [48]
), ArticleFig(id=1212795876733538637, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
方法 原理 优点 缺点 参考文献
挤出法 机械力涂覆 操作简单 难以放大生产 [51]
细胞摄取-分泌法 通过EV涂覆 无需考虑纳米颗粒性质 均一性低 [52]
超声处理法 静电相互作用 操作简单 均匀性差 [53]
微流控电穿孔法 电穿孔 均一性高 设备复杂 [54]
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细胞膜涂覆纳米核心的常用方法

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方法 原理 优点 缺点 参考文献
挤出法 机械力涂覆 操作简单 难以放大生产 [51]
细胞摄取-分泌法 通过EV涂覆 无需考虑纳米颗粒性质 均一性低 [52]
超声处理法 静电相互作用 操作简单 均匀性差 [53]
微流控电穿孔法 电穿孔 均一性高 设备复杂 [54]
), ArticleFig(id=1212795876939059541, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
常用技术 表征内容 原理 参考文献
透射电子显微镜(TEM) 大小和形貌 样品各处紧密性差异 [57]
动态光散射(DLS) 大小 光学 [58]
纳米颗粒追踪分析(NTA) 大小和颗粒浓度 光学 [59]
纳米库尔特(Nano coulter) 大小和颗粒浓度 电学 [57]
原子力显微镜(AFM) 尺寸形状、膜刚度 原子间相互作用力 [60]
), ArticleFig(id=1212795877039722842, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=CN, label=表4, caption=

表征仿生纳米诱饵系统(BNDS)的常用技术

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常用技术 表征内容 原理 参考文献
透射电子显微镜(TEM) 大小和形貌 样品各处紧密性差异 [57]
动态光散射(DLS) 大小 光学 [58]
纳米颗粒追踪分析(NTA) 大小和颗粒浓度 光学 [59]
纳米库尔特(Nano coulter) 大小和颗粒浓度 电学 [57]
原子力显微镜(AFM) 尺寸形状、膜刚度 原子间相互作用力 [60]
), ArticleFig(id=1212795877131997533, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212693249131724812, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
诱饵名称 亲本细胞 用途 疾病
TNF-αR[3,49,64-65] 免疫细胞;HEK 293T细胞 中和促炎细胞因子 炎症
IL-1βR[3,49,64-65] 免疫细胞;HEK 293T细胞 中和促炎细胞因子 炎症
IL-6R[3,49,64-65] 免疫细胞;HEK 293T细胞 中和促炎细胞因子 炎症
CXCR4[20] MSC 中和炎症驱动因子 炎症
PD-1[24] HEK 293T细胞;Raw264.7细胞 阻断PD-1/PD-L1免疫抑制轴 癌症
AchE[8] 红细胞 中和有机磷 有机磷中毒
ACE2[66] HEK 293T细胞;Vero-E6细胞 阻断病毒侵入细胞 病毒感染
硫酸乙酰肝素[67] 红细胞 阻断疟原虫侵入细胞 疟疾
血小板抗原[68] 血小板 中和抗血小板抗体 血小板减少症
CCR2[69] 巨噬细胞 中和趋化因子CCL2 脊髓损伤
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BNDS常见的诱饵及其用途

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诱饵名称 亲本细胞 用途 疾病
TNF-αR[3,49,64-65] 免疫细胞;HEK 293T细胞 中和促炎细胞因子 炎症
IL-1βR[3,49,64-65] 免疫细胞;HEK 293T细胞 中和促炎细胞因子 炎症
IL-6R[3,49,64-65] 免疫细胞;HEK 293T细胞 中和促炎细胞因子 炎症
CXCR4[20] MSC 中和炎症驱动因子 炎症
PD-1[24] HEK 293T细胞;Raw264.7细胞 阻断PD-1/PD-L1免疫抑制轴 癌症
AchE[8] 红细胞 中和有机磷 有机磷中毒
ACE2[66] HEK 293T细胞;Vero-E6细胞 阻断病毒侵入细胞 病毒感染
硫酸乙酰肝素[67] 红细胞 阻断疟原虫侵入细胞 疟疾
血小板抗原[68] 血小板 中和抗血小板抗体 血小板减少症
CCR2[69] 巨噬细胞 中和趋化因子CCL2 脊髓损伤
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仿生纳米诱饵系统在疾病治疗中的研究进展
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杨龙 , 王学清 , 张强 *
中国药学杂志 | 综述 2024,59(19): 1781-1788
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中国药学杂志 | 综述 2024, 59(19): 1781-1788
仿生纳米诱饵系统在疾病治疗中的研究进展
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杨龙, 王学清, 张强*
作者信息
  • 北京大学药学院, 北京 100191

    • 杨龙,男,硕士研究生 研究方向:仿生纳米药物递送系统

通讯作者:

* 张强,男,博士,教授 研究方向:分子药剂学与创新制剂临床转化 Tel: (010)82802791
Advances of Bionic Nano-decoy System in Diseases Treatment
Long YANG, Xueqing WANG, Qiang ZHANG*
Affiliations
  • School of Pharmaceutical Sciences, Peking University, Beijing 100191, China
出版时间: 2024-10-08 doi: 10.11669/cpj.2024.19.001
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摘要
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能够中和多种致病物质的仿生纳米诱饵系统作为一种创新性的疾病治疗策略,近年来在生物医学领域引起了广泛关注。与传统疾病治疗方法相比,仿生纳米诱饵系统展现出了高效的致病分子清除能力、卓越的生物相容性以及可持续或重复给药的特点,这些特性使其在疾病的中和治疗领域具有巨大的应用潜力。笔者综述了仿生纳米诱饵系统的概念和特点、制备技术以及应用范围,旨在为科研人员和医疗专业人士提供设计和开发新型仿生纳米诱饵系统的重要参考,以期推动该领域的发展并加快实现临床应用。

中和  /  仿生  /  纳米诱饵  /  致病物质  /  囊泡

As an innovative diseases treatment strategy, bionic nano-decoy system which can neutralize a variety of pathogenic substances has attracted extensive attention in the biomedical field in recent years. Compared with traditional diseases treatment methods, bionic nano-decoy system shows the characteristics of high-efficiency pathogenic molecule clearance, excellent biocompatibility and sustainable or repeated drug delivery, which make it have great application potential in the field of diseases neutralization and treatment. In this paper, the concept, characteristics, preparation technology and application scope of bionic nano-decoy system are reviewed, aiming to provide important reference for researchers and medical professionals in the design and development of new bionic nano-decoy system, in order to promote the development of this field and ultimately achieve clinical application.

neutralization  /  bionics  /  nano-decoy  /  pathogenic substance  /  vesicle
杨龙, 王学清, 张强. 仿生纳米诱饵系统在疾病治疗中的研究进展. 中国药学杂志, 2024 , 59 (19) : 1781 -1788 . DOI: 10.11669/cpj.2024.19.001
Long YANG, Xueqing WANG, Qiang ZHANG. Advances of Bionic Nano-decoy System in Diseases Treatment[J]. Chinese Pharmaceutical Journal, 2024 , 59 (19) : 1781 -1788 . DOI: 10.11669/cpj.2024.19.001
正文
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中和理念在自身免疫性疾病和感染性疾病等多种疾病治疗中扮演着关键角色。单克隆抗体疗法,一种通过中和致病物质来治疗疾病的手段,已经在多种疾病的治疗中获得批准,并显示出显著的疗效[1]。然而,单抗疗法也面临着挑战,包括抗药抗体的产生、对全身免疫系统的干扰以及抗原靶点的突变,这些因素能导致治疗无效甚至引发严重的毒副作用。为了克服这些挑战,纳米诱饵系统作为一种有前景的替代方案应运而生。该系统通过在纳米颗粒表面模拟宿主细胞的受体或配体,吸引致病物质并与之结合,从而有效治疗疾病。特别的是,基于宿主细胞天然膜成分的仿生纳米诱饵系统,因其低毒性和低免疫原性等独特优势,正受到越来越多的关注[2]。本研究将重点介绍仿生纳米诱饵系统的概念和发展历程,并综合评述其特点、制备方法以及应用范围,旨在为未来相关领域的研究和应用提供参考和启示。
1 仿生纳米诱饵系统的概念和发展历程
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仿生纳米诱饵系统(bionic nano-decoy system, BNDS)中的“仿生”意为模仿生物体;“纳米诱饵”是指用纳米技术递送致病物质的诱饵。该系统将由天然细胞膜衍生的纳米级别囊泡或由天然膜涂覆的纳米颗粒作为载体,将膜上的特定成分(如受体蛋白)作为诱饵,用于“欺骗”并清除致病物质。同时,膜上的其他内源性成分有助于躲避机体网状内皮系统的识别和清除。
BNDS最早可追溯到2011年,Zhang等[2]首次报道了细胞膜仿生技术,该团队采用自上而下的仿生方法,通过用天然红细胞膜包覆可生物降解的聚合物纳米颗粒,能显著延长纳米颗粒的循环半衰期。后来,细胞膜仿生技术在中和领域被广泛研究,已被证明可以将膜上的天然受体或配体作为诱饵,特异性地吸引病原体、病理性蛋白质和化学毒素等致病物质与其结合,在多种疾病模型中观察到了很好的治疗效果[3]。
尽管目前尚无基于BNDS的药物上市,仅有部分研究被FDA获批进入临床前研究[4-6],但可以预见的是,BNDS利用数百万年来生物进化筛选出的特异性相互作用,规避了合成传统中和药物所需要的靶点发现和疗效验证过程,有可能彻底改变疾病治疗的格局[7]。
2 BNDS的特点
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2.1 起效快
BNDS可以直接与致病物质结合,通过物理方式阻止其与宿主细胞的结合,这种直接的物理阻断作用可以迅速减少致病信号的持续传递。
以治疗有机磷中毒为例,常见于杀虫剂和神经毒剂中,可引起乙酰胆碱酯酶 (acetylcholine esterase, AChE) 不可逆的磷酸化和失活,进而导致乙酰胆碱在体内积累,最终导致神经肌肉疾病[8]。传统的药物治疗有机磷中毒主要是通过激活AChE及消除过量乙酰胆碱发挥作用,无法清除体内的有机磷[8]。而BNDS可通过表面携带的天然AChE直接从源头上中和清除有机磷,具备比传统药物更迅速且显著的疗效[8]。
2.2 不良反应少
全身性分布增加了单克隆抗体等传统中和药物剂量限制的毒副作用。相比之下,BNDS表面的成分与宿主细胞膜相似,因此不太可能被免疫系统识别为外来物质,免疫原性较低,具备长期治疗的潜力[9]。
以抗肿瘤为例,肿瘤细胞膜涂覆的纳米颗粒具有优异的同源靶向性,能够依靠半乳糖凝集素-3分子与癌细胞表面的癌胚抗原相互作用,有效靶向癌症部位,显著降低了载体在正常组织中的分布,增加在肿瘤部位的蓄积[10-11]。
2.3 适应性强
传统中和策略需针对致病物质结构进行特异性设计,往往需要经历靶点筛选-中和分子制备-递送系统设计等过程。此外,部分致病物质并没有有效的药理学中和剂,且存在中和靶点突变或耐受等问题,大大增加了治疗的难度[12]。BNDS提供了一种有前景的疾病中和治疗方案,能够依赖表面的天然诱饵分子吸引多种致病物质与其结合,无须考虑致病物质的结构特点。
以治疗严重急性呼吸系统综合征冠状病毒2(SARS-CoV-2)感染为例,尽管单克隆抗体等中和药物对阻断原型病毒感染有效,但对病毒变体的治疗往往无效,需针对变体结构重新设计[13]。由于SARS-CoV和SARS相关冠状病毒(SARSr-CoV)都依赖宿主细胞表面的血管紧张素转化酶2(angiotensin converting enzyme 2, ACE2)受体进入宿主细胞,因此表面携带ACE2受体的BNDS可以有效中和原型病毒及其多种变体,无须考虑变体的结构变化,适应性更强[14]。
2.4 中和效率高
中和一种或有限几种致病物质有时并不足以完全逆转疾病进展,BNDS具备与宿主细胞相似的结合多种致病物质的能力,因此中和效率更高[3]。
Zhuang等[15]用巨噬细胞膜包被了尿酸酶负载的金属有机框架(MΦ-MOF-尿酸酶),在高尿酸血症小鼠模型中静脉给药后,小鼠尿酸迅速降低至基础水平。此外,在尿酸钠晶体诱导的关节炎小鼠模型上,关节内注射MΦ-MOF-尿酸酶后炎症部位的IL-1β/IL-6/TNF-α等多种促炎细胞因子被有效中和,这归因于巨噬细胞膜上丰富的细胞因子受体。
2.5 跨血-脑脊液屏障能力强
跨血-脑脊液屏障(blood-brain barrier,BBB)运送药物和其他活性物质是生物医学面临的主要技术挑战之一,BNDS提供了一种可行的策略,在脑部疾病治疗中被广泛应用[16-17]。
间充质干细胞(mesenchymal stem cell, MSC)等细胞膜上存在整合素α4β1等黏附分子,能够与血管内皮细胞表面的VCAM-1相互作用从而跨过BBB[18]。有研究表明,由MSC分泌的胞外囊泡也能够通过表面的MSC同源分子与BBB细胞表面蛋白相互作用,最终能够有效跨过BBB[19]。Shi等[20]用过表达趋化因子受体CXCR4的间充质干细胞膜涂覆聚多巴胺纳米球,静脉给药后显著改善了纳米颗粒对脑缺血病变的靶向。同时CXCR4作为诱饵,有效吸附和中和了病变部位的炎症驱动因子CXCL12,切断了外周中性粒细胞和单核巨噬细胞的浸润。
3 BNDS中诱饵的分类
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BNDS中用于“欺骗”致病物质的诱饵主要来自细胞天然表达的配体或受体、细胞应激条件下表达的配体或受体以及通过基因工程手段在细胞水平表达的配体或受体等。
3.1 细胞天然表达的配体或受体
多数致病物质通过与免疫细胞等类型的细胞膜上天然表达的丰富配体或受体相互作用促进疾病发生和发展[21]。通过这些细胞衍生的BNDS能够模拟亲本细胞,吸引并结合致病物质。如Toll样受体家族(toll-like receptors, TLR)是广泛存在于免疫细胞表面的一类受体,能够识别结合多种病原体及其分泌的毒素,随后触发细胞内信号转导[22]。有研究表明,由免疫细胞膜包被的纳米颗粒能够通过表面的TLR-4中和脂多糖(lipopolysaccharide, LPS),显著提高了由LPS诱导的内毒素血症小鼠生存率[23]。
3.2 细胞应激条件下表达的配体或受体
部分细胞在施加刺激后也会表达丰富的配体或受体。Hou等[24]使用IFN-γ刺激Raw264.7细胞后,Raw264.7细胞表达了丰富的PD-L1、血管内皮黏附分子配体和多种细胞因子受体,在关节炎和溃疡性结肠炎模型小鼠上验证了IFN-γ激活的Raw264.7细胞膜涂覆的聚乳酸-羟基乙酸共聚物[poly(lactic-co-glycolic acid), PLGA]纳米颗粒能够作为诱饵清除多种促炎细胞因子,并通过程序性细胞死亡配体1(programmed cell death ligand 1,PD-L1)中和游离的程序性细胞死亡蛋白1(programmed cell death protein 1, PD-1),有效介导了免疫抑制。
3.3 通过基因工程手段在细胞水平表达的配体或受体
以上由细胞天然表达或应激条件下表达在膜上的诱饵能够最大程度地保留天然特性,但膜上的其他成分可能会带来安全性问题。因此也有研究通过基因工程手段在FDA批准的工具细胞(如HEK 293T细胞)上过表达诱饵分子,具有更高的安全性。
Zhang等[25]在HEK 293T细胞水平过表达了PD-1蛋白,由此细胞制备的纳米囊泡表面携带PD-1作为诱饵,通过结合肿瘤细胞表面的PD-L1,有效破坏了PD-1/PD-L1免疫抑制轴,增强了癌症免疫治疗效果。
为了增加诱饵分子“欺骗”致病物质的能力,可将天然膜上的诱饵结构进行适当改造,或与一些膜相关蛋白串联表达。Gupta等[26]对TNF-α受体(TNF-αR)的序列进行了优化设计,并在HEK 293T细胞上过表达了TNF-αR与细胞内腔蛋白syntenin-1的融合重组蛋白,由该基因工程细胞分泌的胞外囊泡在溃疡性结肠炎、神经炎症以及全身炎症等多种炎症模型小鼠上观察到很好的抗炎效果。
此外,基因工程手段所表达出的诱饵往往仅针对单一致病物质,通过将不同的诱饵在同一亲本细胞共表达,或将分别表达了不同诱饵的亲本细胞膜进行膜融合,可以制备具有广谱中和能力的BNDS。
肿瘤细胞会过表达CD47和PD-L1,其中PD-L1结合T细胞表面的PD-1以阻断T细胞的免疫激活;CD47结合巨噬细胞表面的信号调节蛋白α(SIRPα)阻断巨噬细胞吞噬。Zhang等[27]将分别通过基因工程过表达SIRPα和PD-1的2种细胞膜融合,由此制备的BNDS可有效阻断肿瘤细胞表面的先天检查点 CD47 和适应性检查点PD-L1。此外,为了使单个纳米诱饵上PD-1和SIRPα的分布更均匀,提高免疫检查点的阻断效率,也可以在同一亲本细胞水平过表达SIRPα和PD-1的融合重组蛋白[27]。
4 BNDS的制备方法
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目前,BNDS主要分为细胞主动分泌的胞外囊泡(extracellular vesicles,EV)、人工制备的细胞衍生纳米囊泡以及具备核心的细胞膜涂覆纳米颗粒三大类(图1)。下面将详细介绍不同类型BNDS的制备方法。
4.1 细胞主动分泌的EV
EV是由细胞主动向外分泌的粒径30~1 000 nm内的膜囊泡,负责细胞间的信号传递等功能。通过对亲本细胞的人工改造可获得工程化的EV,使其表面携带宿主细胞上的天然受体用于吸引病毒、促炎细胞因子和细菌毒素等致病物质[26,28-29]。
通过超速离心、密度梯度离心、沉淀、尺寸排阻色谱、场流分级、微流控及免疫吸附等手段可分离纯化细胞培养液中的EV,表1总结了用于分离纯化EV的常见方法。实际应用时可考虑将不同方法组合使用,以提高EV的分离纯化效率。
Wang等[14]通过切向流过滤和超滤相结合的方法纯化了由人肺球状细胞分泌的EV,在体内外实验中观察到该EV能通过携带的ACE2有效吸引病毒表面的S蛋白,阻断病毒入侵宿主细胞。
Duong等[40]通过聚合物沉淀方法从稳定表达TNF-αR-CD63融合蛋白的HEK 293T细胞条件培养基中分离出表面携带TNF-αR的EV,在体外观察到该EV能够有效阻断TNF-α诱导的促炎信号传导。
4.2 人工制备的细胞衍生纳米囊泡
人工制备的细胞衍生纳米囊泡是指通过挤出、冻融循环、超声、化学诱导、氮气空化、微流控破碎等手段处理亲本细胞后制备出的类EV,在继承EV诸多优势的同时大大提高了产量。表2总结了常见的生产细胞膜囊泡的方法。
Xiao等[49]通过连续挤出中性粒细胞膜的方式获得了表面携带IL-1βR、TNF-αR等多种促炎细胞因子受体的纳米囊泡,该纳米囊泡在脓毒症小鼠模型中给药后表现出出色的生物相容性,显著降低了炎症细胞因子和肝损伤生物标志物的血浆水平,最终提高了脓毒症小鼠的存活率。
Rao等[50]开发了一种用于治疗病毒感染的细胞衍生纳米诱饵,通过超声处理和连续挤出亲本细胞膜相结合的方法制备出了表面携带丰富ACE2和细胞因子受体的纳米诱饵,能够同时中和COVID-19患者体内的冠状病毒和炎性细胞因子,显著抑制病毒复制和感染。
4.3 细胞膜涂覆纳米颗粒
细胞膜涂覆的纳米颗粒是指通过挤出、超声、微流控等方式获得的表面涂覆了细胞膜的纳米颗粒。与前面2种囊泡类的BNDS相比,纳米颗粒核心有助于脂质膜及表面诱饵分子在体内的稳定,不易与下游细胞发生膜融合,因此不会将吸引的病原体或毒素转移到宿主细胞[13]。表3总结了细胞膜涂覆纳米核心的常见方法。
Zhang等[55]用红细胞膜包裹橄榄油纳米液滴,通过红细胞膜上受体的特异性结合和纳米油滴核心的非特异性吸收,有效中和了有机磷中毒模型小鼠组织中的有机磷,显著减轻了有机磷中毒的临床症状,大大提高了小鼠存活率。
但由于传统涂覆过程中膜组分的随机重组导致了膜涂覆纳米颗粒上膜蛋白的空间无序,降低了中和效率[56]。因此仍需开发新的涂覆技术,增加涂覆工艺的稳定性。
5 BNDS的表征方法
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5.1 对BNDS的整体表征
目前尚未建立一种标准化的方法用来全面评估BNDS。多数研究主要表征BNDS的大小和形貌、数量、表面特性、蛋白质含量及其他纳米分子特征。
大小和形貌是BNDS最重要的特征之一,透射电子显微镜(transmission electron microscope, TEM)、动态光散射(dynamic laser scattering, DLS)、纳米颗粒跟踪分析(nanoparticle tracking analysis, NTA)、原子力显微镜(atomic force microscope, AFM)和纳米库尔特技术(nano coulter)在内的多种技术可用于测量BNDS的大小和形貌(表4)。值得注意的是,由于每种方法的原理不完全相同,所以对BNDS的表征结果往往存在一定差异,通常需要相互结合使用。
BNDS的非诱饵成分在其生物学功能中可能起着关键作用,分析这些成分对于了解BNDS的分子组成及功能至关重要。蛋白质免疫印迹、液相色谱-质谱法(LC-MS)、实时定量PCR(qPCR)等各种方法通常用于分析BNDS的脂质、蛋白质及核酸等组成成分[58,60]。
除此之外,对BNDS的细胞摄取、生物分布及等参数的表征也至关重要。通过免疫荧光和流式细胞术等手段[61]可确定BNDS的细胞摄取情况,通过活体成像[14]或组织切片染色[62]可确定BNDS的体内命运。
5.2 对BNDS中诱饵的表征
为了中和具体的致病物质,往往需要评价BNDS中诱饵的负载效率和诱饵的生物活性。
通过免疫荧光、酶联免疫分析(ELISA)、蛋白免疫印迹、纳米流式细胞术及基于磁珠的流式细胞术等手段可以确定BNDS上是否加载了足够的诱饵分子[50],结合BNDS的颗粒浓度可以进一步确定单个纳米诱饵上的平均诱饵丰度。通过生物层干涉、免疫金标记等分析相互作用的技术可以在体外确定诱饵的生物活性[14,50,63]。
6 BNDS的应用
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几乎所有依赖宿主细胞膜上受体发挥作用的致病物质都可以通过BNDS进行中和清除。目前,BNDS已被广泛应用于中和病原体及其分泌的毒素、病理性蛋白质和毒性化学物质等多种致病物质。图2和表5总结了BNDS常见的应用及相应的诱饵分子类型。
6.1 中和病原体及其分泌的毒素
多数细菌、病毒、寄生虫等病原体及其分泌的毒素依赖细胞膜表面的特定受体或其他膜成分发挥功能,因此可通过BNDS进行中和清除。
Carvalho等[70]发现,CD4+T细胞释放的EV具有与亲本T细胞类似的复杂表面蛋白组成,可以在体外以浓度依赖性方式中和HIV-1,降低病毒对CD4+T细胞的感染。基于红细胞膜的BNDS能够有效中和疟疾裂殖子和α-溶血素等细菌毒素,降低病原体对宿主的侵染[69]。且与传统抗生素疗法相比,BNDS伪装成宿主细胞,不太容易导致病原体耐药,在中和病原体及其分泌的毒素应用中具有广泛的应用前景。
6.2 中和病理性蛋白质
炎症性疾病环境中的促炎细胞因子、血小板减少症中的抗血小板抗体及肿瘤细胞表面的PD-L1等病理性蛋白质通过结合下游宿主细胞上的受体或配体发挥功能,促进疾病的发生发展。BNDS可有效吸引这些病理性蛋白质,在这些疾病的治疗中显示出巨大的潜力[3]。
基于免疫细胞、干细胞和上皮细胞等亲本细胞的BNDS在抗炎、病毒中和及阻断PD-L1恢复T细胞免疫等方面发挥了重要作用,能够降低病理性蛋白质对正常细胞的干扰,阻止疾病的发生发展,这主要依赖于BNDS表面丰富的特异性受体或配体[3,14,49,64-66,71-72]。
6.3 中和毒性化学物质
毒性化学物质的多样性使针对结构进行特异性设计的传统治疗理念面临着极大挑战性和低效性[73]。目前大多数毒性化学物质在临床上并没有安全且有效的中和剂,只能针对下游靶点进行治疗[8]。而研究人员开发的BNDS在中和毒性化学物质方面已经取得了显著的解毒效果。
敌敌畏(DDVP)等有机化合物中毒的常用解毒剂往往会导致严重的副作用[74]。Pang等[8]使用红细胞膜涂覆的纳米颗粒(RBC-NPs)保留了膜上的AChE及其活性,能够以剂量依赖性方式结合DDVP,在静脉注射或口服致死剂量DDVP的小鼠中显著提高了小鼠的存活率。
河鲀毒素等动物毒素中毒也可以通过BNDS进行治疗。Wang等[73]基于小鼠神经嵴衍生的神经元细胞膜设计出了一种BNDS,在体外验证了神经元膜涂覆的PLGA纳米粒可以充当河鲀毒素的诱饵,有效地避免了河鲀毒素攻击神经元细胞。
BNDS还可以用于中和一些小分子化疗药物。血小板减少症是与小分子化疗药物ABT-263临床使用相关的主要不良反应,表面涂覆线粒体膜的聚合物纳米粒通过表面的B细胞淋巴瘤蛋白2(B-cell lymphoma protein 2, Bcl-2)结合ABT-263,能够在体外有效抑制ABT-263诱导的细胞死亡和凋亡,并在体内减轻ABT-263诱导的血小板减少症[75]。
6.4 BNDS在生物医学领域的其他应用
BNDS除了“欺骗”并中和致病物质外,还可用于生物成像及疫苗接种等多个领域(图3)[23]。
癌症的早期筛查对提高患者的治愈率至关重要,叶酸修饰的红细胞膜涂覆纳米颗粒可特异性结合肿瘤细胞,显著放大早期肿瘤部位的信号[76]。
肿瘤细胞膜表面具有多种特异性抗原,因此可用于肿瘤疫苗增强机体免疫反应[77]。Johnson等[78]报告了一种急性髓系白血病(acute myeloid leukemia, AML)细胞膜包被的负载佐剂的PLGA纳米颗粒作为疫苗,具备多抗原、个性化、抗原佐剂共定位等特点,在激活 AML 特异性免疫反应和提供长期抗白血病生存获益等方面的效果显著优于全细胞裂解物疫苗。
总的来说,BNDS具有优异的生物模拟能力,是一种极具潜力的技术,有望在生物医学的多个研究领域中发挥越来越重要的作用[23]。
7 总结与展望
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与传统的中和方式相比,BNDS在炎症性疾病、病原体感染及化学物质中毒等多种类型的疾病模型中都具有显著的治疗效果,提供了一种有前景的治疗策略。然而,BNDS中非活性成分带来的不必要的生物学效应、不成熟的生产技术、缺乏统一的表征标准、缺乏优化的细胞膜提取技术以及长期储存问题等因素大大限制了BNDS的临床转化,通过在亲本细胞水平基因敲除潜在的风险分子、优化亲本细胞膜来源以及改进膜提取和涂覆方法将有助于推进BNDS的下一步应用[79-81]。相信随着生物技术的不断发展,BNDS在临床转化方面将取得突破性进展。
基金
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  • 国家重点研发计划资助(2023YFC2605000)
文献
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参考文献 引证文献
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2024年第59卷第19期
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doi: 10.11669/cpj.2024.19.001
  • 接收时间：2024-06-07
  • 首发时间：2025-12-30
  • 出版时间：2024-10-08
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  • 收稿日期：2024-06-07
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国家重点研发计划资助(2023YFC2605000)
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    北京大学药学院, 北京 100191

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* 张强,男,博士,教授 研究方向:分子药剂学与创新制剂临床转化 Tel: (010)82802791
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2种不同金属材料的力学参数
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Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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