A Convergent Bioengineered Platform for Multifunctional Therapeutic Exosomes

多功能治疗性外切体的融合生物工程平台 摘要：Mira应用程序的总体目标是开发一个融合的生物工程平台 制造和设计治疗用外切体。该平台将允许将药物装载到外切体中 高效、高通量地生物制造外切体，并进一步工程化外切体为基础 用于各种疾病的药物输送系统，具有预期的功能，包括靶向输送、跟踪和 综合疗法。胞外体是细胞外小泡的一个子集，直径在50纳米到150纳米之间。 NM，由大多数真核细胞分泌。由于体积小，它们是非常有前途的药物输送工具， 与合成纳米制剂相比，生物相容性、低免疫原性和毒性降低 例如脂质体、树枝状大分子和聚合物。证实外切体中含有抗癌药物的递送 改善体内药代动力学和药效学性质与增强体内抗癌活性的比较 来释放药物分子。将治疗性核酸装载到外切体中可以保护核酸免受 核酸酶和增加细胞摄取和治疗效果由于特定的分子机制 外切体内化。外切体可以穿透血脑屏障并穿透深层组织 与合成纳米载体的疗效进行了比较。此外，它们在癌症转移和转移中起着关键作用。 通过诱导其RNA和蛋白质的转录和表型变化进行再生。因此， 它们可能会被重新设计，以提供基因和蛋白质疗法。然而，仍然有 外切体在临床上的应用面临的基本挑战：i)外切体的载药效率是 非常有限；ii)外切体的生产还没有达到临床试验的足够高的产量，甚至 进一步发展；以及iii)赋予外体多种能力以令人满意的疾病靶向， 追踪和综合疗法要求很高。为了应对这些挑战，公安局建议 1)开发一种高效的手性石墨烯外切体载药技术 2)开发了一种具有刺激压电纳米纤维的外切体生产生物反应器 支架；以及3)工程杂交外切体作为具有靶向性的多功能靶向递送系统 用于近红外成像引导光热肿瘤的配体和功能手性石墨烯量子点 治疗。拟议的研究包含了几种创新的外切体生产、装载和 如果成功和集成，将提供高通量和高效率的外泌体的工程 药物释放的制造平台，并将基于Exosome的药物释放扩展到不同的生物医学和 结合天然外切体和人工合成纳米粒的优点进行临床应用。

A Convergent Bioengineered Platform for Multifunctional Therapeutic Exosomes Abstract: The overall goal of this MIRA application is to develop a convergent bioengineered platform for manufacturing and engineering therapeutic exosomes. The platform will allow the loading of drugs into exosomes with high efficiency, biomanufacturing of exosomes in high throughput, and further engineering exosome-based drug delivery systems for various diseases with desired functions including targeted delivery, tracking, and combinational therapies. Exosomes are a subset of extracellular vesicles, with diameters between 50 nm and 150 nm, secreted by most eukaryotic cells. They are very promising drug delivery vehicles due to their small size, biocompatibility, low immunogenicity, and reduced toxicity in comparison with synthetic nanoscale formulations such as liposomes, dendrimers, and polymers. Delivery of anticancer drugs contained in exosomes demonstrated improved pharmacokinetic and pharmacodynamic properties and enhanced anticancer activity in vivo compared to free drug molecules. Loading of therapeutic nucleic acids into exosomes protects the nucleic acids from nucleases and increases cellular uptake and the therapeutic effect due to specific molecular mechanisms of exosome internalization. Exosomes can cross the blood brain barrier and penetrate deep tissues with improved efficacy compared to that of synthetic nanocarriers. Moreover, they play a key role in cancer metastasis and regeneration by inducing transcriptomic and phenotypic changes with their RNA and protein cargoes. Therefore, they can potentially be reengineered for delivery of gene and protein therapeutics. However, there remain fundamental challenges to the utilization of exosomes in the clinic: i) drug loading efficiency into exosomes is very limited; ii) the production of exosomes has yet to reach sufficiently high throughput for clinical tests or even further development; and iii) endowing exosomes with multiple abilities for satisfactory disease targeting, tracking and combinational therapies is highly demanding. To address these challenges, the PI proposes the following three projects: 1) Developing a high-efficiency exosome drug loading technology with chiral graphene nanoparticles; 2) Developing an exosome production bioreactor with stimulating piezoelectric nanofibrous scaffolds; and 3) Engineering hybrid exosomes as a multifunctional targeted delivery system with targeting ligands and functional chiral graphene quantum dots for near-infrared imaging-guided photothermal cancer therapies. The proposed research contains several innovative approaches of exosome production, loading and engineering that, if successful and integrated, will provide a high-throughput and high-efficiency exosome manufacturing platform for drug delivery, and expand exosome-based drug delivery to diverse biomedical and clinic applications by combining the merits of both the native exosomes and synthetic nanoparticles.