Microfluidic Synthesis of Nanoparticles for Oligonucleotide Delivery

描述（由申请人提供）：治疗性寡核苷酸，特别是微RNA (miR)，有望成为有效的抗癌药物。然而，由于缺乏有效的肠外给药方法，它们的临床应用受到限制。通过多功能纳米颗粒给药是解决这一问题的一种潜在方法。然而，目前的纳米颗粒制造方法是基于多种试剂的大量混合，在临床前和临床试验中只取得了有限的成功。由于颗粒混合在局部环境中是不均匀的，导致纳米颗粒的结构和组成不均匀，因此我们假设通过严格控制合成纳米颗粒的大小、结构和组成，可以实现高效和安全的递送。因此，我们建议采用微流体技术，该技术能够在微米尺度上精确控制混合过程，以合成具有均匀和明确定义的结构和成分的多功能纳米颗粒，用于将治疗性寡核苷酸输送到癌细胞。我们的具体目标是：(1)优化和验证微流控系统，用于合成多功能聚合物和脂质纳米颗粒，这些纳米颗粒含有合成miR（如miR29b）和寡脱氧核糖核苷酸（如G3139）化合物，分别针对黑色素瘤和急性髓性白血病细胞中的特定抗凋亡蛋白，如Bcl-2和Mcl- 1；(2)开展体外和体内临床前研究，以表征微流体基纳米颗粒的递送效率、毒性、生物相容性、药代动力学、生物分布和治疗效果。如果成功，这可能会导致黑色素瘤和白血病等癌症的新治疗方式的发展。

DESCRIPTION (provided by applicant): Therapeutic oligonucleotides, in particular, micro RNA (miR), hold great promise to become effective anticancer agents. Their clinical usage is, however, limited by the lack of efficient methods for parenteral administration. Delivery via multi-functional nanoparticles is a potential solution to this problem. However, current nanoparticle manufacturing methods, which are based on bulk mixing of multiple reagents, have had only limited success in pre-clinical and clinical trials. Since bulk mixing is heterogeneous at local environment and consequently leads to the nanoparticles with poorly defined and non-uniform structures and compositions, we hypothesize that efficient and safe delivery can be achieved by tightly controlling the size, structure, and compositions of the synthetic nanoparticles. We therefore propose to employ microfluidic technology, which is capable of precisely controlling the mixing process at the micrometer scale, to the synthesis of multi-functional nanoparticles with uniform and well-defined structures and compositions for the delivery of therapeutic oligonucleotides to cancer cells. Our specific aims are (1) to optimize and validate microfluidic systems for the synthesis of multi-functional polymer and lipid-based nanoparticles containing synthetic miR (e.g., miR29b) and oligodeoxyribonucleotide (e.g., G3139) compounds targeting specific antiapoptotic proteins e.g., Bcl-2 and Mcl- 1, respectively, in melanoma and acute myeloid leukemia cells; and (2) to conduct in vitro and in vivo preclinical studies to characterize delivery efficiency, toxicity, biocompatibility, pharmacokinetics, biodistribution, and therapeutic efficacy of the microfluidic-based nanoparticles. If successful, this could lead to development of a new treatment modality for cancers such as melanoma and leukemia. Project Narrative: Oligonucleotides (especially micro RNA) hold great potential to become a new class of anticancer drugs, but their clinical applications are limited by a lack of efficient delivery methods. We seek to solve this problem by developing nanoscale, multi-functional, particle-like devices capable of delivering oligonucleotide drugs to cancer cells efficiently. The nanodevices will be assembled through a novel approach based on the precise manipulation of liquid flows at micrometer scale.