Targeted Lipopolyplexes for Oligonucleotide Delivery to AML

描述（由申请人提供）：急性髓性白血病（AML）是成人中最常见的白血病类型，在儿童中第二常见。虽然在治疗几种类型的白血病方面取得了进展，但成人和儿童AML的生存率并未改善。开发载有分子靶向治疗剂的靶向纳米载体代表了改善AML治疗的新策略。GTI-2040是一种针对核糖核苷酸还原酶（RNR）R2亚基的反义寡脱氧核苷酸（ODN），是一种很有前途的克服AML化疗耐药性的药物。GTI-2040的临床功效的增强和副作用的减少可以通过使用由脂聚复合物（ILP）的免疫缀合物组成的多功能纳米载体靶向递送GTI-2040来潜在地实现。因此，本项目的目的是设计、合成和评价用于下调R2和用于AML治疗的抗CD 33 mAb缀合的脂聚复合物（抗CD 33-ILP）。CD 33是一种经常在AML母细胞上表达的骨髓谱系标志物。ILP将通过常规的批量混合（BM）和新型微/纳流体（MF/NF）方法合成，并通过蛋白a接头与抗CD 33缀合。随后将进行R2下调活性的体外评价和药代动力学（PK）特性、药效学（PD）终点和治疗活性的体内评价。本申请的具体目的包括1）设计和优化用于将GTI-2040靶向递送至AML细胞的抗CD 33-ILP制剂; 2）开发用于合成抗CD 33-GTI-2040-ILP的微/纳流体（MF/NF）方法; 3）使用AML细胞系和患者的原发性恶性母细胞体外评估抗CD 33-GTI- 2040-ILP的生物活性; 4）评估抗-CD 33-GTI- 2040-ILP在AML的临床前鼠模型中的药代动力学（PK）特性和药效学（PD）终点和治疗活性。该项目将通过跨学科的方法进行，由具有药物输送（RLee），PK/PD（Chan），纳米工程（LJLee，Guan），生物化学和分子生物学（Jin）和实验治疗学（Marcucci，Byrd，Muthusamy）专业知识的研究人员进行。该项目的成功不仅可能导致AML的新型临床药物的开发，而且可能导致纳米载体设计和合成的技术进步，并在寡核苷酸治疗中具有广泛的应用。 公共卫生相关性：我们寻求开发新的纳米技术来创建用于递送反义寡脱氧核糖核苷酸以治疗急性髓性白血病（AML）的多功能免疫纳米颗粒。该项目的成功不仅可能导致AML的新型临床药物的开发，而且可能导致纳米载体设计和合成的技术进步，并在寡核苷酸治疗中具有广泛的应用。

DESCRIPTION (provided by applicant): Acute myeloid leukemia (AML) is the most common type of leukemia in adults and the second most common in children. While progress has been made in the treatment of several types of leukemia, improvement in survival rate for both adult and pediatric AML has not occurred. Development of targeted nanocarriers loaded with molecularly targeted therapeutics represents a novel strategy for improving treatment of AML. GTI-2040, an antisense oligodeoxyribonucleotide (ODN) against the R2 subunit of ribonucleotide reductase (RNR), is a promising agent for overcoming chemoresistance in AML. Enhancement in clinical efficacy of GTI-2040 and reduction in side effects can potentially be achieved through targeted delivery of GTI-2040 using multifunctional nanovehicles consisting of immunoconjugates of lipopolyplexes (ILPs). The objective of this project, therefore, is to design, synthesize, and evaluate anti-CD33 mAb conjugated lipopolyplexes (anti- CD33-ILPs) for down regulation of R2 and for AML therapy. CD33 is a myeloid lineage marker frequently expressed on AML blast cells. ILPs will be synthesized by both conventional bulk-mixing (BM) and novel micro-/nanofluidic (MF/NF) methods, and conjugated to anti-CD33 via a protein a linker. This will be followed by in vitro evaluation for R2 down regulation activity and in vivo evaluation for pharmacokinetic (PK) properties, pharmacodynamic (PD) endpoints and therapeutic activities. The Specific Aims of this application include to 1) design and optimize anti-CD33-ILP formulation for targeted delivery of GTI-2040 to AML cells; 2) develop micro-/nanofluidic (MF/NF) methods for synthesis of anti-CD33-GTI-2040-ILPs; 3) evaluate in vitro the biological activities of anti-CD33-GTI- 2040-ILPs using AML cell lines and patients' primary malignant blasts; 4) evaluate pharmacokinetic (PK) properties and pharmacodynamic (PD) endpoints and therapeutic activities of anti-CD33-GTI- 2040-ILPs in preclinical murine models of AML. This project will be carried out via an interdisciplinary approach by investigators with expertise in drug delivery (RLee), PK/PD (Chan), nanoengineering (LJLee, Guan), biochemistry and molecular biology (Jin), and experimental therapeutics (Marcucci, Byrd, Muthusamy). Success of this project may lead to not only development of a novel clinical agent for AML, but also technological advances in nanocarrier design and synthesis with broad applications in oligonucleotide therapeutics. PUBLIC HEALTH RELEVANCE: We seek to develop novel nanotechnology to create multifunctional immunonanoparticles for the delivery antisense oligodeoxyribonucleotides to treat acute myeloid leukemia (AML). Success of this project may lead to not only development of a novel clinical agent for AML, but also technological advances in nanocarrier design and synthesis with broad applications in oligonucleotide therapeutics.