Targeted Lipopolyplexes for Oligonucleotide Delivery to AML

描述(申请人提供)：急性髓系白血病(AML)是成人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，但成人和儿童急性髓细胞白血病的存活率并没有改善。开发装载分子靶向治疗药物的靶向纳米载体代表了改善急性髓细胞白血病治疗的新策略。GTI-2040是针对核糖核苷酸还原酶(RNR)R2亚基的反义寡核苷酸(ODN)，是一种有希望克服AML化疗耐药的药物。通过使用由脂多糖免疫结合物(ILPS)组成的多功能纳米载体靶向递送GTI-2040，有可能实现GTI-2040临床疗效的增强和副作用的减少。因此，本项目的目的是设计、合成和评价抗CD33单抗结合脂多糖(抗CD33-ILPS)以下调R2和治疗AML。CD33是一种髓系标记物，常在急性髓系白血病原始细胞上表达。ILPS将通过传统的本体混合(BM)和新颖的微/纳米流体(MF/NF)方法合成，并通过蛋白质a连接物与抗CD33偶联。随后将对R2下调活性进行体外评估，并对药代动力学(PK)特性、药效学(PD)终点和治疗活性进行体内评估。本应用的具体目标包括1)设计和优化靶向GTI-2040靶向AML细胞的抗CD33-ILP制剂；2)开发合成抗CD33-GTI-2040-ILPS的微纳流体(MF/NF)方法；3)利用AML细胞系和患者原代恶性母细胞体外评价抗CD33-GTI-2040-ILPS的生物活性；4)在AML临床前小鼠模型中评价抗CD33-GTI-2040-ILPS的药代动力学(PK)性质、药效学(PD)终点和治疗活性。该项目将由具有药物传递(RLee)、PK/PD(Chan)、纳米工程(LJLee，Guan)、生物化学和分子生物学(JI)以及实验治疗学(Marcucci，Byrd，Muthusamy)专长的研究人员通过跨学科方法进行。该项目的成功不仅可能导致AML新的临床药物的开发，而且还将导致纳米载体设计和合成的技术进步，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多功能免疫纳米颗粒，用于运送反义寡核苷酸治疗急性髓系白血病(AML)。该项目的成功不仅可能导致AML新的临床药物的开发，而且还将导致纳米载体设计和合成的技术进步，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.