Nanoparticle delivery of miRNA-based therapeutics to overcome clinical challenges in triple negative breast cancer

项目摘要 三阴性乳腺癌(TNBC)是所有乳腺癌亚型中病死率最高的。几个 已经确定了乳腺癌治疗的分子靶点，但目前还没有批准的， 广泛适用于TNBC的靶向治疗。通过10年的研究，我们发现延伸率因素 2-激酶(EF2K)的表达是TNBC肿瘤发生和发展的关键驱动因素。我们还发现， 在TNBC患者中，microRNA-22(miR-22)的表达被广泛抑制，并且与EF2K呈负相关 表情。进一步的分析表明，miR-22通过与EF2K特异性结合来抑制肿瘤，EF2K 在多种TNBC模型中抑制EF2K的表达并减少肿瘤生长。考虑到临床 EF2K在TNBC中的意义和潜在的治疗价值，我们因此开发了一种AXL受体靶向 AXL适体包裹的SLNP-miR-22纳米粒系统可将miR-22特异性地输送到TNBC肿瘤中 体内(但不会导致miR-22在正常组织中积聚)。 在此初步工作的基础上，我们假设EF2K是TNBC的有效治疗靶点，并且 使用我们的Axl-apTamer-SLNP-miR-22纳米疗法靶向EF2K可以提供显著的治疗作用 TNBC治疗的疗效。然而，可以理解的是，这个治疗系统是复杂的，而且它已经 难以进一步了解重大影响的潜在生物和物理过程 治疗结果，并确定最佳剂量和最大限度治疗的剂量计划 功效。因此，在这个项目中，我们建议通过整合生物实验来克服这一挑战 数学建模基于潜在的生物和物理机制，涉及 EF2K靶向miR-22中的肿瘤侵袭、药物渗透和药物与癌细胞的相互作用 用于TNBC治疗的纳米疗法。我们的假设将通过实现以下两个具体目标来检验： 1)以EF2K为靶点的miR-22纳米疗法的实验测试(目标1)，以及2)数学模型(目标 2)。在目标1中，我们将重点研究EF2K靶向的体内治疗效果 MIR-22介导的小鼠原位模型治疗。在目标2中，我们将专注于开发、测试和 使用逻辑集成的方法验证EF2K靶向、miR-22为基础的纳米疗法的数学模型 统计和多尺度机械建模方法。来自AIM 1的实验数据将提供给AIM 2用于开发和验证数学模型，目标1中的实验将以发现为指导 从目标2的计算研究中获得。通过这种基于迭代的反馈方法， 将使用数学模型来预测和确定各种参数的影响，包括siRNA 剂量和给药程序，对EF2K靶向miR-22介导性治疗(有或没有)的肿瘤反应 化疗)，并确定最佳药物剂量和给药计划，以优化治疗 功效。该项目的长期目标是证明这种基于miR-22的纳米疗法是安全的和 单独有效，并与标准化疗药物联合作为辅助治疗；以及 完成临床前开发，为TNBC患者未来的潜在临床翻译做准备。

Project Summary Triple-negative breast cancer (TNBC) has the highest patient death rate of all breast cancer subtypes. Several molecular targets have been identified for breast cancer treatment, but currently, there is no approved, broadly applicable targeted therapy for TNBC. Through 10 years of research, we found that elongation factor 2-kinase (EF2K) expression is a critical driver of TNBC tumorigenesis and progression. We also found that microRNA-22 (miR-22) expression is broadly repressed in TNBC patients, and is inversely correlated with EF2K expression. Further analysis revealed that miR-22 suppresses tumors by specifically binding to EF2K, which inhibits EF2K expression and reduces tumor growth in multiple TNBC models. Considering the clinical significance and potential therapeutic value of EF2K in TNBC, we have thus developed an AXL receptor-targeted AXL aptamer-coated SLNP-miR-22 nanoparticle system that can specifically deliver miR-22 to TNBC tumors in vivo (but does not lead to miR-22 accumulation in normal tissues). On the basis of this preliminary work, we hypothesize that EF2K is an effective therapeutic target in TNBC, and that targeting EF2K using our AXL-aptamer-SLNP-miR-22 nanotherapeutics can provide significant therapeutic efficacy in TNBC treatment. However, understandably, this therapeutic system is complex, and it has been difficult to further understand the underlying biological and physical processes that significantly impact treatment outcome, and to identify the optimal doses and dosing schedules for maximizing treatment efficacy. Therefore, in this project, we propose to overcome this challenge by integrating biological experiments with mathematical modeling based on the underlying biological and physical mechanisms that are involved in cancer invasion, drug penetration, and drug-cancer cell interactions in the EF2K-targeted miR-22 nanotherapeutics for TNBC treatment. Our hypothesis will be tested by achieving the following two specific aims: 1) experimental testing of the EF2K-targeted miR-22 nanotherapy (Aim 1), and 2) mathematical modeling (Aim 2). In Aim 1, we will focus on characterizing and determining the in vivo therapeutic efficacy of EF2K-targeted miR-22 mediated therapies in orthotopic mouse models. In Aim 2, we will focus on developing, testing, and validating a mathematical model of EF2K-targeted, miR-22 based nanotherapy, using a logically integrated statistical and multiscale mechanistic modeling approach. Experimental data from Aim 1 will be supplied to Aim 2 for developing and validating the mathematical model, and experiments in Aim 1 will be guided by discoveries obtained from computational investigations in Aim 2. Through this iteration-based feedback approach, the mathematical model will be used to predict and determine the effects of various parameters, including siRNA dose and dosing schedules, on tumor response to EF2K-targeted miR-22 mediated therapies (with or without chemotherapy), and to determine the optimal drug doses and dosing schedules for optimizing therapeutic efficacy. The long-term goal of this project is to demonstrate that this miR-22-based nanotherapy is safe and effective, both alone and in combination with standard chemotherapeutic agents as a co-adjuvant therapy, and to complete preclinical development for potential future clinical translation for TNBC patients.