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患者死亡率最高的。几 已经确定了用于乳腺癌治疗的分子靶点，但是目前，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， 2-激酶（EF2K）表达是TNBC肿瘤发生和进展的关键驱动因素。我们还发现 microRNA-22（miR-22）表达在TNBC患者中被广泛抑制，并与EF2K呈负相关 表情进一步的分析显示，miR-22通过特异性结合EF2K抑制肿瘤， 在多种TNBC模型中抑制EF2K表达并减少肿瘤生长。考虑临床 EF2K在TNBC中的意义和潜在的治疗价值，因此我们开发了一种AXL受体靶向的 AXL适体包被的SLNP-miR-22纳米颗粒系统，其可以特异性地将miR-22递送至TNBC肿瘤， 体内（但不导致miR-22在正常组织中积累）。 在此初步工作的基础上，我们假设EF2K是TNBC的有效治疗靶点， 使用我们的AXL-适体-SLNP-miR-22纳米治疗剂靶向EF2K可以提供显著的治疗效果， TNBC治疗的效果。然而，可以理解的是，这种治疗系统是复杂的， 很难进一步了解潜在的生物和物理过程， 治疗结果，并确定最佳剂量和给药方案，以最大限度地提高治疗效果 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 数学模型的基础上的基本生物和物理机制，涉及 EF2K靶向miR-22中的癌症侵袭、药物渗透和药物-癌细胞相互作用 用于TNBC治疗的纳米治疗剂。我们的假设将通过实现以下两个具体目标来检验： 1)靶向EF2K的miR-22纳米疗法的实验测试（Aim 1），和2）数学建模（Aim 2）。在目标1中，我们将专注于表征和确定EF2K靶向的靶向药物的体内治疗功效。 原位小鼠模型中的miR-22介导的疗法。在目标2中，我们将专注于开发、测试和 验证EF2K靶向的、基于miR-22的纳米治疗的数学模型，使用逻辑整合的 统计和多尺度机械建模方法。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Aim 2中的计算研究中获得。通过这种基于迭代的反馈方法， 数学模型将用于预测和确定各种参数的影响，包括siRNA 剂量和给药方案，对肿瘤对靶向EF2K的miR-22介导的治疗（有或没有）的反应 化疗），并确定最佳药物剂量和给药方案，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， 单独使用和与标准化疗剂联合作为辅助治疗均有效，并且 以完成临床前开发，为TNBC患者提供潜在的未来临床转化。

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.