Reprogramming Tumor-Associated Macrophages in PDAC with MicroRNA Nano-Vectors

临床前和临床证据表明，肿瘤相关巨噬细胞(TAMs)是 主要是M2表型，支持免疫抑制、细胞侵袭、血管生成、 转移和治疗耐药。 本项目的主要目标是评估microRNA(MiR)-125b在双重(CD44和CD44)中的传递 M2pep)靶向透明质酸(HA)自组装纳米颗粒影响肿瘤相关 巨噬细胞()从M2表型向M1表型的复极化改善治疗效果 在KrasLSL-G12D/+，p53fl/fl基因工程小鼠(KPC-GEM)模型中建立胰腺导管 腺癌(PDAC)。 在我们的初步研究中，我们已经表明，功能化的HA块形成了一个模块化的自我 组装用于稳定包裹核酸结构的纳米平台，包括小干扰 核糖核酸。这些以CD44为靶点的核壳纳米颗粒显示出有效的siRNA递送和向下- 体内抗细胞凋亡基因的调控。Cieslewicz等人的研究成果。已经表明，M2pep具有优先 与其他细胞相比，小鼠M2极化的细胞具有结合、快速内化和蓄积的特点。在……里面 我们的初步研究，我们已经观察到M2pep序列与HA自身的偶联 组装纳米颗粒显示这些纳米颗粒在体外M2中优先积聚 巨噬细胞。 该项目的具体目标如下：AIM-1研究将侧重于拟订和 HA-PEI/HA-PEG/HA-m2pep纳米组装体中miR-125b双链包裹的表征。AIM-2 研究将针对纳米组件的体外评估，以重新极化巨噬细胞和 它们调节促炎细胞因子的能力。AIM-3研究将涉及抗肿瘤的评估 HA-PEI/HA-PEG/HA-PEI/HA-PEI包埋miR-125b的疗效及对转移的抑制作用 在PDAC的KPC-GEM模型中，m2pep联合吉西他滨+NAB-紫杉醇。 这项研究在评估巨噬细胞复极(从主要的M2到 作为治疗PDAC和使用巨噬细胞靶向HA的治疗策略 纳米粒子传递用于完成复极化的microRNA。创新的基于HA的模块化 纳米平台的用途非常广泛，可以提供高效的核酸封装、有效载荷保护和 体内特异性地传递给进行高效复极。

Preclinical and clinical evidence suggests that tumor-associated macrophages (TAMs) are predominantly of the M2 phenotype that supports immuno-suppression, cellular invasion, angiogenesis, metastasis, and therapeutic resistance. The main objective of this project is to evaluate microRNA (miR)-125b delivery in a dual (CD44 and M2pep) targeted hyaluronic acid (HA)-based self-assembling nanoparticles to affect tumor associated macrophage (TAM) repolarization from a predominant M2 to M1 phenotype for improved therapeutic effect in in a KrasLSL-G12D/+, p53fl/fl genetically-engineered mouse (KPC-GEM) model of pancreatic ductal adenocarcinoma (PDAC). In our preliminary studies, we have shown that functionalized HA blocks form a modular self- assembling nano-platform for stable encapsulation of nucleic acid constructs, including small interfering RNA. These CD44-targeted core-shell nanoparticles have shown effective siRNA delivery and down- regulation of anti-apoptotic genes in vivo. Cieslewicz, et al. have shown that M2pep has preferential binding, rapid internalization, and accumulation in murine M2-polarized TAM's compared to other cells. In our preliminary investigation, we have observed that conjugation of M2pep sequence to the HA self- assembling nanoparticles shows preferential accumulation of these nanoparticles in vitro in M2 macrophages. The specific aims of the project are as follows: Aim-1 studies will focus on formulation and characterization of miR-125b duplex encapsulated in HA-PEI/HA-PEG/HA-m2pep nano-assemblies. Aim-2 studies will be directed towards in vitro evaluation of the nano-assemblies to re-polarize macrophages and their capacity to regulate pro-inflammatory cytokines. Aim-3 studies will address evaluation of the anti-tumor therapeutic response and inhibition of metastasis of miR-125b duplex encapsulated in HA-PEI/HA-PEG/HA- m2pep in combination with gemcitabine plus nab-Paclitaxel in a KPC-GEM model of PDAC. This study is highly significant in evaluating macrophage repolarization (from predominant M2 to M1) as a therapeutic strategy for the treatment of PDAC and using macrophage targeted HA-based nanoparticles to deliver microRNA for accomplishing repolarization. The innovative modular HA-based nano-platform is extremely versatile to afford efficient nucleic acid encapsulation, payload protection, and specific delivery to TAM in vivo for efficient repolarization.