Targeting tumor and its microenvironment using nanotherapeutics for pancreatic cancer

使用纳米疗法治疗胰腺癌靶向肿瘤及其微环境

• 批准号: 10223249
• 负责人: Satyanarayana Rachagani
• 金额: $ 37.23万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10223249
• 关键词: Address; Biodistribution; CASP3 gene; CASP7 gene; Cancer cell line; Cells; Cleaved cell; Clinic; Clinical; Clinical Trials; Combined Modality Therapy; Complex; Cytotoxic agent; Data; Development; Device Designs; Devices; Diagnosis; Diagnostic; Diffusion; Disease-Free Survival; Distant; Dose; Dose-Limiting; Down-Regulation; Drug Combinations; Drug Kinetics; Drug resistance; Ectopic Expression; Electrostatics; Encapsulated; Epithelial; Extracellular Matrix; Future; Genes; Goals; Growth; HIF1A gene; Human; In Vitro; KPC model; MUC4 mucin; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Mesenchymal; MicroRNAs; Modeling; Molecular; Mus; Nano delivery; Neoplasm Metastasis; Normal Cell; Organ; Organoids; Pancreatic Intraepithelial Neoplasia; Pathogenesis; Pathologic; Pathway interactions; Patients; Perfusion; Pharmaceutical Preparations; Play; Prognosis; Property; Reporting; Resistance; Role; SHH gene; Signal Transduction; Signaling Molecule; Site; Survival Rate; System; Temperature; Testing; Therapeutic; Therapeutic Agents; Tissues; Toxic effect; Treatment Efficacy; Up-Regulation; Vascular Endothelium; Work; Xenograft procedure; aldehyde dehydrogenase 1; angiogenesis; base; beta catenin; c-myc Genes; cancer cell; cancer stem cell; cancer therapy; cell motility; cell stroma; chemosensitizing agent; chemotherapy; clinical application; cohort; copolymer; drug release kinetics; drug sensitivity; efficacious treatment; epithelial to mesenchymal transition; gemcitabine; humanized mouse; improved; in vivo; liposome vector; lymph nodes; mouse model; nanoscale; nanotherapeutic; novel; novel therapeutics; pancreatic cancer cells; pancreatic cancer patients; pancreatic neoplasm; pancreatic stellate cell; pre-clinical; prognostic; restoration; side effect; stem cell survival; success; symptomatic improvement; synergism; therapeutic evaluation; therapeutic miRNA; therapeutic target; therapy outcome; therapy resistant; tumor; tumor growth; tumor progression; tumor xenograft

ABSTRACT Pancreatic cancer (PC) is lethal with a five-year survival rate of less than 9.2 % and a median survival of 5-6 months. The limited efficacy of mono-therapies has led to the exploration of combination therapies with limited success because of challenges associated with dose-limiting side effects, drug-associated toxicities, drug resistance, and poor pharmacokinetics. Importantly, these past approaches have not attempted the concurrent targeting of the pancreatic tumor and its stroma and PC stem cells. Our proposed work addresses these challenges by determining functional and clinic-pathological significance of miR-345 as well as developing a dual delivery nanoscale device (DDND) for combined delivery of miR-345 and GEM for the treatment of PC. Our preliminary studies have shown that miR-345 targets several important genes, including sonic hedgehog (Shh), Kras, MUC4 mucin and its downstream targets, genes-associated with cancer stem cells (ALDH1, ESA, Hif1α, and Oct/3/4), and causes up regulation of cleaved caspase-3, -7, and PARP. The Kras, Shh and MUC4-signaling play critical roles in tumor growth and metastasis by promoting epithelial to mesenchymal transition (EMT), PC stem cells, angiogenesis, desmoplasia, which limit the delivery and efficacy of chemotherapy. MiR-345 targeting Kras, Shh and MUC4, which makes miR-345 is an excellent candidate for diagnostic/prognostic and therapeutic targets in PC. We hypothesize that downregulation of miR-345 contributes to PC pathogenesis by upregulation of Kras, SHH, and MUC4; Its restoration, combination with GEM through the DDND, enhances GEM sensitivity in PC through modulation of SHH/Kras/MUC4 pathways, resulting in inhibition of desmoplasia, pancreatic stellate cells, and PC stem cells leading to an improved therapeutic outcome of GEM in PC through improving its tumor perfusion. The DDND is based on temperature and pH responsive pentablock copolymers electrostatically complexed with miR-345 and subsequently self-assembled with GEM encapsulated layers. The DDND design allows effective co-incorporation of miRNA/GEM combination; facilitates cellular entry; enhances stability compared to liposomal carriers; provides miRNA protection; allows targeting by selectively facilitating endosomal escape in cancer cells as opposed to normal cells by exploiting intracellular pH differences; and allows dose- sparing of the cytotoxic drugs. Aim 1 will focus to determine functional role and clinico-pathological significance of miR-345/Shh/Kras/MUC4 axis in highly aggressive and metastatic PC. Aim 2 will focus on the development of DDND loaded miR-345/GEM as a novel therapeutic agent against lethal PC by evaluating their therapeutic efficacy in vitro. In the final Aim 3, we will evaluate therapeutic efficacy of DDND loaded miR-345/GEM alone or in combination in mouse models. Altogether, the proposed work decipher the clinic-pathological significance of miR-345 and expected to significantly advance the goal of combining GEM and miR-345 delivery for treatment of PC patients, enhance understanding of the synergistic mechanisms involved, and will provide a novel DDND design for delivery of other therapeutics as well in the future.

抽象的 胰腺癌 (PC) 具有致命性，五年生存率低于 9.2%，中位生存期为 5-6 几个月。单一疗法的疗效有限，导致了对联合疗法的探索，但联合疗法的效果有限。 成功是因为与剂量限制副作用、药物相关毒性、药物相关的挑战相关 耐药性和药代动力学较差。重要的是，这些过去的方法并没有尝试并发 靶向胰腺肿瘤及其基质和 PC 干细胞。我们提出的工作解决了这些问题 通过确定 miR-345 的功能和临床病理意义以及开发双重挑战 用于联合递送 miR-345 和 GEM 来治疗 PC 的纳米级递送装置 (DDND)。我们的 初步研究表明，miR-345靶向几个重要基因，包括音猬因子（Shh）、 Kras、MUC4 粘蛋白及其下游靶标、与癌症干细胞相关的基因（ALDH1、ESA、Hif1α、 和 Oct/3/4)，并导致 cleaved caspase-3、-7 和 PARP 的调节上调。 Kras、Shh 和 MUC4 信号传导 通过促进上皮间质转化 (EMT)、PC 在肿瘤生长和转移中发挥关键作用 干细胞、血管生成、结缔组织增生，这些限制了化疗的输送和疗效。 MiR-345 靶向 Kras、Shh 和 MUC4，这使得 miR-345 成为诊断/预后和治疗的优秀候选者 PC 中的目标。我们假设 miR-345 的下调通过上调促进 PC 发病机制 Kras、SHH 和 MUC4；其修复体通过DDND与GEM结合，增强GEM灵敏度 在 PC 中，通过调节 SHH/Kras/MUC4 通路，抑制结缔组织增生、胰星状细胞 细胞和 PC 干细胞通过改善 PC 的肿瘤来改善 GEM 的治疗效果 灌注。 DDND 基于静电响应温度和 pH 值的五嵌段共聚物 与 miR-345 复合，随后与 GEM 封装层自组装。 DND设计 允许 miRNA/GEM 组合有效共掺入；促进细胞进入；增强稳定性 与脂质体载体相比；提供 miRNA 保护；允许通过选择性促进内体进行靶向 与正常细胞不同，癌细胞利用细胞内 pH 差异进行逃逸；并允许剂量- 避免使用细胞毒性药物。目标 1 将重点确定功能作用和临床病理意义 miR-345/Shh/Kras/MUC4 轴在高度侵袭性和转移性 PC 中的作用。目标2将侧重于发展 通过评估 DDND 负载 miR-345/GEM 作为针对致命性 PC 的新型治疗剂的治疗效果 体外疗效。在最终目标 3 中，我们将评估单独加载 miR-345/GEM 的 DDND 或 结合小鼠模型。总而言之，拟议的工作破译了临床病理意义 miR-345 有望显着推进 GEM 和 miR-345 联合递送治疗的目标 PC 患者的研究，增强对所涉及协同机制的理解，并将提供一种新的 DDND 设计用于将来提供其他疗法。