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: 10383703
• 负责人: Satyanarayana Rachagani
• 金额: $ 36.52万
• 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 2022-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10383703
• 关键词: Address; Biodistribution; CASP3 gene; CASP7 gene; Cancer cell line; Cells; 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.

摘要 胰腺癌（PC）是致命的，5年生存率低于9.2%，中位生存率为5-6 个月单一疗法的有限功效导致了对具有有限疗效的联合疗法的探索。 成功是因为与剂量限制性副作用、药物相关毒性、药物 耐药性和不良的药代动力学。重要的是，这些过去的方法并没有尝试同时 靶向胰腺肿瘤及其基质和PC干细胞。我们提出的工作解决这些问题 通过确定miR-345的功能和临床病理学意义以及开发一种双重的 递送纳米级装置（DDND），用于组合递送miR-345和GEM以治疗PC。我们 初步研究表明，miR-345靶向几个重要基因，包括音刺猬（Shh）， Kras，MUC 4粘蛋白及其下游靶点，与癌症干细胞相关的基因（ALDH 1，ESA，Hif 1 α， 和Oct/3/4），并引起裂解的caspase-3、-7和PARP的上调。Kras、Shh和MUC 4信号转导 通过促进上皮细胞向间质细胞转化（EMT），PC在肿瘤生长和转移中起关键作用 干细胞、血管生成、结缔组织增生，这限制了化疗的输送和功效。MiR-345靶向 Kras、Shh和MUC 4使miR-345成为诊断/预后和治疗的极好候选者。 PC中的目标我们假设miR-345的下调通过上调miR-345的表达而促进PC的发病。 Kras、SHH和MUC 4;其恢复，通过DDND与GEM组合，增强GEM敏感性 在PC中，通过调节SHH/Kras/MUC 4通路，导致结缔组织增生、胰腺星状细胞增生和胰腺癌的抑制。 细胞和PC干细胞，通过改善其肿瘤，改善GEM在PC中的治疗结果 灌注。DDND是基于温度和pH响应五嵌段共聚物静电 与miR-345复合，随后与GEM包封层自组装。DDND设计 允许miRNA/GEM组合的有效共掺入;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靶向 通过利用细胞内pH差异在癌细胞中逃逸，而不是在正常细胞中逃逸;并允许剂量- 节省细胞毒性药物。目的1将着重于确定功能作用和临床病理意义 miR-345/Shh/Kras/MUC 4轴在高侵袭性和转移性PC中的表达。目标2将侧重于发展 DDND负载的miR-345/GEM作为一种新的治疗药物，通过评估其治疗作用， 体外药效在最后的目标3中，我们将评估DDND负载的miR-345/GEM单独或联合应用的治疗功效。 在小鼠模型中联合使用。总而言之，拟议的工作破译的临床病理意义， miR-345，并有望显著推进GEM和miR-345联合递送治疗的目标 的PC患者，增强了对所涉及的协同机制的理解，并将提供一种新的DDND 设计用于将来输送其他治疗药物。