Application of mucus modulating multipurpose bromelain nanoparticles to overcome the mucus barrier in appendiceal pseudomyxoma peritonei

项目摘要 腹膜假性粘液瘤(PMP)是一种潜在的但致命的恶性肿瘤，指的是腹膜转移。 阑尾粘液性肿瘤(AMN)。PMP的特征是大量的细胞外粘液积聚 并在癌细胞周围形成保护性屏障，阻碍化疗药物 送货。我们假设细胞外粘液的破坏(粘液溶解)和/或调制(破坏) 结构是一种很有前途的治疗策略，可以克服细胞外的保护性粘液屏障 PMP中的粘液，使化疗药物输送成为可能。在我们之前的研究中，我们使用了患者来源的 异种移植(PDX)模型的PMP，以展示一种新型的腹膜内(IP)给药 粘液溶解药物的组合，菠萝酶(BRO；一种半胱氨酸蛋白酶)和N-乙酰半胱氨酸(NAC，一种还原 打破二硫键的试剂)，成功地溶解细胞外粘液(粘液溶解)，并增强 向癌细胞输送化疗药物。这项提议的目的是利用纳米技术来 提高BROO+NAC的IP保留时间和瘤内(IT)渗透率，以优化粘液溶解和 PMP中的化疗药物输送。为此，我们已经生成了初步数据，表明它 有可能成功地设计出粘液调节多用途BRO纳米颗粒(MBN)，该纳米颗粒包括 四个组分：(A)用于酶粘液分解的BrO纳米颗粒核心；(B)偶联L-半胱氨酸，用于 通过二硫键形成的粘液粘连；(C)结合的NAC，通过减少 二硫键；和(D)共轭阿霉素(DOX)，用于癌症治疗。此外，我们还成功地 合成了包覆氢离子和谷胱甘肽的氧化铁纳米团簇(InC-H+G)，用于共沉积。 与我们的MBN一起传递，增加粘液的还原潜力，增强MBN的功能。这个 这项研究计划的具体目的是：(A)评估MBN调节粘液结构和 使用患者衍生的PMP体外模型增强化疗药物的释放；和(B)观察其效果 MBN对PMP PDX模型粘液性肿瘤生长的影响我们希望证明拟议的 MBN在药代动力学和药效学方面优于游离粘液/细胞毒 毒品。值得注意的是，建议的MBN也是由生物兼容和可生物降解的材料合成的， 增加了该方法的可译性。我们在开展临床前研究方面具有独特的优势。 鉴于我们已经建立了AMN/PMP的体外和体内模型，我们的 拥有自己的实验室，我们的机构是主要的国际患者管理转诊中心之一 使用PMP。

PROJECT ABSTRACT Pseudomyxoma peritonei (PMP), an insidious but lethal malignancy, refers to peritoneal metastases from appendix mucinous neoplasms (AMN). PMP is characterized by abundant extracellular mucus that accumulates in the abdominal cavity and forms a protective barrier around cancer cells, hindering chemotherapeutic drug delivery. We hypothesize that destruction (mucolysis) and/or modulation (disruption) of extracellular mucus structure is a promising therapeutic strategy to overcome the protective mucus barrier posed by extracellular mucus in PMP, enabling chemotherapeutic drug delivery. In our prior research we utilized patient derived xenograft (PDX) models of PMP in order to demonstrate that intraperitoneal (IP) administration of a novel combination of mucolytic drugs, bromelain (BRO; a cysteine protease) and N-acetylcysteine (NAC, a reducing agent that breaks disulfide bonds), successfully dissolves extracellular mucus (mucolysis) and enhances chemotherapeutic drug delivery to cancer cells. The aim of this proposal is to leverage nanotechnology to enhance IP retention time and intratumoral (IT) penetration by BRO+NAC in order to optimize mucolysis and chemotherapeutic drug delivery in PMP. To this end, we have generated preliminary data demonstrating that it is possible to successfully engineer mucus modulating multipurpose BRO nanoparticles (MBN) comprised of four components; (a) a BRO nanoparticle core, for enzymatic mucolysis; (b) conjugated L-cysteine, for mucoadhesion through disulfide bond formation; (c) conjugated NAC, for mucus disruption through reduction of disulfide bonds; and (d) conjugated doxorubicin (DOX), for cancer therapy. In addition, we have also successfully synthesized iron oxide nanoparticle clusters coated with hydrogen ions and glutathione (INC-H+G), for co- delivery with our MBN, to increase the reduction potential of mucus and enhance the function of the MBN. The specific aims of this research proposal are to (a) assess the ability of MBN to modulate mucus structure and enhance chemotherapeutic drug delivery using patient-derived in vitro models of PMP; and (b) observe the effect of MBN on mucinous tumor growth in PDX models of PMP. We expect to demonstrate that the proposed MBN will provide a pharmacokinetic and pharmacodynamic advantage over free mucolytic/cytotoxic drugs. Notably, the proposed MBN are also synthesized from biocompatible and biodegradable materials, increasing the translatability of the approach. We are uniquely positioned to conduct the preclinical studies in this proposal given that we have already developed the proposed in vitro and in vivo models of AMN/PMP in our own laboratory and our institution is one of the major international referral centers for the management of patients with PMP.