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

项目摘要 腹膜假粘液瘤（PMP）是一种隐匿但致命的恶性肿瘤，是指腹膜转移， 阑尾粘液性肿瘤（AMN）。PMP的特征是大量的细胞外粘液， 并在癌细胞周围形成保护屏障，阻碍化疗药物 交付.我们假设细胞外粘液的破坏（粘液溶解）和/或调节（破坏） 结构是一种很有前途的治疗策略，以克服由细胞外基质引起的保护性粘液屏障。 PMP中的粘液，使得化疗药物递送成为可能。在我们之前的研究中，我们使用了患者来源的 PMP异种移植（PDX）模型，以证明腹膜内（IP）给予新的 粘液溶解药物菠萝蛋白酶（BRO;半胱氨酸蛋白酶）和N-乙酰半胱氨酸（NAC，还原半胱氨酸蛋白酶）的组合 断裂二硫键的试剂），成功溶解细胞外粘液（粘液溶解）并增强 化疗药物递送至癌细胞。这项提案的目的是利用纳米技术， 通过BRO+NAC增强IP保留时间和瘤内（IT）渗透，以优化粘液溶解， PMP中的化疗药物递送。为此，我们已经生成了初步数据，表明它 可以成功地工程化粘液调节多用途BRO纳米颗粒（MBN），其包含 四种组分;（a）BRO纳米颗粒核心，用于酶促粘液溶解;（B）缀合的L-半胱氨酸，用于 通过二硫键形成的粘膜粘附;（c）缀合的NAC，用于通过减少 二硫键;和（d）缀合的阿霉素（DOX），用于癌症治疗。此外，我们还成功地 用氢离子和谷胱甘肽（INC-H+G）包覆合成的氧化铁纳米颗粒簇，用于共 与我们的MBN一起输送，以增加粘液的还原潜力并增强MBN的功能。的 本研究建议的具体目标是（a）评估MBN调节粘液结构的能力， 使用患者来源的PMP体外模型增强化疗药物递送;和（B）观察效果 MBN对PMP的PDX模型中粘液性肿瘤生长的影响。我们希望证明， MBN将提供优于游离粘液溶解/细胞毒性的药代动力学和药效学优势 毒品值得注意的是，所提出的MBN也是由生物相容性和可生物降解的材料合成的， 增加了方法的可翻译性。我们在开展临床前研究方面具有独特的优势， 这一建议是因为我们已经在我们的研究中开发了AMN/PMP的体外和体内模型。 我们拥有自己的实验室，我们的机构是管理患者的主要国际转诊中心之一 关于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.