TUMOR SPECIFIC DELIVERY OF VERTICILLIN A OVERCOMES EPIGENETIC SILENCING RESPONSIBLE FOR DRUG RESISTANCE

摘要 该提案描述了一种创新的解决方案，以治疗间皮瘤失败的主要原因：药物 阻力解决方案是控制和靶向递送一种新的表观遗传修饰剂- 轮枝青霉素A或其类似物之一-在化疗中重建化疗敏感性 通过表观遗传学沉默的肿瘤抑制基因的再表达来治疗耐药肿瘤。与其他小型 分子表观遗传因子，包括FDA批准的药物，这是全球DNA甲基化和组蛋白 脱乙酰酶抑制剂Verticillin A选择性靶向H3 K9 me 2/3。一种刺激响应型微粒给药系统， 使用采用独特的基于材料的靶向策略的系统，以确保 化疗剂定位并集中在腹膜肿瘤中。这种pH响应的药物输送 系统利用肿瘤的基本病理生理学特性（例如，弱酸性胞外 环境和高代谢率）以诱导组成和结构变化（例如，颗粒溶胀 0.1至1微米），导致肿瘤特异性积累。这种基于材料的目标选择方法 克服了传统策略的局限性（例如，通过增强的渗透性进行被动靶向， 保留效应和通过基于抗体的亲和力的主动靶向），同时提供平台以递送 疏水性和亲水性药物分别通过单乳剂和双乳剂。拟议 实验将检验这样的假设， （作为单一药剂或与包封或游离药物的共制剂的组合疗法）， 间皮瘤肿瘤将克服耐药性和延长生存相比，交付的 或单独使用化疗药物。重要的是，关键的初步数据支持拟议的 研究，良好的特性材料和严格的实验设计建立，和必要的交叉- 学科协作和专门知识已经到位，以解决这一假设。这五个具体目标- 今年的目标是：1。评价轮枝菌素A及其五种类似物在人体中的表观遗传活性 间皮瘤细胞（MSTO-211 H）和从患者胸腔积液收集的肿瘤细胞以及 通过肝毒性、心脏毒性和线粒体毒性的体外测定的毒性特征。目标二。优化 两种维他西林与紫杉醇、顺铂或培美曲塞的联合制剂， 耐药间皮瘤细胞系和从患者胸膜收集的肿瘤细胞中的细胞毒性作用 积液目标3：评估健康小鼠中的毒性以及优化制剂在健康小鼠中的PK和PD /疗效。 耐药间皮瘤的患者来源的异种移植（PDX）鼠模型。

ABSTRACT This proposal describes an innovative solution to a primary cause of treatment failure in mesothelioma: drug resistance. The solution is the controlled and targeted delivery of a novel epigenetic modifying agent – verticillin A or one of its analogs – which re-establishes chemotherapeutic susceptibility in chemotherapeutic resistant tumors via re-expression of epigenetically silenced tumor suppressor genes. Unlike other small molecule epigenetic agents, including FDA-approved drugs, which are global DNA methylation and histone deacetylase inhibitors, Verticillin A selectively targets H3K9me2/3. A stimuli-responsive particle drug delivery system employing a unique materials-based targeting strategy is used in order to ensure that the verticillin and chemotherapeutic agents localize and concentrate in peritoneal tumors. This pH-responsive drug delivery system leverages fundamental pathophysiological properties of tumors (e.g., mildly acidic extracellular environment and high metabolic rate) to induce compositional and architectural changes (e.g., particle swelling from 0.1 to 1 micron) resulting in tumor-specific accumulation. This materials-based targeting approach overcomes limitations of traditional strategies (e.g., passive targeting via the enhanced permeability and retention effect, and active targeting via antibody-based affinity) while providing a platform to deliver both hydrophobic and hydrophilic drugs via single- and double-emulsions, respectively. The proposed experiments will test the hypothesis that delivery of a verticillin agent with a chemotherapeutic (combination therapy either as single agents or co-formulation with encapsulated or free drug) to mesothelioma tumors will overcome drug resistance and extend survival compared to the delivery of verticillin or the chemotherapeutic alone. Importantly, key preliminary data are in support of the proposed studies, well-characterized materials and rigorous experimental designs are established, and essential cross- disciplinary collaborations and expertise are in place to address this hypothesis. The specific aims of this five- year proposal are: Aim 1. Evaluate the epigenetic activity of verticillin A and five of its analogs in human mesothelioma cells (MSTO-211H) and tumor cells collected from patient pleural effusions as well as the toxicity profile via in vitro assays for hepatotoxicity, cardiotoxicity, and mitochondrial toxicity. Aim 2. Optimize the co-formulation of the two verticillin agents with paclitaxel, cisplatin, or pemetrexed to achieve the maximum cytotoxic effect in drug resistant mesothelioma cell lines and tumor cells collected from patient pleural effusions. Aim 3. Assess toxicity in healthy mice and PK and PD / efficacy of the optimized formulation in patient-derived xenograft (PDX) murine models of drug resistant mesothelioma.