Heterocellular 3D ovarian tumor arrays for imaging and mechanistic combinations

描述(由申请者提供)：晚发现和耐药性使卵巢癌(OvCa)的严峻统计数据保持了几十年的稳定。假设使用机械上不同成分的联合疗法的新方法是最有效的。这项研究的长期目标是开发、集成和验证关键平台技术，以筛选基于机制的联合方案和光动力疗法(PDT)治疗残留和复发的Ovca。异质细胞3D打印肿瘤阵列融合了OvCa生物学的关键决定因素(内皮细胞和间皮细胞与巨噬细胞和成纤维细胞)，以及用于同时对多个生物标志物进行定量成像的高光谱显微镜，将在高通量平台上提供对OvCa生长和治疗反应的特殊洞察。为了解决导致OvCa相关死亡的令人精疲力竭的毒性和频繁的复发，我们利用我们的纳米技术计划制造纳米结构，用于靶向抑制剂的细胞内递送，并提供与PDT的合理组合方案，PDT是FDA批准的一种基于光化学的治疗方法，已显示出治疗OvCa的临床前景。PDT对化疗和辐射耐药细胞有效，并与化疗和生物制剂协同作用，从而提高疗效。在异质细胞3D阵列中优化的治疗结果将在体内和体外患者组织衍生3D培养中得到严格验证，以展示基于生物和成像的筛查平台的预测能力。这些目标将在3个具体目标中实现：1)开发和表征用于微转移OvCa的异质细胞3D打印肿瘤阵列，以及用于细胞内治疗药物输送的纳米结构。2)部署异种细胞肿瘤阵列，以评估对两级治疗方法的细胞毒性和分子反应-i)一线PDT+残留疾病的化疗，然后ii)二线PDT+靶向生物疗法治疗化疗耐药复发疾病。3)验证3D肿瘤阵列在体内的治疗反应，以及在3D培养的体外组织来源细胞中的治疗反应。这项建议的主要成果将是：i)用于多药物细胞内输送的异细胞3D OvCa肿瘤阵列和纳米结构，ii)减少再生的最佳组合输送策略和条件，以及iii)在临床相关的转移性OvCa小鼠模型和患者组织衍生3D培养的背景下对异细胞肿瘤阵列数据的严格验证。这项研究的结果将影响晚期(II-IV期)和耐药Ovca患者以及接受挽救治疗的患者的预后。通过这种高度集成的方法开发的基础设施将创建一个新的框架，以快速评估和优化新的治疗策略，这些策略将适用于广泛的转移性肿瘤和分子靶点。由于分子表达和反应可能是特殊的，拟议的对生物标记物表达和治疗诱导的生物标记物变化的快速监测为未来患者定制治疗创造了可能性。

DESCRIPTION (provided by applicant): Late detection and drug resistance have maintained the grim statistics for ovarian cancer (OvCa) steady over decades. New approaches using combination therapies with mechanistically distinct components are hypothesized to be most effective. The long term goal of this research is to develop, integrate and validate key platform technologies to screen mechanism-based combination regimens with photodynamic therapy (PDT) for residual and recurrent OvCa. Heterocellular 3D printed tumor arrays that incorporate critical determinants of OvCa biology (endothelial and mesothelial cells with macrophages and fibroblasts) along with hyperspectral microscopy for simultaneous quantitative imaging of multiple biomarkers will provide exceptional insight into OvCa growth and treatment response on a high throughput platform. To address the grueling toxicities and frequent recurrence that cause OvCa-related deaths, we leverage our nanotechnology program to fabricate nanoconstructs for intracellular delivery of targeted inhibitors, and deliver rational combination regimens with PDT, an FDA approved photochemistry-based treatment that has shown clinical promise for OvCa. PDT is effective on chemo and radiation resistant cells and synergizes with chemotherapeutic and biologic agents resulting in improved efficacy. Treatment results optimized in the heterocellular 3D arrays will be rigorously validated in vivo and in ex vivo patient tissue-derived 3D cultures to demonstrate the predictive capabilities of the bio- and imaging-based screening platform. The goals will be realized in 3 specific aims: 1) Develop and characterize heterocellular 3D printed tumor arrays for micrometastatic OvCa, and nanoconstructs for intracellular delivery of therapeutic agents. 2) Deploy heterocellular tumor arrays for assessment of cytotoxic and molecular responses to a two-tiered therapeutic approach- i) first-line PDT + chemotherapy of residual disease followed by ii) second-line treatment of chemoresistant recurrent disease with PDT + targeted biological therapies. 3) Validate treatment response of 3D tumor arrays in vivo, and in ex-vivo tissue derived cells in 3D culture. Major deliverables of this proposal will be i) Heterocellular 3D OvCa tumor arrays and nanoconstructs for multi-agent intracellular delivery, ii) Optimal combination delivery strategies and conditions to mitigate regrowth, and iii) Rigorous validation of the heterocellular tumor array data in the context of both a clinically-relevant murine model for metastatic OvCa, and in patient tissue-derived 3D cultures. The findings from this study will impact outcomes for patients with advanced (stage II-IV) and resistant OvCa and those receiving salvage therapy. The infrastructure developed through this highly integrated approach will create a new framework to rapidly evaluate and optimize new therapeutic strategies that will be adaptable to a broad array of metastatic tumors and molecular targets. Because molecular expressions and responses can be idiosyncratic, the proposed rapid monitoring of biomarkers expression and treatment-induced biomarkers changes creates the possibility of patient-customized treatments in the future.