Heterocellular 3D ovarian tumor arrays for imaging and mechanistic combinations

描述（由申请人提供）：晚期检测和耐药性使卵巢癌（OvCa）的严峻统计数据保持了数十年的稳定。新的方法，使用联合治疗与机械不同的组件被假设为是最有效的。本研究的长期目标是开发、整合和验证关键平台技术，以筛选基于机制的联合方案与光动力疗法（PDT），用于治疗残留和复发性OvCa。异质细胞3D打印肿瘤阵列结合了OvCa生物学的关键决定因素（内皮细胞和间皮细胞以及巨噬细胞和成纤维细胞），沿着高光谱显微镜，用于多种生物标志物的同时定量成像，将在高通量平台上提供对OvCa生长和治疗反应的特殊见解。为了解决导致OvCa相关死亡的剧烈毒性和频繁复发，我们利用我们的纳米技术计划来制造用于细胞内递送靶向抑制剂的纳米结构，并提供与PDT的合理组合方案，PDT是FDA批准的基于光化学的治疗，已显示出OvCa的临床前景。PDT对化疗和放射抗性细胞有效，并与化疗剂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离体患者组织衍生的3D培养物中进行严格验证，以证明基于生物和成像的筛选平台的预测能力。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的异细胞3D打印肿瘤阵列，以及用于治疗剂细胞内递送的纳米结构。2)部署异质细胞肿瘤阵列，以评估对两层治疗方法的细胞毒性和分子反应- i）一线PDT +残留疾病化疗，然后ii）PDT +靶向生物治疗对化疗耐药复发性疾病的二线治疗。3)在3D培养物中观察体内3D肿瘤阵列和离体组织衍生细胞的治疗反应。该提案的主要可交付成果将是i）用于多药剂细胞内递送的异细胞3D OvCa肿瘤阵列和纳米构建体，ii）减轻再生长的最佳组合递送策略和条件，以及iii）在转移性OvCa的临床相关鼠模型和患者组织衍生的3D培养物中对异细胞肿瘤阵列数据进行严格验证。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结局。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异质的，因此所提出的对生物标志物表达和治疗诱导的生物标志物变化的快速监测创造了未来患者定制治疗的可能性。

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.