Optimization of High Frequency Irreversible Electroporation (H-FIRE) for tumor ablation and immune system activation in pancreatic cancer applications

项目摘要：胰腺癌约占美国所有癌症的3% 约占所有与癌症相关的死亡人数的7%。迫切需要新的治疗模式。新兴 肿瘤消融技术已经显示出巨大的前景。该提案将重点放在高频 不可逆电穿孔(H-FIRE)，通过插入的电极传递一系列电脉冲 直接进入肿瘤，在靶细胞膜上产生结构缺陷，导致癌细胞死亡。 这项建议的目的是利用我们的小鼠和新的猪临床前动物模型来扩展 本提案中提供的初步数据并生成关键的机械、安全和功效数据 以支持未来胰腺癌患者的H-FIRE临床试验。我们最重要的假设 H-Fire将有效地减轻生理和临床相关的胰腺的异质性 肿瘤，治疗导致关键组织结构附近的连续消融区。我们进一步 假设H-Fire的好处最终将扩展到局部肿瘤消融并产生 可预测、可调节的全身性抗肿瘤免疫反应可减少转移负担和预防 复发。具体目标1将描述胰腺癌细胞的生物物理反应 纸巾呼叫H-FIRE。这一目标将评估H-Fire脉冲参数可调至 实现高度相关的不同细胞死亡结果(细胞凋亡、下垂、坏死性下垂或坏死) 对肿瘤消融、肿瘤微环境、抗肿瘤免疫反应的影响。一致地，我们将评估 使用傅立叶分析光谱学(FAST)的实时治疗反馈消融进展。我们 希望确定哪些参数(即脉冲宽度、通电时间、相间/脉冲间延迟)起作用 在调节相关癌细胞系和体外组织中的细胞死亡方面发挥重要作用。特定目标 2将建立胰腺癌的H-Fire治疗策略，以优化肿瘤消融和 全身性抗肿瘤免疫反应。使用Pan02小鼠模型，这一目标将检验以下假设 H-FIRE是体内精确、彻底切除胰腺肿瘤的一种有效的治疗方法。我们也 假设由于H-fire介导的细胞死亡和由此导致的肿瘤变化的独特特征 微环境下，局部肿瘤消融将产生可预测和可调节的全身抗肿瘤宿主免疫 减轻转移负担和防止复发的反应。具体目标3将定义H-FIRE 利用生理和临床相关的治疗参数并确定其安全性 猪胰腺癌模型。这一目标将检验H-Fire可以有效烧蚀的假设 生理和临床相关原位条件下的原位胰腺肿瘤。为了测试这一点 假设，我们将利用新的原位猪胰腺癌模型，该模型具有不同范围的 临床相关的物理特性，预测会影响人类患者的H-FIRE疗效。

PROJECT SUMMARY: Pancreatic cancer accounts for approximately 3% of all cancers in the United States and approximately 7% of all cancer related deaths. New treatment paradigms are direly needed. Emerging tumor ablation techniques have shown significant promise. This proposal will focus on High-Frequency Irreversible Electroporation (H-FIRE), which delivers a series of electric pulses through electrodes inserted directly into the tumor to produce structural defects in the target cell membrane resulting in cancer cell death. The objective of this proposal is to utilize our mouse and novel pig preclinical animal models to expand upon the preliminary data presented in this proposal and generate critical mechanistic, safety, and efficacy data necessary to support future H-FIRE clinical trials in pancreatic cancer patients. Our overarching hypothesis is that H-FIRE will effectively mitigate heterogeneity in physiologically and clinically relevant pancreatic tumors, with treatments leading to contiguous zones of ablation near critical tissue structures. We further postulate that the benefits of H-FIRE will ultimately extend beyond focal tumor ablation and generate a predictable, tunable systemic anti-tumor immune response reducing metastatic burden and preventing recurrence. Specific Aim 1 will characterize the biophysical response of pancreatic cancer cells and tissues to H-FIRE. This Aim will evaluate the hypothesis that H-FIRE pulse parameters can be tuned to achieve different cell death outcomes (apoptosis, pyroptosis, necroptosis, or necrosis) that are highly relevant to tumor ablation, the tumor microenvironment, and anti-tumor immune responses. In concert, we will assess ablation development with real time treatment feedback using Fourier Analysis Spectroscopy (FAST). We expect to determine which parameters (i.e. pulse width, energized time, interphase/interpulse delay) play significant roles in tuning cell death elicited within relevant cancer cell lines and ex vivo tissues. Specific Aim 2 will establish H-FIRE treatment strategies for pancreatic cancer that optimize tumor ablation and systemic anti-tumor immune responses. Using Pan02 mouse models, this Aim will test the hypothesis that H-FIRE is an effective treatment modality for precise and complete pancreatic tumor ablation in vivo. We also postulate that due to the unique features of H-FIRE mediated cell death and resultant changes in the tumor microenvironment, focal tumor ablation will result in predictable and tunable systemic anti-tumor host immune responses reducing metastatic burden and preventing recurrence. Specific Aim 3 will define H-FIRE treatment parameters and determine its safety profile utilizing physiologically and clinically relevant porcine models of pancreatic cancer. This Aim will test the hypothesis that H-FIRE can effectively ablate orthotopic pancreatic tumors under physiologically and clinically relevant in situ conditions. To test this hypothesis, we will utilize novel, orthotopic, porcine pancreatic cancer models featuring a diverse range of clinically relevant physical properties that are predicted to impact H-FIRE efficacy in human patients.