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

在美国，胰腺癌约占所有癌症的3%。 占所有癌症相关死亡的7%。迫切需要新的治疗模式。新兴 肿瘤消融技术已经显示出显著的前景。该提案将重点关注高频 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。 本提案的目的是利用我们的小鼠和新型猪临床前动物模型， 本提案中提供初步数据，并生成关键的机制、安全性和有效性数据 这是支持未来在胰腺癌患者中进行H-FIRE临床试验的必要条件。我们的首要假设是 H-FIRE将有效地减轻生理和临床相关胰腺组织中的异质性， 肿瘤，其中治疗导致在关键组织结构附近的连续消融区。我们进一步 假设H-FIRE的益处最终将超出局灶性肿瘤消融， 可预测的、可调节的全身性抗肿瘤免疫应答， 复发特异性目标1将表征胰腺癌细胞的生物物理反应， H-FIRE。本目标将评估H-FIRE脉冲参数可以调整到 实现高度相关的不同细胞死亡结果（凋亡、焦亡、坏死性凋亡或坏死） 肿瘤消融、肿瘤微环境和抗肿瘤免疫反应。我们将共同评估 使用傅立叶分析光谱学（FAST）的具有真实的时间治疗反馈的消融发展。我们 期望确定哪些参数（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离体组织内引起的细胞死亡中的重要作用。具体目标 2将为胰腺癌建立H-FIRE治疗策略，优化肿瘤消融， 全身性抗肿瘤免疫应答。使用Pan 02小鼠模型，本目标将检验以下假设： H-FIRE是一种有效的治疗方式，可在体内进行精确和完全的胰腺肿瘤消融。我们也 假设由于H-FIRE介导的细胞死亡的独特特征和肿瘤中的结果变化， 局部肿瘤消融将导致可预测和可调的全身抗肿瘤宿主免疫应答。 减少转移负担和预防复发的反应。具体目标3将定义H-FIRE 治疗参数，并利用生理学和临床相关信息确定其安全性 胰腺癌的猪模型。本目标将检验H-FIRE可有效消融的假设 原位胰腺肿瘤在生理和临床相关的原位条件下。为了验证这一 假设，我们将利用新的，原位，猪胰腺癌模型，具有不同的范围， 临床相关的物理性质，预计会影响H-FIRE在人类患者中的疗效。

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.