Project 3: Immune Checkpoint Inhibition Therapy Enhanced by Integrated Photodynamic Treatment and Image Guidance in Preclinical Models of Pancreatic Cancer

尽管联合治疗取得了进展，但胰腺导管腺癌(PDAC)的统计数据仍然令人沮丧。 化疗。它在很大程度上是耐化疗和耐辐射的，而手术是唯一的治愈选择。 仅对约20%的患者有效。免疫检查点抑制(ICI)带来了希望，但在PDAC中，即使是 成功(抗PD1：pembrolizumab)仅限于1-3%表现为微卫星不稳定的患者。这个 大多数肿瘤缺乏有效的ICI所需的免疫渗透。根据我们的数据和文献， 我们假设光动力启动(PDP)是光动力疗法(PDT)的一个结果。 而我们的主要研究重点是改变肿瘤微环境，使其增敏，从而加强ICI治疗。PDP 诱导免疫原性细胞死亡并促进肿瘤浸润性淋巴细胞(TIL)迁移， PDP诱导的肿瘤通透性增加。我们利用这种启动效应来设计一种非经验性方法 进行PDT-ICI联合治疗。在这方面，我们最新的高光谱成像创新给了我们很大的帮助。 首次能够在活的荷瘤动物身上同时监测6个生物标记物；它允许我们 以量化TIL(和亚集)以及PD1/PD-L1表达的变化，以产生免疫核心(IS)。 我们假设，当肿瘤已经被PDP刺激为 “最热”，从而将ICI剂量和相关毒性降至最低，类似于观察到的PDP化疗 组合。这一假设将在3个目标中得到验证。目标1将在小鼠原位肿瘤中鉴定PDP- 剂量学和IS调制的最佳时机确定抗PD1治疗的最大益处的时间 (增加存活率，减少转移)和增加耐受性。目标2，由目标1通知，将使用这两个 PDP-ICI原位和双侧皮下注射PDAC小鼠模型的建立 心理治疗。最佳时机、单剂ICI治疗与多剂量联合应用的比较 单免疫检查点阻断(抗PD1)与双阻断(抗PD1/抗PD-L1)的比较 也被建立起来。AIM 3将利用患者来源的肿瘤免疫器官(PDIO)，概括 异质性和评估联合治疗的结果。PDP诱导的IS将在最佳时间通知 PDIO的抗PD1/抗PD-L1治疗指导。相关性和影响1)它使非免疫性变得敏感 有反应的肿瘤和扩大符合条件的PDAC患者(目前只有3%)。2)A 引入了非经验法。3)结合成像启用的抗PD1剂量的减少，它减少了 免疫药物与化疗和受体酪氨酸激酶抑制剂的毒性报道。4)它提供 广泛的成像平台，用于在体内以细胞分辨率实时提供多标记信息，并充当 通过允许同时可视化几个分子靶点(例如，肿瘤)的通用护理点工具 免疫细胞表型和细胞因子/基质/代谢信号)，用于设计新的治疗方法 其他疾病，并有可能扩大使用细针抽吸活组织检查PDAC ICI剂量。

The statistics for pancreatic ductal adenocarcinoma (PDAC) remain dismal despite advances in combination chemotherapies. It is largely chemo and radiation resistant, and surgery, the only curative option, is available to only ~20% of patients. Immune checkpoint inhibition (ICI) has brought hope but in PDAC even the modest success (anti-PD1: Pembrolizumab) is limited to only 1-3% of patients displaying microsatellite instability. The majority of tumors lack the immune infiltration necessary for effective ICI. Based on our data and the literature, we hypothesize that Photodynamic Priming (PDP), a process that is a fallout of photodynamic therapy (PDT) and our main research focus, alters the tumor microenvironment to sensitize it to enhance ICI therapy. PDP induces immunogenic cell death and enhances tumor infiltrating lymphocyte (TIL) migration, augmented by PDP-induced higher tumor permeability.. We capture this priming effect for designing a non-empiric approach for a PDT-ICI combination. We are greatly helped in this by our recent innovation of hyperspectral imaging capable, for the first time, of monitoring 6 biomarkers simultaneously in live tumor bearing animals; it allows us to quantify TILs (and subsets) along with PD1/PD-L1 expression changes to generate an immunoscore (IS). We posit that ICI administration will be most beneficial when the tumor has been PDP-stimulated to be the “hottest” thus minimizing ICI dose and associated toxicities, similar to that observed for PDP-chemotherapy combinations. The hypothesis will be tested in 3 aims. Aim 1 will identify, in orthotopic murine tumors, PDP- dosimetry and optimal timing for IS modulation to establish the time for maximal benefits of anti-PD1 therapy (increased survival, decreased metastasis) and increased tolerability. Aim 2, informed by aim 1, will use both orthotopic and a bilateral subcutaneous PDAC murine model to determine abscopal effects of PDP-ICI therapy. A comparison of optimally timed, single dose ICI therapy with multiple dosages along with a comparison of single immune checkpoint blockade (anti-PD1) with dual blockade (anti-PD1/anti-PD-L1) will also be established. Aim 3 will utilize patient derived tumor immune organoids (PDIO), to recapitulate heterogeneity and evaluate the combination therapy outcomes. PDP induced IS will inform optimally timed anti-PD1/anti-PD-L1 therapy guidance for PDIOs. Relevance and Impact 1) it sensitizes non-immune responsive tumors to responsive ones and expands the eligible PDAC patients (currently only at 3%). 2) A non-empiric approach is introduced. 3) Combined with imaging-enabled dose reduction of anti-PD1 it reduces toxicities of immune drugs as reported with chemotherapy and receptor tyrosine kinase inhibitors. 4) It provides a broad imaging platform for in vivo real-time multiple-marker information at cellular resolution and serves as a general point-of-care tool by enabling simultaneous visualization of several molecular targets (e.g., tumor immune cell phenotypes and cytokines/stromal/metabolic signatures) for designing new therapies for many other diseases, and with potential to expand the use of fine needle aspirate biopsy for PDAC ICI dosimetry.