Project 1: Combining PARP inhibition with radiation to sensitize HR proficient pancreatic cancers to immunotherapy

项目总结/摘要（项目1） 我们发现DDR（DNA损伤反应）抑制剂增强辐射诱导的T1 IFN（I型 干扰素）介导的先天免疫和随后的适应性免疫。我们还表明，PARP 抑制剂是辐射增敏剂，具有在辐射诱导的DNA损伤中诱导损伤的独特性质 网站.在本申请中，我们研究了它们与辐射的组合作为诱导先天免疫的策略 并使PDAC（胰腺导管腺癌）对免疫疗法敏感。我们确认，与A 临床研究表明，辐射本身是一种弱免疫刺激剂。相反，辐射与 PARP抑制剂奥拉帕尼在PDAC中强烈诱导T1 IFN介导的先天免疫，导致致敏 局部和全身肿瘤的免疫治疗。这些数据共同支持我们提出的临床研究。 在LAPC（局部晚期胰腺癌）患者中联合奥拉帕尼、放疗和durvalumab的试验。 该提案的总体目标是在临床前开发一种将PARP抑制剂与放射治疗相结合的策略 通过促进肿瘤细胞DNA损伤使PDAC对ICB（免疫检查点阻断）敏感， 免疫原性，将转化为LAPC患者的临床试验。我们将在三个具体的 目标。特异性目的1将阐明先天免疫调节及其T1 IFN依赖性的新机制。 PDAC细胞中PARP抑制剂与辐射的表型结果。初步数据显示， 与单独的PARP抑制剂相比，PARP抑制剂奥拉帕尼与辐射协同作用， 并在HR（同源重组）熟练的PDAC中诱导T1 IFN先天免疫应答。目标1将 定义新的机制T1干扰素的产生及其随后的表型后果。我们预计 定义由奥拉帕尼和辐射诱导的DNA启动的新型模式识别受体（PRR）途径 损伤或复制应激导致肿瘤先天免疫增加。具体目标2将决定 PARP抑制剂、放疗和ICB联合治疗的治疗获益和分子终点 PDAC。我们将确定局部和全身抗肿瘤疗效和毒性的高或低剂量每部分 辐射（即，SBRT，立体定向体部放射或标准分割放射）和最佳时间表， 奥拉帕尼和抗PD-L1。我们将在肿瘤和血液中建立反映DNA损伤、肿瘤先天性 和适应性免疫，以及更广泛的免疫概况（CyTOF）。我们希望能取得一个有利的 SBRT的治疗指数与肿瘤DNA损伤和T1 IFN介导的先天性和 适应性免疫在具体目标3中，我们将进行一项奥拉帕尼、放疗和杜伐鲁单抗的临床试验， LAPC患者我们将使用至事件发生时间连续再评估方法递增奥拉帕尼的剂量 （TiTE-CRM）联合SBRT（除非目标2另有说明）和PD-L1阻断 LAPC患者中的durvalumab抗体。我们预计，我们将实现一个安全和有效的 奥拉帕尼与放射和durvalumab的组合，其促进最大免疫原性， 激发了一项随机2期试验，以最终提高LAPC患者的生存率。

PROJECT SUMMARY/ABSTRACT (PROJECT 1) We discovered that inhibitors of the DDR (DNA damage response) enhance radiation-induced T1IFN (Type I interferon)-mediated innate immunity and, subsequently, adaptive immunity. We have also shown that PARP inhibitors are radiation sensitizers with unique properties in inducing lesions at radiation-induced DNA damage sites. In this application, we investigate their combination with radiation as a strategy to induce innate immunity and sensitize PDAC (pancreatic ductal adenocarcinoma) to immunotherapy. We confirm, in agreement with a clinical study, that radiation alone is a weak immune stimulator. In contrast, the combination of radiation with the PARP inhibitor olaparib strongly induces T1IFN-mediated innate immunity in PDAC resulting in sensitization of both local and systemic tumors to immunotherapy. Taken together these data support our proposed clinical trial combining olaparib, radiation and durvalumab in patients with LAPC (locally advanced pancreatic cancer). The overall goal of this proposal is to preclinically develop a strategy combining PARP inhibitors with radiation for sensitizing PDAC to ICB (immune checkpoint blockade) by promoting tumor cell DNA damage and immunogenicity that will be translated to a clinical trial in LAPC patients. We will accomplish this in 3 specific aims. Specific Aim 1 will elucidate novel mechanisms of innate immune modulation and their T1IFN-dependent phenotypic consequences by PARP inhibitors with radiation in PDAC cells. Our preliminary data show, in contrast to PARP inhibitor alone, that the PARP inhibitor olaparib synergizes with radiation to reduce survival and induce a T1IFN innate immune response in HR (homologous recombination) proficient PDAC. Aim 1 will define novel mechanisms of T1IFN generation and its subsequent phenotypic consequences. We anticipate defining novel pattern recognition receptor (PRR) pathways initiated by olaparib and radiation-induced DNA damage or replication stress that result in increased tumor innate immunity. Specific Aim 2 will determine the therapeutic benefit and molecular endpoints of combined therapy with PARP inhibitor, radiation and ICB in PDAC. We will define the local and systemic anti-tumor efficacy and toxicity of high or lower dose per fraction radiation (i.e., SBRT, stereotactic body radiation or standard fractioned radiation) and the optimal schedule with olaparib and anti-PD-L1. We will establish endpoints in tumors and blood reflecting DNA damage, tumor innate and adaptive immunity, as well as the broader immune profile (CyTOF). We expect to achieve a favorable therapeutic index with SBRT that is associated with tumor DNA damage and T1IFN-mediated innate and adaptive immunity. In Specific Aim 3, we will conduct a clinical trial of olaparib, radiation, and durvalumab in patients with LAPC. We will dose escalate olaparib using the Time-to-Event Continual Reassessment Method (TiTE-CRM) with the combination of SBRT (unless otherwise informed by Aim 2) and the PD-L1 blocking antibody durvalumab in patients with LAPC. We anticipate that we will achieve a safe and efficacious combination of olaparib with radiation and durvalumab that promotes maximal immunogenicity and motivates a randomized phase 2 trial to ultimately improve survival in patients with LAPC.