Project 2: Local immunotherapy corrects chemokine patterns in ovarian cancer

肿瘤微环境中高数量的CD8+CTL和低数量的调节性T(REG)细胞 预测卵巢癌(OvCa)患者的生存时间延长和PD-1阻滞剂在 其他癌症。我们观察到tlr配体、干扰素α和cox2阻滞剂之间的强烈协同作用。 在临床前OvCa模型中诱导CTL趋化因子，但抑制Treg趋化因子。 Rintatolimod(TLR3-配体)、干扰素α和塞来昔布(趋化因子调节方案)联合应用 促进肿瘤特异性CTL在小鼠卵母细胞TME中的聚集，并与PD-1拮抗剂协同作用 携带PD-1抗性ID8肿瘤的小鼠，导致50%的动物长期存活(&gt；100天)。我们 启动I/II阶段试验(NCT02432378)以测试本地(I.P.)Rintatolimod/干扰素α用于指导临床安全有效 αDc1疫苗诱导CXCR3+/CCR5+CTL产生OvCaTME。我们观察到该药耐受性良好。CKM和 CD8、PD-1、PD-L1、PD-L2局部升高。我们还观察到CKM能吸引CTL和NK细胞 诱导两条独立的(PD-1/PD-L1和COX2依赖)免疫抑制通路， 为它们同时靶向以实现长期治疗效果提供了理论依据。我们建议： 目的1：确定腹腔注射CKM的安全性及其促进局部治疗的有效性 αDc1疫苗免疫卵巢癌患者TME中CTL的积聚接受第二期治疗的病人 NCT02432378将接受新辅助化疗和αDc1疫苗接种，有或没有局部(Ip)。CKM。 我们将评估I.P.的影响。CKM对局部CTL流入的幅度和持续时间的影响。我们将获得 CKM/αdc1疫苗治疗对pFS和OS潜在临床影响的初步数据。 目的2：探讨人卵母细胞TME“继发性”抑制的诱导机制。 对免疫攻击的反应。我们将检验CKM优先吸引CTL、NK和Th1的假设 细胞(与MDSCs和Treg细胞相比)，但由这些效应细胞产生的干扰素γ/肿瘤坏死因子α触发两个独立的 PD-1/PD-L1/PD-L2系统和COX2/PGE2-系统介导的“二次”抑制途径。 目的3：确定同时靶向PD-1和COX2是否导致持续治疗 αDc1疫苗的效果。我们将测试αDc1/CKM激活的CTL是否激活PD-1/PD-L1和COX2/PGE_2- 在OvCa荷瘤小鼠中的途径以及它们的互补靶向是否产生治疗协同效应。指导原则 这些结果，我们将最终确定设计，并将测试CKMDc1[肿瘤]/(α联合PD-1阻断将 诱导不能切除卵巢癌患者的客观临床反应(IRECIST)。 规划作用：项目2的独特作用是评估地方管理的纵向影响 CKM、疫苗接种、CKM和PD-1阻断之间的相互作用以及PD-1/PD-L1/PD-L2系统的作用 与COX2/PGE2系统相比，作为TME“二次抑制”的互补介质。

High numbers of CD8+ CTLs and low numbers of regulatory T(reg) cells in tumor microenvironments (TME) predicts prolonged survival of ovarian cancer (OvCa) patients and the clinical effectiveness of PD-1 blockers in other cancers. We observed strong synergy between TLR ligands, IFNα and COX2 blockers in specifically inducing the CTL-attracting chemokines, but suppressing Treg attractants in preclinical OvCa models. Combination of rintatolimod (TLR3-ligand), IFNα and celecoxib (chemokine-modulating regimen; CKM) promoted accumulation of tumor-specific CTLs in mouse OvCa TME, and synergized with PD-1 blockade in mice with PD-1 resistant ID8 tumors, resulting in long-term (>100 days) survival in 50% of the animals. We initiated a phase I/II trial (NCT02432378) to test if local (i.p.) rintatolimod/IFNα is safe and effective in guiding αDC1 vaccine-induced CXCR3+/CCR5+ CTLs to OvCa TME. We observed good tolerability of i.p. CKM and local elevation of CD8, PD-1, PD-L1 and PD-L2. We also observed that CKM-attracted CTLs and NK cells induce two separate (PD-1/PD-L1- and COX2-dependent), pathways of “secondary” immune suppression, providing rationale for their simultaneous targeting to achieve long-lasting therapeutic effects. We propose to: Aim 1: Determine the safety of i.p-administered CKM and its effectiveness in promoting local accumulation of CTLs in the TME of αDC1-vaccinated OvCa patients. Patients on phase II of NCT02432378 will receive neoadjuvant chemotherapy and αDC1 vaccination, with or without local (i.p.) CKM. We will evaluate the impact of i.p. CKM on the magnitude and duration of local CTL influx. We will obtain preliminary data on potential clinical impact of the CKM/αDC1-vaccine treatment on PFS and OS. Aim 2: Determine the mechanism of induction of “secondary” suppression in human OvCa TME in response to immune attack. We will test the hypothesis that CKM preferentially attracts CTLs, NK & Th1 cells (vs. MDSCs & Tregs), but that IFNγ/TNFα-production by these effector cells triggers two independent pathways of “secondary” suppression, mediated by PD-1/PD-L1/PD-L2 system and COX2/PGE2-system. Aim 3: Determine whether simultaneous targeting of PD-1 and COX2 results in sustained therapeutic effects of αDC1 vaccines. We will test if αDC1/CKM-activated CTLs activated PD-1/PD-L1 and COX2/PGE2- pathways in OvCa-bearing mice and if their complementary targeting results in therapeutic synergy. Guided by these results, we will finalize the design and will test if αDC1[tumor]/(CKM combined with PD-1 blockade will induce objective clinical responses (iRECIST) in patients with unresectable OvCa. Programmatic Role: The unique role of Project 2 is to evaluate the longitudinal effects of locally-administered CKM, interplay between vaccination, CKM and PD-1 blockade, and the roles of the PD-1/PD-L1/PD-L2 system vs. COX2/PGE2 systems, as complementary mediators of “secondary suppression” in TME.