Project 1: Combinatorial adjuvants promote uniform and selective intratumoral CTL infiltration in colorectal cancer

项目1：摘要 肿瘤微环境(TME)的CTL浸润可预测结直肠癌患者的生存时间延长 癌症(CRC)。它还区分了一小部分患有微卫星不稳定的患者(5%)。 高[MSI-H]CRC，肿瘤内CTL水平高，对PD-1阻断有反应 结直肠癌患者无反应。我们的初步数据表明，a)基于TLR3的佐剂诱导 CTL选择性地吸引肿瘤间质中的趋化因子，但不能在非肿瘤组织周围；b) TLR3配体(如Rintatolimod)与干扰素α协同诱导高水平的CTL- 诱导剂在所有肿瘤病变中均匀存在；c)COX2阻滞剂的加入增强了CKM在 促进CTL吸引，但抑制Treg吸引。三组分趋化因子的调节作用 方案(CKM rintatolimod，干扰素α塞来昔布)增加CTL在肿瘤内的积聚，延长生存期和 与PD1-和PD-L1阻滞剂在诱导治疗(&gt；150天存活)IP小鼠方面有协同作用。MC38肿瘤， 单独抵抗PD-1封锁。我们完成了对肝脏患者系统性(静脉)CKM的I期评估。 转移性结直肠癌(NCT01545141)，观察其很好的耐受性和CTL/Treg比率的改善 TME中的标志物(与我们仅接受标准护理的患者相比)。我们建议： 目的1.评价静脉注射CKM促进局部免疫的住院效果 IIa期试验NCT03403634中肝转移结直肠癌病灶中CTL的积聚。比较之前与 治疗后12例肝转移癌患者的肿瘤活检，我们将测试系统的CKM是否会 消除TME异质性，均匀增加TME内CTL数，但不增加周围组织CTL数。 目的：评价CKM和CKM序贯给药与周期给药的免疫和抗肿瘤作用。 PD1阻断和额外免疫的优势，对持久的抗肿瘤有好处。在……里面 临床前研究，我们将检验CKM吸引的DC、NK细胞和T细胞将a)促进 局部和全身肿瘤特异性免疫和b)将放大CKM启动的肿瘤内CTL的产生 干扰素γ和肿瘤坏死因子α依赖机制中的引诱剂，导致TME对 即使在没有额外疫苗接种的情况下，PD1的持续抗肿瘤活性也会被阻断。 目的3.进行一项I/II期试验，以测试CKM联合PD-1阻断的临床活性 微卫星稳定(MSS)结直肠癌患者。在I/II期试验中，我们将评估临床疗效(iORR； 对19例传统免疫治疗耐药的MSS-CRC患者进行CKM/抗PD-1治疗。 规划角色：项目1的独特作用是评估有效性、一致性和肿瘤- 系统应用CKM的选择性，开发具有持续抗癌作用的基于CKM的治疗方法。 它的成功将为我们提供一种工具，将免疫疗法的治疗益处扩大到特别大的 目前对检查站封锁无反应的MSS-CRC和其他癌症患者组。

PROJECT 1: ABSTRACT CTL infiltration of tumor microenvironments (TME) predicts prolonged survival of patients with colorectal cancer (CRC). It also differentiates between the small subset of patients (<5%) with microsatellite instability- high [MSI-H] CRC, who show high levels of intratumoral CTLs and respond to PD-1 blockade from the rest of CRC patients who do not respond. Our preliminary data demonstrate that a) TLR3-based adjuvants induce CTL-attracting chemokines selectively in tumor stroma, but not surrounding non-tumor tissues; b) that combination of TLR3 ligands (such as rintatolimod) with IFNα synergistically induce high levels of CTL- attractants uniformly in all tumor lesions; and c) that inclusion of COX2 blockers enhances specificity of CKM in promoting CTL attraction but suppressing Treg attraction. The three-component chemokine-modulatory regimen (CKM rintatolimod, IFNα celecoxib) enhanced intratumoral CTL accumulation, prolonged survival and synergized with PD1- and PD-L1 blockers in inducing cures (> 150 day survival) in mice with i.p. MC38 tumors, resistant to PD-1 blockade alone. We completed phase I evaluation of systemic (i.v) CKM in patients with liver- metastatic CRC (NCT01545141), observing its very good tolerability and improved ratios of CTL-to-Treg markers in TME (compared to our patients receiving standard care only). We propose to: Aim 1. Evaluate the in-patient immunologic effectiveness of i.v.- administered CKM to promote local CTL accumulation in liver-metastatic CRC lesions in phase IIa trial NCT03403634. Comparing pre- versus post-treatment tumor biopsies of 12 patients with liver-metastatic CRC, we will test if systemic CKM will abrogate the TME heterogeneity and uniformly increase CTL numbers in TMEs, but not in surrounding tissues. Aim 2. Evaluate the immune and antitumor effects of sequential versus cyclic application of CKM and PD1 blockade and the advantage of additional immunization for long-lasting anti-tumor benefit. In preclinical studies, we will test the hypotheses that the CKM-attracted DCs, NK cells and T cells will a) promote local and systemic tumor-specific immunity and b) will amplify the CKM-initiated intratumoral production of CTL attractants in an IFNγ and TNFα-dependent mechanism, resulting in sustained conditioning of the TME for continued antitumor activity of PD1 blockade, even in the absence of additional vaccination. Aim 3. Perform a phase I/II trial to test the clinical activity of CKM combined with PD-1 blockade in patients with microsatellite-stable (MSS) CRC. In phase I/II trial, we will evaluate the clinical efficacy (iORR; iRESIST) of CKM/anti-PD-1treatment in 19 patients with MSS-CRC, traditionally resistant to immunotherapy. Programmatic Role: The unique role of Project 1 is to evaluate the effectiveness, uniformity and tumor- selectivity of systemically-applied CKM and develop CKM-based treatments with sustained anticancer effect. Its success will provide us with a tool to extend the therapeutic benefit of immunotherapy to a particularly large group of patients with MSS-CRC and other cancers currently non-responsive to checkpoint blockade.