First-in-class TREM-1 inhibitors in combination therapy for pancreatic cancer

项目摘要/摘要 胰腺癌，或胰腺癌(PC)，是癌症相关死亡的第四大原因 在美国。根据美国癌症协会的数据，2016年预计会有55,300个新病例。 尽管在治疗方面取得了进展，但5年存活率还不到4%。目前对PC的治疗包括 手术、放射治疗、化疗和免疫治疗，但它们都只能略微延长生命或 缓解PC患者的症状。吉西他滨(GEM)是治疗晚期PC的一线疗法，仅 在随机临床试验中疗效适中，中位生存期约为6个月。 不同抗癌剂的宝石相比，并没有显示出显著的生存优势 只有宝石一家。现有治疗方法在疗效方面的这些局限性突出表明需要新的治疗方法。 众所周知，胰腺炎症会增加患PC的风险。高巨噬细胞在肿瘤中的侵袭 肿块与促进肿瘤生长和转移发展密切相关。触发受体 表达在髓样细胞(TREM-1)上，是一种炎症放大因子，在PC的进展中发挥作用。 PC和PC患者肿瘤相关巨噬细胞(TAM)上TREM-1的表达上调 与疾病的严重程度相关。最近，我们证明了一种一流的TREM-1抑制肽 GF9以自由形式存在并结合到巨噬细胞靶向脂肽复合体上，模拟人的高密度 脂蛋白(GF9-HDL)抑制PC动物模型中的肿瘤生长。我们还表明，封锁 在这些动物模型中，TREM-1抑制细胞因子和M-CSF的释放。 该项目的主要假设是包括TREM-1抑制剂和TERM-1的联合疗法 抗癌药物和靶点癌症相关炎症和肿瘤细胞直接可以协同作用 提高PC患者的存活率。我们还假设，这种效应在PC中会特别明显 瘤内巨噬细胞高度浸润者。我们的初步研究有力地支持了这一点 假设。拟议项目的长期目标是开发一种新的组合方法。 要有效地瞄准PC。第一阶段研究的主要目标是证明特定的灭活 一流抑制肽与GEM或纳米白蛋白(NAB)结合的TREM-1- 结合紫杉醇(NAB-PTX)，另一种有希望的直接靶向癌细胞的药物，被广泛 被批准用于治疗转移性乳腺癌(BC)，协同抑制PC肿瘤 在动物模型系统中取得进展，并提高存活率。第一阶段的具体目标是：1)评估效果 以及GF9-GEM和GF9-NaB-PTX组合的体外作用机制；2)检测GF9-GEM和GF9-NAB-PTX。 NAB-PTX在两种异种PC小鼠模型中的联合应用。无毒多肽GF9，它采用了新的， 不依赖配体的TREM-1抑制机制，预计副作用较轻。在……里面 为了增加多肽的溶解度、生物利用度和靶向性，我们将利用SignaBlok的 基于高密度脂蛋白的专有纳米系统，用于巨噬细胞靶向输送水，不溶性水和劣质水 可溶药物。我们将使用体外巨噬细胞摄取实验来阐明A 推测受体介导的GF9靶向传递给巨噬细胞的过程。我们将优化GF9 基于它们的稳定性、GF9含量和巨噬细胞体外摄取的制剂。我们将使用In 体外细胞毒试验和免疫印迹分析检测BxPC-3和ASPC-1细胞的增殖以及 GEM、NaB-PTX及其组合对磷酸化Stathmin和α-微管蛋白表达的影响 使用GF9配方。我们将使用BxPC-3和ASPC-1小鼠异种移植模型来检测GF9- GEM和GF9-NaB-PTX联合应用协同抑制肿瘤进展和促进生存 与GF9、GEM、NaB-PTX单独比较。自由GF9和GF9-高密度脂蛋白将被测试。全面 将进行组织学和免疫组织化学研究，以分析肿瘤内血管生成 巨噬细胞的渗透，以及对器官/组织的潜在非特异性毒性。 预计拟议的研究将确定一种新的抗癌组合方法， 将为PC的新的靶向联合疗法的开发奠定基础，从而导致 患者存活率较高。如果成功，第一阶段之后将在第二阶段进行毒理学， 吸收/处置/代谢/排泄(ADME)、药理学和化学/制造/ 控制(CMC)研究，向美国食品和药物管理局提交调查性新药(IND)申请 美国食品和药物管理局(FDA)和随后的人体评估。最终产品将是稳定的TREM-1- 靶向脂肽制剂，可用于PC患者的联合治疗，以延长其生存时间 生死存亡。重要的是，我们最近的数据表明，阻断TREM-1可以抑制体内的进展 不仅是PC，还包括非小细胞肺癌(NSCLC)。因此，第一阶段的成功完成将 提供可能适用于各种炎症的假说的概念证明- 相关肿瘤，如NSCLC、BC、结肠癌等。

Project Summary/Abstract Carcinoma of the pancreas, or pancreatic cancer (PC), is the fourth leading cause of cancer-related death in the United States. According to the American Cancer Society, 55,300 new cases are expected in 2016. Despite advances in therapy, the 5-year survival rate is less than 4%. Current treatments of PC include surgery, radiation therapy, chemotherapy, and immunotherapy but they all only slightly prolong survival or relieve symptoms in patients with PC. Gemcitabine (GEM), first line therapy for advanced PC, is only modestly effective with a median survival of about 6 months in randomized clinical trials, The combination of GEM with different anticancer agents does not show significant survival advantage as compared with GEM alone. These limitations in efficacy of available treatments highlight the need for new treatments. Pancreatic inflammation is known to increase the risk of PC. High macrophage infiltration into the tumor mass correlates with the promotion of tumor growth and metastasis development. Triggering receptor expressed on myeloid cells (TREM-1), an inflammation amplifier, plays a role in PC progression. Expression of TREM-1 on tumor-associated macrophages (TAMs), is upregulated in patients with PC and correlates to disease severity. Recently, we demonstrated that a first-in-class TREM-1 inhibitory peptide GF9 in free form and bound to macrophage-targeted lipopeptide complexes that mimic human high density lipoproteins (GF9-HDL) inhibits tumor growth in animal models of PC. We also showed that blockade of TREM-1 inhibits release of cytokines and M-CSF in these animal models. The main hypothesis of this project is that a combination therapy that includes TREM-1 inhibitors and anticancer agents and targets cancer-related inflammation and tumor cells directly can synergistically improve survival of PC patients. We also hypothesize that this effect will be especially pronounced in PC patients with high intratumoral macrophage infiltration. Our preliminary studies strongly support this hypothesis. The long-term objective of the proposed project is to develop a novel combinatorial approach to efficiently target PC. The major goal of the Phase I study is to demonstrate that specific inactivation of TREM-1 with first-in-class inhibitory peptides in combination with GEM or nanoparticle albumin (nab)- bound paclitaxel (nab-PTX), another promising agent that directly targets cancer cells and is widely approved for the treatment of metastatic breast cancer (BC), synergistically suppresses PC tumor progression in animal model system and improves survival. Phase I specific aims are to: 1) evaluate effects and mechanisms of GF9-GEM and GF9-nab-PTX combinations in vitro, and 2) test GF9-GEM and GF9- nab-PTX combinations in two xenograft mouse models of PC. Non-toxic peptide GF9, which employs novel, ligand-independent mechanisms of TREM-1 inhibition, is anticipated to have less severe side effects. In order to increase peptide solubility, bioavailability and targeting to TAMs, we will utilize SignaBlok's proprietary HDL-based nanosystem for macrophage-targeted delivery of water insoluble and poorly water soluble drugs. We will use in vitro macrophage uptake assay to elucidate the molecular mechanisms of a putative receptor-mediated process of targeted delivery of GF9 to macrophages. We will optimize GF9 formulations based upon their stability, GF9 content, and macrophage uptake in vitro. We will use an in vitro cytotoxicity assay and immunoblot analysis to test proliferation of BxPC-3 and AsPC-1 cells as well as expression of phospho-stathmin and alpha-tubulin in the presence of GEM, nab-PTX or their combinations with GF9 formulations. We will use BxPC-3 and AsPC-1 mouse xenograft models to test the ability of GF9- GEM and GF9-nab-PTX combinations to synergistically inhibit tumor progression and promote survival as compared with GF9, GEM, and nab-PTX alone. Free GF9 and GF9-HDL will be tested. Comprehensive histology and immunohistochemistry studies will be performed to analyze angiogenesis, intratumoral macrophage infiltration, and potential non-specific toxicity for organ/tissues. It is anticipated that the proposed research will identify a novel anticancer combination approach that will set the stage for the development of new targeted combination therapies of PC, thereby leading to a higher survival rate of the patients. If successful, the Phase I will be followed in the Phase II by toxicology, absorption/ disposition/ metabolism/ excretion (ADME), pharmacology and chemistry/ manufacturing/ control (CMC) studies, filing an Investigational New Drug (IND) application with the US Food and Drug Administration (FDA) and subsequent evaluation in humans. Final product will be the stable TREM-1- targeted lipopeptide formulation that can be used in combination therapies of PC patients to prolong their survival. Importantly, our recent data demonstrate that blockade of TREM-1 suppresses in vivo progression of not only PC but also non-small cell lung cancer (NSCLC). Thus, successful completion of Phase I will provide the proof of concept of the hypothesis that might be applicable to a variety of inflammation- associated tumors such as NSCLC, BC, colon cancer, and others.