IDO-silencing Salmonella therapy for the treatment of primary and metastatic PDAC

IDO 沉默沙门氏菌疗法用于治疗原发性和转移性 PDAC

• 批准号: 8595122
• 负责人: Don J Diamond
• 金额: $ 18.27万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-08 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8595122
• 关键词: Affinity; Apoptosis; Apoptotic; Area; Attenuated; Bacteria; Blood Vessels; Cancer Etiology; Cancerous; Cell Death; Cessation of life; Characteristics; Chemicals; Clinical; Clinical Trials; Collaborations; Combined Modality Therapy; Desmoplastic; Diagnosis; Disease; Disease Progression; Distal; Erlotinib; Evaluation; Flow Cytometry; Frequencies; Generations; Genetically Engineered Mouse; Goals; Growth; Human; Human Cell Line; Human Characteristics; Hyaluronan; Hyaluronidase; Hypoxia; Immunity; Immunosuppression; Immunosuppressive Agents; Incidence; Infiltration; Luciferases; Malignant neoplasm of pancreas; Measurement; Microscopy; Modeling; Mus; Neoplasm Metastasis; Neutrophil Infiltration; Operative Surgical Procedures; Organ; Oxidative Stress; Pancreas; Pancreatic Adenocarcinoma; Pancreatic Ductal Adenocarcinoma; Patients; Penetration; Pharmaceutical Preparations; Play; Production; Property; Protein Array; Protein Array Analysis; Reactive Oxygen Species; Recombinants; Recruitment Activity; Recurrence; Role; Salmonella; Salmonella typhimurium; Signal Pathway; Specificity; Specimen; Staining method; Stains; Surgical Oncologist; Sutent; Techniques; Technology; Testing; Therapeutic; Therapeutic Effect; Tissues; Toxic effect; Transgenic Organisms; Translations; Transplantation; Tryptophan 2,3 Dioxygenase; Tumor Cell Line; Tumor Tissue; Tumor Volume; Tumor-Derived; Vascular Permeabilities; Work; Xenograft Model; Xenograft procedure; base; cancer stem cell; cell motility; chemotherapy; cytokine; design; effective therapy; gemcitabine; gemzar; improved; inhibitor/antagonist; intravital imaging; killings; mouse model; neoplastic cell; neutrophil; novel; overexpression; plasmid DNA; prevent; public health relevance; response; small hairpin RNA; stellate cell; treatment strategy; tumor; tumor growth; tumor microenvironment; tumor specificity

DESCRIPTION (provided by applicant): Advanced pancreatic ductal adenocarcinoma (PDAC) is often inoperable, and is only transiently responsive to existing therapies. Overexpression of indoleamine 2,3-dioxygenase (IDO) in PDAC plays a major role in accelerating disease progression by suppressing antitumor immunity. Current IDO inhibitors inadequately reverse immunosuppression while systemic off target effects contribute to their toxicity. ShIDO-ST is a novel Salmonella typhimurium (ST)-based therapy that expresses a small hairpin (sh)RNA to specifically silence tumor-derived IDO with decreased toxicity. ST as an shRNA delivery vehicle offers superior penetration against desmoplastic PDAC tissue and anti-metastatic function due to its motility and affinity for poorly vascularized, hypoxic tissue. he combination of IDO silencing and abundant ST tumor colonization increases intratumoral local reactive oxygen species through the recruitment and activation of polymorphonuclear neutrophils (PMN), which can cause oxidative stress-induced apoptosis of tumor cells, cancer stem cells (CSC), and vascular stroma to inhibit PDAC progression. PMN are also known to have anti-metastatic function, thus providing additional protection against tumor spread. The proposed work will provide detailed analysis of signaling pathways influencing PMN recruitment and activation upon shIDO-ST treatment. Cytokines involved in PMN recruitment will be evaluated using protein array analysis and flow cytometry of PDAC tumors from mice treated with shIDO-ST versus scrambled sequence control (shScr-ST), which does not silence IDO. The polarization and apoptotic state of infiltrating PMN will also be evaluated using flow cytometry, as IDO activity is known to regulate PMN function. The finding that both ST and PMN provide primary and anti-metastatic protection supports evaluation of shIDO-ST in more aggressive models of PDAC. The KrasG12D;Trp53R127H;Pdx1-Cre genetically engineered mouse model (KPC-GEMM) recapitulates desmoplasia and metastasis characteristic of human PDAC. Exclusive access to fresh surgical PDAC tissue, by virtue of our close collaboration with surgical oncologists, will allow for evaluation of human (Hu)shIDO-ST in xenograft models to provide greater support for clinical translation. Combination therapy with Sutent(tm) (Pfizer), which induces vascular collapse in PDAC, will be tested because it increases the hypoxic area of the tumor to augment colonization by ST and the recruitment of PMN. To improve penetration of tumors by shIDO-ST and Sutent, we propose using PEGylated human recombinant hyaluronidase (PEGPH20, Halozyme Inc.), which depletes hyaluronan abundant in PDAC tissue and increases vascular permeability. The effects of proposed treatments on primary and metastatic burden, CSC frequency, and ST colonization will be evaluated in transgenic and xenograft settings through cross sectional or longitudinal measurements of tumor volume, metastatic burden, CSC markers, and bacterial enumeration. The long-term objective is to develop shIDO-ST into a suitable, effective therapy for the treatment of patients with advanced inoperable PDAC.

描述（由申请人提供）：晚期胰腺导管腺癌（PDAC）通常无法手术，并且对现有疗法仅短暂有效。 PDAC 中吲哚胺 2,3-双加氧酶 (IDO) 的过度表达通过抑制抗肿瘤免疫在加速疾病进展中发挥着重要作用。目前的 IDO 抑制剂不足以逆转免疫抑制，而全身脱靶效应则导致其毒性。 ShIDO-ST 是一种基于鼠伤寒沙门氏菌 (ST) 的新型疗法，它表达小发夹 (sh)RNA，以特异性沉默肿瘤源性 IDO，同时降低毒性。 ST 作为 shRNA 递送载体，由于其对血管化不良、缺氧组织的运动性和亲和力，对促纤维增生性 PDAC 组织具有卓越的渗透性和抗转移功能。 IDO 沉默和丰富的 ST 肿瘤定植相结合，通过招募和激活多形核中性粒细胞 (PMN) 增加肿瘤内局部活性氧，从而导致氧化应激诱导的肿瘤细胞、癌症干细胞 (CSC) 和血管基质细胞凋亡抑制 PDAC 进展。 PMN 还具有抗转移功能，从而提供额外的保护以防止肿瘤扩散。拟议的工作将详细分析影响 shIDO-ST 治疗后 PMN 募集和激活的信号通路。将使用蛋白质阵列分析和流式细胞术对经 shIDO-ST 处理的小鼠 PDAC 肿瘤与不沉默 IDO 的乱序序列对照 (shScr-ST) 处理的小鼠的 PDAC 肿瘤进行评估，从而评估参与 PMN 募集的细胞因子。渗透性 PMN 的极化和凋亡状态也将使用流式细胞术进行评估，因为已知 IDO 活性可调节 PMN 功能。 ST 和 PMN 均提供主要保护和抗转移保护的发现支持在更具侵袭性的 PDAC 模型中评估 shIDO-ST。 KrasG12D;Trp53R127H;Pdx1-Cre 基因工程小鼠模型 (KPC-GEMM) 概括了人类 PDAC 的结缔组织形成和转移特征。凭借我们与外科肿瘤学家的密切合作，独家获得新鲜的手术 PDAC 组织将允许在异种移植模型中评估人 (Hu)shIDO-ST，从而为临床转化提供更大的支持。将测试与Sutent(tm)（辉瑞公司）的联合疗法，该疗法可诱导PDAC 中的血管塌陷，因为它增加了肿瘤的缺氧面积，从而增强了ST 的定植和PMN 的募集。为了提高 shIDO-ST 和 Sutent 对肿瘤的渗透，我们建议使用聚乙二醇化人重组透明质酸酶（PEGPH20，Halozyme Inc.），它可以消耗 PDAC 组织中丰富的透明质酸并增加血管通透性。将通过肿瘤体积、转移负荷、CSC标记物和细菌计数的横断面或纵向测量，在转基因和异种移植环境中评估拟议治疗对原发性和转移性负担、CSC频率和ST定植的影响。长期目标是将 shIDO-ST 开发为一种合适、有效的疗法，用于治疗晚期无法手术的 PDAC 患者。