Oncolytic Viruses with Therapeutic Genes in the Treatment of Breast Cancer

具有治疗基因的溶瘤病毒治疗乳腺癌

• 批准号: 8384916
• 负责人: Danuta B Kozbor
• 金额: $ 19.31万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8384916
• 关键词: Affect; Aftercare; Apoptosis; Autologous; Behavior; Blood Vessels; Blood flow; Breast Cancer Treatment; Breast Carcinoma; CSF3 gene; CXCL12 gene; CXCR; CXCR4 Receptors; CXCR4 gene; Cell Communication; Cell Differentiation process; Cells; Clinical; Combined Modality Therapy; Cytolysis; Development; Education; Effectiveness; Epithelial; Event; Excision; Exhibits; Fibroblasts; Fluorescence; Goals; Growth; Human; IFN consensus sequence binding protein; ITGAM gene; Inbred BALB C Mice; Infection; Infiltration; Inflammatory; Interleukin-10; Interleukin-12; Knock-out; Knockout Mice; Knowledge; Ligands; Lung; Malignant Epithelial Cell; Malignant Neoplasms; Malignant Stromal Cell; Mammary Neoplasms; Measures; Modeling; Mus; Myelogenous; Myeloid Cells; Neoplasm Metastasis; Oncolytic; Oncolytic viruses; Pathogenesis; Perfusion; Peripheral; Phenotype; Population; Primary Neoplasm; Process; Production; Proteins; Receptor Signaling; Relative (related person); Role; SCID Mice; Signal Pathway; Specimen; Spleen; Stromal Cell-Derived Factor 1; Stromal Cells; Therapeutic; Transgenic Mice; Tumor-Derived; Vaccinia virus; Vascular Endothelial Growth Factors; Virus; Xenograft procedure; adaptive immunity; cancer cell; cell transformation; chemokine; comparative; cytokine; density; design; granulocyte; killings; macrophage; malignant breast neoplasm; neoplastic cell; neutrophil; preclinical study; progenitor; research study; therapeutic gene; tumor; tumor growth; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): Oncolytic viruses are designed to eliminate tumors by direct infection and lysis of cancer cells. Recently, we and others have observed that in addition to a direct tumor killing by the virus there is also an indirect cytopathic effect due to vascular collapse associated with a loss of blood flow to the interior of the tumor and infiltration by peripheral Gr1+CD11b+ neutrophils. Furthermore, our preliminary results demonstrated that oncolytic vaccinia virus (OVV) expressing an antagonist of the CXCR4 receptor for CXCL12 chemokine (OVV- CXCR4-A-Fc) was more effective in inhibiting growth of triple-negative (TN) breast carcinomas 4T1 in syngeneic mice compared to the treatment with OVV expressing the enhanced green fluorescence protein (OVV-EGFP). The inhibition of primary and metastatic tumors by OVV-CXCR4-A-Fc was associated with i) an increased expression of interferon regulatory factor-8 (IRF8), a putative determinant of protumorigenic behavior of MDSCs, in tumor-infiltrating granulocytes, ii) reduced percentage of FAP+ cancer associated fibroblasts (CAFs), iii) decreased levels of CXCL12, and iv) collapse of the tumor vasculature. These changes within the tumor microenvironment were either not observed (such as elevated expression of IRF8) or were less pronounced after treatment with OVV-EGFP. The mechanism by which OVV-CXCR4-A-Fc affects interaction between CAFs and MDSCs and expression of IRF8 is unclear. We hypothesize that the enhanced expression of IRF8 in MDSCs and promotion of their differentiation toward type 1 phenotype with antitumorigenic activity is regulated by the CXCL12/CXCR4 ligand/receptor signaling pathway in CAFs. We propose to identify the key events of the CAF/MDSC interaction that regulates expression of IRF8 by analyzing changes in the production of tumor- derived factors (TDFs) in 4T1-bearing syngeneic mice and human breast carcinoma MDA-MB-23 xenografts in SCID mice. The Specific Aims are: 1). We will compare quantitative and qualitative changes in CAFs, myeloid cells, epithelial tumor cells, TDFs, and tumor vasculature induced by OVV-EGFP and OVV-CXCR4-A-Fc treatments. 2). Using IRF8 knockout and IRF8 transgenic mice and autologous AT-3 breast carcinoma cells, we will explore the role of IRF8 in modulation of myeloid differentiation after blocking the CXCL12/CXCR4 ligand/receptor signaling pathway by OVV-CXCR4-A-Fc. 3). Using the established 4T1 breast tumor model for spontaneous lung metastasis, we will compare the reciprocal dynamics in colonization of a metastatic niche in the lungs by IRF8+ MDSCs and mature neutrophils after OVV-CXCR4-A-Fc and OVV-EGFP treatments. We anticipate that understanding the activation of inflammatory cells can be used for enhancing the effectiveness of OV therapeutics for treatment of the highly aggressive tumors. PUBLIC HEALTH RELEVANCE: We propose to investigate changes in the dynamic interaction between malignant and stromal cells that facilitate tumor-free survival and provide a clear understanding of the role of the CXCR4/SDF-1 signaling pathway in breast cancer pathogenesis. The goal of this application is to target CXCR4 expression in triple- negative breast carcinomas by an oncolytic virotherapy expressing CXCR4 antagonist and elucidate the mechanism of primary and metastatic tumor growth inhibition in preclinical studies in mice. We anticipate that a combination therapy that attacks both malignant cells and the 'other half' of the tumor mass (that is, the stromal cells) could be more effective and might also elicit long-lasting adaptive immunity to the transformed cells.

描述（由申请人提供）：溶瘤病毒旨在通过直接感染和裂解癌细胞来消除肿瘤。最近，我们和其他人观察到，除了病毒直接杀死肿瘤外，由于与肿瘤内部血流减少和外周 Gr1+CD11b+ 中性粒细胞浸润相关的血管塌陷，还产生间接细胞病变效应。此外，我们的初步结果表明，表达 CXCR4 趋化因子 CXCR4 受体拮抗剂 (OVV-CXCR4-A-Fc) 的溶瘤痘苗病毒 (OVV) 在同基因中更有效地抑制三阴性 (TN) 乳腺癌 4T1 的生长。小鼠与表达增强型绿色荧光蛋白 (OVV-EGFP) 的 OVV 治疗进行比较。 OVV-CXCR4-A-Fc 对原发性和转移性肿瘤的抑制与 i) 肿瘤浸润粒细胞中干扰素调节因子 8 (IRF8) 的表达增加有关，IRF8 是 MDSC 促肿瘤行为的假定决定因素，ii) FAP+癌症相关成纤维细胞 (CAF) 的百分比降低，iii) CXCL12 水平降低，以及 iv) 肿瘤脉管系统崩溃。肿瘤微环境内的这些变化要么没有观察到（例如 IRF8 表达升高），要么在 OVV-EGFP 治疗后不太明显。 OVV-CXCR4-A-Fc 影响 CAF 和 MDSC 之间的相互作用以及 IRF8 表达的机制尚不清楚。我们假设 MDSC 中 IRF8 表达的增强以及促进其向具有抗肿瘤活性的 1 型表型分化是由 CAF 中的 CXCL12/CXCR4 配体/受体信号通路调节的。我们建议通过分析SCID中携带4T1的同基因小鼠和人乳腺癌MDA-MB-23异种移植物中肿瘤衍生因子（TDF）产生的变化来确定调节IRF8表达的CAF/MDSC相互作用的关键事件老鼠。具体目标是：1）。我们将比较 OVV-EGFP 和 OVV-CXCR4-A-Fc 治疗诱导的 CAF、骨髓细胞、上皮肿瘤细胞、TDF 和肿瘤脉管系统的定量和定性变化。 2）。使用IRF8敲除和IRF8转基因小鼠以及自体AT-3乳腺癌细胞，我们将探讨通过OVV-CXCR4-A-Fc阻断CXCL12/CXCR4配体/受体信号通路后IRF8在调节骨髓分化中的作用。 3）。使用已建立的自发性肺转移的 4T1 乳腺肿瘤模型，我们将比较 OVV-CXCR4-A-Fc 和 OVV-EGFP 治疗后 IRF8+ MDSC 和成熟中性粒细胞在肺部转移微环境中定植的相互动态。我们预计，了解炎症细胞的激活可用于增强 OV 疗法治疗高度侵袭性肿瘤的有效性。 公共健康相关性：我们建议研究恶性细胞和基质细胞之间动态相互作用的变化，以促进无肿瘤生存，并清楚地了解 CXCR4/SDF-1 信号通路在乳腺癌发病机制中的作用。本申请的目标是通过表达CXCR4拮抗剂的溶瘤病毒疗法来靶向三阴性乳腺癌中的CXCR4表达，并阐明小鼠临床前研究中原发性和转移性肿瘤生长抑制的机制。我们预计，一种既攻击恶性细胞又攻击肿瘤细胞“另一半”的联合疗法 肿瘤块（即基质细胞）可能更有效，并且还可能引发对转化细胞的持久适应性免疫。