Systemic delivery of an oncolytic adenovirus targeting TGFβ to enhance anti-PD-1 and anti-CTLA-4 therapy for triple negative breast cancer

全身递送靶向 TGFβ 的溶瘤腺病毒，以增强三阴性乳腺癌的抗 PD-1 和抗 CTLA-4 治疗

• 批准号: 10117795
• 负责人: Weidong Xu
• 金额: $ 39.51万
• 依托单位: NORTHSHORE UNIVERSITY HEALTHSYSTEM
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10117795
• 关键词: 4T1; Abraxane; Adenoviruses; Amino Acids; Antibodies; Antibody titer measurement; Antitumor Response; Attenuated; Binding; Biological Assay; Biological Markers; Breast Cancer Cell; Breast Cancer Model; Breast Cancer Patient; Breast Cancer Treatment; CTLA4 gene; Cell Death; Cells; Clinical; Clinical Trials; Combination immunotherapy; Combined Modality Therapy; Complementarity Determining Regions; Cytometry; Development; FDA approved; Fiber; Future; Genes; Genetic Transcription; Goals; Hepatic; Homing; Human; Immune; Immune checkpoint inhibitor; Immunoglobulin G; Immunologics; Immunology procedure; Immunosuppressive Agents; Inflammatory; Liver; Lymphatic; Malignant Neoplasms; Mammary Neoplasms; Mediating; Metastatic Neoplasm to the Bone; Modeling; Mus; Neoplasm Metastasis; Normal tissue morphology; Oncogenes; Oncolytic viruses; Osteolytic; Paclitaxel; Pathway interactions; Patients; Peptides; Phenotype; Population; Prevalence; RNA; Research; Resistance; Role; Safety; Signal Pathway; Signal Transduction; Spleen; TGFB1 gene; Technology; Testing; Therapy Clinical Trials; Therapy trial; Toxic effect; Transforming Growth Factor beta; Transforming Growth Factor beta Receptors; Tumor Immunity; Viral; Virus; Woman; anti-CTLA-4 therapy; anti-CTLA4; anti-PD-1; anti-PD-1/PD-L1; anti-PD-L1; anti-PD-L1 antibodies; anti-PD1 antibodies; anti-tumor immune response; antitumor agent; base; bone; cytokine; gene panel; immune activation; immunogenicity; improved; in vivo; interest; macrophage; molecular subtypes; nano-string; neoplastic cell; neutralizing antibody; novel; novel therapeutics; oncolytic adenovirus; peripheral blood; programmed cell death ligand 1; programmed cell death protein 1; receptor; research clinical testing; response; subcutaneous; systemic toxicity; therapy resistant; transcriptome; transcriptome sequencing; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor microenvironment; tumor specificity; uptake

PROJECT SUMMARY The development of novel therapies for the treatment of breast cancer is a major unmet need. In recent years, immune checkpoint inhibitors including anti-PD-1, anti-PD-L1 and anti-CTLA-4 have shown promise as antitumor agents, and are approved for the treatment of several malignancies. Clinical trials in breast cancer patients have shown that about 20% of triple negative breast cancers (TNBC) respond favorably to anti-PD-1 antibodies and Atezolizumab, an anti-PD-L1 antibody in combination with nab-paclitaxel (Abraxane) is now FDA approved for advanced stage TNBC patients with positive PDL-1 expression. However, many TNBC patients are resistant to anti-PD-1/PDL-1 and anti-CTLA-4 treatments which could be due to weak immunogenicity of the tumors and poor inflammatory but highly immune suppressive tumor microenvironment. In recent years TGFβ has been shown to be a strong immune suppressor and can potentially produce a tumor microenvironment that is resistant to anti-PD-1 and anti-CTLA-4 therapy. To overcome resistance to anti-PD-1 and anti-CTLA-4 we have developed adenoviruses (Ad) expressing sTGFβRIIFc (soluble TGFβ receptor II fused with human IgG Fc). sTGFβRIIFc acts as a TGFβ decoy, and can inhibit TGFβ pathways. Initially, we created Ad5 based Ad.sT expressing sTGFβRIIFc. To reduce hepatic/systemic toxicity associated with systemic delivery of Ad.sT, we created mHAd.sT, a liver-detargeted virus, by replacing hypervariable regions (HVRs) (1-7) of Ad.sT with Ad48 HVRs. To enhance tumor specificity, we have now created mHAdLyp.sT by introducing LyP-1 peptide, a 9-amino acid long tumor homing-cell peptide, into the HI loop of the mHAd.sT fiber. In this proposal, we will test the hypotheses that systemic administration of mHAdLyp.sT in mice bearing 4T1 triple negative mammary tumors will result in reduced hepatic/systemic toxicity but produce high levels of sTGFβRIIFc and inhibit TGFβ pathways. This will alter the tumor microenvironment, induce tumor immunity, and overcome resistance to anti-PD-1 and anti-CTLA-4., and examine the expression profiles of TGFβ-1, TGFβ-1 regulated genes, and PD-1 and CTLA-4 signaling pathways. We will examine immuno-phenotypes in tumors, peripheral blood and spleen (Aim 1). Next, we will examine mHAdLyp.sT, anti-PD-1 and anti-CTLA-4 combination therapies in mouse tumor models. We will conduct RNA-Seq analysis of the whole transcriptomes, and examine the role of immune activation in mediating the anti-tumor responses (Aim 2). We will test the hypothesis that systemic administration of mHLypAd.sT, in combination with anti-PD-1 and anti- CTLA-4 in mice with pre-established metastases will be effective (Aim 3). To guide us for the combination therapy trials with mHAdLyp.sT, anti-PD-1 and anti-CTLA-4, we will examine human TNBC tumors by Nanostring technology for RNA profiling, and will further examine the TGFβ-1 and other relevant biomarkers and TILS in human TNBC tumors. We will also screen the human population for the Ad neutralizing antibodies titers (Aim 4). We believe that our research described here is critical to bring forward our oncolytic virus mHAdLyp.sT targeting TGFβ in combination with anti-PD-1 and anti-CTLA-4 for clinical evaluation in TNBC patients.

项目摘要 用于治疗乳腺癌的新型疗法的发展是一种主要的未满足需求。最近几年， 免疫检查点抑制剂在内 抗肿瘤剂，并获得批准用于治疗几种恶性肿瘤。乳腺癌的临床试验 患者表明，约有20％的三重负乳腺癌（TNBC）对抗PD-1的反应有利 抗体和atezolizumab，一种抗PD-L1抗体，结合使用 NAB-列酰胺（Abraxane） 现在 FDA批准了PDL-1表达阳性的晚期TNBC患者。但是，许多TNBC 患者对抗PD-1/PDL-1和抗CTLA-4治疗具有抗性，这可能是由于弱 肿瘤的免疫原性和炎症但高度免疫抑制的肿瘤微环境。 近年来，TGFβ已被证明是强烈的免疫抑制剂，可能会产生肿瘤 对抗PD-1和抗CTLA-4治疗具有抗性的微环境。克服对抗PD-1的抗性 和抗CTLA-4我们开发了表达STGFβRIIFC的腺病毒（AD）（可溶性TGFβ受体II 与人类IgG FC融合。 STGFβRIIFC充当TGFβ诱饵，可以抑制TGFβ途径。最初，我们 创建了基于AD5的AD.ST表达STGFβRIIFC。降低肝/全身毒性 AD.ST的系统性输送，我们通过更换高变量区域创建了一种肝脏定位的病毒Mhad.st AD.ST的（1-7）AD48 HVRS（1-7）。为了提高肿瘤特异性，我们现在创建了Mhadlyp.st 将LYP-1肽（一种9-氨基酸长肿瘤归巢肽）引入MHAD.ST的HI环中 纤维。在此提案中，我们将检验以下假设，即在轴承的小鼠中系统地给予mhadlyp.st 4T1三重阴性乳腺肿瘤将导致肝/全身毒性降低，但产生高水平 STGFβRIIFC并抑制TGFβ途径。这将改变肿瘤微环境，诱导肿瘤免疫学， 并克服对抗PD-1和抗CTLA-4。的耐药性，并检查TGFβ-1的表达曲线， TGFβ-1调控基因，PD-1和CTLA-4信号通路。我们将检查中的免疫表型 肿瘤，外周血和脾脏（AIM 1）。接下来，我们将检查mhadlyp.st，anti-pd-1和anti-ctla-4 小鼠肿瘤模型中的组合疗法。我们将对整个进行RNA-seq分析 转录组，并检查免疫激活在介导抗肿瘤反应中的作用（AIM 2）。我们 将检验以下假设，即Mhlypad.ST的全身给药结合抗PD-1和抗 - 具有预先建立转移的小鼠中的CTLA-4将是有效的（AIM 3）。指导我们组合 通过Mhadlyp.ST，抗PD-1和抗CTLA-4进行的治疗试验，我们将检查人类TNBC肿瘤 用于RNA分析的纳米弦技术，并将进一步检查TGFβ-1和其他相关的生物标志物 和人类TNBC肿瘤中的TIL。我们还将筛选人类的广告中和抗体 滴度（目标4）。我们认为，这里描述的研究对于提出溶瘤病毒至关重要 Mhadlyp.ST靶向TGFβ与抗PD-1和抗CTLA-4结合使用，用于TNBC中的临床评估 患者。