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: 10357928
• 负责人: Weidong Xu
• 金额: $ 37.44万
• 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/10357928
• 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 开发用于治疗乳腺癌的新疗法是一个主要的未满足的需求。近年来， 包括抗PD-1、抗PD-L1和抗CTLA-4在内的免疫检查点抑制剂已显示出作为 抗肿瘤剂，并且被批准用于治疗几种恶性肿瘤。乳腺癌临床试验 患者已经表明，约20%的三阴性乳腺癌（TNBC）对抗PD-1有良好的反应， 抗体和Atezolizumab，一种抗PD-L1抗体与 白蛋白结合型紫杉醇（Abraxane） 现在 FDA批准用于PDL-1表达阳性的晚期TNBC患者。然而，许多TNBC 患者对抗PD-1/PDL-1和抗CTLA-4治疗具有抗性，这可能是由于弱的 肿瘤的免疫原性和不良的炎性但高度免疫抑制的肿瘤微环境。 近年来，TGFβ已被证明是一种强免疫抑制剂，并可能产生肿瘤 在一些实施方案中，所述微环境对抗PD-1和抗CTLA-4疗法具有抗性。为了克服抗PD-1抗体的耐药性 和抗CTLA-4，我们已经开发了表达sTGFβRIIFc（可溶性TGFβ受体II）的腺病毒（Ad 与人IgG Fc融合）。sTGFβRIIFc作为TGFβ诱饵，可抑制TGFβ通路。最初我们 构建了表达sTGFβRIIFc的基于Ad 5的Ad.sT。为了减少与以下相关的肝脏/全身毒性： 系统性递送Ad.sT，我们通过替换高变区创建了mHAd.sT，一种肝脏去靶向病毒 （HVR）（1-7）的Ad.sT与Ad 48 HVR。为了增强肿瘤特异性，我们现在已经通过以下方式创建了mHAdLyp.sT： 将LyP-1肽（一种9个氨基酸长的肿瘤归巢细胞肽）引入mHAd的HI环中。 光纤在该提议中，我们将测试在携带mHAdLyp.sT的小鼠中全身施用mHAdLyp.sT的假设。 4 T1三阴性乳腺肿瘤将导致肝/全身毒性降低，但产生高水平的 sTGFβRIIFc和抑制TGFβ通路。这将改变肿瘤微环境，诱导肿瘤免疫， 并克服对抗PD-1和抗CTLA-4的耐药性，并检测TGFβ-1的表达谱， TGFβ-1调节基因，PD-1和CTLA-4信号通路。我们将检查免疫表型， 肿瘤、外周血和脾脏（Aim 1）。接下来，我们将检查mHAdLyp.sT、抗PD-1和抗CTLA-4 在小鼠肿瘤模型中的联合疗法。我们将进行RNA-Seq分析， 转录组，并检查免疫活化在介导抗肿瘤应答中的作用（目的2）。我们 将检验mHLypAd.sT与抗PD-1和抗PD-2抗体的组合的全身给药的假设。 CTLA-4在具有预先建立的转移的小鼠中将是有效的（目的3）。来指导我们 在使用mHAdLyp.sT、抗PD-1和抗CTLA-4的治疗试验中，我们将通过以下方法检查人TNBC肿瘤： Nanostring技术用于RNA分析，并将进一步检查TGFβ-1和其他相关生物标志物 和TILS。我们还将筛选人群中的Ad中和抗体 滴度（Aim 4）。我们相信，我们在这里描述的研究对于提出我们的溶瘤病毒至关重要。 靶向TGFβ的mHAdLyp.sT与抗PD-1和抗CTLA-4联合用于TNBC的临床评价 患者