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.

项目总结 开发治疗乳腺癌的新疗法是一个尚未得到满足的主要需求。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良好的前景 抗肿瘤药物，并被批准用于治疗几种恶性肿瘤。乳腺癌的临床试验 患者已经表明，大约20%的三阴性乳腺癌(TNBC)对抗PD-1有良好的反应 抗体和抗PD-L1抗体Atezolizumab与 NaB-紫杉醇(亚伯拉索) 现 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治疗具有抵抗力，这可能是由于 肿瘤免疫原性差，炎性但高度免疫抑制的肿瘤微环境。 近年来，转化生长因子β已被证明是一种强大的免疫抑制因子，并有可能导致肿瘤。 对抗PD-1和抗CTLA-4治疗耐药的微环境。克服对抗PD-1抗体的耐药性 和抗CTLA-4，我们已经研制出了表达sTGFRIIFc(可溶性ββ受体II)的腺病毒 与人Ig G Fc融合)。转化生长因子βRIIFc可作为转化生长因子β的诱饵，抑制转化生长因子β途径。最初，我们 创建了基于Ad5的Ad.sT，表达了sTGFβRIIFc。减少与以下物质相关的肝脏/全身毒性 通过系统传递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 (HVRs)(1-7)Ad.sT，其中Ad48 HVRs。为了增强肿瘤特异性，我们现在通过以下方式创建了mHAdLyp.sT 在mHAd.sT的HI环中引入LYP-1肽，这是一种由9个氨基酸组成的长肿瘤归巢细胞多肽 纤维。在这项提案中，我们将测试mHAdLyp.sT在小鼠体内系统性给药的假设 4T1三阴性乳腺肿瘤将导致肝脏/全身毒性降低，但会产生高水平的 并抑制转化生长因子β的β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，检测转化生长因子β-1的表达谱， 转化生长因子β-1调控基因，以及PD-1和CTLA-4信号通路。我们将检测免疫表型 肿瘤、外周血和脾(目标1)。接下来，我们将检测mHAdLyp.sT、抗PD-1和抗CTLA-4 小鼠肿瘤模型的联合治疗。我们将对整个病毒进行RNA-Seq分析 转录本，并检测免疫激活在介导抗肿瘤反应中的作用(目标2)。我们 将检验mHLypAd.sT与抗PD-1和抗-PD-1联合全身应用的假设 CTLA-4对预先确定转移的小鼠将是有效的(目标3)。为我们的组合提供指引 用mHAdLyp.sT、抗PD-1和抗CTLA-4进行治疗试验，我们将通过 纳米串技术用于核糖核酸分析，并将进一步检测转化生长因子β-1和其他相关生物标志物 和TIL在人TNBC肿瘤中的表达。我们还将对人群进行Ad中和抗体的筛查 效价(目标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是至关重要的 靶向转化生长因子β的mHAdLyp.sT联合抗PD-1和抗CTLA-4治疗肿瘤的临床评价 病人。