Aptamer mediated silencing of the spliceosome machinery to increase the immunogenicity of metastatic breast cancer

适体介导的剪接体机制沉默以增加转移性乳腺癌的免疫原性

基本信息

批准号：
10286216
负责人：
Paolo Serafini
金额：
$ 17.94万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-04 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10286216
关键词：
4T1 Affect Affinity Alternative Splicing Antigens Antitumor Response Award Binding Biodistribution Breast Cancer Cell Breast cancer metastasis Cells Chemicals Chimera organism Clinic Clinical Trials Containment Disease Disseminated Malignant Neoplasm Drug Kinetics Epitopes FDA approved Future Genes Genetic Genetic Transcription Human Immune Immune checkpoint inhibitor Immune system Immunocompetent Immunotherapeutic agent Immunotherapy In Vitro Intelligence Intervention Introns Ligands Malignant Neoplasms Measures Mediating Metastatic breast cancer Mus Mutation Neoplasm Metastasis Oligonucleotides Oncoproteins Paclitaxel Pathway interactions Patients RNA Reagent Resistance Ribonucleases Role Safety Side Small Interfering RNA Somatic Mutation Spliceosomes Structure Testing Therapeutic Therapeutic Effect Tissues Toxic effect Transfection Translations Tumor Immunity Up-Regulation Vaccination Vertebral column Viral Vector anti-PD-1 aptamer bioinformatics tool cancer cell cancer therapy checkpoint inhibition checkpoint therapy clinically relevant efficacy testing experimental study high reward high risk immune checkpoint blockade immunogenicity immunological intervention improved in vivo mRNA Precursor malignant breast neoplasm mouse model nanoparticle neoantigens neoplastic cell preclinical development programmed cell death ligand 1 receptor response screening siRNA delivery side effect success targeted delivery targeted treatment three dimensional structure transcriptome sequencing triple-negative invasive breast carcinoma tumor tumor microenvironment tumor progression

项目摘要

Even with the FDA approval of Atezolizumab in combination with nab-Paclitaxel for PDL1 positive triple negative breast cancer, metastatic breast cancer is still a deadly disease. The modest impact of checkpoint inhibitor therapy in this disease can be partially explained by the relatively low tumor mutation burden, by the low expression of neoantigens and the overall low immunogenicity of metastatic breast cancer compare to other malignancies in which checkpoint inhibitors are highly effective. Recent RNA sequencing and MHC-peptidome analyses of tumors from patients undergoing checkpoint inhibition therapy revealed the important contribution of intron-derived epitopes in the neoantigen pools. In breast cancer these intron-derived neoantigens are poorly expressed because of the constitutive upregulation of the spliceosome machinery. Here we will test the hypothesis that aptamer mediated targeted silencing snRPE, a key component of the spliceosome machinery, can increase breast cancer immunogenicity and synergize with anti-PD1 in the eradication of metastatic breast cancer. Toward this targeted therapy, we have identified two RNA aptamers that can recognize and mouse and human metastatic breast cancer cells in vitro and in vivo. RNA aptamers are small oligonucleotides that because of their 3D structure can recognize their ligand with high affinity. Aptamers are chemically synthetized with a backbone that confer RNAse resistance and lack of immunogenicity and can be modified for improved pharmacokinetic and for therapeutic delivery. We have conjugated our aptamer with siRNA against snRPE and showed their in vitro efficacy. In this study, we will employ two aggressive model of mouse breast cancer: the 4T1 and the E0771 in two different genetic backgrounds. First, we will characterize the role of the immune system in the antitumor response using NSG and immune competent mice, depletion experiments, and by characterizing the tumor microenvironment and the anti-tumor immunity. Then, we will measure the therapeutic effect of aptamer chimera as monotherapy or in conjunction with checkpoint inhibition therapy and vaccination against intron derived neoepitopes for the treatment of metastatic breast cancer. We expect that this treatment will be well tolerated and will increase tumor immunogenicity and the efficacy of checkpoint inhibition therapy. Since our aptamers recognize both mouse and human metastatic cancers, the translation of positive finding in the clinic might should be facilitated and might have, in the future, an important impact on metastatic breast cancer allowing its eradication and/or containment with minimal side effect.

即使 FDA 批准 Atezolizumab 联合白蛋白结合型紫杉醇治疗 PDL1 阳性三联疗法阴性乳腺癌，转移性乳腺癌仍然是一种致命的疾病。检查点的影响不大这种疾病的抑制剂治疗可以部分解释为相对较低的肿瘤突变负担、较低的与其他癌症相比，转移性乳腺癌的新抗原表达和总体免疫原性较低检查点抑制剂对恶性肿瘤非常有效。最近的 RNA 测序和 MHC 肽组对接受检查点抑制治疗的患者肿瘤的分析揭示了新抗原池中内含子衍生的表位。在乳腺癌中，这些内含子衍生的新抗原的作用很差由于剪接体机制的组成性上调而表达。在这里，我们将检验适体介导的靶向沉默 snRPE 的假设，snRPE 是剪接体机制，可以增加乳腺癌免疫原性并与抗 PD1 协同作用根除转移性乳腺癌。针对这种靶向治疗，我们已经鉴定出两种 RNA 适体可以在体外和体内识别小鼠和人类转移性乳腺癌细胞。 RNA 适体是小寡核苷酸，由于其 3D 结构，可以以高亲和力识别其配体。适体是化学合成的，具有赋予 RNAse 抗性和缺乏免疫原性的主链，并且可以进行修饰以改善药代动力学和治疗递送。我们已经将我们的适体与针对 snRPE 的 siRNA 并显示了其体外功效。在这项研究中，我们将采用两种侵袭性小鼠乳腺癌模型：4T1 和 E0771 两种不同的遗传背景。首先，我们将描述免疫系统在抗肿瘤中的作用使用 NSG 和免疫活性小鼠、耗竭实验以及通过表征肿瘤来进行反应微环境与抗肿瘤免疫。然后，我们将测量适配体嵌合体的治疗效果作为单一疗法或与检查点抑制疗法和内含子衍生疫苗联合使用用于治疗转移性乳腺癌的新表位。我们预计这种治疗将具有良好的耐受性，并会增加肿瘤的免疫原性和检查点抑制疗法的疗效。由于我们的适体可识别小鼠和人类的转移瘤癌症，临床上积极发现的转化可能应该得到促进，并且可能在未来，对转移性乳腺癌具有重要影响，可以以最小的副作用根除和/或遏制它影响。