Targeting FOXP3 mRNA splicing for breast cancer immunotherapy

靶向 FOXP3 mRNA 剪接用于乳腺癌免疫治疗

• 批准号: 10717185
• 负责人: Xiongbin Lu
• 金额: $ 58.52万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-27 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10717185
• 关键词: Adoptive Cell Transfers; Adverse effects; Alternative Splicing; Asthma; Autoimmune Diseases; Autoimmunity; Bioinformatics; Birth; Breast Cancer Model; Breast Cancer Treatment; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cancer Biology; Cell physiology; Cells; Clinical; Data; Development; Disease; Duchenne muscular dystrophy; Elements; Exhibits; Exons; FDA approved; FOXP3 gene; Gene Expression; Homeostasis; Human; Hypersensitivity; IL2 gene; Immune; Immune checkpoint inhibitor; Immune system; Immunity; Immunology; Immunosuppression; In Vitro; Infiltration; Inflammation; Injections; Interferon Type II; Knock-in; Knock-out; Length; Mammary Neoplasms; Mediating; Messenger RNA; Modeling; Mus; Mutant Strains Mice; Organoids; Outcome; Patients; Pharmaceutical Preparations; Play; Protein Isoforms; Public Health; Publications; RNA Splicing; Regulatory Element; Regulatory T-Lymphocyte; Research; Research Personnel; Resistance; Role; Science; Severities; Systemic Lupus Erythematosus; T cell therapy; T-Cell Development; T-Lymphocyte; Testing; The Cancer Genome Atlas; Toxic effect; Transgenic Organisms; Tumor Immunity; Tumor Promotion; Wild Type Mouse; anti-tumor immune response; cancer genomics; cancer immunotherapy; cancer therapy; cancer type; clinical development; directed differentiation; efficacy evaluation; efficacy testing; human disease; in vivo; mRNA Expression; malignant breast neoplasm; mouse model; neoantigens; neoplastic cell; novel; novel therapeutic intervention; patient derived xenograft model; patient subsets; precision medicine; programs; success; targeted treatment; transcriptomics; tumor; tumor growth

Project Abstract The development of cancer immunotherapy including immune checkpoint inhibitors and adoptive cell transfer (ACT)-based therapies revolutionized cancer treatment. The success, however, is limited to a relatively small subset of patients and cancer types. Regulatory T cells (Tregs) play a central role in maintaining immune system homeostasis and negatively regulate immune-mediated inflammation such as autoimmune diseases, asthma and allergy. However, tumor infiltrating Tregs also suppress effective anti-tumor immunity. FOXP3 is a master regulator of Treg development and function. Human FOXP3 gene encodes two major isoforms through mRNA alternative splicing – a long full-length isoform (FOXP3L) and a shorter isoform lacking exon 2 region (FOXP3S). To study the function of FOXP3S in Tregs, we generated a Foxp3S mouse line expressing only the Foxp3S isoform. Our preliminary studies with orthotopic mouse breast tumor models demonstrate that the Foxp3S mice are completely resistant to tumor development, while the littermate wildtype mice expressing Foxp3L exhibit significant tumor growth. Intratumoral injection of Foxp3S-promoting morpholino drug (MO) into the breast tumors in wildtype mice markedly increases IFN-γ+ CD4 and CD8 T cells in the tumors and thus suppresses tumor growth. Interestingly, bioinformatics analysis of transcriptomic data from The Cancer Genome Atlas (TCGA) reveals that FOXP3S mRNA expression in human breast cancer is positively correlated with clinical outcomes. Given that over 60% of tumor-infiltrating CD4 T cells are Tregs and the most dominant TCRs from intratumoral Tregs were tumor-reactive and recognize specific tumor neoantigens, we hypothesize that promoting FOXP3S isoform expression will reprogram tumor-reactive Tregs and convert these immune suppressive Tregs to tumor-specific T helpers thus promoting antitumor immunity. We propose to use unique mouse lines, FOXP3S-promoting morpholinos and patient-derived organoids to test our hypothesis. We will determine the mechanisms for FOXP3S-mediated antitumor immunity and further define the efficacy of FOXP3S- promoting morpholinos in breast cancer treatment. Successful completion of this study will identify a novel therapeutic strategy for breast cancer immunotherapy.

项目摘要 癌症免疫疗法的发展，包括免疫检查点抑制剂和过继性细胞移植 基于 ACT 的疗法彻底改变了癌症治疗。然而，成功仅限于相对较小的 患者子集和癌症类型。调节性 T 细胞 (Treg) 在维持免疫系统中发挥着核心作用 体内平衡并负面调节免疫介导的炎症，例如自身免疫性疾病、哮喘 和过敏。然而，肿瘤浸润性Treg细胞也会抑制有效的抗肿瘤免疫。 FOXP3是高手 Treg 发育和功能的调节因子。人类 FOXP3 基因通过 mRNA 编码两种主要亚型 选择性剪接 – 长的全长异构体 (FOXP3L) 和缺少外显子 2 区域的较短异构体 (FOXP3S)。 为了研究 FOXP3S 在 Tregs 中的功能，我们生成了仅表达 Foxp3S 的 Foxp3S 小鼠品系 同工型。我们对原位小鼠乳腺肿瘤模型的初步研究表明，Foxp3S 小鼠 完全抵抗肿瘤发展，而表达 Foxp3L 的同窝野生型小鼠表现出 显着的肿瘤生长。将 Foxp3S 促进吗啉药物 (MO) 瘤内注射到乳房中 野生型小鼠的肿瘤显着增加肿瘤中的 IFN-γ+ CD4 和 CD8 T 细胞，从而抑制 肿瘤生长。有趣的是，对癌症基因组图谱转录组数据进行生物信息学分析 (TCGA)揭示人类乳腺癌中FOXP3S mRNA的表达与临床表现呈正相关 结果。鉴于超过 60% 的肿瘤浸润 CD4 T 细胞是 Tregs，并且最主要的 TCR 是来自 瘤内 Tregs 具有肿瘤反应性并识别特定的肿瘤新抗原，我们假设 促进 FOXP3S 同工型表达将重新编程肿瘤反应性 Tregs 并将这些免疫细胞转化为 抑制Tregs对肿瘤特异性T辅助细胞的影响，从而促进抗肿瘤免疫。我们建议使用独特的 小鼠品系、促进 FOXP3S 的吗啉和源自患者的类器官来检验我们的假设。我们将 确定 FOXP3S 介导的抗肿瘤免疫机制，并进一步确定 FOXP3S- 的功效 促进吗啉在乳腺癌治疗中的应用。成功完成这项研究将确定一种新颖的 乳腺癌免疫治疗的治疗策略。