Evaluation of lead FANA oligos targeting FOXP3, with and without anti-PD-1 mAb, as an approach for cancer immunotherapy

对靶向 FOXP3 的先导 FANA 寡核苷酸（含或不含抗 PD-1 mAb）作为癌症免疫治疗方法的评估

• 批准号: 10546343
• 负责人: Veenu Aishwarya
• 金额: $ 40万
• 依托单位: AUM LIFETECH, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10546343
• 关键词: Adverse effects; Affect; Antisense Oligonucleotides; Autoimmunity; Award; CD8-Positive T-Lymphocytes; CD8B1 gene; Cancer Patient; Cancer Vaccines; Cause of Death; Cessation of life; Chemicals; Chemotherapy and/or radiation; Clinical; Clinical Research; Clinical Trials; Complex; Data; Development; Effectiveness; Effector Cell; Elements; Evaluation; FOXP3 gene; Funding; Future; Gene Expression; Generations; Goals; Granzyme; Host resistance; Human; IL2RA gene; Immune checkpoint inhibitor; Immune system; Immunocompetent; Immunodeficient Mouse; Immunotherapy; Impairment; In Vitro; Infiltration; Injections; Interferon Type II; Interphase Cell; Lead; Legal patent; Liver; Lung Neoplasms; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Messenger RNA; Modeling; Modification; Molecular; Monoclonal Antibodies; Monoclonal Antibody Therapy; Mus; Myeloid-derived suppressor cells; Oligonucleotides; Operative Surgical Procedures; Patients; Peripheral Blood Mononuclear Cell; Pharmacologic Substance; Phase; Preparation; Process; Production; Prognosis; RNA-targeting therapy; Radiation; Regulatory T-Lymphocyte; Resistance; Rest; Services; Small Interfering RNA; Source; Surface; Survival Rate; T cell response; Technology; Testing; Therapeutic; Toxic effect; Transcriptional Regulation; Transgenic Mice; Tumor Immunity; Tumor-infiltrating immune cells; Vaccine Therapy; Validation; Woman; anti-PD-1; anti-PD1 antibodies; anti-tumor immune response; base; cancer immunotherapy; cancer survival; cancer therapy; cancer type; checkpoint inhibition; checkpoint therapy; chemotherapy; clinical application; commercialization; cytotoxic CD8 T cells; effective therapy; effector T cell; efficacy study; gene network; humanized mouse; immunoregulation; in vivo; knock-down; men; mortality; mouse model; neoplasm immunotherapy; neoplastic cell; next generation; novel; novel strategies; nuclease; pharmacokinetics and pharmacodynamics; preclinical study; prevent; reconstitution; research and development; response; transcription factor; tumor; tumor growth; tumor microenvironment; tumor progression

PROJECT SUMMARY/ABSTRACT Lung cancer accounts for >25% of cancer-related deaths in both men and women, making it the largest source of cancer-related mortality. Despite advancements in surgical intervention, radiation, chemotherapy, and immunotherapy, the 5-year survival rate for all types of lung cancer is 19%. The advent of immune checkpoint inhibitor therapy is associated with remarkable efficacy is some patients with lung cancers, but >75% of patients do not achieve meaningful clinical responses. Currently, T-regulatory (Treg) T cells are regarded as one of the major obstacles to the successful clinical application of tumor immunotherapy. Tregs contribute to the early establishment and progression of tumors, dampen effector cell responses by various mechanisms, and promote and support pro-tumoral myeloid-derived suppressor cell development and function. Tregs are thereby key to the immunosuppressive state of the tumor microenvironment that prevents effective anti-tumor immune responses. Pre-clinical studies show targeting of Tregs, by their depletion or functional modulation, can have significant therapeutic benefit, alone or in combination with other immunomodulatory approaches. We and others have described the significant accumulation of intratumoral Treg in lung cancer patients, and their increased intratumoral numbers correlate with poor prognosis in many types of cancers. Tregs are defined by their expression of the transcription factor FOXP3, which controls Treg development, stability, and function. FOXP3 is the key element of multiple Treg-associated molecular complexes that regulate the Treg-specific gene network and mechanisms of Treg-mediated immune regulation. FOXP3 is located intracellularly, and previous efforts to target Tregs using their CD25 surface marker have unintentionally co-targeted CD25+ activated T effector cells, causing a lack of robust anti-tumoral efficiency in pre-clinical and clinical studies, along with severe adverse effects and toxicity. On the other hand, Foxp3-specific depletion of Treg using tumor bearing DEREG transgenic mice, in combination with anti-tumor vaccine therapy, resulted in robust antitumor immunity and an absence of overt autoimmunity. Thus, there is an urgent and unmet need for therapies that can selectively target Treg cells, allowing the body to mount its normal defenses against the cancer, without negative influences on the rest of the immune system. We have developed third generation FANA antisense oligonucleotides (FANA ASOs) that selectively inhibit Tregs by knockdown of FOXP3 expression. This next generation of ASO differs from their predecessors by chemical modifications that allow their efficient (gymnotic) self-delivery, even to resting cells, and make them resistant to degradation by endogenous nucleases. In preliminary studies, anti-Foxp3 FANA ASOs reduced Foxp3 expression and suppressive functions of murine Tregs, and in 50% of syngeneic mice bearing lung tumors, daily injection of FANA ASOs showed complete destruction of the tumors as compared to scrambled controls and without overt toxicity. We now propose validation of newly developed human FOXP3 FANAs, alone or in conjunction with anti-PD-1 monoclonal antibody therapy. Aim 1 will assess their ability to knockdown FOXP3 and promote anti-tumor immunity in humanized mice in vivo. Aim 2 will evaluate their activity against actual human lung cancer associated Treg cells.

项目总结/摘要 肺癌占男性和女性癌症相关死亡的25%以上，是最大的癌症。 癌症相关死亡率的来源。尽管外科手术、放疗、化疗取得了进步， 和免疫疗法，所有类型肺癌的5年生存率为19%。免疫系统的出现 检查点抑制剂治疗与一些肺癌患者的显著疗效相关，但 >75%的患者没有获得有意义的临床反应。目前，T调节性（Treg）T细胞是 被认为是肿瘤免疫治疗成功应用于临床的主要障碍之一。Tregs 有助于肿瘤的早期建立和进展，通过各种途径抑制效应细胞反应， 机制，并促进和支持促肿瘤骨髓源性抑制细胞的发展， 功能因此，T细胞是肿瘤微环境免疫抑制状态的关键， 阻止有效的抗肿瘤免疫反应。临床前研究显示，通过其 单独或与以下物质组合，可以具有显著的治疗益处： 其他免疫调节方法。我们和其他人已经描述了 肺癌患者肿瘤内Treg数量的增加与肿瘤内Treg的不良反应相关。 许多类型癌症的预后。 调节性T细胞通过其转录因子FOXP 3的表达来定义，FOXP 3控制Treg发育， 稳定性和功能。FOXP 3是多种Treg相关分子复合物的关键元件， 调节Treg特异性基因网络和Treg介导的免疫调节机制。foxp 3是 位于细胞内，先前使用其CD 25表面标记物靶向TcB的努力已经取得了进展。 无意中共靶向的CD 25+活化的T效应细胞，导致缺乏强有力的抗肿瘤治疗。 在临床前和临床研究中的有效性，沿着严重的副作用和毒性。另 另一方面，使用携带肿瘤的DEREG转基因小鼠，与 抗肿瘤疫苗治疗，导致强大的抗肿瘤免疫和明显的自身免疫的缺乏。 因此，迫切需要能够选择性靶向Treg细胞的疗法，从而允许治疗。 身体对癌症进行正常防御，而不会对其他免疫系统产生负面影响。 系统 我们开发了第三代FANA反义寡核苷酸（FANA ASO），选择性地 通过敲低FOXP 3表达来抑制TcP。下一代阿索与他们的 通过化学修饰，使其有效的（裸）自我交付，甚至休息的前辈， 细胞，并使它们抵抗内源性核酸酶的降解。在初步研究中，抗Foxp 3 FANA反义寡核苷酸降低了小鼠T细胞Foxp 3的表达和抑制功能，在50%的小鼠中， 对于携带肺肿瘤的同系小鼠，每天注射FANA ASO显示完全破坏了肺肿瘤的细胞。 肿瘤相比，乱对照，没有明显的毒性。我们现在建议确认新的 开发了人FOXP 3 FANA，单独或与抗PD-1单克隆抗体疗法联合。 目的1将评估它们在人源化小鼠中敲低FOXP 3和促进抗肿瘤免疫的能力 in vivo.目的2将评价它们对实际人肺癌相关Treg细胞的活性。