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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/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。尽管手术干预、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%。免疫的出现检查点抑制剂治疗对一些肺癌患者具有显着疗效，但是 >75% 的患者未达到有意义的临床反应。目前，调节性 T (Treg) T 细胞被认为是肿瘤免疫治疗成功临床应用的主要障碍之一。调节性T细胞有助于肿瘤的早期形成和进展，抑制各种效应细胞的反应机制，促进和支持促肿瘤骨髓源性抑制细胞的发育和功能。因此，Tregs 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。临床前研究表明，Tregs 的靶向作用是通过其消耗或功能调节，可以单独或联合使用具有显着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瘤内 Treg 数量增加，且瘤内 Treg 数量增加与较差的相关性许多类型癌症的预后。 Treg 的定义是转录因子 FOXP3 的表达，FOXP3 控制 Treg 的发育，稳定性、功能。 FOXP3 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。 FOXP3 是位于细胞内，之前使用 CD25 表面标记物靶向 Tregs 的努力已经无意中共同靶向 CD25+ 激活的 T 效应细胞，导致缺乏强大的抗肿瘤作用临床前和临床研究的效率，以及严重的不良反应和毒性。另一方面另一方面，使用携带肿瘤的 DEREG 转基因小鼠结合 Foxp3 特异性消除 Treg 抗肿瘤疫苗治疗产生了强大的抗肿瘤免疫力并且没有明显的自身免疫。因此，迫切需要能够选择性靶向 Treg 细胞的疗法，从而使身体对癌症建立正常的防御能力，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 FOXP3 表达来抑制 Tregs。下一代 ASO 与他们的不同通过化学修饰，使其能够有效（体操）自我分娩，甚至在休息时细胞，并使它们抵抗内源核酸酶的降解。在初步研究中，抗 Foxp3 FANA ASO 降低了 Foxp3 的表达和小鼠 Tregs 的抑制功能，并且在 50% 的小鼠中患有肺肿瘤的同基因小鼠，每天注射 FANA ASOs 显示肺肿瘤被完全破坏与乱序对照相比，肿瘤没有明显的毒性。我们现在建议验证新的开发了人 FOXP3 FANA，单独或与抗 PD-1 单克隆抗体疗法联合使用。目标 1 将评估其敲低 FOXP3 并促进人源化小鼠抗肿瘤免疫力的能力体内。目标 2 将评估它们针对实际人类肺癌相关 Treg 细胞的活性。