"Modulation of Immune Checkpoints by Self-Deliverable RNAi for Adoptive Cell Transfer"

“通过自传递 RNAi 调节免疫检查点以进行过继细胞转移”

• 批准号: 9254537
• 负责人: Alexey Wolfson
• 金额: $ 23.15万
• 依托单位: ADVANCED RNA TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2019-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9254537
• 关键词: Adoptive Cell Transfers; Adoptive Transfer; Antibodies; B-Cell Lymphomas; Biochemical; Biological Assay; Biotechnology; CAR receptor; CBL gene; CD19 gene; Cell Line; Cell Therapy; Cells; Chemicals; Chemistry; Clinic; Clinical; Collaborations; Custom; Data; Development; Disease remission; Drug resistance; Electroporation; Engineering; Flow Cytometry; Formulation; Future; Gene Silencing; Genes; Goals; Hematologic Neoplasms; Human; Immune; Immune checkpoint inhibitor; Immunosuppression; Immunosuppressive Agents; In Vitro; Individual; Lead; Length; Lipids; Longevity; Malignant neoplasm of ovary; Mediating; Methods; Modification; Monoclonal Antibodies; Natural Killer Cells; Ovarian Carcinoma; PDCD1LG1 gene; Pathway interactions; Patients; Pattern; Peripheral; Phase; Pre-Clinical Model; Property; Protocols documentation; RNA Interference; Reagent; Research; Series; Small Business Innovation Research Grant; Small Interfering RNA; Solid Neoplasm; Structure; T-Lymphocyte; Technology; Technology Transfer; Therapeutic; Therapeutic Effect; Toxic effect; Transfection; Tumor-Infiltrating Lymphocytes; Variant; Viral Vector; Western Blotting; Work; Xenograft procedure; antitumor agent; base; cancer immunotherapy; cell growth; cell type; cellular engineering; combinatorial; comparative efficacy; cytotoxicity; efficacy study; immune checkpoint; immune checkpoint blockade; immunoregulation; improved; in vivo; inorganic phosphate; knock-down; melanoma; mesothelin; mouse model; neoplastic cell; novel; phase 2 study; phosphorothioate; preclinical efficacy; prevent; safety study; scaffold; success; tumor; tumor microenvironment

Two of the most promising approaches to cancer immunotherapy are immune checkpoint blockade by monoclonal antibodies and the adoptive transfer of immune cells. The recent successes in treating melanoma with anti-PD1 and anti-CTLA antibodies as well as the extremely successful application of T cells engineered to express the CAR receptor targeting CD19 to treat B cell lymphomas initiated a waterfall of research in identifying new indications and targets for both antibodies and immune cells. While highly effective in treating hematological malignancies, adoptive transfer of natural or engineered T cells is much more challenging to implement for the treatment of solid tumors, in large part due to the immunosuppressive tumor microenvironment. Advirna is proposing to improve the efficacy of adoptive cell transfer therapies by pre- treatment of immune cells ex vivo with RNAi reagents to prevent immunosuppression. Our approach is based on the use of unique, self-deliverable RNAi (sdRNAi) molecules that have been chemically modified, conferring them stability and the inherent ability to enter immune cells, without the need for exogenous reagents or methods. Because of its simplicity, self-deliverable RNAi technology is perfectly compatible with existing cell growth protocols (sdRNAs are used as media component) and is very easy to implement. sdRNAi high efficiency of delivery (>99%), lack of toxicity and strong and long-lasting effect of gene suppression enables knockdown of multiple immune checkpoint and/or regulatory molecules involved in immunosuppression. This may protect the adoptively transferred immune cells in the tumor microenvironment and enhance their efficacy as antitumor agents. sdRNAs can be applied to a broad variety of ACT technologies (CAR-T, engineered TCR, TIL, NK). The unique properties of sdRNAi technology makes it a method of choice for multiplexed immune check point modulation in CAR-T cells. We have generated lead sdRNA compounds targeting a number of immune checkpoints (PD1, LAG3, TIGIT etc.).Our preliminary results in tumor infiltrating lymphocytes (TILs) (enhancing their cytotoxicity to melanoma tumor cells) and in mesothelin targeting CAR-T cells (increasing their efficacy in a mouse model of ovarian cancer) show promise for the success of the approach. We propose to further optimize the existing chemical modification pattern of sdRNAs to improve the knockdown efficiency and longevity of the RNAi effect, and to evaluate the effect of simultaneous knockdown of multiple immune checkpoints in vitro and in vivo using mesothelin CAR-T cells. Our overall goal is to create a series of optimized sdRNAs targeting most of the known immune checkpoints to allow customizable combinatorial RNAi “cocktails” for specific tumor and ACT cell types to protect immune cells and increase their efficacy. Future in vivo studies in collaboration with academic groups and biotech companies will further evaluate this concept with a goal to move this technology to clinic as a general approach for enhancement of efficacy of any cell-based therapies to treat solid tumors.

两种最有希望的癌症免疫治疗方法是通过以下方式阻断免疫检查点 单抗和免疫细胞的过继转移。黑色素瘤治疗的最新进展 随着抗PD1和抗CTLA抗体以及T细胞工程的极其成功地应用 表达靶向CD19的CAR治疗B细胞淋巴瘤启动了瀑布式的研究 识别抗体和免疫细胞的新适应症和靶点。虽然在治疗中非常有效 血液系统恶性肿瘤，过继转移天然或工程T细胞更具挑战性 用于治疗实体肿瘤的工具，很大程度上是由于免疫抑制肿瘤 微环境。Advirna正在提议通过预先治疗来提高过继细胞转移疗法的疗效 用RNAi试剂体外治疗免疫细胞以防止免疫抑制。我们的方法是基于 关于使用经过化学修饰的独特的自我递送RNAi(SdRNAi)分子， 它们的稳定性和固有的进入免疫细胞的能力，不需要外源试剂或 方法：研究方法。由于其简单性，可自行交付的RNAi技术与现有细胞完全兼容 生长协议(sdRNA用作媒体组件)并且非常易于实施。SdRNAi高 传递效率(&gt；99%)、无毒性和强大而持久的基因抑制效应使 击倒参与免疫抑制的多个免疫检查点和/或调节分子。这 可保护肿瘤微环境中过继转移的免疫细胞，提高其疗效 作为抗肿瘤药物。SdRNAs可应用于多种ACT技术(CAR-T，工程化 TCR、TIL、NK)。SdRNAi技术的独特特性使其成为多路传输的首选方法 免疫检查点在CAR-T细胞中的调节。 我们已经产生了针对许多免疫检查点(PD1、LAG3、TIGIT)的先导sdRNA化合物 我们在肿瘤浸润性淋巴细胞(TIL)(增强其对黑色素瘤的细胞毒性)方面的初步结果 肿瘤细胞)和以CAR-T细胞为靶点(提高其在卵巢癌小鼠模型中的疗效) 癌症)显示了这种方法的成功前景。我们建议进一步优化现有的化学品 SdRNAs的修饰模式，以提高RNAi效应的击倒效率和寿命，并 体内外同时敲除多个免疫检查点的效果评价 中草药CAR-T细胞。我们的总体目标是创建一系列针对大多数 已知的免疫检查点允许针对特定肿瘤和ACT的可定制组合RNAi“鸡尾酒” 保护免疫细胞并提高其效力的细胞类型。未来的活体研究与 学术团体和生物技术公司将进一步评估这一概念，目标是推动这项技术 将其作为提高任何基于细胞的治疗实体瘤疗效的一般方法应用于临床。