MSC-Derived Exosomes and MicroRNA in Glioma Therapy

MSC 衍生的外泌体和 MicroRNA 在神经胶质瘤治疗中的应用

• 批准号: 10305631
• 负责人: FREDERICK F LANG
• 金额: $ 35.87万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-11 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10305631
• 关键词: Address; Adult; Biology; Blood; Brain; Brain Neoplasms; Clinic; Data; Disease; Dose; Down-Regulation; Fibroblasts; Gene Expression; Genes; Glioblastoma; Glioma; Goals; Grant; Growth; Home; Homing; Human; In Vitro; Lentivirus; Light; Literature; Malignant neoplasm of brain; Mass Spectrum Analysis; Mediating; Mesenchymal Stem Cells; Methods; MicroRNAs; Mus; New Agents; Patients; Persons; Primary Brain Neoplasms; Proteins; Proteomics; Public Health; Regulator Genes; Reporter; Safety; Schedule; Surface; System; Testing; Tetracyclines; The Cancer Genome Atlas; Therapeutic; Therapeutic Uses; Time; Translations; Tumor Suppressor Proteins; Untranslated RNA; base; bioluminescence imaging; blood-brain tumor barrier; bone marrow mesenchymal stem cell; delivery vehicle; differential expression; effective therapy; efficacy testing; exosome; high throughput screening; improved; in vivo; in vivo bioluminescence imaging; nanovesicle; novel; novel strategies; novel therapeutic intervention; overexpression; prevent; restoration; stem cell exosomes; stem-like cell; therapeutic miRNA; therapeutically effective; therapy resistant; treatment response

Glioblastoma (GBM), the most common and deadly adult primary brain tumor, is notoriously resistant to therapy, not only because of its unique biology, including Glioma Stem-like Cells (GSCs), but also because of the delivery challenges imposed by the blood-brain and blood-tumor barriers. Thus, there is an urgent need to identify effective GBM-specific therapeutics and to elucidate strategies for delivering these new agents. A promising new approach is treatment with microRNAs (miRs), which are small, noncoding RNAs that are powerful regulators of gene expression. We have shown that restoration of tumor suppressor miRs is capable of killing GSCs and, therefore, treatment with miRs is potentially a new paradigm in GBM therapy. However, it is currently unknown which miRs will be most effective against GBMs and how these miRs should be delivered. This grant is significant because it directly addresses both of these problems by identifying the most effect anti-GBM miRs and by developing exosomes derived from bone marrow mesenchymal stem cells (MSCs) as a new delivery strategy for these miRs. Exosomes are naturally occurring nanovesicles that function as intercellular transport vehicles. We hypothesize that miRs can be used as effective therapeutics against GBMs, and that exosomes derived from cultured MSCs can be used to systemically deliver these antiglioma miRs. Aim 1 will identify miRs that are most effective against GSCs. In an in vitro high-throughput screen (HTS) of 578 miRs against a panel of GSCs, we have already identified 50 antiglioma miRs that effectively kill GSCs in vitro. This aim will use an in vivo HTS to determine which of the 50 top candidates is most effective against orthotopic GSCs in mice, and it will define the most effective miR combinations. Mechanistic studies will identify the target(s) responsible for the anti-GBM effects. Aim 2 will develop MSC-derived exosomes as novel delivery vehicles of antiglioma miRs. We have shown that after transduction with lentiviruses containing antiglioma miRs, MSCs package the miRs into exosomes (called Exos-miRs) and secrete the Exo-miRs into the culture medium. These Exo-miRs can be isolated and delivered systemically, after which they home to GSCs and inhibit target genes. This aim will define the optimal dose/schedule of Exos-miRs using bioluminescence imaging and a novel TET-reporter system. This dose/schedule will be used to test the efficacy and safety of the best antiglioma miRs or miR combinations identified in Aim 1. Aim 3 will elucidate the mechanisms underlying the ability of Exos-miRs to home to GBMs. By using proteomic analyses to compare strongly homing exosomes (MSC-Exos) with poorly homing ones (fibroblast-Exos), we have identified candidate homing proteins. In this aim, we will perform functional analyses to determine the extent to which the identified proteins mediate the homing of Exo-miRs to GBMs. Successful completion of this grant will result in a new and integrated agent/delivery paradigm that uses miRs to treat GBMs and MSC-derived exosomes to deliver these miRs, and that has the potential to improve the survival of the estimated 15,000 people who die of GBM each year due to the current lack of effective therapy for this disease.

胶质母细胞瘤（GBM）是最常见和致命的成人原发性脑肿瘤，众所周知对治疗有抵抗力， 不仅因为其独特的生物学特性，包括神经胶质瘤干细胞（GSC），还因为交付 血脑屏障和血肿瘤屏障带来的挑战。因此，迫切需要识别 有效的 GBM 特异性疗法并阐明提供这些新药物的策略。一个有前途的新 方法是用 microRNA (miR) 进行治疗，这是一种小的非编码 RNA，是强有力的调节剂 基因表达。我们已经证明，肿瘤抑制 miR 的恢复能够杀死 GSC，并且， 因此，miR 治疗可能成为 GBM 治疗的新范例。但目前尚不清楚 哪些 miR 对 GBM 最有效，以及如何传递这些 miR。这笔补助意义重大 因为它通过识别最有效的抗 GBM miR 并通过 开发源自骨髓间充质干细胞（MSC）的外泌体作为新的递送策略 对于这些 miR。外泌体是天然存在的纳米囊泡，充当细胞间运输工具。 我们假设 miR 可用作针对 GBM 的有效疗法，并且源自 培养的 MSC 可用于系统性递送这些抗神经胶质瘤 miR。目标 1 将识别最常见的 miR 对 GSC 有效。在针对一组 GSC 的 578 个 miR 的体外高通量筛选 (HTS) 中，我们 已经鉴定出 50 种抗神经胶质瘤 miR 可以在体外有效杀死 GSC。该目标将使用体内 HTS 确定 50 个顶级候选药物中哪一个对小鼠原位 GSCs 最有效，并且它将定义 最有效的 miR 组合。机制研究将确定负责抗 GBM 的靶标 影响。目标 2 将开发 MSC 衍生的外泌体作为抗神经胶质瘤 miR 的新型递送载体。我们有 结果表明，用含有抗神经胶质瘤 miR 的慢病毒转导后，MSC 将 miR 包装成 外泌体（称为 Exos-miR）并将 Exo-miR 分泌到培养基中。这些 Exo-miR 可以是 分离并系统递送，然后它们归巢于 GSC 并抑制靶基因。这一目标将定义 使用生物发光成像和新型 TET 报告系统确定 Exos-miR 的最佳剂量/时间表。这 剂量/时间表将用于测试最佳抗神经胶质瘤 miR 或 miR 组合的功效和安全性 目标 1 中确定。目标 3 将阐明 Exos-miR 归巢于 GBM 的能力的机制。 通过使用蛋白质组学分析来比较强归巢外泌体 (MSC-Exos) 和弱归巢外泌体 （成纤维细胞-Exos），我们已经确定了候选归巢蛋白。为此，我们将进行功能分析 以确定所鉴定的蛋白质介导 Exo-miR 归巢至 GBM 的程度。成功的 此项资助的完成将产生一种新的综合药物/递送范例，使用 miR 来治疗 GBM 和 MSC 衍生的外泌体来传递这些 miR，这有可能提高细胞的存活率 由于目前缺乏有效的治疗方法，估计每年有 15,000 人死于 GBM。