Project 4: Inhibiting Novel Autophagy Mediator ATG4B for Treating Glioblastoma

项目4：抑制新型自噬介质ATG4B治疗胶质母细胞瘤

基本信息

批准号：
10224127
负责人：
Shi-Yuan Cheng
金额：
$ 22.51万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10224127
关键词：
Animals Apoptosis Autophagocytosis Brain Brain Neoplasms Catabolic Process Cell model Cells Chemotherapy and/or radiation Chloroquine Clinical Research Clinical Trials Combined Modality Therapy Cytotoxic Chemotherapy Data Development Development Plans Developmental Therapeutics Program Diagnosis Disease Effectiveness Enzymes Epigenetic Process Excision Exhibits FRAP1 gene Foundations Genetic Glioblastoma Glioma Goals Growth Homeostasis In Vitro Individual Intracranial Neoplasms Malignant - descriptor Malignant Neoplasms Malignant neoplasm of brain Mass Spectrum Analysis Mediating Mediator of activation protein Modeling Molecular Molecular Profiling Mus Newly Diagnosed Nude Mice Nutrient Organelles Patient-Focused Outcomes Patients Pharmacologic Substance Pharmacology Phosphorylation Plasma Preclinical Testing Primary Brain Neoplasms Process Prognosis Radiation therapy Recurrence Recycling Safety Serine Signal Transduction Specificity Testing Therapeutic Tissues Toxic effect Transgenes Tumorigenicity antitumor effect base blood-brain barrier penetration cancer cell cancer therapy chemoradiation clinical development combat energy balance experimental study genotoxicity improved in vivo in vivo Model inhibitor/antagonist interest knock-down mTOR inhibition mutant neoplastic cell novel novel therapeutics patient derived xenograft model pre-clinical preclinical study protein aggregation radiation effect response small molecule inhibitor stem-like cell symposium temozolomide therapy resistant translational approach treatment effect tumor tumor growth tumor metabolism tumor progression tumorigenic

项目摘要

PROJECT 4: PROJECT SUMMARY Enhancing GBM Cytotoxic Therapy Through Inhibition of Key Autophagy Mediator ATG4B Glioblastoma (GBM) is the most common and malignant primary brain tumor. Despite treatment consisting of surgical removal, radiation and chemotherapy, most patients with GBM die within 14 to 16 months after diagnosis, underscoring the urgent need for new therapies to combat this deadly disease. Autophagy is a conserved catabolic process that maintains homeostasis by regulating the energy balance of the cell. Cancer cells use autophagy to remove damaged organelles and aggregated proteins, and to recycle nutrients in high demand to support tumor growth. Radiochemotherapy (RT-TMZ) is the front-line treatments against GBM, but also activate the autophagic response in tumor cells, thus protecting the cells from undergoing apoptosis. Inhibition of mTOR signaling is a common target in cancer therapy. However, mTOR inhibition also induces autophagy in cancer cells. Consequently, there is immense interest in inhibiting this protective mechanism while treating cancer. Non-specific autophagy inhibitors like chloroquine (CQ) and hydroxy-CQ (HCQ) are being investigated in a large number of clinical trials. However, the lack of specificity of these compounds is , associated with toxicity and may diminish its efficacy. We discovered that ATG4B, an enzyme that converts LC3 to LC3-I/II, which is required for autophagy process in cells, is phosphorylated at serine residue 383 (p-S383) in patient-derived glioma stem-like cells (glioma initiating cells or GICs). We also found that ATG4B S383 phosphorylation increases GIC autophagic activity, and that intratumoral expression of p-S383 ATG4B correlates with poor prognosis in GBM patients. In contrast, knockdown of ATG4B, or expression of a non- phosphorylatable ATG4B mutant transgene (S383A), inhibits GSC autophagic response and tumorigenicity when engrafted in the brains of athymic mice. Furthermore, pharmacologic inhibition of ATG4B, using a NCI compound NSC185058 that inhibits ATG4B enzymatic activity and GBM tumorigenicity, markedly enhanced RT inhibition on GBM tumor growth and increases the survival of animals with intracranially engrafted GIC. Additionally, NSC185058 also markedly enhanced inhibitory effects by a catalytic mTORC inhibitor AZD2014 on GICs. Based on these strong data and our established multiple PDX GBM models of distinct genetic + epigenetic profiles, in this project, we propose to determine the anti-GBM efficacy of ATG4B inhibitor NSC185058, as monotherapy and in combination with RT-TMZ; investigate the therapeutic potential of combining inhibition of ATG4B, mTOR and RT and develop NSC185058 for use in patients, and test the ATG4B inhibitor, both as a single agent and in combination with cytotoxic therapy, in a clinical trial for treating patients with recurrent GBM. This project has a strong molecular mechanistic foundation and associated therapeutic hypothesis that, if proven correct, has the potential for positive impact on the treatment of GBM, and perhaps other malignancies.

项目 4：项目摘要通过抑制关键自噬介质 ATG4B 增强 GBM 细胞毒性治疗胶质母细胞瘤（GBM）是最常见和恶性的原发性脑肿瘤。尽管治疗包括手术切除、放疗和化疗后，大多数 GBM 患者在术后 14 至 16 个月内死亡诊断，强调迫切需要新疗法来对抗这种致命疾病。自噬是一种保守的分解代谢过程，通过调节细胞的能量平衡来维持体内平衡。癌症细胞利用自噬去除受损的细胞器和聚集的蛋白质，并在高浓度下回收营养物质支持肿瘤生长的需求。放化疗 (RT-TMZ) 是针对 GBM 的一线治疗方法，但是还激活肿瘤细胞的自噬反应，从而保护细胞免于凋亡。 mTOR 信号传导的抑制是癌症治疗的常见目标。然而，mTOR 抑制也会诱导癌细胞中的自噬。因此，人们对抑制这种保护机制非常感兴趣，同时治疗癌症。非特异性自噬抑制剂如氯喹 (CQ) 和羟基 CQ (HCQ) 正在被研究经过大量的临床试验研究。然而，这些化合物缺乏特异性，与毒性有关，并可能降低其功效。我们发现 ATG4B，一种将 LC3 转化为细胞自噬过程所需的 LC3-I/II 在丝氨酸残基 383 (p-S383) 处被磷酸化患者来源的神经胶质瘤干细胞样细胞（神经胶质瘤起始细胞或 GIC）。我们还发现ATG4B S383 磷酸化增加 GIC 自噬活性，并且 p-S383 ATG4B 的瘤内表达与 GBM 患者的不良预后相关。相反，ATG4B 的敲除或非- 可磷酸化的 ATG4B 突变转基因 (S383A)，抑制 GSC 自噬反应和致瘤性当植入无胸腺小鼠的大脑时。此外，使用 NCI 对 ATG4B 进行药理学抑制化合物 NSC185058 抑制 ATG4B 酶活性和 GBM 致瘤性，显着增强 RT 抑制 GBM 肿瘤生长并提高颅内植入 GIC 的动物的存活率。此外，NSC185058 还显着增强了催化 mTORC 抑制剂 AZD2014 的抑制作用关于担保投资证。基于这些强有力的数据和我们建立的多个具有不同遗传+的PDX GBM模型表观遗传图谱，在这个项目中，我们建议确定 ATG4B 抑制剂的抗 GBM 功效 NSC185058，作为单一疗法并与 RT-TMZ 联合使用；研究治疗潜力结合ATG4B、mTOR和RT的抑制，开发NSC185058用于患者，并测试ATG4B 抑制剂，既作为单一药物，又与细胞毒疗法联合使用，用于治疗患者的临床试验患有复发性 GBM。该项目具有强大的分子机制基础和相关的治疗方法假设如果被证明正确，可能会对 GBM 的治疗产生积极影响，也许其他恶性肿瘤。