A Phase 0 Trial of Hydroxychloroquine in Patients with Stage III and IV Resectabl

羟氯喹治疗 III 期和 IV 期可切除患者的 0 期试验

基本信息

批准号：
7892776
负责人：
Janice M. Mehnert
金额：
$ 32.02万
依托单位：
UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-07 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7892776
关键词：
Address Adverse effects Affect Aftercare Allografting Animal Model Antineoplastic Agents Ants Apoptosis Apoptotic Autophagocytosis Autophagosome Binding Proteins Biological Assay Biopsy Biopsy Specimen Blood Catabolic Process Cell Death Cell Line Cell Survival Cells Chloroquine Cleaved cell Clinical Clinical Trials Clinical Trials Design Connective Tissue Diseases Correlative Study Detection Development Digestion Dose Electron Microscopy Evaluation Excision FDA approved Future Glucose Goals Gold Homeostasis Human Hydroxychloroquine Imagery Immunohistochemistry In Situ Nick-End Labeling In Vitro Individual Knowledge Laboratories Lysosomes Malaria Malignant Neoplasms Measures Mediating Melanoma Cell Membrane Metabolic stress Methods Mitotic Modeling Molecular Chaperones Mus Necrosis Organelles Pathway interactions Patients Pharmaceutical Preparations Phase Polyubiquitin Process Proteins Recycling Sampling Skin Skin Tissue Specimen Staging Staining method Stains Stimulus Stress Sum Surrogate Markers Techniques Testing Time Tissue Sample Tumor Tissue Vesicle Western Blotting cancer cell cancer therapy caspase-3 cytotoxic drug efficacy glucose-regulated protein 170 human tissue in vivo inhibition of autophagy inhibitor/antagonist melanocyte melanoma molecular marker neoplastic cell new therapeutic target novel palliative programs protein degradation public health relevance research study response standard measure stress tolerance therapeutic development treatment effect tumor

项目摘要

DESCRIPTION (provided by applicant): Autophagy enables tumors to survive metabolic stress in a dormant state for prolonged periods of time and as such, this pathway is an emerging target for the creation of new anticancer therapies. Hydroxychloroquine (HCQ) is a drug that blocks autophagy, raising intralysosomal pH and inhibiting the final step of this pathway, impairing autophagic protein degradation. This results in autophagosome accumulation, potentially disturbing the survival of tumor cells reliant on autophagy and shunting them toward apoptotic and/or necrotic cell death. Preliminary results in melanoma cell lines indicate that treatment with chloroquine (CQ), a similar drug to HCQ used readily in the laboratory, increases autophagosome accumulation, and increased cell death was noted from treatment with CQ at varying concentrations. Additionally, CQ induced autophagosome accumulation in a mammalian tumor allograft model. Given that the autophagy pathway may be active in melanoma, we hypothesize that administration of HCQ can modulate the process of autophagy in melanoma tumor tissue samples in humans in vivo. To test this hypothesis, a phase 0 trial of HCQ in patients with stage III or IV melanoma planned for palliative or curative resection is proposed. Specifically, we will look for signs of autophagy inhibition by HCQ, comparing the mean number of autophagic vesicles per cell seen on electron microscopy (EM) in pre- versus post- treatment tumor specimens. Pre- and post- treatment skin samples and PBMCs will similarly be assayed to explore the potential of these samples as surrogate markers for treatment effect. While autophagosome accumulation on EM is a reliable method for detection of autophagy inhibition, it is expensive, tedious and requires a skilled individual to perform and hence, is not widely applicable to clinical specimens to detect autophagy inhibition. Thus, as part of this trial we will examine several proteins shown to detect inhibition of autophagy in laboratory models and in some preliminary studies of human tumor tissue, with the hope that these simpler assays will facilitate autophagy detection in human samples. Markers that will be examined include Beclin1, an essential autophagy regulator, and LC3, which is cleaved into LC3-I and LC3- II, with the latter translocated to the autophagosome membrane during autophagy. In addition, our group has discovered that autophagy deficient tumors under stress accumulate p62, a polyubiquitin binding protein that delivers aggregates to the lysosome for degradation, as well as several chaperone proteins including glucose related protein 70 (GRp170). It is expected that treatment with HCQ in melanomas with functional autophagy will result in increased autophagosome formation on EM and increased levels of LC3-II, p62 and GRp170 by immunohistochemistry and Western blot. Potential proapoptotic and/or ant proliferative effects of HCQ will also be explored through the use of pre- and post-treatment activated caspase-3, TUNEL, Ki-67 and mitotic rate assays. The goal of this proposal is to modulate autophagy for the first time in human tumors, using results from these experiments to guide development of markers of autophagy in future clinical trials. PUBLIC HEALTH RELEVANCE: We propose a preliminary trial in humans with melanoma (a Phase O trial) to see if administering hydroxychloroquine (HCQ), a FDA approved drug with little to no side effects, can block the process of autophagy, a survival mode for several different types of tumor cells. We will use a method called electron microscopy to study if HCQ is inhibiting autophagy in melanoma tumor tissue and blood and skin samples, but since this method is expensive and laborious we will also examine four proteins -- Beclin1, LC3, p62 and GRp170 -- which detect autophagy inhibition in cell lines, animal models, and some preserved human tissue samples to demonstrate that these simpler tests may also detect inhibition of autophagy in human beings. Our hope is that we will be able to use these results to better understand and detect autophagy, making it easier to develop future drugs against this pathway in cancer.

描述（由申请人提供）：自噬使肿瘤能够在休眠状态下长时间生存代谢应激，因此，该途径是创建新抗癌疗法的新兴目标。羟氯喹 (HCQ) 是一种阻断自噬的药物，可提高溶酶体内 pH 值并抑制该途径的最后一步，从而损害自噬蛋白的降解。这导致自噬体积聚，可能扰乱依赖自噬的肿瘤细胞的生存，并使它们走向凋亡和/或坏死性细胞死亡。黑色素瘤细胞系的初步结果表明，用氯喹（CQ）（一种与实验室中常用的 HCQ 类似的药物）进行处理，可以增加自噬体的积累，并在不同浓度的 CQ 处理中观察到细胞死亡增加。此外，CQ 在哺乳动物肿瘤同种异体移植模型中诱导自噬体积累。鉴于自噬途径可能在黑色素瘤中活跃，我们假设施用 HCQ 可以调节人体黑色素瘤肿瘤组织样本中的自噬过程。为了检验这一假设，建议对计划进行姑息性或治愈性切除的 III 期或 IV 期黑色素瘤患者进行 HCQ 0 期试验。具体来说，我们将寻找 HCQ 抑制自噬的迹象，比较治疗前和治疗后肿瘤标本中电子显微镜 (EM) 上看到的每个细胞的自噬囊泡的平均数量。治疗前和治疗后的皮肤样本和 PBMC 也将进行类似的分析，以探索这些样本作为治疗效果替代标志物的潜力。虽然自噬体在电镜上的积累是检测自噬抑制的可靠方法，但其昂贵、繁琐并且需要熟练的个人来执行，因此不能广泛应用于临床标本来检测自噬抑制。因此，作为本试验的一部分，我们将在实验室模型和人类肿瘤组织的一些初步研究中检查几种显示可检测自噬抑制的蛋白质，希望这些更简单的测定将有助于人类样本中自噬的检测。将要检查的标记物包括 Beclin1（一种重要的自噬调节因子）和 LC3（它被裂解为 LC3-I 和 LC3-II，后者在自噬过程中易位至自噬体膜）。此外，我们的研究小组还发现，自噬缺陷的肿瘤在应激下会积累p62（一种多聚泛素结合蛋白，可将聚集物输送到溶酶体进行降解）以及几种伴侣蛋白，包括葡萄糖相关蛋白70 (GRp170)。通过免疫组织化学和蛋白质印迹预计，用 HCQ 治疗具有功能性自噬的黑色素瘤将导致 EM 上自噬体形成增加，并增加 LC3-II、p62 和 GRp170 水平。还将通过使用治疗前和治疗后激活的 caspase-3、TUNEL、Ki-67 和有丝分裂率测定来探索 HCQ 的潜在促凋亡和/或蚂蚁增殖作用。该提案的目标是首次在人类肿瘤中调节自噬，利用这些实验的结果指导未来临床试验中自噬标记物的开发。公共健康相关性：我们建议对患有黑色素瘤的人类进行一项初步试验（O 期试验），看看服用羟氯喹 (HCQ)（一种 FDA 批准的几乎没有副作用的药物）是否可以阻止自噬过程，自噬是黑色素瘤的一种生存模式。几种不同类型的肿瘤细胞。我们将使用一种称为电子显微镜的方法来研究 HCQ 是否抑制黑色素瘤肿瘤组织以及血液和皮肤样本中的自噬，但由于这种方法昂贵且费力，我们还将检查四种蛋白质 - Beclin1、LC3、p62 和 GRp170 -检测细胞系、动物模型和一些保存的人体组织样本中的自噬抑制，以证明这些更简单的测试也可以检测人类自噬的抑制。我们希望能够利用这些结果更好地理解和检测自噬，从而更容易开发未来针对癌症这一途径的药物。