Secondary Prevention through Surveillance and Intervention

项目摘要 尽管早期发现和辅助治疗，乳腺癌仍然是癌症的主要原因。 妇女的死亡率，主要是由于几年甚至几十年后出现的遥远的、无法治愈的复发 原发肿瘤的治疗。这些复发、转移的肿瘤起源于残留的局部和 在初次治疗后存活并保持在宿主体内假定处于休眠状态的播散性肿瘤细胞(DTC) 州政府。事实上，治疗后骨髓DCs的存在独立地与 大大增加了复发的风险。然而，目前，使残留肿瘤细胞能够 维持休眠并最终复发的机制知之甚少，DTC指导的监测和治疗 方法是不存在的。因此，对生物特征、准确测量和 靶向潜伏的DTCs将是预防复发的一种变革性的新方法。 我们假设，禁用休眠的DTCs所使用的生存机制将减少肿瘤 复发，从而提高存活率。使用基因工程小鼠模型，忠实地 总结肿瘤的休眠和复发，我们发现自噬和mTOR信号是 每一个都对DTC的生存至关重要，并且抑制这些途径的药物耗尽了休眠的储存库 残留肿瘤细胞，从而防止肿瘤复发。这项提议的目标是将这些 生物学见解和临床前治疗数据，以产生介入方法，所需的实验室 检测和演示靶向DTCs的可行性、安全性和临床疗效，这将是 大规模、确凿的临床试验和监测研究。 本申请的具体目的是：1)执行伊波利莫斯的概念验证临床试验(Eve， 靶向mTOR)和羟基氯喹(HCQ，靶向自噬)在术后可检测到DTC的女性中的应用 初级治疗；以及2)利用临床前小鼠模型同时优化治疗方法 以及根除DTC的先进发现。AIM 1中的随机开放标签试点试验将 探讨HCQ、EVE或联合应用的可行性和安全性，以及它们对DTC负荷的影响。我们 还将改进和验证一种新的流式细胞仪分析，以提高检测、计数的灵敏度 以及DTC生物标记物的分子表征。在目标2中，在小鼠身上进行的联合临床试验将优化 HCQ、EVE及其组合的影响，调查这些影响的关键参数，以告知 临床试验，并扩展这些模型以更接近地反映临床治疗。此外，分子 残留肿瘤细胞的表型鉴定将为未来的试验发现更多的靶点。归根结底，有能力 识别、列举和治疗靶向DTCs具有改变监测和治疗的潜力 为乳腺癌幸存者提供选择，并防止妇女死于这一致命疾病。

Project Summary Despite early detection and adjuvant therapy, breast cancer remains the leading cause of cancer mortality in women, largely due to distant, incurable recurrences arising years, or even decades, after treatment of the primary tumor. These recurrent, metastatic tumors arise from the pool of residual local and disseminated tumor cells (DTCs) that survive primary treatment and remain in the host in a presumed dormant state. Indeed, the presence of bone marrow DTCs following treatment is independently associated with a substantially increased risk of recurrence. At present, however, the mechanisms enabling residual tumor cells to maintain dormancy and ultimately recur are poorly understood, and DTC-directed surveillance and treatment approaches are non-existent. Consequently, the ability to biologically characterize, accurately measure and therapeutically target dormant DTCs would be a transformational new approach to preventing recurrence. We hypothesize that disabling the survival mechanisms employed by dormant DTCs will reduce tumor recurrence and thereby improve survival. Using genetically engineered mouse models that faithfully recapitulate tumor dormancy and recurrence, we have discovered that autophagy and mTOR signaling are each critical to the survival of DTCs, and that agents inhibiting these pathways deplete the reservoir of dormant residual tumor cells, thereby preventing tumor recurrence. The objective of this proposal is to translate these biological insights and preclinical therapeutic data to generate the interventional approach, requisite laboratory assays, and demonstration of feasibility, safety and clinical efficacy of targeting DTCs that will be required for large-scale, definitive clinical trials and surveillance studies. The specific aims of this application are to: 1) Perform a proof-of-concept clinical trial of everolimus (EVE, targeting mTOR), and hydroxychloroquine (HCQ, targeting autophagy) in women with detectable DTCs after primary treatment; and 2) Employ preclinical mouse models to concurrently optimize therapeutic approaches and advance discoveries for the eradication of DTCs. The randomized, open-label pilot trial in Aim 1 will investigate the feasibility and safety of HCQ, EVE or the combination, and their effects on DTC burden. We will also refine and validate a novel flow cytometric assay to improve the sensitivity of detection, enumeration and molecular characterization of the DTC biomarker. In Aim 2, a co-clinical trial in mice will optimize the effects of HCQ, EVE, and their combination, investigate critical parameters of these effects necessary to inform clinical trials, and extend these models to more closely reflect clinical treatment. In addition, molecular phenotyping of residual tumor cells will uncover additional targets for future trials. Ultimately, the ability to identify, enumerate and therapeutically target DTCs has the potential to transform surveillance and treatment options for breast cancer survivors and prevent women from succumbing to this deadly disease.