All-Human Microphysical Model of Metastasis Therapy

描述(申请人提供)：转移治疗的全人类微观物理模型成功根除转移疾病仍然是降低实体肿瘤死亡率的重大挑战。虽然消融治疗方法很少，但全身化疗通常是抑制进展和延长生存时间的唯一可行的选择，尽管它很少治愈。我们对化疗药物为什么无法消除转移的理解，以及我们创造更有效治疗策略的能力，都受到以下两方面的限制：我们在分子和细胞水平上解剖肿瘤-宿主相互作用方面的缺陷，以及缺乏相关的模型系统来筛选新的治疗方法；以及缺乏以相关方式做到这一点的所有人类系统。有证据表明，肿瘤细胞受到转移微环境的影响，对化疗产生更强的抗药性，面对转移的疾病，化疗新陈代谢发生了变化。我们提出了一种系统，它不仅将提供一个完全与人类相关的转移微环境，而且还将与药物代谢和肝脏的正常生理功能密切相关，这可能会阻碍或增强治疗的有效性或毒性。与任何其他常见的转移部位相比，肝脏在一天中经历了代谢和荷尔蒙状态的剧烈波动。这些波动对转移性肿瘤的恶性行为和化疗反应的影响程度尚不清楚。在这个项目中，我们在体外捕捉到了这种情况的复杂性，采用了一种适合于药物开发流水线的格式，使用了多孔板格式的3D微灌流器官型肝脏。我们的方法旨在促进功能性宿主肝组织中相对较大的、临床相关的转移性结节(&gt；0.5毫米)的发展。我们将(I)确定与标准培养相比，宿主肝组织内递送到肿瘤细胞的激素、细胞因子和营养物质的周期性/日间变化是否改变了肿瘤细胞的表型行为，如增殖、侵袭特性和特定肿瘤标记物的表达；(Ii)利用肝转移微环境中的一组人类肿瘤细胞，以及普通化疗药物(代谢和非代谢药物)和靶向生物治疗药物(激酶抑制物类)，确定化疗药物对转移肿瘤的疗效是否受日常代谢和激素控制的影响，如果是，这些是否与可以现场测量的肿瘤的性质有关(Iii)评估肝脏的化疗毒性是否因转移灶的侵袭或肝脏流出的周期性/昼夜变化而改变(Iv)检验肝脏轻度炎症状态是否会刺激肿瘤生长和改变化疗药物的疗效。 与公共卫生相关：癌症的发病率和死亡率绝大多数是由于肿瘤扩散到最初的发育点以外造成的，这不仅是为了避免直接的手术和放射切除，而且似乎比原发肿瘤对系统治疗更具抵抗性。由于我们对肿瘤生物学和化疗如何在转移的利基中代谢，特别是对激素、营养物质和炎症信号的循环和昼夜节律做出反应的知识匮乏，这些播散性癌细胞显然需要开发新的方法。目前的体外模型未能捕捉到微环境的复杂性，整个动物模型不能实时和连续地监测关键建立的第一个月期间的事件和细胞行为。生物反应器为这一时期提供了一个独特的窗口；我们建议开发下一代全人类肝脏生物反应器，它提供对人类情况的生理模拟，以帮助药物开发和转移癌症的治疗方法。

DESCRIPTION (provided by applicant): All-Human Microphysical Model of Metastasis Therapy Successful eradication of metastatic disease remains the grand challenge in reducing mortality from solid tumors. Although ablative approaches are infrequently possible, systemic chemotherapy usually is the only feasible option for inhibiting progression and increasing survival time, though it is rarely curative. Our understanding of why chemotherapeutic agents fail to eliminate metastases, and our ability to create more effective therapeutic strategies, is limited by both our deficit in dissecting the tumor-host interactions at a molecular and cellular level, and a lack of relevant model systems to screen novel therapies; and a dearth of all human systems to do this in a relevant manner. There is evidence that the tumor cells are affected by the metastatic micro-environment to become more resistant to chemotherapy and that chemotherapeutic metabolism is altered in the face of metastatic disease. We propose a system that will not only provide an all human contextual metastatic micro-environment, but one that is intimately linked to drug metabolism and to normal physiological functions of liver that may hinder or augment the efficacy or toxicities of therapies. More than any other common site of metastasis, the liver experiences dramatic swings in metabolic and hormonal state throughout the day. The extent to which these fluctuations influence malignant behaviors and chemotherapy responses in metastatic tumors is unknown. In this project we capture the complexity of this situation in vitro in a format amenable to incorporation in the drug development pipeline, using a 3D micro- perfused organotypic liver in a multiwell plate format. Our approach is designed to foster development of relatively large, clinically relevant metastatic nodules (>0.5 mm) in functional host liver tissue. We will (i) determine whether cyclic/diurnal changes in hormones, cytokines and nutrients delivered to tumor cells within host liver tissue alters the phenotypic behavior of the tumor cells compared to standard culture, such as proliferation, invasive properties, and expression of specific tumor markers; (ii) Determine whether the efficacy of chemotherapy agents against metastatic tumors is influenced by diurnal control of metabolism and hormones, using a panel of human tumor cells within the liver metastatic microenvironment and both general chemotherapeutics (metabolized and non-metabolized agents) and a targeted bio-therapeutics (in the kinase inhibitor class) and if so, if these are related to properties of the tumor that can be measured in situ (iii) Assess whether the chemotherapeutic toxicities on the liver are altered by metastatic involvement or by cyclical/diurnal variations in the liver affluent (iv) Test the hypothesis that mild inflammatory states of liver stimulate tumor growth and alter efficacy of chemotherapeutics. PUBLIC HEALTH RELEVANCE: Carcinoma morbidity and mortality result overwhelmingly from the dissemination of the tumor beyond the initial site of development, to not only avoid directed surgical and radiological removal but also appear as more resistant to systemic therapies than the primary tumor. Development of new approaches that are obviously needed for these disseminated carcinoma cells is stymied by our dearth of knowledge concerning the tumor biology and how chemotherapy is metabolized in the metastatic niche, particularly in response to cyclic and diurnal rhythms of hormones, nutrients and inflammatory signals. Current in vitro models fail to capture the complexity of the microenvironment, and whole animal models do not allow for real time and continuous monitoring of the events and cell behaviors during the critical first month of establishment. Bioreactors offer a unique window into this period; we propose to develop a next generation all human liver bioreactor that provides physiologic mimicry of the human situation to aid in drug development and therapeutic approaches to metastasized cancers.