FemKube, the human female reproductive tract-on-a-chip, as a platform for studying high grade serous ovarian cancer and developing novel cancer chemotherapeutics

项目摘要/摘要 高级别浆液性卵巢癌(HGSOC)是最致命的妇科恶性肿瘤，通常 在发生远处转移后诊断，化疗耐药性使目前的治疗短暂。 该领域存在两个主要的知识空白：我们缺乏对早期病变和发展的了解 新的抗癌药物。HGSOC很难研究和建模，因为对最常见的 细胞起源，现在被认为是输卵管上皮(FTE)，因为月经 周期在HGSOC的发展中起着重要作用。以前没有这种疾病的模型存在，其中包括 输卵管和卵巢都能重现人类的月经周期。FemKube，第一个 芯片上的女性生殖道，是通过多机构合作创造的 伊利诺伊大学西北分校和德雷珀实验室支持原代人类输卵管组织和 小鼠卵巢被设计成驱动生理上准确的28天人类月经周期，在 微流控流量的设置。我们将利用这项创新技术通过以下方式解决这两个领域的问题 利用它研究输卵管早期致癌事件并增强临床前 开发一种有前景的新型天然产物化疗药物--叶下珠(Phys)。 我们的合作团队已经证明了FemKube支持人类输卵管生长的能力 精确复制月经周期长度的组织。假设FTE已损坏 通过卵泡期(月经周期的前半部分)卵巢产生的分泌因子 包括卵泡成熟)，这是在卵泡分泌的孕激素影响下恢复的 黄体晚期(月经周期的后半段)的黄体(排卵后剩余的卵泡)。 我们的第一个目标是通过绘制DNA图谱来研究周期卵巢如何影响FTE中HGSOC的启动 损伤、增殖和凋亡。我们将使用月经周期激素和新生激素的抑制剂 致癌介质，如已知的DNA突变物、炎症和生长因子、活性氧物种 中和剂和肿瘤抑制剂，从力学上研究FemKube系统中HGSOC的启动。 我们的第二个目标是将我们在FemKube培养原代人体组织的能力结合起来 系统进入临床前药物开发流水线。在俄亥俄州立大学我们的合作者的帮助下 大学，我们已经开发出一类很有前途的化合物，来自天然产品，phys，与 体外培养HGSOC细胞株的纳摩尔效力。我们将确认PHY的凋亡和抗癌能力 体外培养。我们将在FemKube系统中展示它们对肿瘤的体外疗效，并对我们的发现进行基准测试 对照金标准在小鼠体内的化疗试验。总体而言，FemKube的引入 技术将回答以前无法回答的关于HGSOC启动的问题，并增强药物 开发流水线，从而填补当前卵巢癌研究中的关键空白。

PROJECT SUMMARY/ABSTRACT High grade serous ovarian cancer (HGSOC), the most lethal gynecologic malignancy, is typically diagnosed after distant metastasis has occurred, and chemoresistance renders current treatments short-lived. Two major knowledge gaps exist in the field: we lack an understanding of early lesions and development of new anticancer drugs. HGSOC has been difficult to research and model due to debate over the most common cell of origin, which is now accepted to be the fallopian tube epithelium (FTE), and because the menstrual cycle plays a role in HGSOC development. No models of the disease previously existed, which incorporate both the fallopian tube and an ovary capable of recapitulating the human menstrual cycle. FemKube, the first female reproductive tract-on-a-chip, was created through a multi-institutional collaboration between the University of Illinois, Northwestern, and Draper Labs to support primary human fallopian tube tissues and murine ovaries, which are engineered to drive a physiologically accurate 28-day human menstrual cycle, in the setting of microfluidic flow. We will leverage this innovative technology to address both issues in the field by utilizing it to investigate early oncogenic events in the fallopian tube and to enhance the preclinical development of a promising new class of natural product chemotherapeutics, Phyllanthusmins (PHYs). Our collaborative team has demonstrated the ability of FemKube to support growth of human fallopian tissues for the length of an accurately reproduced menstrual cycle. It is hypothesized that the FTE is damaged by secreted factors produced by the ovary during the follicular phase (first half of the menstrual cycle that encompasses follicle maturation), which is restored under the influence of progestins secreted from the corpus luteum (what remains of the follicle after ovulation) in the late luteal phase (second half of the menstrual cycle). Our first aim is to investigate how the cycling ovary impacts HGSOC initiation in the FTE by mapping DNA damage, proliferation, and apoptosis. We will use inhibitors of menstrual cycle hormones and nascent oncogenic mediators, such as known DNA mutators, inflammatory and growth factors, reactive oxygen species neutralizers and tumor suppressors, to mechanistically study HGSOC initiation in the FemKube system. Our second aim seeks to incorporate our ability to culture primary human tissues in the FemKube system into the preclinical drug development pipeline. With the help our collaborators at the Ohio State University, we have developed a promising class of compounds derived from natural products, PHYs, with nanomolar potency on HGSOC cell lines in vitro. We will confirm PHY's apoptotic and anticancer abilities in vitro. We will demonstrate their efficacy on tumors ex vivo in the FemKube system and benchmark our findings against gold standard in vivo chemotherapeutic assays in mice. Overall, the introduction of FemKube technology will answer previously inaccessible questions on HGSOC initiation and enhance the drug development pipeline, thereby addressing key gaps in current ovarian cancer research.