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

项目总结/摘要 高级别浆液性卵巢癌（HGSOC），最致命的妇科恶性肿瘤，通常是 在远处转移发生后才被诊断，并且化学抗性使得目前的治疗短暂。 该领域存在两个主要的知识差距：我们缺乏对早期病变和 新的抗癌药物。HGSOC一直难以研究和建模，因为对最常见的 细胞的起源，这是现在接受的输卵管上皮（FTE），因为月经 循环在HGSOC发展中起作用。以前没有这种疾病的模型， 输卵管和卵巢都能重现人类的月经周期。FemKube，第一个 女性生殖道芯片，是通过多机构合作， 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， 小鼠卵巢，这是工程驱动生理上准确的28天人类月经周期，在 微流体流动的设置。我们将利用这一创新技术解决这两个问题， 利用它来研究输卵管中的早期致癌事件， 开发一类有前途的新的天然产物化学治疗剂，叶下珠毒素（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。 组织的长度准确再现月经周期。假设FTE受损 通过卵泡期（月经周期的前半期， 包括卵泡成熟），其在从体分泌的孕激素的影响下恢复 黄体（排卵后卵泡的剩余部分）在黄体后期（月经周期的后半部分）。 我们的第一个目的是通过绘制DNA图谱来研究周期性卵巢如何影响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儿 致癌介质，如已知的DNA突变体、炎症和生长因子、活性氧物质 中和剂和肿瘤抑制剂，以机械地研究FemKube系统中的HGSOC起始。 我们的第二个目标是将我们培养原代人体组织的能力纳入FemKube 系统进入临床前药物开发管道。在我们俄亥俄州的合作者的帮助下 大学，我们已经开发出一类有前途的化合物，来自天然产物，PHY， 在体外对HGSOC细胞系的纳摩尔效力。我们将证实PHY的凋亡和抗癌能力， 体外我们将在FemKube系统中证明它们对肿瘤的体外疗效，并对我们的发现进行基准测试。 对照小鼠体内化疗测定的金标准。总的来说，FemKube的介绍 这项技术将回答以前无法回答的关于HGSOC启动的问题， 开发管道，从而解决目前卵巢癌研究的关键差距。

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.