Imaging mass spectrometry methodologies for studying the metabolites of cancer metastasis

高级别浆液性卵巢癌（HGSC）是最常见和最致命的卵巢癌形式。 新出现的证据表明，这些肿瘤发生在输卵管上皮（FTE），因此，它们的 卵巢中的存在代表原发转移。我们的初步数据表明， 靠近卵巢有助于疾病的侵袭性。卵巢定植后，肿瘤 细胞侵入腹膜器官，主要是网膜。我们假设， 原发性和继发性HGSC转移部分是由HGSC之间的化学通讯介导的。 癌细胞和转移器官我们的建议旨在定义代谢物和生物分子， 促使输卵管来源的高级别浆液性癌转移到卵巢和网膜。本 最后，我们的团队优化了卵巢和输卵管来源的肿瘤模型的3D共培养， 这就需要成像质谱技术来识别代谢组学驱动的通信， 在卵巢的原发性定殖期间和在向网膜的继发性转移期间。利用这种新兴的 技术，我们确定了几种代谢物，增强高级别浆液性肿瘤的迁移，侵袭， 粘附在卵巢上。目标1的重点是揭示允许FTE致瘤细胞 劫持卵巢产生的NE，以增加它们在原发性卵巢中侵入和粘附卵巢的能力。 转移目的1将明确NE介导的细胞侵袭和粘附过程中的信号通路， 卵巢，然后证实NE在体内的重要性，使用鼠和人细胞模型来源于 FTE。将使用CRISPR删除关键的肾上腺素能受体，以确认该途径在 转移将用β肾上腺素能受体拮抗剂治疗荷瘤模型，以尝试 将这些发现转化为阻断卵巢定植的新策略。目标2的重点是 一种新的输卵管致瘤分泌蛋白的鉴定和特性分析 卵巢癌是卵巢癌的一种重要类型，它是卵巢癌细胞的重要组成部分，并负责卵巢去甲肾上腺素的产生，从而驱动肿瘤细胞的侵袭和粘附。 我们将使用蛋白质组学来确认分泌蛋白的身份，然后通过基因缺失来确定分泌蛋白的基因。 蛋白质，以研究在卵巢定植中的作用。Aim 3将建立在我们现有的技术基础上 3D器官和肿瘤细胞通信模型，并扩展到继发性转移。我们现在已经 优化了我们的技术，用于网膜与肿瘤细胞模型的共培养， 代谢物，这是独特的，不包括去甲肾上腺素。相反，发现了一种新的代谢物， 当肿瘤细胞与网膜一起生长时， 叶酸，叶酸受体的配体，在肿瘤细胞中过度表达。总的来说，我们的 创新性实验方法将产生新的途径和靶点，以减轻原发性高转移癌 卵巢恶性浆液性癌

High grade serous ovarian cancer (HGSC) is the most common and deadliest form of ovarian cancer. Emerging evidence indicates that these tumors arise in the fallopian tube epithelium (FTE), and thus their presence in the ovary represents the primary metastasis. Our preliminary data identified that xenografting in close proximity to the ovary contributes to the aggressiveness of the disease. After ovarian colonization, tumor cells invade the peritoneal organs, primarily the omentum. We hypothesize that the biological problem of primary and secondary HGSC metastasis is partially mediated by chemical communication between the cancer cells and the metastatic organ. Our proposal seeks to define metabolites and biomolecules that drive the metastasis of fallopian tube derived high grade serous cancers to the ovary and the omentum. To this end, our teams optimized a 3D co-culture of the ovary and fallopian tube derived tumor models and adapted this to imaging mass spectrometry technology to identify the metabolomics-driven communication that occurs during primary colonization of the ovary and during secondary metastasis to the omentum. Using this emerging technology, we identified several metabolites that enhanced high grade serous tumor migration, invasion, and adhesion to the ovary. The focus of Aim 1 is to uncover the mechanisms allowing FTE tumorigenic cells to hijack NE produced by the ovary to increase their ability to invade and adhere to the ovary during primary metastasis. Aim 1 will define the signaling pathways mediated by NE during invasion and adhesion to the ovary and then confirm the importance of NE in vivo using both murine and human cell models derived from FTE. The key adrenergic receptor will be deleted using CRISPR to confirm the importance of this pathway in metastasis. Tumor bearing models will be treated with beta adrenergic receptor antagonists in an attempt to translate these findings for a new strategy to block ovarian colonization. The focus of Aim 2 is on the identification and characterization of a newly identified protein that is secreted from tumorigenic fallopian tube cells and is responsible for the production of ovarian norepinephrine driving tumor cell invasion and adhesion. We will use proteomics to confirm the identity of the secreted protein, followed by genetic deletion of the protein from FTE models to study the role in ovarian colonization. Aim 3 will build upon our existing technology of 3D organ and tumor cell communication models and expand into secondary metastasis. We have now optimized our technology for co-culture of the omentum together with tumor cell models and have an inventory of metabolites, which are unique and did not include norepinephrine. Instead a novel metabolite found to be produced in significantly more abundance when tumor cells were grown with the omentum corresponded to folate, the ligand for the folic acid receptor that is overexpressed in the tumor cells. Taken together, our innovative experimental approach will yield new pathways and targets to mitigate primary metastasis of high grade serous cancer to the ovary.