Development of quantitative mass spectrometry assays and imaging for cancer metastasis

开发癌症转移的定量质谱分析和成像

• 批准号: 10533035
• 负责人: Joanna E Burdette
• 金额: $ 8.58万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10533035
• 关键词: 3-Dimensional; Adhesions; Adrenergic Antagonists; Adrenergic Receptor; Automobile Driving; Award; Biological; Biological Assay; Cell Communication; Cell model; Cells; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Communication; Data; Development; Disease; Emerging Technologies; Epithelial; Equipment and supply inventories; Exhibits; Folic Acid; Genetic; Human; Invaded; Ligands; Malignant Neoplasms; Malignant neoplasm of ovary; Mammalian Oviducts; Mass Spectrum Analysis; Mediating; Modeling; Mus; Neoplasm Metastasis; Norepinephrine; Omentum; Organ; Organ Culture Techniques; Ovarian; Ovary; Parents; Pathway interactions; Peritoneal; Production; Proteins; Proteomics; Role; Serous; Signal Pathway; Signal Transduction; Site; System; Technology; Translating; Tumor Cell Invasion; Tumor-Derived; Xenograft procedure; beta-adrenergic receptor; cancer cell; cancer imaging; folate-binding protein; in vivo; innovation; mass spectrometric imaging; metabolomics; migration; neoplastic cell; novel; overexpression; tool; tumor; tumorigenic

Parent award abstract. 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.

家长奖摘要。高级别浆液性卵巢癌（HGSC）是卵巢癌中最常见和最致命的形式。 卵巢癌新出现的证据表明，这些肿瘤发生在输卵管上皮（FTE）， 因此它们在卵巢中的存在代表原发性转移。我们的初步数据表明， 在卵巢附近的异种移植有助于疾病的侵袭性。排卵后 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转移的生物学问题部分由化学物质介导， 癌细胞和转移器官之间的联系。我们的建议旨在定义代谢物 以及促使输卵管来源的高级别浆液性癌转移到卵巢的生物分子 和网膜为此，我们的团队优化了卵巢和输卵管的3D共培养， 肿瘤模型，并将其应用于成像质谱技术，以识别代谢组学驱动的 在卵巢的原发性定殖期间和在向卵巢的继发性转移期间发生的通信。 网膜利用这项新兴技术，我们鉴定了几种代谢物， 肿瘤迁移、侵袭和粘附到卵巢。目标1的重点是揭示机制 允许FTE致瘤细胞劫持卵巢产生的NE以增加它们的侵袭能力， 在原发转移时粘附于卵巢。目的1将定义NE介导的信号通路， 侵袭和粘附卵巢，然后使用小鼠和 来自FTE的人细胞模型。将使用CRISPR删除关键的肾上腺素能受体，以确认 这一途径在转移中的重要性。荷瘤模型将接受β肾上腺素能受体治疗 拮抗剂，试图将这些发现转化为阻断卵巢定植的新策略。重点 目的2是鉴定和表征一种新鉴定的蛋白质， 致瘤性输卵管细胞，并负责卵巢去甲肾上腺素驱动肿瘤的产生 细胞侵袭和粘附。我们将使用蛋白质组学来确认分泌蛋白的身份，然后 从FTE模型中遗传缺失蛋白质以研究在卵巢定殖中的作用。目标3将建立在 我们现有的3D器官和肿瘤细胞通信模型技术，并扩展到二级 转移我们现在已经优化了网膜与肿瘤细胞共培养的技术 模型，并有代谢物的库存，这是唯一的，不包括去甲肾上腺素。而是 发现当肿瘤细胞在含有抗肿瘤药物的培养基中生长时， 网膜对应于叶酸，叶酸受体的配体在肿瘤细胞中过表达。 综上所述，我们的创新实验方法将产生新的途径和目标，以减轻主要的 高级别浆液性癌转移到卵巢。