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。高级浆液卵巢癌（HGSC）是最常见，最致命的形式 卵巢癌。新兴的证据表明，这些肿瘤出现在输卵管上皮（FTE）中， 因此，它们在卵巢中的存在代表了主要转移。我们的初步数据确定 与卵巢紧密的Xenografting有助于该疾病的侵略性。卵巢之后 定殖，肿瘤细胞侵入腹膜器官，主要是脑器官。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介导 癌细胞与转移器官之间的通信。我们的建议旨在定义代谢物 和驱动输卵管转移的生物分子，衍生出卵巢的高级浆液癌 和omentum。为此，我们的团队优化了卵巢和输卵管的3D共同培养 肿瘤模型并将其改编为成像质谱技术，以识别代谢组驱动的 卵巢初级定植和继发转移期间发生的交流 omentum。使用这种新兴技术，我们确定了几种增强高级浆液的代谢产物 肿瘤迁移，侵袭和对卵巢的粘附。目标1的重点是揭示机制 允许卵巢产生的FTE肿瘤细胞劫持其侵入能力和 在原发转移期间粘附于卵巢。 AIM 1将定义NE介导的信号通路 对卵巢的侵袭和粘附，然后确认使用鼠和使用鼠类体内的NE的重要性 源自FTE的人类细胞模型。关键的肾上腺素能受体将使用CRISPR删除以确认 这种途径在转移中的重要性。肿瘤轴承模型将用β肾上腺素能受体处理 反对者试图将这些发现转化为一种新的策略来阻止卵巢殖民化。重点 AIM 2的鉴定和表征是新近鉴定的蛋白质，该蛋白质分泌 肿瘤性输卵管细胞，负责产生卵巢去甲肾上腺素驱动肿瘤 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 FTE模型中蛋白质的遗传缺失研究了卵巢定植中的作用。 AIM 3将基于 我们现有的3D器官和肿瘤细胞通信模型的技术，并扩展为次要 转移。现在，我们已经优化了我们的技术与肿瘤细胞共同培养的技术 模型并具有代谢物的清单，这些代谢物是独一无二的，不包括去甲肾上腺素。而是 当肿瘤细胞与 omentum对应于叶酸，叶酸是叶酸受体过表达的叶酸受体的配体。 综上所述，我们创新的实验方法将产生新的途径和目标，以减轻初级 高级浆液性癌症转移到卵巢。