Imaging mass spectrometry methodologies for studying the metabolites of cancer metastasis

研究癌症转移代谢物的成像质谱方法

• 批准号: 10622483
• 负责人: Joanna E Burdette
• 金额: $ 38.34万
• 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/10622483
• 关键词: 3-Dimensional; Adhesions; Adrenergic Antagonists; Adrenergic Receptor; Adrenergic beta-Antagonists; Anatomy; Automobile Driving; Biological; Biological Assay; Biological Models; Biology; Cell Communication; Cell model; Cells; Cessation of life; Chemicals; Chemistry; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Communication; Consumption; Data; Detection; Development; Disease; Emerging Technologies; Epithelial Cells; Epithelium; Equipment and supply inventories; Event; Exhibits; Folic Acid; Freezing; Genetic; Glycogen; Human; Invaded; Ions; Knock-out; Ligands; Lipids; Location; Malignant Neoplasms; Malignant neoplasm of ovary; Mammalian Oviducts; Measures; Mediating; Metastatic Malignant Neoplasm to the Ovary; Methodological Studies; Modeling; Mus; Mutation; Names; Neoplasm Metastasis; Newly Diagnosed; Norepinephrine; Omentum; Organ; Organ Culture Techniques; Ovarian; Ovary; Pathway interactions; Patient-Focused Outcomes; Peritoneal; Peritoneum; Process; Production; Protein Secretion; Proteins; Proteomics; Protocols documentation; Role; Route; Sampling; Series; Serous; Signal Pathway; Signal Transduction; Site; Source; Spatial Distribution; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; System; Techniques; Technology; Testing; Time; Tissues; Translating; Tumor Cell Invasion; Tumor Cell Migration; Tumor Expansion; Work; Xenograft procedure; beta-adrenergic receptor; cancer cell; folate-binding protein; in vivo; innovation; mass spectrometric imaging; metabolome; metabolomics; migration; mortality; neoplastic cell; novel; overexpression; small molecule; therapeutic target; three dimensional cell culture; tool; tumor; tumorigenic

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.

高级别浆液性卵巢癌（HGSC）是最常见和最致命的卵巢癌。 新的证据表明这些肿瘤出现在输卵管上皮 (FTE) 中，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，异种移植 靠近卵巢会导致疾病的侵袭性。卵巢定植后，肿瘤 细胞侵入腹膜器官，主要是大网膜。我们假设生物学问题 原发性和继发性 HGSC 转移部分是由化学通讯介导的 癌细胞和转移器官。我们的提案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至卵巢和大网膜。对此 最后，我们的团队优化了卵巢和输卵管衍生肿瘤模型的 3D 共培养，并进行了调整 这通过成像质谱技术来识别发生的代谢组学驱动的通信 在卵巢的原发定植期间和在大网膜的继发转移期间。利用这一新兴的 技术，我们鉴定了几种增强高级别浆液性肿瘤迁移、侵袭和侵袭的代谢物 与卵巢粘连。目标 1 的重点是揭示 FTE 致瘤细胞的机制 劫持卵巢产生的 NE，以增加其在初级阶段侵入和粘附卵巢的能力 转移。目标 1 将定义 NE 在侵袭和粘附过程中介导的信号通路 卵巢，然后使用来源于小鼠和人类的细胞模型确认 NE 在体内的重要性 全职员工。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。荷瘤模型将用 β 肾上腺素能受体拮抗剂治疗，试图 将这些发现转化为阻止卵巢定植的新策略。目标 2 的重点是 致瘤性输卵管分泌的新鉴定蛋白质的鉴定和表征 细胞并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，然后对 来自 FTE 模型的蛋白质用于研究卵巢定植中的作用。目标 3 将建立在我们现有技术的基础上 3D器官和肿瘤细胞通讯模型并扩展到继发性转移。我们现在有 优化了我们的网膜与肿瘤细胞模型共培养技术并拥有库存 代谢物，其独特且不包括去甲肾上腺素。相反，发现了一种新的代谢物 当肿瘤细胞与对应的网膜一起生长时，产生的丰度显着增加 叶酸，叶酸受体的配体，在肿瘤细胞中过度表达。综合起来，我们的 创新的实验方法将产生新的途径和目标，以减轻高危原发性转移 卵巢浆液性癌。