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.

高级浆液卵巢癌（HGSC）是卵巢癌最常见，最致命的形式。 新兴的证据表明，这些肿瘤在输卵管上皮（FTE）中出现，因此 卵巢中的存在代表主要转移。我们的初步数据确定了Xenografting 与卵巢的近距离接近疾病的侵略性。卵巢定植后肿瘤后 细胞侵入腹膜器官，主要是大脑器官。我们假设 原发性和继发性HGSC转移部分是通过化学通信介导的 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 驱动输卵管的转移，衍生出高级浆液癌和卵巢。对此 结束时，我们的团队优化了卵巢和输卵管衍生的肿瘤模型的3D共同培养 这是对成像质谱技术的成像，以识别发生代谢组学驱动的通信 在卵巢的一级定殖和次生转移期间。使用此新兴 技术，我们确定了几种代谢产物，可以增强高级浆液肿瘤迁移，侵袭和 对卵巢的粘附。目标1的重点是发现允许FTE肿瘤细胞的机制 卵巢产生的劫持NE增加了他们在初级期间入侵和粘附的能力 转移。 AIM 1将定义NE在入侵期间介导的信号传导途径，并粘附到 卵巢，然后使用源自鼠和人类细胞模型来确认体内NE的重要性 fte。关键的肾上腺素能受体将使用CRISPR删除，以确认该途径在 转移。肿瘤轴承模型将用β肾上腺素能受体拮抗剂处理，以尝试 将这些发现转化为一种新的策略来阻止卵巢殖民化。 AIM 2的重点是 鉴定和表征新鉴定的蛋白质，该蛋白质是从肿瘤的输卵管分泌 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确认分泌蛋白的身份，然后是遗传缺失 来自FTE模型的蛋白质来研究卵巢定殖的作用。 AIM 3将建立在我们现有技术的基础上 3D器官和肿瘤细胞通信模型，并扩展为继发转移。我们现在有 优化了我们用于肿瘤细胞模型共同培养的技术，并具有库存 代谢产物，它们是独一无二的，不包括去甲肾上腺素。相反，一种新型的代谢物是 当肿瘤细胞与大脑对应于 叶酸，在肿瘤细胞中过表达的叶酸受体的配体。总的来说，我们的 创新的实验方法将产生新的途径和靶标，以减轻高的原发转移 卵巢等级浆液癌。