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: 10393491
• 负责人: Joanna E Burdette
• 金额: $ 36.05万
• 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/10393491
• 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; Diagnosis; Disease; Emerging Technologies; Epithelial; Epithelial Cells; 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; Norepinephrine; Omentum; Organ; Organ Culture Techniques; Ovarian; Ovary; Pathway interactions; Patient-Focused Outcomes; Peritoneal; Peritoneum; Process; Production; 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; Tumor-Derived; Work; Xenograft procedure; base; 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。 新出现的证据表明，这些肿瘤发生在输卵管上皮(FTE)，因此他们的 卵巢中的存在代表了原发转移。我们的初步数据表明异种移植在 靠近卵巢有助于该病的侵袭性。卵巢定植后，肿瘤 细胞侵入腹膜器官，主要是大网膜。我们假设生物问题 原发和继发HGSC转移部分由HGSC与HGSC之间的化学通讯所介导 癌细胞和转移器官。我们的提案试图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大网膜。对这件事 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，并对 这要用成像质谱学技术来识别发生的代谢组学驱动的沟通 在卵巢的原发定植期间和在大网膜的继发转移期间。利用这一新兴技术 技术，我们确定了几种代谢产物，它们促进了高级别浆液性肿瘤的迁移、侵袭和 粘连在卵巢上。目标1的重点是揭示允许FTE致瘤细胞 劫持卵巢产生的去甲肾上腺素，以增强它们在初选期间侵袭和附着卵巢的能力 转移。目标1将确定NE在侵袭和黏附过程中介导的信号通路。 然后用小鼠和人的细胞模型证实去甲肾上腺素在体内的重要性 全时当量。关键的肾上腺素能受体将利用CRISPR被删除，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移居的新策略。《目标2》的重点是 一种新发现的致瘤输卵管分泌蛋白的鉴定与鉴定 并负责卵巢去甲肾上腺素的产生，推动肿瘤细胞的侵袭和黏附。 我们将使用蛋白质组学来确认分泌蛋白的身份，然后基因缺失 从FTE模型中提取蛋白质，研究其在卵巢定植中的作用。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，并扩展为继发性转移。我们现在有了 优化了我们与肿瘤细胞模型共培养大网膜的技术，并有一个清单 代谢物，这是独一无二的，不包括去甲肾上腺素。相反，一种新的代谢物被发现是 当肿瘤细胞生长时，与相应的大网膜相对应的产生显著更多的 叶酸，叶酸受体的配体，在肿瘤细胞中过度表达。总而言之，我们的 创新的实验方法将产生新的途径和靶点来减轻高血压病患者的原发转移 卵巢浆液性癌级别。