Profiling the Fluid Assisted Dissemination of Pre-malignant cells in Fallopian Tubes

分析输卵管癌前细胞的液体辅助传播

• 批准号: 10718158
• 负责人: Geeta Mehta
• 金额: $ 61.89万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-07 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10718158
• 关键词: 3-Dimensional; Abdominal Cavity; Acceleration; Affect; Anatomy; Animal Model; BRCA1 gene; Bioinformatics; Biological Assay; Biology; Biophysical Process; Bioreactors; CDK4 gene; Carcinoma; Cell Culture Techniques; Cell Death; Cell Line; Cells; Collagen; Complement; Custom; Cytoskeleton; DNA Sequence Alteration; Data; Detection; Development; Disease; Disease Progression; Early Diagnosis; Environment; Epithelial Cells; Epithelium; Exposure to; F-Actin; Genes; Genetic; Genetic Variation; Goals; Greater sac of peritoneum; Growth; Harvest; Human; Immune; Immune system; Immunocompetent; Immunocompromised Host; Immunodeficient Mouse; Immunologic Deficiency Syndromes; Immunotherapy; In Vitro; Intraperitoneal Injections; Invaded; Lesion; Link; Liquid substance; Location; Malignant neoplasm of ovary; Mammalian Oviducts; Mechanics; Microfluidic Microchips; Microfluidics; Modeling; Molecular; Molecular Profiling; Movement; Mus; Mutate; Mutation; Neoplasm Metastasis; Omentum; Organ; Ovarian; Ovary; Pathway interactions; Patient-Focused Outcomes; Patients; Perfusion; Peritoneal; Peritoneum; Pharmaceutical Preparations; Play; Polysaccharides; Premalignant Cell; Prevention; Proliferating; Proteomics; Publishing; RB1 gene; Randomized; Recurrent disease; Regulation; Research; Role; Scaffolding Protein; Screening for Ovarian Cancer; Secretory Cell; Sepharose; Serous; Signal Pathway; Signal Transduction; Site; Stage at Diagnosis; Structure; Surface; Survival Rate; TP53 gene; Testing; Tube; Tumor Burden; Validation; Woman; Work; cancer cell; cancer diagnosis; cancer initiation; cancer therapy; cell growth; clinical diagnosis; clinical translation; design and construction; differential expression; driver mutation; early detection biomarkers; effective therapy; fluid flow; gain of function; genetic signature; implantation; improved; in vivo; in vivo Model; intraperitoneal; loss of function; malignant breast neoplasm; mechanical signal; mechanical stimulus; mechanotransduction; migration; mortality; mouse model; neoplastic cell; novel; screening; shear stress; sobriety; statistics; stem; stemness; subcutaneous; trait; transcriptome sequencing; tumor; tumor initiation; tumor progression; viscoelasticity

PROJECT SUMMARY The high mortality rate in ovarian cancers is explained in part, by late-stage clinical diagnosis where intraperitoneal tumor burden is already prevalent and widespread. Shedding and implantation of transformed secretory cells, originating from the fallopian tube, is considered one of the main initiators of ovarian cancer. During the early stages of this disease, secretory fallopian tube cells gain mutations that support migration (against the direction of fluid flow) to the ends of the fallopian tubes (fimbriae). At the fimbriae, the mutated fallopian tube cells form small early precursor lesions. Both the ovaries and fallopian tubes are suspended within the abdominal cavity, where the environment further exposes early precursor tumor cells to dynamic shear stresses. We therefore hypothesize that the fluidic shear stress stimulates the early precursor lesions in the fallopian tubes and modulates their dissemination to the ovary and peritoneal organs. In order to test this hypothesis, we will utilize microfluidic devices and bioreactors to circulate cell growth medium around transformed human and mouse fallopian tube cell lines that are supported on an agarose-collagen, polysaccharide-protein scaffold. Human immortalized fallopian tube cells with varying degrees of genetic mutation will be probed for changes in cell replication, migration, invasion, cell death, and genetic variation after stimulation of shear stress. Our preliminary data suggests a robust increase in the expression of GPRC5A in fallopian tube secretory epithelial cells (FTSEC) under shear stress stimulation. Therefore, we will validate this discovery in FTSEC cell lines with driver mutations and patient-derived cells, and investigate the GPRC5A molecular pathway and its components that are activated in FTSEC under shear stress stimulation, by utilizing gain-of-function and loss-of-function assays. Shear stressed and static control mutated fallopian tube cells will be tested for stemness and the capacity to initiate tumors in immunodeficient mice. Stimulated transformed fallopian tube cells will also be injected into immunocompromised mice to investigate the cell’s ability to colonize and form tumors. These studies will also be performed in mice with intact immune systems in order to assess whether immune cells impact growth and dissemination. Given that no published studies have yet identified the role of shear stresses in dissemination of early precursor lesions in ovarian cancers, the proposed work can potentially have much broader impact. For example, with our dynamic microfluidic and 3D bioreactor models, mechanotransduction and immunotherapy drugs can be screened for development of effective cancer therapies. The components of the shear stress-induced mechanotransduction that are identified in our proposed work, could also be utilized in early detection of ovarian cancers. As a result, the important role of shear stresses in the fluidic niches of ovarian cancers will be established. Lastly, our study on the mechanical regulation of transformed epithelial cells will be highly relevant to fundamental biology and clinical translational alike.

项目概要 卵巢癌的高死亡率部分是由于晚期临床诊断造成的。 腹膜内肿瘤负担已经普遍且广泛。转化体的脱落和植入 起源于输卵管的分泌细胞被认为是卵巢癌的主要引发者之一。 在这种疾病的早期阶段，分泌性输卵管细胞获得支持迁移的突变 （与流体流动方向相反）到输卵管（菌毛）的末端。在菌毛处，突变的 输卵管细胞形成小的早期前体病变。卵巢和输卵管都悬在里面 腹腔，环境进一步将早期前体肿瘤细胞暴露于动态剪切力下 压力。因此，我们假设流体剪切应力刺激了早期前体病变 输卵管并调节它们向卵巢和腹膜器官的传播。 为了验证这一假设，我们将利用微流体装置和生物反应器来循环细胞生长培养基 围绕由琼脂糖胶原支持的转化的人类和小鼠输卵管细胞系， 多糖-蛋白质支架。具有不同程度遗传性的人类永生化输卵管细胞 突变后将探测细胞复制、迁移、侵袭、细胞死亡和遗传变异的变化 剪切应力的刺激。我们的初步数据表明 GPRC5A 的表达在 剪切应力刺激下的输卵管分泌上皮细胞（FTSEC）。因此，我们将验证这一点 在具有驱动突变的 FTSEC 细胞系和患者来源的细胞中发现，并研究 GPRC5A FTSEC 在剪切应力刺激下被激活的分子途径及其成分，通过利用 功能获得和功能丧失检测。剪切应力和静态控制突变的输卵管细胞将 测试干性和在免疫缺陷小鼠中引发肿瘤的能力。刺激转化 输卵管细胞也将被注射到免疫功能低下的小鼠体内，以研究细胞的定植能力 并形成肿瘤。这些研究还将在具有完整免疫系统的小鼠中进行，以评估 免疫细胞是否影响生长和传播。鉴于尚未发表的研究已确定 剪切应力在卵巢癌早期前驱病变传播中的作用，拟议的工作可以 可能产生更广泛的影响。例如，利用我们的动态微流体和 3D 生物反应器模型， 可以筛选机械传导和免疫治疗药物以开发有效的癌症疗法。 在我们提出的工作中确定的剪切应力引起的机械传导的组成部分， 也可用于卵巢癌的早期检测。因此，剪应力在 卵巢癌的流体生态位将被建立。最后，我们对机械调节的研究 转化的上皮细胞将与基础生物学和临床转化高度相关。