Developing Adhesome Technology as a Physical Marker of Highly Metastatic Cells

开发粘附体技术作为高度转移细胞的物理标记

• 批准号: 9922219
• 负责人: Adam J Engler
• 金额: $ 23.63万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9922219
• 关键词: Address; Adhesions; Adhesives; Biological Assay; Biological Markers; Biophysics; Biopsy; Blood; Cations; Cell Count; Cell Line; Cells; Clinic; Clinical; Data; Detection; Devices; Disease; Disease Progression; Engineering; Ensure; Environment; Epithelial Cells; Estrogen Receptors; Extravasation; Fatty acid glycerol esters; Fibroblasts; Flow Cytometry; Focal Adhesions; Heterogeneity; Hour; Impairment; In Vitro; Invaded; Label; Lead; Lung; Lung Neoplasms; Malignant neoplasm of lung; Mammary Neoplasms; Mammary gland; Measures; Metastatic breast cancer; Methods; Molecular; NOD/SCID mouse; Needle biopsy procedure; Neoplasm Metastasis; Nonmetastatic; Pancreas; Patient Monitoring; Patients; Phenotype; Population; Prevalence; Primary Neoplasm; Process; Prognostic Marker; Prostate; Prostatic Neoplasms; Relapse; Research; Risk; Solid Neoplasm; Sorting - Cell Movement; Speed; TACSTD1 gene; Technology; Testing; Time; Tissues; Tumor Markers; Tumor-Derived; Woman; base; cancer cell; cancer therapy; clinical predictors; diagnostic biomarker; improved; in vivo; innovation; malignant breast neoplasm; metastatic process; migration; neoplastic cell; physical property; physical state; prognostic; prostate cancer cell line; screening; stem-like cell; technology validation; trait; tumor; tumor xenograft

Project Summary Diagnostic markers have improved our ability to identify tumors at earlier stages, but secondary metastases are still exceedingly common and aggressive. Metastases result from primary tumors shedding a small subpopulation of stem-like cells that can migrate through adjacent stroma and disseminate throughout the body unlike those cells remaining in the primary tumor. Progress towards identifying molecular prognostic markers of these stem-like cells that could assess metastatic risk has been limited; very recently however, evidence has suggested that a tumor's physical state may act as an alternative prognostic marker . We have observed that one such physical marker–adhesion strength–varies in metastatic epithelial cell lines from mammary, prostate, and lung tumors due to differences in focal adhesion assembly in stromal-like conditions, e.g. low Mg+2 and Ca+2. Thus strongly adherent sub-populations are slower and less processive than their unselected counterparts. Our hypothesis is that the converse population, i.e. weakly adherent tumor cells in the stroma, represents the most metastatic sub-population, which can be purified based on their adhesion strength and counted to determine metastatic risk. To isolate this converse population of weakly adherent cells, we have developed an “adhesome” flow chamber assay that will subject cells to low shear (<10 dynes/cm2) in stromal-like environments and rapidly (<10 min) sort 107 cells. Using this technology for proof-of-concept in Aim 1, we will validate its sorting efficiency and fidelity and observe the potential of weakly adherent cells to detach, migrate, and invade engineered microenvironments that resemble adjacent tumor stroma. In Aim 2, we will test the adhesome flow chamber's ability to predict breast cancer metastatic risk in vivo by injecting selected cells into mammary fat pads. We will then assess the propensity of weakly or strongly adherent cells to form or not form metastases, respectively. Cells from primary tumor stroma will be directly assessed to identify cells with a weakly adhesive signature and correlate that with secondary metastasis formation as would occur in the clinic. By validating across tumor types, this Innovative Molecular and Cellular Analysis Technologies R21 proposal will establish an adhesome flow chamber technology and validate our hypothesis that weakly adherent tumor cells are capable of migrating and forming secondary metastases, i.e. establish metastatic risk. Thus this technology should provide clinicians with addition prognostic assessment of a tumor's metastatic risk.

项目概要 诊断标记物提高了我们在早期阶段识别肿瘤的能力，但继发性转移 仍然非常常见且具有攻击性。转移是由于原发肿瘤脱落少量细胞所致 干细胞样细胞亚群，可以迁移通过邻近的基质并在整个组织中传播 身体不同于原发肿瘤中残留的细胞。确定分子预后的进展 这些可以评估转移风险的干细胞样细胞标记物是有限的；然而最近， 有证据表明肿瘤的物理状态可以作为替代的预后标志。我们有 观察到这样一种物理标记——粘附强度——在转移性上皮细胞系中存在差异 由于间质样条件下粘着斑组装的差异，导致乳腺、前列腺和肺肿瘤， 例如低 Mg+2 和 Ca+2。因此，强烈依附的亚群比它们的亚群速度更慢，处理能力也更弱。 未选定的对应方。我们的假设是相反的群体，即弱粘附的肿瘤细胞 基质，代表最具转移性的亚群，可以根据其粘附强度进行纯化 并计数以确定转移风险。为了分离这种相反的弱贴壁细胞群，我们 开发了一种“粘附”流动室测定，可以使细胞在低剪切力（<10达因/厘米2）下进行 基质样环境并快速（<10 分钟）分选 107 个细胞。使用该技术进行概念验证 目标1，我们将验证其分选效率和保真度，并观察弱贴壁细胞的潜力 分离、迁移和侵入类似于邻近肿瘤基质的工程微环境。在目标 2 中，我们 将测试粘附体流动室通过注射预测体内乳腺癌转移风险的能力 选定的细胞进入乳腺脂肪垫。然后我们将评估弱或强贴壁细胞的倾向 分别形成或不形成转移。来自原发性肿瘤基质的细胞将被直接评估 识别具有弱粘附特征的细胞，并将其与继发性转移形成相关联 发生在诊所。通过跨肿瘤类型验证，这种创新的分子和细胞分析 技术 R21 提案将建立粘附流室技术并验证我们的假设 弱粘附的肿瘤细胞能够迁移并形成继发性转移，即建立 转移风险。因此，这项技术应该为临床医生提供肿瘤的额外预后评估 转移风险。