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.

项目摘要 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 仍然是非常普遍和咄咄逼人的。转移是由于原发肿瘤脱落了一小部分 类干细胞亚群，可通过邻近的基质迁移并扩散到整个 身体不像那些留在原发肿瘤中的细胞。识别分子预后的研究进展 这些干细胞样细胞可以评估转移风险的标记物一直有限；然而，最近， 有证据表明，肿瘤的物理状态可能会作为一种替代的预后标志。我们有 观察到这样一个物理标记-黏附强度-在转移性上皮细胞系中有所不同 乳腺、前列腺和肺肿瘤，由于基质样条件下局部粘连组装的不同， 例如，低Mg+2和低Ca+2。因此，强粘附性亚群比它们的 未选择的对应物。我们的假设是相反的群体，即弱附着的肿瘤细胞 间质，代表转移最多的亚群，可根据其黏附强度进行纯化 并进行计数以确定转移风险。为了分离这种反向的弱黏附细胞群，我们 已经开发出一种“黏附”的流室分析方法，它将使细胞承受低剪切(&lt；10dynes/cm2)。 像基质一样的环境和快速(10分钟)分拣107个细胞。使用此技术进行概念验证 目的1，验证其分选效率和保真度，并观察弱黏附细胞对 分离、迁移和入侵类似于邻近肿瘤间质的工程化微环境。在目标2中，我们 将通过注射在体内测试粘性流室预测乳腺癌转移风险的能力 选定的细胞放入乳房脂肪垫中。然后我们将评估弱黏附或强黏附细胞的倾向。 形成或不形成转移。来自原发肿瘤间质的细胞将被直接评估为 鉴定具有弱黏附特征的细胞，并将其与继发转移形成相关联 发生在临床上。通过跨肿瘤类型的验证，这种创新的分子和细胞分析 技术R21提案将建立一种粘性流室技术，并验证我们的假设 弱附着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对肿瘤预后的额外评估 转移风险。