Stimulated Brillouin Flow Cytometry for biomechanical assessment of metastatic potential

受激布里渊流式细胞仪用于转移潜能生物力学评估

基本信息

批准号：
10358051
负责人：
Konstantinos Konstantopoulos
金额：
$ 28.44万
依托单位：
UNIV OF MARYLAND, COLLEGE PARK
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-14 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10358051
关键词：
Acoustics Address Adopted Adoption Benchmarking Biological Assay Biomechanics Breast Cancer Cell Cancer Biology Cell Line Cell Nucleus Cell Separation Cells Cellular biology Cessation of life Chemicals Classification Clinical Collaborations Communities Cytometry Cytoskeleton DNA Data Detection Development Diagnosis Drug Screening Extracellular Matrix Flow Cytometry Frequencies Gene Expression Goals In Vitro Label Lasers Lead Light Malignant - descriptor Malignant Neoplasms Measurement Mechanical Stimulation Mechanics Microfluidic Microchips Microfluidics Microscope Microscopy Modulus Names Neoplasm Circulating Cells Neoplasm Metastasis Optics Population Primary Neoplasm Process Property Rapid screening Research Resolution Risk Role Sampling Scanning Screening procedure Sorting - Cell Movement Speed Surface Techniques Technology Translations Tumor Biology Validation Viscosity base biomechanical test cost density field study improved in vivo instrument interest light scattering mammary mechanical properties mechanical stimulus migration mouse model neoplastic cell new technology physical state prognostic value programs submicron technology development tool tumor tumor progression validation studies viscoelasticity

项目摘要

ABSTRACT While our ability to detect and treat primary tumors has significantly increased in the past decade, it remains difficult to diagnose the origination of metastasis which remains responsible for nearly 90% of cancer-related deaths. In this respect, recently, the critical role of the mechanical state of tumors and tumor cells has been recognized for tumor progression, malignancy transformation and metastasis but it remains poorly exploited due to the lack of suitable measurement tools. Many microfluidic deformability approaches have recently emerged, but they only give an average value of the overall cell mechanical properties, while it would be important to separate nucleus vs cytoskeleton contributions, and they need mechanical stimulation to probe properties, which is deleterious since cells strongly react to mechanical stimuli. In the past few years, we have been developing an all-optical approach to this challenge, named Brillouin microscopy, and strongly established it in tumor biology to characterize cell mechanics during metastatic cascade. However, current technology is inherently limited in speed (~50ms/point), as it relies on spontaneous Brillouin interaction, and thus is not suitable for rapid screening/sorting of tumor cells. Here, we will develop stimulated Brillouin cytometry which 1) increases speed by ~100-fold and 2) provides additional contrast mechanisms such as viscosity and mass density with micron-scale resolution. Based on this breakthrough, we will develop and validate a flow cytometry/cell sorting platform using elastic modulus, viscosity, and density as label-free contrast mechanisms (Aim 1). We will then validate our mechanical assessment of metastatic cells against microfluidic assays to assess migration and proliferation and in mice models to assess cell’s metastatic ability in vivo (Aim 2). The rigorous process of technology development, validation, benchmarking and field-testing will yield a platform with unprecedented capabilities to characterize and sort cells based on their mechanical properties within an instrument compatible with traditional flow cytometry, thus ready to be widely adopted by the cancer biology community. The proposal features the collaboration between optical technology experts (UMD) and cancer metastasis pioneers in both in vitro (JHU) and in vivo (UMB) settings with an established track record of fruitful collaboration.

抽象的虽然我们检测和治疗原发性肿瘤的能力在过去十年中显着增强，但仍然存在难以诊断转移的起源，而转移的起源仍然是近 90% 癌症相关的原因在这方面，最近，肿瘤和肿瘤细胞的机械状态发挥了关键作用。其在肿瘤进展、恶性肿瘤转化和转移中的作用已得到认可，但仍未得到充分利用由于缺乏合适的测量工具，最近出现了许多微流体变形方法。出现了，但它们只给出了整体电池机械性能的平均值，而对于分离细胞核与细胞骨架的贡献很重要，并且它们需要机械刺激来探测性质，这是有害的，因为细胞对机械刺激有强烈反应，在过去的几年里，我们已经发现了这一点。一直在开发一种全光学方法来应对这一挑战，称为布里渊显微镜，并且强烈在肿瘤生物学中建立了它来表征转移级联过程中的细胞力学。技术本质上在速度上受到限制（~50ms/点），因为它依赖于自发的布里渊相互作用，并且因此不适合肿瘤细胞的快速筛选/分选。在这里，我们将开发受激布里渊法。细胞计数法，1) 将速度提高约 100 倍，2) 提供额外的对比机制，例如基于这一突破，我们将开发和实现微米级分辨率的粘度和质量密度。使用无标记的弹性模量、粘度和密度验证流式细胞术/细胞分选平台然后我们将验证我们对转移细胞的机械评估。微流体测定法评估迁移和增殖，并在小鼠模型中评估细胞的转移能力体内（目标 2）严格的技术开发、验证、基准测试和现场测试过程。将产生一个具有前所未有的能力的平台，可以根据细胞的机械特性对细胞进行表征和分类仪器内的特性与传统流式细胞术兼容，因此可以被广泛采用该提案的特点是光学技术专家之间的合作。（UMD）和体外（JHU）和体内（UMB）环境中的癌症转移先驱，具有既定的卓有成效的合作记录。