Harmonic Acoustics for Neighboring cell Dynamic studies(HANDs)

项目摘要 动态的细胞间相互作用对于健康的细胞行为和适当的细胞间通讯至关重要。 受损的细胞间通讯已经牵涉到各种疾病的病理学，包括 癌症、神经退行性疾病、细菌和病毒感染、自身免疫性疾病和心血管疾病 疾病因此，探测细胞间的相互作用对生物医学研究的许多领域都是必不可少的。它可以导致 更详细地了解各种疾病和开发新的治疗策略， 个性化免疫肿瘤学。然而，目前的单细胞分析技术是缓慢的，并且需要 与感兴趣的细胞的潜在有害的物理接触，阻碍了阐明这些现象的进展。 最近，我们发明了用于相邻细胞动态研究的谐波声学（HANDs），一种声学- 基于，自动化，无接触，细胞-细胞配对技术，克服了与 现有的技术。在这个R 01项目中，我们将开发和验证HANDs平台， 特点：（1）无接触性和高生物相容性：无需直接与固体接触 在衬底或珠的情况下，所提出的HANDs技术是非接触式方法。此外，与其暴露于 大的剪切力，强大的压力，或强大的光学，这可能会导致生理损伤，细胞在 我们的装置是用低功率声波轻轻操纵的。建议的HANDs平台允许长时间- 长期（>24小时）细胞-细胞相互作用研究。使用现有的最先进的技术， 原子力光谱等技术。(2)高通量可逆的细胞-细胞相互作用和 单细胞水平的精确定量分析：HANDs技术的多重捕获性质 能够以单细胞精度同时和并行研究大量（> 20，000）细胞对。现有 单细胞技术要么局限于在任何给定时间研究单对细胞，要么缺乏精确度， 需要控制细胞配对和分离，以及精确的定量分析。(3)自动化操作： 与需要复杂的程序和工具来实现操作的现有细胞配对技术不同， 所提出的技术使用声阱自动对准细胞-细胞对。此外，一旦控制 信号被指定，细胞可以以任何自动化和规定的接触方式接触和分离 图案是测试所需的。(4)高分辨率（~100 nm）：使用单相单向传感器和 谐波频率调制，我们将提高我们的HANDs技术的空间分辨率从~1 μm到 ~100 nm。我们将在两个成熟的模型中验证我们的HANDs平台的性能： T细胞和癌细胞之间的相互作用，以及干细胞和巨噬细胞之间的相互作用。在这 在这方面，我们的目标是展示HANDs的深远潜力，以改善以下领域的研究： 从基础生物学到个性化免疫肿瘤学和药物发现。

PROJECT SUMMARY Dynamic cell-cell interactions are crucial for healthy cell behavior and proper intercellular communication. Impaired intercellular communication has been implicated in the pathologies of various diseases including cancer, neurodegenerative diseases, bacterial and viral infections, autoimmune diseases, and cardiovascular diseases. As a result, probing cell-cell interactions is essential to many areas of biomedical research. It can lead to a more detailed understanding of various diseases and the development of novel therapeutic strategies, such as personalized immuno-oncology. However, current single-cell analysis techniques are slow and require potentially harmful physical contact with cells of interest, hindering the progress in elucidating these phenomena. Recently, we invented Harmonic Acoustics for Neighboring cell Dynamic studies (HANDs), an acoustic- based, automated, contact-free, cell-cell-pairing technology, which overcomes the key obstacles associated with the existing technologies. In this R01 project, we will develop and validate the HANDs platform with the following features: (1) Contactless nature and high biocompatibility: Instead of requiring direct contact with solid substrates or beads, the proposed HANDs technology is a contactless method. In addition, rather than exposure to large shear forces, strong pressures, or powerful optics, which can cause physiological damage, the cells in our setup are manipulated gently with low-power acoustic waves. The proposed HANDs platform allows long- term (>24 hours) cell-cell interaction studies. This feat cannot easily be achieved using existing state-of-the-art technologies such as atomic force spectroscopy. (2) High-throughput reversible cell-cell interactions and precise quantitative analysis at the single-cell level: The multi-trapping nature of the HANDs technology enables the simultaneous and parallel study of numerous (>20,000) cell pairs with single-cell precision. Existing single-cell techniques are either limited to studying a single pair of cells at any given time or lack the precision needed to control cell pairing and separation, and precise quantitative analysis. (3) Automated operation: Unlike existing cell pairing technologies which require complicated procedures and tools to achieve operation, the proposed technology automatically aligns cell-cell pairs using acoustic traps. Additionally, once the control signal is specified, cells can be brought into contact and separated in whatever automated and prescribed contact pattern is desired for testing. (4) High resolution (~100 nm): Using single-phase unidirectional transducers and harmonic frequency modulation, we will improve the spatial resolution of our HANDs technology from ~1 μm to ~100 nm. We will validate the performance of our HANDs platform across two well-established models: interactions between T cells and cancer cells, and interactions between stem cells and macrophages. In this regard, we aim to demonstrate the far-reaching potential of HANDs to enable improved research in areas ranging from fundamental biology to personalized immuno-oncology and drug discovery.