Developing Nanosensor Chemical Cytometry (NCC) to Support the Development of Cellular Therapeutics

生物如何在细胞水平上对环境作出反应在人类健康和疾病中起着重要作用。例如，相同类型和来源的细胞在功能上通常在不同细胞之间存在显著差异。虽然以前的方法是将细胞群体作为一个群体来研究，但越来越清楚的是，单独测量细胞可以提供重要的信息，有助于预测它们作为生物体的一部分的行为。因此，该项目的目标是开发一种新的平台，以快速的方式在单细胞水平上以无损的方式探测细胞的功能特性（即细胞细胞术）。该平台将利用新型纳米传感器，通过检测近红外光照射下单个细胞的反射来确定微通道中单个细胞的特性。该平台的潜在好处包括早期检测慢性疾病，如糖尿病和癌症，因为许多最初的变化始于细胞群的功能变化，以及加强对细胞作为治疗手段的新研究。因为这些细胞是从患者身上提取的，所以开发技术在单细胞水平上检查它们的质量，以确保它们按预期功能是很重要的。该项目还将为大波士顿地区的本科生、高中生和少数族裔提供更多的教育机会，特别是通过为教师、高中和本科少数族裔学生提供指导实习项目。新平台将通过将新的细胞传感平台连接到现有平台来增强正在进行的纳米孔测序实验室模块，使学生能够研究单个细胞的遗传和物理特性。该项目的目标是利用高通量微流控通道内荧光纳米传感器阵列的界面，以便在单细胞水平上对科学相关的无标签细胞群体进行无损研究。这项工作建立在研究的基础上，这些研究将推动麻省理工学院研究者实验室开创的一类新的生物光子监测平台。该传感器由近红外（nIR）荧光单壁碳纳米管（SWNT）纳米传感器组成，沿着微流体通道引导流动细胞。这些纳米传感器可以被设计成对生物相关化学信号的局部浓度做出光学响应。初步结果表明，可以利用流动细胞本身作为高斯透镜放大纳米传感器发射信号，并实时提取每个细胞的丰富信息。由于研究者的成像数据收集方法，提取的生物分子信息可以与单个细胞的物理性质交叉相关。作为这个项目的一部分，首先，这样一个系统的能力，以适应多个目标纳米传感器的多路单细胞检测将被研究。接下来，将研究细胞透镜的物理现象，包括不同细胞类型和实验条件的影响。最后，新开发的传感器平台将用于研究细胞治疗中常用的细胞类型，包括：单核细胞、巨噬细胞、t细胞或神经干细胞。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

How biology responds to the environment at the cellular level play an important role in human health and diseases. For example, it is common for cells of the same type and origin to be functionally significantly different from one cell to another. While previous methods have studied cell populations as a group, it is becoming increasingly clear that measuring cells individually provides important information that helps to predict their behavior as part of organisms. Thus, the goal of this project is to develop a new platform for probing the functional properties of cells (i. e., cell cytometry) in a nondestructive way at the single cell level in a rapid manner. This platform will make use of novel nanosensors that can determine the properties of single cells flowing in a microchannel by detecting their reflections when illuminated with near infrared light. Potential benefits of this platform include earlier detection of chronic conditions such as diabetes and cancer, as many of the initial changes responsible start as functional shifts within a cell population, and enhancement of new research into the use of cells as therapeutics. Because these cells are extracted from patients, it is important to develop techniques to examine their quality at the single-cell level to ensure that they function as intended. The project will also further educational opportunities for undergraduates, high school students, and underrepresented minorities in the greater Boston area specifically through guided internship programs for teachers and high school and undergraduate underrepresented minority students. The new platform will enhance an ongoing Nanopore Sequencing Laboratory Module by connecting the new cell sensing platform to the existing platform, allowing students to study both the genetic and physical properties of a single cell.The goal of this project is to utilize the interface of fluorescent nanosensor arrays within high throughput microfluidic channels such that scientifically relevant label free cellular populations can be studied nondestructively at the single cell level. The work builds on studies that will advance a new class of biophotonic monitoring platform pioneered by the investigator’s Lab at MIT. The sensor consists of near-infrared (nIR) fluorescent single-walled carbon nanotube (SWNT) nanosensors along a microfluidic channel through which flowing cells are guided. These nanosensors can be engineered to be optically responsive to local concentrations of biologically relevant chemical signals. Preliminary results showed that one can utilize the flowing cell itself as a Gaussian lens to amplify the nanosensor emission signal and extract rich information on a per cell basis in real-time. Due to the investigator’s method of imaging data collection, the biomolecular information extracted can be cross-correlated with individual cellular physical properties. As part of this project, first, the capabilities of such a system to accommodate multiple targeted nanosensors for multiplexed single cell detection will be studied. Next, the physical phenomenon of cellular lensing, including the effects of different cell types and experimental conditions will be studied. Finally, the newly developed sensor platform will be utilized to study commonly used cell types used in cell therapy, including: monocytes, macrophages, T-cells or neural stem cells.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.