Cell Control via Spatiotemporal Microenvironmental pH Modulation

通过时空微环境 pH 调节进行细胞控制

• 批准号: 10713388
• 负责人: Jinglei Ping
• 金额: $ 37.92万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10713388
• 关键词: Address; Advanced Development; Behavior; Bicarbonates; Biomedical Engineering; Buffers; Carbon Dioxide; Cardiac; Cardiac Myocytes; Cardiovascular system; Cell model; Cell physiology; Cells; Cellular biology; Chemicals; Communication; Development; Devices; Diffusion; Drug Delivery Systems; Failure; Goals; Malignant Neoplasms; Metabolism; Methods; Microelectrodes; Microscopic; Mission; Morphogenesis; Morphology; Optics; Outcome; Pathogenesis; Positioning Attribute; Property; Public Health; Reaction Time; Regenerative Medicine; Regulation; Resolution; Signal Transduction; System; Techniques; Testing; Therapeutic; Time; Tissue Engineering; Transducers; United States National Institutes of Health; anti-cancer; cell behavior; disability; graphene; improved; meter; nanomaterials; neoplastic cell; pH gradient; spatiotemporal; tool; two-dimensional

Microenvironmental pH is a key factor in cell functioning and pathogenesis. To control the function and behavior of cells by modulating pH microenvironments is critical to advancing the development of cell biology and tissue engineering and enabling applications in drug delivery and regenerative medicine. However, pH-based cell control remains a challenge due to the lack of means to real-time, spatioselective modulation of microenvironmental pH. While pH microenvironments in cell systems are highly heterogeneous in time and space, known pH-modulation methods are through CO2/HCO3− buffering and H+ diffusion, which are slow, isotropic, and nonspecific. An urgent need, therefore, is to modulate pH microenvironments in a spatiotemporally specific manner. Failure to do so means that pH, an essential factor that determines cell fate and function, is not in good control. The PI’s long-term goal is microenvironmental pH–based closed-loop regulation of cell function, metabolism, and morphogenesis. The overall goal of this project, a critical step towards the long-term goal, is to control cells by real-time, spatioselective modulation of pH microenvironments. The hypothesis is that cell function and behavior can be regulated with ultra-high spatiotemporal resolutions (10–100 µm, <50 s), compared to conventional, diffusion-based methods (>103 µm, >103 s), in pH microenvironments that are modulated nanoelectrochemically by microelectrodes based on graphene, a two-dimensional nanomaterial with unique outstanding bio-transduction properties that address the primary challenge of on-chip pH modulation of living cell systems for typical microelectrode materials. The approach to test this hypothesis is to quantify real-time responses of model cell systems to arrayed pH microenvironment generated by an array of bidirectional graphene-microelectrode transducers that are optically transparent to allow microscopic characterization and communicate with cellular systems through electrical signal interrogation and rapid nanoelectrochemical microenvironmental-pH modulation. The following milestone goals will be reached in this project: (1) to create densely arrayed pH microenvironment by developing an array of bidirectional graphene-microelectrode transducers and (2) to control the function and behavior of model cell systems (cardiomyocytes and tumor cells) via spatiotemporal microenvironmental pH modulation using the graphene transducer array. The PI is uniquely positioned to conduct the project due to the ability of the PI’s lab to create graphene microelectrodes integrable in a fluidic device for interfacing cellular systems, interrogating electrical/chemical cell signals, and controlling cell behavior by generating microscale pH gradients. To harness and combine these techniques allows the development of arrays of bidirectional graphene transducers for selective, real-time pH-microenvironment modulation and cell control. The expected outcome of the project is pH-based cell-control tools with over two- orders-of-magnitude enhanced spatiotemporal resolutions compared to conventional methods. This outcome is to generate positive impact on bioengineering development, regenerative medicine, and synthetic morphology.

微环境pH值是细胞功能和发病机制中控制功能和行为的关键因素。 通过调节 pH 微环境来调节细胞对于促进细胞生物学和组织的发展至关重要 然而，基于 pH 的细胞在药物输送和再生医学中的工程和应用。 由于缺乏实时、空间选择性调制的手段，控制仍然是一个挑战 微环境 pH 值虽然细胞系统中的 pH 微环境在时间和时间上具有高度异质性。 空间，已知的 pH 调节方法是通过 CO2/HCO3− 缓冲和 H+ 扩散，这些方法很慢， 因此，迫切需要在时空上调节 pH 微环境。 如果不这样做，就意味着决定细胞命运和功能的重要因素 pH 值不存在。 PI 的长期目标是基于微环境 pH 值的细胞功能闭环调节， 该项目的总体目标是实现长期目标的关键一步。 通过实时、空间选择性调节 pH 微环境来控制细胞。 相比之下，功能和行为可以通过超高时空分辨率（10–100 µm，<50 s）进行调节 传统的基于扩散的方法（>103 µm，>103 s），在调节的 pH 微环境中 通过基于石墨烯的微电极进行纳米电化学，石墨烯是一种具有独特功能的二维纳米材料 出色的生物转导特性，可解决生物芯片上 pH 调节的主要挑战 测试这一假设的方法是实时量化。 模型细胞系统对由一系列双向阵列产生的阵列 pH 微环境的响应 石墨烯微电极传感器具有光学透明性，可以进行微观表征和 通过电信号询问和快速纳米电化学与细胞系统通信 该项目将达到以下里程碑目标：（1）创建微环境-pH调节。 通过开发双向石墨烯微电极阵列来形成密集排列的 pH 微环境 (2) 控制模型细胞系统（心肌细胞和肿瘤细胞）的功能和行为 通过使用石墨烯传感器阵列进行时空微环境 pH 调节，PI 是独一无二的。 由于 PI 实验室有能力制造可集成的石墨烯微电极，因此有能力开展该项目 在流体装置中用于连接细胞系统、询问电/化学细胞信号以及控制 通过产生微尺度 pH 梯度来控制细胞行为。利用和结合这些技术可以实现 开发用于选择性、实时 pH 微环境的双向石墨烯传感器阵列 该项目的预期成果是基于 pH 的细胞控制工具，具有超过两种功能： 与传统方法相比，这个结果提高了几个数量级的时空分辨率。 对生物工程发展、再生医学和合成形态学产生积极影响。