Modulating 3D Cellular Connectivity Via Spatially-Controlled Programmable Bonding

项目摘要 组织内细胞的层级排列在决定功能方面起着重要作用。作为这项工作的一部分 分层排列，不同单元类型以如下方式彼此接触地在空间上排列 传递重要的信号信号，引导大量不同功能和功能障碍的细胞 反应，如基因表达改变、迁移、代谢物共享和生存。一个更伟大的 了解细胞排列如何影响这些行为将对宿主产生重要影响 发育过程包括干细胞分化，癌症转移，疤痕组织形成， 免疫学和血管生成。虽然空间调节的分层单元排列的重要性是 已经确立的、以高精度再现这种复杂性的方法仍然有限。按照惯例， 模式生物已经了解了许多关于这些过程的知识，但通常不允许 持续的直接观察和控制。快速发展的3D打印方法极大地增强了我们的能力 将单元放置在具有不同材质库的衬底上。然而，这些技术不允许人们 精确地将单个细胞相互接触，以了解不同的排列是如何驱动的 高度异质性细胞群体中的生物学过程。同样，促进细胞间接触的技术 放置不容易实现具有多种不同单元类型的3D控制。这项建议旨在建立 解决这一生物医学技术需求的技术的可行性。具体地说，它评估使用 使用寡核苷酸(短DNA序列)来精确控制细胞在可互换和在线的 时尚飞翔。本提案的目标1旨在建立新的方法和设计规则，以 将多个不同的单元顺序添加到高保真和空间控制的曲面上。《目标2》寻求 开发一种使用可编程DNA构建空间控制的3D细胞组件的新方法。 这项提案的完成将为这项技术在未来生物医学中的应用奠定可行性 学习。

Project Summary The hierarchical arrangement of cells within tissue plays an important role in determining function. As part of this hierarchical arrangement, different cell types are spatially arranged in contact with one another in a way that transmits important signaling cues that direct a multitude of different functional and dysfunctional cellular responses, such as altered gene expression, migration, metabolite sharing, and survival. A greater understanding of how cell arrangement impacts these behaviors would have important repercussions for a host of developmental processes that include stem cell differentiation, cancer metastasis, scar tissue formation, immunology, and angiogenesis. While the importance of spatially-regulated hierarchical cell arrangements is well established, methods for reproducing this complexity with high precision remain limited. Conventionally, model organisms have informed much of what is understood about these processes, but often do not allow constant direct observation and control. Rapidly evolving 3D printing methods have greatly enhanced our ability to place cells on substrates with libraries of different materials. However, these technologies do not allow one to precisely place individual cells in contact with each other in order to understand how different arrangements drive biological processes in highly heterogenous cell populations. Likewise, techniques that facilitate cell-cell contact placement do not readily enable 3D control with multiple different cell types. This proposal seeks to establish the feasibility of technology that would address this biomedical technological need. Specifically, it evaluates the use of oligonucleotide (short DNA sequences) to precisely control cell placement in an interchangeable and on-the- fly fashion. Aim 1 of this proposal seeks to establish new methods and design rules for dynamically and sequentially adding multiple different cells to a surface with high fidelity and spatial control. Aim 2 seeks to develop a new approach to building spatially controlled 3D cell assemblies using programmable DNA. Completion of this proposal will establish feasibility of this technology for future applications in biomedical studies.