Modulating 3D Cellular Connectivity Via Spatially-Controlled Programmable Bonding

项目摘要 组织内细胞的分级排列在决定功能方面起着重要作用。作为其中的一部分 在分层布置中，不同的细胞类型在空间上布置成彼此接触， 传递重要的信号线索，指导许多不同的功能和功能失调的细胞 反应，如改变基因表达，迁移，代谢物共享和生存。更大 了解细胞排列如何影响这些行为将对宿主产生重要影响 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小区布置的重要性是显而易见的， 已经确立的用于以高精度再现这种复杂性的方法仍然有限。按照惯例， 模式生物已经提供了有关这些过程的大部分信息，但通常不允许 持续的直接观察和控制。快速发展的3D打印方法大大增强了我们的能力， 将细胞放置在具有不同材料库的基板上。然而，这些技术并不允许人们 精确地将单个细胞相互接触，以了解不同的排列如何驱动 高度异质性细胞群体中的生物过程。同样，促进细胞间接触的技术 放置不容易实现对多种不同细胞类型的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。具体而言，它评估了使用 寡核苷酸（短DNA序列），以精确控制细胞在可互换和上的位置， 飞行时尚。本提案的目标1旨在建立新的方法和设计规则， 以高保真度和空间控制将多个不同的细胞顺序地添加到表面。目标2旨在 开发一种新的方法，使用可编程DNA构建空间控制的3D细胞组件。 该提案的完成将确立该技术在生物医学领域未来应用的可行性 问题研究

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.