Synthetic Adhesome Cells: Engineering Interfaces Between Synthetic and Live Cells for Controlled Delivery

合成粘附体细胞：合成细胞和活细胞之间的工程界面以实现受控递送

• 批准号: 2218467
• 负责人: Brian Belardi
• 金额: $ 150万
• 依托单位: University of Texas at Austin
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2218467&HistoricalAwards=false
• 关键词: Synthetic; Adhesome; Cells; Engineering; Interfaces

Multicellular organisms represent a remarkable transition in evolution, wherein single cells began working together to form a cohesive and coherent unit known as tissue. This transition was made possible by the emergence of cell-cell adhesion. Adhesions within tissue allow cells to communicate with their neighbors, both directly and indirectly, creating an integrated multi-cell circuit. In one form of communication, cells can directly transfer material from one cell to another, which is a grand challenge for engineers that are trying to emulate cellular processes for delivery purposes. This research team will focus on user-defined transfer of material between synthetic cells and living cells, which will ultimately lead to new delivery mechanisms to alter biological behavior for biotechnology, chemical sensing, and biomedicine. Finally, the project seeks to explore ethical issues with the proposed technology by collaborating with an ethicist and a variety of students from diverse backgrounds and to define design laws surrounding the use of synthetic cells. Synthetic cells offer the promise of combining complex cellular machinery with precise molecular triggers and logic-gates to control cell-like processes for a variety of applications in biotechnology, chemical sensing and production, and biomedicine. A key capability of live cells, which has not been recapitulated by synthetic cells, is organization into functional multi-cellular tissue – the defining characteristic of all metazoans. Cells within multi-cellular tissue communicate and contact one another through cell-cell junctions, such as adhesive junctions and junctions that enable diffusion of solutes from cell to cell, which are called gap junctions. The Belardi lab has developed synthetic cells that form functional adhesive junctions. This work makes use of adhesive and regulatory proteins – both natural and synthetic – to engineer multi-synthetic cell systems. In parallel, the Stachowiak lab has shown that synthetic cells containing gap junction proteins can be used to transfer molecular cargo directly into the cytosol of live cells. Here, the project seeks to combine these two nascent capabilities to solve a key bottleneck in biotechnology – regulated delivery of biomolecules to the interior of cells. Specifically, the collaboration will build upon the natural ability of adhesive junctions to regulate the assembly of gap junctions, allowing precise, engineered control over molecular transfer between synthetic and live cells. By providing regulatory control – a feature that native cells possess - this work will enable synthetic cells to reach their transformative and safe potential. The objective in this proposal is to take components and biophysical principles from adhesive cells and create “Synthetic Adhesome Cells”, with new functionalities that depend on optical triggers, co-adhesive partners and ligation-dependent logic-gates. The overarching goal with the Synthetic Adhesome Cell platform is two-fold: i) to create systems with user-defined control over intracellular delivery and ii) to shed light on the biophysical mechanisms underlying junction formation and communication at cell-cell interfaces.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.

多细胞生物体代表了进化中的一个显着转变，其中单细胞开始一起工作，形成一个被称为组织的有凝聚力和连贯性的单位。细胞间粘附的出现使这种转变成为可能。组织内的粘附允许细胞直接和间接地与它们的邻居通信，从而创建集成的多细胞电路。在一种形式的通信中，细胞可以直接将材料从一个细胞转移到另一个细胞，这对于试图模拟细胞过程以实现交付目的的工程师来说是一个巨大的挑战。该研究团队将专注于合成细胞和活细胞之间的用户定义的材料转移，这将最终导致新的传递机制，以改变生物技术，化学传感和生物医学的生物行为。最后，该项目旨在通过与伦理学家和来自不同背景的各种学生合作，探索拟议技术的伦理问题，并定义围绕使用合成细胞的设计法律。合成细胞提供了将复杂的细胞机制与精确的分子触发器和逻辑门相结合的前景，以控制生物技术，化学传感和生产以及生物医学中各种应用的细胞样过程。活细胞的一个关键能力是组织成功能性的多细胞组织，这是所有后生动物的定义特征，但还没有被合成细胞概括。多细胞组织内的细胞通过细胞-细胞连接（例如粘附连接和能够使溶质从细胞扩散到细胞的连接，其被称为间隙连接）彼此通信和接触。Belardi实验室已经开发出形成功能性粘附连接的合成细胞。这项工作利用天然和合成的粘附蛋白和调节蛋白来设计多合成细胞系统。与此同时，Stachowiak实验室已经证明，含有间隙连接蛋白的合成细胞可以用于将分子货物直接转移到活细胞的胞质溶胶中。在这里，该项目寻求将这两种新生能力联合收割机结合起来，以解决生物技术调节的生物分子向细胞内部输送的一个关键瓶颈。具体而言，该合作将建立在粘附连接调节间隙连接组装的天然能力的基础上，从而对合成细胞和活细胞之间的分子转移进行精确的工程控制。通过提供调节控制-天然细胞具有的特征-这项工作将使合成细胞能够实现其转化和安全的潜力。该提案的目标是从粘附细胞中提取成分和生物物理原理，并创建“合成粘附体细胞”，其新功能取决于光学触发器，共粘附伴侣和连接依赖性逻辑门。合成粘附体细胞平台的总体目标是双重的：i）创建对细胞内递送具有用户定义控制的系统，ii）阐明细胞-细胞界面处连接形成和通信的生物物理机制。该奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。