From synthetic bacterial adhesions to synthetic bacterial materials

从合成细菌粘附到合成细菌材料

基本信息

批准号：
10707441
负责人：
Hans Ingmar Riedel-Kruse
金额：
$ 31.1万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-20 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10707441
关键词：
3-Dimensional Adhesions Anabolism Area Bacteria Bacterial Adhesins Bacterial Adhesion Basic Science Biochemical Biocompatible Materials Biophysics Cell Adhesion Cell surface Cell-Cell Adhesion Cells Deposition Development Diagnostic Diagnostic Equipment Drug Delivery Systems Engineering Escherichia coli Foundations Future Gene Expression Regulation Genetic Models Goals Growth Health Image Industry Investigation Kinetics Length Literature Logic Medial Medical Medicine Methodology Microbial Biofilms Microfluidics Microscopic Modeling Molecular Morphology Natural Compound Outcome Pathway interactions Pattern Pharmaceutical Preparations Play Porosity Property Proteins Publications Research Resolution Signal Transduction Specificity Structure Surface System Testing Work bacterial community biophysical model biophysical properties cell growth design diagnostic assay diagnostic strategy experimental study in vivo infancy innovation instrumentation meter novel optogenetics predictive modeling self assembly small molecule support tools synthetic biology three dimensional structure tool viscoelasticity

项目摘要

Engineering of bacterial synthetic multicellular systems and materials hold promise for many health- relevant applications such as modular drug biosynthesis, living diagnostic devices, and synthetic bioﬁlm research models. To date, bacterial synthetic biology has largely focused on the scales of molecules and single cells. Equivalent work on bacterial synthetic consortia is much less advanced, in signiﬁcant part due to the previous lack of suitable synthetic and genetically encoded cell-cell adhesion tools to control the assembly, development, and functionality of multicellular systems. We recently developed the ﬁrst such synthetic cell-cell adhesion toolbox, as well as tools for optogenetically controlling cell-surface deposition and patterning. The speciﬁc objectives of this research are to signiﬁcantly advance these synthetic cell-adhesion tools, and to develop design principles and predictive modeling tools that enable consortia engineering and patterning that integrate all relevant length scales (i.e., molecular, cellular, and multicellular), and ultimately pave the way for medially relevant applications. Our main hypothesis is that we can signiﬁcantly advance our control over the strength, speciﬁcity, and subcellular localization of synthetic adhesion proteins in Escherichia coli, which will allow rational tuning of consortium-level biophysical properties such as porosity and viscoelasticity, and which will ultimately enable versatile multicellular consortium engineering and patterning. This work will constitute a foundation for various biomedical applications such as biocompatible materials, multicellular plug-and-play pathway engineering, targeted in-vivo drug delivery, and living diagnostic devices. Our interdisciplinary methodology combines synthetic biology, biophysics, instrumentation and modeling. All experiments will be done in a quantitative manner. The proposed investigations include three independent yet synergistic Speciﬁc Aims motivated by our hypothesis: (Aim 1) Advance the functionality of the synthetic adhesin toolkit at the subcellular level; (Aim 2) Achieve engineering control over synthetic consortium properties such as viscoelasticity and porosity at the scale of 10-100 µm; and (Aim 3) Achieve higher-level consortium patterning on the scale of centimeters and demonstrate potential for medical applications. The PI (Prof. Riedel-Kruse) and his team are well-suited for this project as we have signiﬁcant expertise in synthetic biology, biophysics, instrumentation (e.g., microﬂuidics, imaging), and modeling genetic circuits and biophysical systems across scales. We developed the ﬁrst synthetic cell-cell and optogenetic cell-surface adhesion toolboxes in bacteria. Multiple collaborators provide additional domain expertise in key areas. Overall, this project's innovation lies in establishing synthetic adhesins as an essential and integral component of the synthetic circuit-engineering toolbox and in establishing a novel paradigm for modular engineering of multicellular living materials. Accordingly, this project will broadly impact the engineering of synthetic consortia for basic research as well as enable a dynamic spectrum of future applications in health.

细菌合成多细胞系统和材料的工程为许多健康带来了希望相关应用，例如模块化药物生物合成、活体诊断设备和合成生物膜研究迄今为止，细菌合成生物学主要集中在分子和单细胞的尺度上。细菌合成联合体的等效工作远不那么先进，这在很大程度上是由于之前的缺乏合适的合成和基因编码的细胞间粘附工具来控制组装、发育和我们最近开发了第一个这样的合成细胞-细胞粘附工具箱，如以及光遗传学控制细胞表面沉积和图案化的工具。这项研究的具体目标是显着推进这些合成细胞粘附工具，并且开发设计原则和预测建模工具，使联盟工程和图案化能够整合所有相关的长度尺度（即分子、细胞和多细胞），并最终为我们的主要假设是我们可以显着提高对疾病的控制。大肠杆菌中合成粘附蛋白的强度、特异性和亚细胞定位，这将允许合理调整联盟级别的生物物理特性，例如孔隙率和粘弹性，并且最终将实现多功能多细胞联合体工程和图案化。为各种生物医学应用奠定了基础，例如生物相容性材料、多细胞即插即用通路工程、靶向体内药物输送和活体诊断设备。我们的跨学科方法结合了合成生物学、生物物理学、仪器和建模。实验将以定量的方式进行，建议包括三个独立的研究。由我们的假设激发的协同具体目标：（目标 1）提高合成粘附素的功能亚细胞水平的工具包；（目标 2）实现对合成联合体特性的工程控制，例如 10-100 µm 范围内的粘弹性和孔隙率；以及（目标 3）实现更高水平的联合图案化在厘米尺度上，并展示了医疗应用的潜力。 PI（Riedel-Kruse 教授）和他的团队非常适合这个项目，因为我们有重要的合成生物学、生物物理学、仪器（例如微流体、成像）和遗传建模方面的专业知识我们开发了第一个合成细胞-细胞和光遗传学细胞表面。细菌粘附工具箱。多个合作者在关键领域提供了额外的专业知识。该项目的创新在于将合成粘合剂建立为粘合剂的重要组成部分。合成电路工程工具箱并建立多细胞模块化工程的新范例因此，该项目将广泛影响基础合成联合体的工程。研究并实现未来健康领域的动态应用。