EAGER: Magnetothermal Control of Cell Fates and Function

远程指导细胞功能和指导衰老或受损细胞和组织再生的潜力将在生物医学领域产生广泛的影响。细胞是生命的基本组成部分，具有不可思议的能力；开发控制细胞功能的能力将使重建患病和受损的组织、替换老化的器官或指导细胞寻找和消除癌症成为可能。作为个体，细胞具有巨大的能力，细胞也具有协同工作的天然能力，因此，如果对这种团队努力进行适当的控制，细胞的能力可以被放大；想象一下，你的肝脏有1亿个细胞，它们共同帮助你消化食物，将营养物质分配给身体，储存能量，净化血液。细胞的功能是由基因和蛋白质组成的回路调节的，这些回路可以通过生物开关来控制。然而，迄今为止，一种将一些开关转移到细胞并从远处控制它们的方法尚未被描述。该项目旨在开发具有多个开关的生物遥控器，可用于指导细胞执行功能并协同工作。这些被称为“工程细胞器”的远程控制模块可以通过磁场进行操作。利用磁控工程细胞器，磁场的快速闪烁可以加热工程细胞器，并打开或关闭控制细胞功能的生物开关。该开关设计具有多路复用功能，因此可以远程控制多种功能。工程细胞器同样被设计用于控制细胞内的多种生物功能并指导它们协同工作。该项目的多学科性质为生物工程、物理科学、合成生物学和再生医学领域的下一代STEM研究人员提供了独特的培训机会。研究团队邀请来自代表性不足群体的学生通过密歇根州立大学（MSU）成功的ENSURE（工程暑期本科生研究经验）项目参与暑期研究。此外，该小组还为高中科学教师举办了关于工程生物遥控器的讲习班，使他们能够更新他们的知识，并用最先进的研究更新他们的课程。为了提高公众对工程细胞器所能完成的工作的理解和欣赏，研究小组参加了密歇根州立大学科学节，迎合了更大的兰辛地区。项目成果将纳入本科课程，并纳入生物工程和合成生物学研究生课程，教授利用工程细胞器指导生物学的新原理。该项目旨在模拟内共生并修饰原核细胞，以创建可使用交变磁场（AMF）控制的工程磁内共生体（eMEs）。这涉及工程铁颗粒包裹或含铁原核生物，趋磁细菌（MTB），作为磁热调节的伪细胞器。磁热控制可激活eME，使其表达指向宿主细胞核的哺乳动物转录因子（txn），从而控制宿主基因的表达。利用光学报告基因作为读出器，将遗传热开关设计到MTB中，并将细胞质中eME的报告基因传递到宿主细胞核。报告基因的使用将有助于指导编码哺乳动物txn因子的eME的发育，从而使分化的巨噬细胞重编程为iPSCs，然后再重编程为肝细胞。一种化学（甘露糖）调控的ipsc生成操纵子将被设计表达用于细胞重编程的Oct3/4、Sox2、KLF4、c-Myc、txn因子，一种热调控的肝细胞生成操纵子表达txn因子HNF4A、HNF1A。FoxA1和FoxA3。一旦在EES中得到证实，操作子将被转移到MTB中，并使用这些eMEs将巨噬细胞编程为iPSCs，然后通过磁热控制转化为肝细胞。该项目由分子和细胞生物科学部的系统和合成生物学集群以及化学、生物工程、环境和运输系统部的工程生物学和健康集群资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The potential to guide cellular functions from a distance and direct the regeneration of aged or damaged cells and tissues will have broad impacts in biomedicine. Cells are the basic building blocks of life, and have incredible capacity; developing the ability to control cellular functions will enable rebuilding diseased and damaged tissues, replacing aged organs or directing cells to hunt and eliminate cancer. As individuals, cells have tremendous capability, and cells also have a natural ability to work together such that with proper control of this team effort the abilities of cells can be amplified; think of all 100 million single cells of your liver working together to help you digest food, distribute nutrients to the body, store energy and purify your blood. The functions of cells are regulated by circuits comprised of genes and proteins that can be controlled with biological switches. However, to date, a means of transferring a number of switches to cells and controlling them from a distance has not been described. This project seeks to develop biological remote controls with multiple switches that can be used to direct cells to perform functions and work together. These remote control modules that are called, “engineered organelles”, can be operated with magnetic fields. With magnetically controlled engineered organelles, rapid flashes of magnetic fields can warm the engineered organelle and turn on or off biological switches that control the cells function. The switches are designed with multiplexing capability so multiple functions can be controlled remotely. Engineered organelles are similarly designed for controlling multiple biological functions within a cell and direct them to work together. The multidisciplinary nature of the project offers unique training opportunities for the next generation of STEM researchers in the fields of bioengineering, physical sciences, synthetic biology and regenerative medicine. The research team invites students from underrepresented groups to participate in summer research via the successful ENSURE (EngiNeering Summer Undergraduate Research Experience) program at Michigan State University (MSU). In addition, the team develops workshops on engineered biological remote controls for high school science teachers to enable them to update their knowledge and refresh their curricula with state-of-the-art research. To enhance public understanding and appreciation of what engineered organelles can accomplish, the research team participates in the MSU Science Festival which caters to the greater Lansing area. Results from the project will be integrated into undergraduate course curricula and be incorporated into graduate courses in bioengineering and synthetic biology to teach the new principle of using engineered organelles to guide biology.This project aims at mimicking endosymbiogenesis and modify prokaryotic cells to create engineered magnetoendosymbionts (eMEs) that can be controlled using alternating magnetic fields (AMF). This involves engineering iron particle-coated or iron-containing prokaryotes, magnetotactic bacteria (MTB), to act as magnetothermally regulated pseudo-organelles. Magnetothermal control will activate eME to express mammalian transcription (txn) factors directed to host cell nuclei for controlling host gene expression. Genetic thermal switches will be engineered into MTB using optical reporter genes as a readout, and deliver reporters from eME in the cytoplasm to the nucleus of the host cell. The use of reporter genes will help guide development of eME that encode mammalian txn factors to enable reprogramming of differentiated macrophages into iPSCs and then into hepatocytes. A chemically (mannose) regulated iPSC-generating operon will be engineered expressing Oct3/4, Sox2, KLF4, c-Myc, txn factors for cellular reprogramming and a thermally regulated hepatocyte-generating operon expressing txn factors HNF4A, HNF1A. FoxA1 and FoxA3. Once demonstrated in the EES the operons will be transferred to MTB and used these eMEs to program macrophages to iPSCs and then to hepatocytes with magnetothermal control.This project is funded by the Systems and Synthetic Biology Cluster in the Division of Molecular and Cellular Biosciences and the Engineering Biology and Health Cluster in the Division of Chemical, Bioengineering, Environmental and Transport Systems.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.