Molecular Imaging for Detection of Synthetic Biology Circuits, Oscillators and Toggle Switches in Regenerative Medicine

用于检测再生医学中的合成生物学电路、振荡器和拨动开关的分子成像

• 批准号: 10176612
• 负责人: Assaf A Gilad
• 金额: $ 33.24万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10176612
• 关键词: Adipocytes; Amplifiers; Animals; Biological; Biological Products; Biomedical Engineering; Brain; Calcium; Catfish; Cell Culture System; Cell Differentiation process; Cell Fate Control; Cell membrane; Cell surface; Cells; Chronic; Clinical Paths; Cultured Cells; Data; Devices; Diabetes Mellitus; Diagnostic; Disease; Dopamine; Electromagnetic Fields; Electromagnetics; Electronics; Engineered Gene; Engineering; Eukaryotic Cell; FOS gene; Future; Gene Expression; Gene Family; Genes; Genetic; Glass; Goals; Herpesvirus 1; Hormone secretion; Hormones; Human; Image; Immediate-Early Genes; In Situ; Induced pluripotent stem cell derived neurons; Injections; Insulin; Lab-On-A-Chips; Lead; Ligand Binding; Magnetic Resonance Imaging; Malignant Neoplasms; Mammalian Cell; Membrane Proteins; Microfluidic Microchips; Microscopy; Molds; Monitor; Mus; Neurons; Neurotransmitters; Nuclear; Oocytes; Optical reporter; Outcome; Parkinson Disease; Pathway interactions; Pattern; Periodicity; Pharmaceutical Preparations; Population; Positron-Emission Tomography; Process; Production; Proteins; Pump; Regenerative Medicine; Reporter Genes; Rodent; Specificity; Surface; System; Technology; Testing; Therapeutic; Thyrotropin; Time; Tissues; Translating; Transplantation; Visualization; Wireless Technology; cancer therapy; clinical translation; controlled release; cytokine; deoxyribonucleoside kinases; design; ferrite; glucagon-like peptide 1; imaging detection; in vivo; in vivo imaging; innovation; member; microdevice; microorganism; molecular imaging; nanoparticle; neurotropic; novel; pluripotency; promoter; remote control; stem cell fate; stem cell therapy; stem cells; success; synthetic biology; therapeutic gene; thymidine kinase 1; tissue regeneration; tool

Harnessing the power of therapeutic stem cells holds great promises for the discovery of new treatments to many chronic and uncured diseases. Despite few milestone studies, the overall success has been limited. Studies around the world showed that it is safe to use such cells, however, those cells must be first engineered so they can preformed the desired task, similar to the way that electronic component are engineered for preforming computational tasks. In synthetic biology, much like in electronics, we can use biological components to build a circuit that can preform a very specific function. Instead of using electronic parts we can used biological parts or “bio-parts” that interact with the outmost specificity one with another. Here we seek to use three “bio-parts” that will be sufficient to control stem cell activity and fate in situ. This is extremely important because to date, there is no way to remote control cells, inside the body, without invasively penetrating the body tissue. To that end, we will construct a biological circuit inside stem cells. For the “switch” we will use a novel protein, encoded by a gene that was discovered in our lab. This novel protein can sense and respond to an external electromagnetic field, i.e., can be switch on and off by a static magnet or an electronic device. Once it is activated it lead to release of calcium. A calcium sensitive promoter is used as an “amplifier”. The third bio-part is a reporter gene that we will use for visualization of the activity and in the future can be replace by a therapeutic gene. In the first aim we will establish a cell culture system that can express all the bio-parts of the “device” in stem cells. Specifically in iPSCs-derived neurons and adipocytes derived stem cells (ADSCs). In the second Aim we will test two modes of cell activation. The first one is with ferromagnetic (not to be confused with paramagnetic) nanoparticles (FMNPs). Those FMNPs will be functionalized with ligands that bind to the stem cell membrane. This will result in continues activation (“off”→”on”) of genes. This is relevant to cases where we need to induce cell differentiation either to specific linage or even to reverse pluripotency. The second mode of activation is oscillations, induced by an electromagnet, that can we alternately switch on and off for brief time periods. This can create an oscillatory pattern that can result in cyclic production of metabolites, hormones and drugs. This is relevant, for example, for disease such as diabetes where it can replace the daily insulin injection or in cancer where are controlled drug release is required. In the third Aim, we will transplant the bioengineered stem cells in the rodent brain and will monitor both the Toggle switch and the oscillator in vivo using a reporter gene engineered for both MRI and PET. Thus, this innovative study is a unique approach to transition synthetic biology strategies from microorganism to mammalian system with clear path for clinical translation.

利用治疗性干细胞的力量具有发现新疗法的巨大承诺 许多慢性疾病。尽管有很少的里程碑研究，但总体成功还是有限的。 世界各地的研究表明，使用此类细胞是安全的，但是，这些单元必须首先进行设计 因此，他们可以预定所需的任务，类似于电子组件的设计方式 预制计算任务。在合成生物学中，就像电子产品一样，我们可以使用生物学 组件要构建可以预先形成非常特定功能的电路。而不是使用电子零件，我们可以 使用的生物学零件或“生物零件”与最大特异性相互作用。我们在这里寻求 使用三个“生物零件”，足以控制干细胞活性并原位命运。这是非常 重要的是因为迄今为止，没有远程控制单元在体内，而没有侵入性 穿透身体组织。 为此，我们将在干细胞内构建生物回路。对于“开关”，我们将使用一种新型蛋白质， 由在我们实验室中发现的基因编码。这种新颖的蛋白质可以感知并回应外部 可以通过静态磁铁或电子设备打开和关闭电子场，即可以打开和关闭电子场。一旦是 激活它导致钙的释放。钙敏感启动子用作“放大器”。第三个生物部分 是一个记者基因，我们将用于可视化活动，将来可以用 治疗基因。 在第一个目标中，我们将建立一个可以表达茎中“设备”的所有生物零件的细胞培养系统 细胞。特别是在IPSC衍生的神经元和脂肪细胞衍生的干细胞（ADSC）中。在第二个目标中 将测试两种细胞激活模式。第一个是用铁磁磁性的（不要与顺磁相混淆） 纳米颗粒（FMNP）。这些FMNP将用与干细胞膜结合的配体功能化。 这将导致基因的继续激活（“ OFF”→“”）。这与我们需要诱导的情况有关 细胞分化与特定的线条，甚至逆转多能。第二种激活方式是 由电子磁体引起的振荡，我们可以在短时间内打开和关闭。这 可以创建一种振荡模式，可以导致代谢物，激素和药物的环状产生。这 例如，与糖尿病等疾病相关，可以替代每日胰岛素注射或 癌症是需要控制药物释放的地方。在第三个目标中，我们将移植生物工程的 啮齿动物大脑中的干细胞，将使用记者在体内监测开关和振荡器 基因为MRI和PET设计。那是这项创新研究是一种过渡合成的独特方法 从微生物到哺乳动物系统的生物学策略，具有清晰的临床翻译路径。

项目成果

Development of a Synthetic Biosensor for Chemical Exchange MRI Utilizing In Silico Optimized Peptides.

利用计算机优化的肽开发用于化学交换 MRI 的合成生物传感器。

• DOI: 10.1101/2023.03.08.531737
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Fillion,AdamJ;Bricco,AlexanderR;Lee,HarveyD;Korenchan,David;Farrar,ChristianT;Gilad,AssafA
• 通讯作者: Gilad,AssafA

Bioengineering of Genetically Encoded Gene Promoter Repressed by the Flavonoid Apigenin for Constructing Intracellular Sensor for Molecular Events.

• DOI: 10.3390/bios11050137
• 发表时间: 2021-04-28
• 期刊: Biosensors
• 作者: Desmet NM;Dhusia K;Qi W;Doseff AI;Bhattacharya S;Gilad AA
• 通讯作者: Gilad AA

Proposed three-phenylalanine motif involved in magnetoreception signalling of an Actinopterygii protein expressed in mammalian cells.

• DOI: 10.1098/rsob.230019
• 发表时间: 2023-11
• 期刊: Open biology
• 影响因子: 5.8

A Novel Protein for the Bioremediation of Gadolinium Waste.

用于钆废物生物修复的新型蛋白质。

• DOI: 10.1101/2023.01.05.522788
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Lee,HarveyD;Grady,ConnorJ;Krell,Katie;Strebeck,Cooper;Good,NathanM;Martinez-Gomez,NCecilia;Gilad,AssafA
• 通讯作者: Gilad,AssafA

Non-invasive neuromodulation using rTMS and the electromagnetic-perceptive gene (EPG) facilitates plasticity after nerve injury.

• DOI: 10.1016/j.brs.2020.10.006
• 发表时间: 2020-11
• 期刊: Brain stimulation
• 影响因子: 7.7
• 作者: Cywiak C;Ashbaugh RC;Metto AC;Udpa L;Qian C;Gilad AA;Reimers M;Zhong M;Pelled G
• 通讯作者: Pelled G