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为发现新疗法带来了巨大希望 许多慢性和未治愈的疾病。尽管里程碑式的研究很少，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，但必须首先对这些细胞进行工程改造 这样他们就可以执行所需的任务，类似于电子元件的设计方式 执行计算任务。在合成生物学中，就像在电子学中一样，我们可以使用生物 组件来构建可以执行非常特定功能的电路。我们可以不使用电子零件 使用生物部件或“生物部件”，它们彼此之间具有最外在的特异性。在这里我们寻求 使用三个足以原位控制干细胞活性和命运的“生物部件”。这是极其 重要的是，到目前为止，还没有办法在不侵入性的情况下远程控制体内的细胞 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， 由我们实验室发现的基因编码。这种新颖的蛋白质可以感知并响应外部 电磁场，即可以通过静磁铁或电子设备打开和关闭。一旦是 激活它会导致钙的释放。钙敏感启动子被用作“放大器”。生物第三部分 是一个报告基因，我们将使用它来可视化活动，将来可以用 治疗基因。 第一个目标是建立一个细胞培养系统，可以在干细胞中表达“装置”的所有生物部分。 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，我们 将测试两种细胞激活模式。第一个是铁磁（不要与顺磁混淆） 纳米颗粒（FMNP）。这些 FMNP 将通过与干细胞膜结合的配体进行功能化。 这将导致基因持续激活（“关”→“开”）。这与我们需要诱导的情况相关 细胞分化为特定谱系，甚至逆转多能性。第二种激活方式是 由电磁体引起的振荡，我们可以在短时间内交替打开和关闭。这 可以产生振荡模式，导致代谢物、激素和药物的循环产生。这 例如，对于糖尿病等疾病，它可以代替每日注射胰岛素，或者在 需要控制药物释放的癌症。第三个目标，我们将移植生物工程 啮齿动物大脑中的干细胞，将使用报告器监测体内的切换开关和振荡器 针对 MRI 和 PET 进行基因工程改造。因此，这项创新研究是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，具有明确的临床转化路径。

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