Revolutionary Technology for Efficient Derivation of Human iPSCs with Messenger R

利用 Messenger R 高效衍生人类 iPSC 的革命性技术

• 批准号: 8590253
• 负责人: Jiwu Wang
• 金额: $ 19.99万
• 依托单位: ALLELE BIOTECHNOLOGY AND PHARMACEUTICALS
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8590253
• 关键词: Adult; Area; Benchmarking; Biological Assay; Biomedical Research; Cell Count; Cell Culture Techniques; Cell Density; Cells; Clinical; Custom; Dependence; Derivation procedure; Development; Disease; Disease model; Dose; Drug Addiction; Drug abuse; Elements; Event; Family; Functional disorder; Funding; Future; Generations; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Genome; Genomics; Goals; Healthcare Systems; Human; Immune; Immune response; In Vitro; Interferons; Marketing; Messenger RNA; Methamphetamine; Methods; Modeling; Modification; Molecular; Molecular Neurobiology; Mutation; National Institute of Drug Abuse; Neurons; Patients; Performance; Pharmaceutical Preparations; Population Density; Procedures; Process; Production; Property; Proteins; Protocols documentation; Publishing; Quality Control; Reagent; Recombinant Proteins; Reporting; Research; Research Personnel; Research Proposals; Retroviridae; Ribonucleotides; Running; Sendai virus; Small Business Innovation Research Grant; Social Problems; Solutions; Source; Specialist; Stem cells; System; Technology; Testing; Time; Variant; Viral; Virus; Withdrawal; Xeno; addiction; base; cell type; cost; design; dopamine transporter; dopaminergic neuron; immunogenicity; improved; induced pluripotent stem cell; neurotransmission; novel; programs; public health relevance; small molecule; substance abuser; tool; vesicular monoamine transporter

DESCRIPTION (provided by applicant): The goal of the proposed project is to develop a commercial kit that will allow drug addiction researchers with ongoing projects using cell-based models to be able to successfully and conveniently produce human induced pluripotent stem cells (iPSCs). Addiction to drugs destroys lives of substance abusers and their families, creating enormous social problems and exerting cost on the nation's health care system. Molecular mechanisms that contribute to the initiation and establishment of addiction are largely unknown, partially due to the lack of neural cell types from the patients. The technologies that made possible of human iPSCs opened the doors to new areas of direct interrogation of the pathophysiology of addiction by using patient-specific cell sources. Currently, the NIDA is funding projects that rely on iPS technologies to: 1) characterize dopaminergic neurotransmission function of neurons derived from iPSCs with genomic aberrations, such as chromosomal polymorphism regions containing dopamine transporter (DAT) and vesicular monoamine transporter (VMAT2) genes; 2) identify genetic variations that can be functionally correlated to addiction in patient cell-derived neurons via iPSCs; 3) make dopamine neurons from patient iPSCs and expose them to methamphetamine and withdrawal in order to analyze the electrophysiological properties of patient-specific neurons versus controls, etc. Even though it is considered a technology with great potential in studies of addiction or other diseases, reprogramming of adult human cells to iPSCs is still an inefficient, tedious and costly process to most researchers. The currently prevailing methods of reprogramming have additional problems such as alteration of genomes or introduction of viruses that can complicate the interpretation of results from the cell-based assays. This project aims to develop a turn-key tool kit that will enable many biomedical researchers to generate iPSCs not only with 10 to 100 folds higher efficiency than other methods, in less than 2 weeks, but also through a feeder-free, xeno-free and footprint-free procedure.

描述（由申请人提供）：拟议项目的目的是开发一种商业套件，该套件将允许使用基于细胞的模型进行正在进行的项目的吸毒研究人员能够成功，方便地生产人类诱导的多能干细胞（IPSC）。对毒品的成瘾破坏了滥用药物及其家人的生活，造成了巨大的社会问题并为国家的医疗保健系统施加成本。有助于成瘾的启动和建立的分子机制在很大程度上是未知的，部分原因是患者缺乏神经细胞类型。通过使用患者特异性细胞来源，使人IPSC成为可能的技术为直接询问成瘾的病理生理学的新领域打开了大门。目前，NIDA是依靠IPS技术的资金项目：1）表征来自具有基因组畸变的IPSC的多巴胺能神经传递功能，例如染色体多态性区域，含有多巴胺转运剂（DAT）和Vesicular Monoamine Monoamine Monoamine Transporter（Vmatportorter）（Vmat2）Geness（Vmat2）; 2）确定可以通过IPSCS与患者细胞衍生的神经元成瘾功能相关的遗传变异； 3）从患者IPSC中制作多巴胺神经元，并将其暴露于甲基苯丙胺和戒断中，以分析患者特异性神经元与对照组的电生理特性，即使它被认为是在成瘾或其他疾病研究中具有巨大潜力的技术，但对成人人类细胞的研究很大，对IPSC的重新编程仍然是对IPSC的重新处理，并且对大多数效率进行了成本，并且成本又是成本的。当前流行的重编程方法还有其他问题，例如基因组的改变或引入病毒，这些病毒可能使基于细胞的测定结果解释结果复杂化。该项目旨在开发一个交钥匙工具套件，该工具包将使许多生物医学研究人员在不到2周内的效率上，不仅可以比其他方法高10到100倍，而且还可以通过无馈物，无XENO和足迹的免费程序来生成IPSC。