Expressing humanized bacterial luciferase in stem cells: Moving beyond firefly luciferase to expand the informational capacity of animal models for regenerative medicine

在干细胞中表达人源化细菌荧光素酶：超越萤火虫荧光素酶，扩大再生医学动物模型的信息能力

• 批准号: 8981338
• 负责人: Dan Morrison
• 金额: $ 15万
• 依托单位: 490 BIOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8981338
• 关键词: Address; Adipose tissue; Animal Experiments; Animal Model; Animal Welfare; Animals; Bacterial Luciferases; Biological Models; Bioluminescence; Businesses; Cell Line; Cell Lineage; Cell Therapy; Cell physiology; Cells; Chemicals; Clinical Trials; Color; Complement; Data; Data Collection; Dependence; Discrimination; Disease; Electroporation; Engineering; Engraftment; Evaluation; Fireflies; Firefly Luciferases; Fluorescence; Gene Delivery; Gene Transfer; Genes; Genome; Green Fluorescent Proteins; Healed; Health; Human; Image; Imaging technology; Implant; In Vitro; Injection of therapeutic agent; Legal patent; Life; Light; Longevity; Luc Gene; Luciferases; Marketing; Measurement; Mediating; Medical center; Mesenchymal Stem Cells; Methods; Monitor; Morphologic artifacts; Noise; Organ; Osteogenesis; Outcome; Output; Performance; Phase; Phenotype; Physiological; Play; Procedures; Production; Protocols documentation; Reagent; Regenerative Medicine; Renilla; Reporter; Reporter Genes; Research; Research Personnel; Role; Safety; Signal Transduction; Small Business Technology Transfer Research; Stem cells; Subfamily lentivirinae; Surface; Techniques; Technology; Technology Transfer; Tennessee; Therapeutic; Therapeutic Effect; Time; Tissue Engineering; Tissues; Transduction Gene; Transfection; Transplantation; Transposase; United States National Institutes of Health; Universities; Variant; base; bioimaging; clinical practice; consumer demand; data acquisition; expression vector; healing; human stem cells; imaging platform; implantation; improved; in vivo; in vivo imaging; innovation; instrumentation; light emission; matrigel; migration; mouse model; novel; public health relevance; research and development; response; scaffold; self-renewal; stem cell fate; stem cell therapy; treatment strategy; two-dimensional

 DESCRIPTION (provided by applicant): This Small Business Technology Transfer (STTR) Phase I project proposes to develop autonomously bioluminescent human stem cells for continuous, reagent-free, and real-time bioimaging to address the National Institutes of Health's request for new techniques for non-invasive, long-term tracking of stem cell survivability, engraftment, and migration following in vivo implantation. The ability of stem cells to self- renew and differentiate into other cell lineages has emerged as a valuable therapeutic approach to functionally heal previously irreparable tissues and organs. However, for the regenerative medicine field to effectively transition toward translational and clinical practice outcomes, a strong dependence on animal models will be required to fully understand the capabilities and complexities of stem cells. 490 BioTech proposes to expand the informational capacity of animal models by creating stem cell lines that self- generate bioluminescent light via expression of a 'humanized' bacterial luciferase, thereby enabling stem cells to be continuously imaged throughout their lifetime as they physiologically function within their animal host. This differs significantly from the current market of bioluminescent imaging technologies that rely on a firefly luciferase gene construct that must be provided with a chemical substrate to activate its light emission response, resulting in only marginally informative single time point snapshots of cell function in tandem with repetitive animal injections that invoke unknown and potentially interfering interactions and adversely effects animal welfare. In partnership with the University o Tennessee Medical Center, the specific objectives of this R&D effort are to develop piggyBac transposition and lentiviral transduction methods for streamlined integration of the bioluminescent phenotype into adipose-derived mesenchymal stem cell lines followed by performance evaluation in in vitro 3D scaffolds and in vivo mouse models to demonstrate proficiency toward uninterrupted imaging and enriched data flows that far exceed that of existing firefly luciferase methods. With no change in instrumentation or fundamental bioluminescent protocols necessary, researchers can seamlessly transition from firefly luciferase to 490 BioTech's humanized bacterial luciferase technology to advance their in vivo experimental R&D to more informative endpoints with fewer animals required. The contribution of this innovative imaging platform to the field of regenerative medicine will provide more physiologically relevant and representative data critical to predicting the efficacy and safety of treatment strategies as they precede to clinical trials.