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Reprogramming Non-myocytes to Cardiomyocytes in vivo

在体内将非心肌细胞重编程为心肌细胞

基本信息

批准号：
9493517
负责人：
RICHARD T LEE
金额：
$ 42.78万
依托单位：
HARVARD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9493517
关键词：
Animal Experiments Animals Biology Brain Stem Cardiac Myocytes Cell Lineage Cells Complement Congestive Heart Failure Creatine Effector Cell Engraftment Epidemic Extravasation Fibroblasts Future Generations Genetic Heart Heart failure Human Human Volunteers Image Injury Isotope Labeling Isotopes Label Laboratories Mammals Mass Spectrum Analysis Metabolic Methods Mus Muscle Cells Myocardial Myocardial Infarction Natural regeneration Organ Other Genetics Patients Phosphocreatine Physiologic pulse Publications Research Personnel Resolution Skeletal Muscle Stem cells Techniques Technology Testing Therapeutic Thymidine Tissues Tracer United States base cardiogenesis cell type experimental study heart cell heart function imaging approach improved in vivo interest mouse model new technology non-genetic novel strategies progenitor promoter recombinase stable isotope stem cell biology stem cell differentiation stem cell niche transdifferentiation uptake

项目摘要

There is intense interest in approaches to generating new cardiomyocytes, including not only through laboratory generation of cardiomyocytes but also by promoting cardiomyocyte formation therapeutically through endogenous cardiogenesis. The endogenous cardiogenesis approaches would not need delivery of cells with issues of engraftment and survival, and thus could have advantages. One of these exciting endogenous cardiogenesis approaches is direct reprogramming of non-cardiac cells to cardiomyocyte. To study reprogramming in vivo, the inducible cre approach in mice is the most widely used method for genetic fate-mapping of cells. However, inducible cre and other genetic lineage mapping approaches may be limited by even very transient leakage of promoters or spontaneous recombinase activity in the absence of the inducer molecule, and these studies can currently only be performed in mice. To gain confidence in the study of endogenous cardiogenesis, approaches that complement genetic fate mapping could provide compelling evidence that our field is headed toward the best regeneration strategy. We have now developed an entirely new approach to marking the identity of cells in vivo using non-radioactive isotopes in a cell-specific metabolic compound. This “Metabolic Fate-Mapping” approach utilizes an isotope-enriched metabolic tracer, specifically creatine, which is taken up by muscle cells, phosphorylated, and utilized in the cytoplasmic phosphocreatine shuttle. Cellular uptake of creatine by muscle cells is rapid, while subsequent turnover of creatine is slow, making it a suitable metabolic label for myocytes. Cells that are creatine-positive can then be identified via Multi-Isotope Imaging Mass Spectrometry (MIMS), a high resolution approach that we have adapted for myocardial biology. We have previously demonstrated usage of labeled thymidine in vivo to demonstrate rare proliferation of cardiomyocytes over months, and we have also demonstrated the stable isotope imaging approach in human volunteers. We will use this new Metabolic Fate-Mapping approach to cell lineage mapping along with an inducible cardiomyocyte cre mouse and an inducible fibroblast cre mouse to study reprogramming of non-myocytes to cardiomyocytes in vivo. Unlike genetic fate-mapping strategies, the stable isotope lineage mapping approach is amenable to any species on any genetic background, and this will enable future large animal experiments of reprogramming. Finally, because this approach can be applied with stable, non-radioactive isotopes such as 13C and 15N, which have been widely used in humans and are regarded by the FDA as safe, studies of human regeneration in diverse tissues could be enabled with Metabolic Fate- Mapping by identification of cellular labels specific to cell type.

人们对产生新心肌细胞的方法有着浓厚的兴趣，不仅包括通过