Evaluation of Human Adipose Tissue Stromal Heterogeneity in Metabolic Disease Using Single Cell RNA-Seq
Technical Report,01 Aug 2016,31 Aug 2017
Beth Israel Deaconess Medical Center Boston United States
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We have developed a robust protocol to generate single cell transcriptional profiles from subcutaneous adipose tissue samples of both human and mouse subjects using Drop-seq, a recently developed, cost efficient method of highly parallel genome-wide expression profiling using nanoliter droplets. We have collected subcutaneous adipose tissue samples from greater than 15 human subjects and generated transcriptional profiles for over 46,000 individual cells. Additionally, we have generated profiles from approximately 10,000 cells from mouse subcutaneous and epididymal depots. In both data sets, our analyses demonstrate expression profiles can be used to cluster individual cells into distinct cell types in an unbiased fashioned. We identify most cell types known to be contained within adipose tissue SVF and have uncovered a number of cell types and subtypes that have not previously been described. We determine novel markers for many cell types with higher specificity than current markers and characterize many species and depot-specific cell types and markers. These data provide a comprehensive transcriptional atlas of subcutaneous adipose tissue cell types that will provide molecular handles to understanding and manipulating each cell types function. These results are hypothesis-generating, and provide the foundation for future studies that will 1 define functional roles for individual genes and cell types in development of obesity and insulin resistance and 2 examine novel targets against which we can design therapies to target specific pathogenic or health-promoting cell types.
- Medicine and Medical Research