Transcriptomic, metabolomic, proteomic and epigenetic studies have greatly enhanced our understanding of cell and organ biology. However, the vast majority of these studies have been conducted in bulk tissues derived from humans or animals with each tissue type comprised of multiple different cells, each with its own unique transcriptomic, metabolomic, proteomic and epigenetic profile. The huge body of knowledge gained from these studies has suffered, nevertheless, from a lack of cell-type-specific fidelity making it nearly impossible to attribute biological features and signatures to their cell-type of origin. In addition, significant differences in transcriptomic, metabolomic, proteomic and epigenetic profiles of different cell types have been missed because they have been overshadowed by profiles in other cell types, and, perhaps more significantly, heterogeneous high and low expression profiles of significance to function and disease are obscured by buffering to their mean when different cells of different types are homogenized together. These limitations reduce the scope of discoveries, including those of high significance to our Veterans, since significant disease-specific changes in transcriptomic, metabolomic, proteomic and epigenetic profiles may not be appreciable in bulk tissue homogenates that average expression levels across cell types. To mitigate these limitations the established, funded JJ Peters VA scientists participating in this ShEEP request (10 Major Users and 3 Minor Users) have begun to shift aspects of their research to cell-type-specific studies using high throughput fluorescence activated cell sorting (FACS). These efforts have been limited and significantly hampered by the lack of local access to a sophisticated high-throughput multi-cell type sorting capable FACS instrument. Hindrances have included having to transport fragile and precious samples across 7.5 miles to the affiliate core facilities; competition for instrument time with hundreds of affiliate core facility users and inability to FACS process fresh specimens from Veteran donors. To overcome these limitations and hindrances we propose to use ShEEP and already secured in-kind resources to purchase and operate a state-of-the-art FACS instrument of sufficient sophistication to permit its use locally at the JJ Peters VAMC in the context of the funded Veteran-centric studies of high priority diseases and conditions to the VA, such as suicide, TBI, SCI, PTSD, Alzheimer?s disease, serious mental illness and cancer.
The huge body of knowledge gained from these studies has suffered, nevertheless, from a lack of cell-type-specific fidelity making it nearly impossible to attribute biological features and signatures to their cell-type of origin. These limitations reduce the scope of discoveries, including those of high significance to our Veterans (e.g., suicide, TBI, SCI, PTSD, Alzheimer?s disease, serious mental illness and cancer), since significant disease-specific changes in cell- specific profiles may not be appreciable in bulk tissue homogenates that average expression levels across cell types. To overcome these limitations the established, funded JJ Peters VA scientists participating in this ShEEP request (10 Major Users and 3 Minor Users) propose to ShEEP resources to purchase and operate a state-of-the-art FACS instrument of sufficient sophistication to permit its use locally at the JJ Peters VAMC.
