Illuminating, profiling and manipulating perivascular plasticity in microvasculature
I worked as a postdoctoral fellow in Dr. Gary K Owens's laboratory to examine the smooth muscle cells utilizing unique smooth muscle lineage tracing mouse model and how they change in response to diet induced obesity (DIO).
Using scRNAseq, we identified 8 clusters (13-20) with predominantly perivascular derived cells, among a total of 20 clusters.
Importantly, diet induced obesity increased the abundance of cells in cluster 13 (“lymphatic muscle cells”) by more than 2-fold. Pathway analysis of cluster 13 revealed enrichment of not only classical smooth muscle cell contractile genes but also cardiac and skeletal muscle contractile genes relevant to lymphatic muscle cells.
Another lineage tracing model I have used was Myh11-Lgals3 dual lineage tracing system. This mouse model labels smooth muscle cells that have undergone activation of Lgals3 gene. Similar model systems can be devised to trace lymphatic smooth muscle cells as well, although that would be pretty expensive :)
Recently, I worked on human blood monocyte derived macrophage (hBMDM) culture and manipulations to test hypotheses relevant to lung diseases and protease Neutrophil Elastase. I have effectively simplified and implemented hBMDM generation method from healthy donor Buffy Coat (purchased from RedCross), Cystic Fibrosis and other lung disease patient samples.
This system is highly physiologically relevant as we are using human donors and easy to initiate and apply to various scientific questions. In addition hBMDM culture provides a valuable platform to test molecular, cellular mechanisms related to macrophage biology.
It is tempting to propose to test my obesity research relevant ideas on human blood monocyte derived macrophages. For example, Dr. Voynow's laboratory demonstrated that macrophages, similar to neutrophils, release Macrophage Extracellular Traps (METs) in response to neutrophil elastase, which is highly present in chronic lung inflammation.
One potential avenue could be to test, whether adipose tissue inflammation also causes MET release. This could be tested using conditions mimicking healthy, overweight and obese adipose tissue microenvironment and by quantifying and imaging MET release using PicoGreen assay and confocal microscopy.