Exosomal miRNA Profiles of Endothelial Cells and Pericytes in Pulmonary Niche on an Organ-on-a-chip Model



Description

Mehmet O. Ozen1,2, Elya A. Shamskhou2,3,4, Abinaya Nathan2,3,4, Ananya Chakraborty2,3,4, Ke Yuan2,3,4, Mark E. Orcholski2,3,4, Vinicio A. de Jesus Perez2,3,4 and Utkan Demirci1,2

1Dept of Radiology, Canary Center for Early Cancer Detection, 2Stanford Cardiovascular Institute, 3Division of Pulmonary & Critical Care Medicine, 4Wall Center for Pulmonary Vascular Research, Stanford University, CA.

Pulmonary arterial hypertension (PAH) is a disorder effecting pulmonary circulation. PAH is a result of reduced blood flow due to partial or entire elimination of microvessels because of attenuated recruitment of pericytes (Pc), in addition to genetic aberrations and environmental factors including endothelial cell (EC) death. To understand the cross-talk between cells, extracellular vesicles (EVs), are being increasingly investigated since their shedding from cells occurs in response to global and local changes in their environment and can be used to monitor cargo molecules carried by them, in response to these conditions in vitro and in vivo. Exosomes involve in this communication, which will impact the recipient cell’s fate, packed with microRNAs (miRNAs).

Motivated by this strong relationship between exosomes and their regulatory behaviors, we hypothesize that shear stress on ECs can trigger exosome mediated Pc recruitment during PAH progression through WNT family. The objective of this work is to determine molecular changes during Pc recruitment of ECs leading to PAH via deciphering exosomal miRNA profiling.

We cultured patient derived healthy and PAH EC and Pc cell lines in static conditions as monocultures to draw a baseline of exosomal miRNA profiles. We profiled secreted exosomes from these cultures. And collected RNA for sequencing, where PAH EC and Pc cultures had more exosomes and exosomal RNAs, compared to healthy donor cultures (n=3). We will report on cultures in static co-culture models, and on an in-house developed dynamic organ-on-a-chip model to decode their exosomal cross-talk through exosomal miRNA profiling.

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