S.OWP3.09 = LBF05.Alterations within the miRNA cargo of HIV-infected macrophage-derived extracellular vesicles promote pulmonary smooth muscle proliferation Himanshu Sharma; Navneet K. Dhillon; Mahendran Chinnappan; Stuti Agarwal; Pranjali Dalvi University of Kansas Healthcare Center, Kansas City, USABackground: Our preceding research consistently demonstrate enhanced pulmonary vascular remodelling in HIV-1-infected people, simian immunodeficiency virus-infected macaques and HIV-transgenic rats exposed to illicit drugs. We reported considerable perivascular inflammation around the remodelled vessels; on the other hand, the exact function of theseThursday, 03 Mayinflammatory cells in the development of pulmonary vascular remodelling remains unknown. Our current in vitro findings revealed that HIV-1infected and cocaine (H+C)-treated human monocyte-derived macrophages (MDMs) secrete larger quantity of extracellular vesicles (EVs) compared to mono-treatments. We now hypothesize that dual hit of HIV-1 and cocaine may alter miRNA cargo of macrophage-derived EVs in a way that promotes smooth muscle proliferation. Methods: EVs were isolated by ultracentrifugation from supernatants collected from HIV-1Bal infected and cocaine (H+C)-treated MDMs at four days post-infection and applied for evaluation of miRNA expression. We chosen five PI3/AKT signalling-associated miRNAs for evaluation based on small RNA seq findings. Human primary pulmonary arterial smooth muscle cells (HPASMCs) were treated with EVs or MDM supernatants followed by proliferation assay. Final Mineralocorticoid Receptor Proteins Accession results: We observed significant enhance within the expression of miR130a and 27a in EVs derived from H+C-treated MDMs in comparison to untreated group with significantly elevated miR130a levels in H+C EVswhen in comparison to only HIV-1 or only cocaine mono-treatments. Examining the effect of EVs on HPASMCs showed that both mRNA and protein expression of PTEN, TSC-1 and TSC-2 had been substantially reduced in cells exposed to H+C EVs and this corresponded to increased activation of PI3K-AKT signalling and proliferation of smooth muscle cells. Additionally, inhibition of miRNA130a in HPASMCs with antagomir-130a blocked the EV-mediated lower in PTEN mRNA expression, thus confirming direct function of miR130a in modulating PTEN expression and hence potentiating the PI3/AKT signalling-mediated cell proliferation. Summary/conclusion: In summary, our findings suggest a pivotal part of EVs derived from HIV-1-infected and cocaine-treated macrophages in modulating pulmonary smooth proliferation and this may play a important part in development of HIV-associated pulmonary arterial hypertension. Funding: R01DA034542, R01DA042715 and R01HL129875.ISEV 2018 abstract bookBlood EV’s Roadmap Auditorium 16:307:15 Meet the Protein tyrosine phosphatases Proteins Recombinant Proteins Journal Editors Room 5 Chair: Hector Peinado; Marca Wauben 16:307:15 Meet the National Societies and Outreach Strategies Space 6 Chair: Isabel Guerrero 18:300:00 Meet the Specialist Session: RNA and EVs: What, Why and Where of their Interaction Auditorium Chair: Andrew Hill 18:300:00 Meet the Specialist Session: Regulatory Elements of EVs to Attain the Clinic Area 5 Chair: Susmita Sahoo 18:300:Thursday, 03 MayPoster Session PT01: EVs, Pathogens and Cross-organism Communication Parasitic Infections Chairs: Martin Jaular Lorena; Elena Mercade Location: Exhibit Hall 17:158:PT01.GP63-enriched Leishmania exosomes concur to cutaneous leishmaniasis improvement Alonso da Silva Lira Filho; Pauline Clement; Martin Olivier McGill University, Montreal, CanadaBackgr.
