PMID- 34499621 OWN - NLM STAT- MEDLINE DCOM- 20220311 LR - 20220311 IS - 2379-3708 (Electronic) IS - 2379-3708 (Linking) VI - 6 IP - 21 DP - 2021 Nov 8 TI - Complement-containing small extracellular vesicles from adventitial fibroblasts induce proinflammatory and metabolic reprogramming in macrophages. LID - e148382 [pii] LID - 10.1172/jci.insight.148382 [doi] AB - Pulmonary hypertension (PH) is a severe cardiopulmonary disease characterized by complement-dependent, fibroblast-induced perivascular accumulation and proinflammatory activation of macrophages. We hypothesized that, in PH, nanoscale-sized small extracellular vesicles (sEVs), released by perivascular/adventitial fibroblasts, are critical mediators of complement-dependent proinflammatory activation of macrophages. Pulmonary adventitial fibroblasts were isolated from calves with severe PH (PH-Fibs) and age-matched controls (CO-Fibs). PH-Fibs exhibited increased secretion of sEVs, compared with CO-Fibs, and sEV biological activity was tested on mouse and bovine bone marrow-derived macrophages (BMDMs) and showed similar responses. Compared with sEVs derived from CO-Fibs, sEVs derived from PH-Fibs (PH-Fib-sEVs) induced augmented expression of proinflammatory cytokines/chemokines and metabolic genes in BMDMs. Pharmacological blockade of exosome release from PH-Fibs resulted in significant attenuation of proinflammatory activation of BMDMs. "Bottom-up" proteomic analyses revealed significant enrichment of complement and coagulation cascades in PH-Fib-sEVs, including augmented expression of the complement component C3. We therefore examined whether the PH-Fib-sEV-mediated proinflammatory activation of BMDMs was complement C3 dependent. Treatment of PH-Fibs with siC3-RNA significantly attenuated the capacity of PH-Fib-sEVs for proinflammatory activation of BMDMs. PH-Fib-sEVs mediated proglycolytic alterations and complement-dependent activation of macrophages toward a proinflammatory phenotype, as confirmed by metabolomic studies. Thus, fibroblast-released sEVs served as critical mediators of complement-induced perivascular/microenvironmental inflammation in PH. FAU - Kumar, Sushil AU - Kumar S AD - Cardiovascular Pulmonary Research Laboratories, Departments of Pediatrics and Medicine, School of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. FAU - Frid, Maria G AU - Frid MG AD - Cardiovascular Pulmonary Research Laboratories, Departments of Pediatrics and Medicine, School of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. FAU - Zhang, Hui AU - Zhang H AD - Cardiovascular Pulmonary Research Laboratories, Departments of Pediatrics and Medicine, School of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. FAU - Li, Min AU - Li M AD - Cardiovascular Pulmonary Research Laboratories, Departments of Pediatrics and Medicine, School of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. FAU - Riddle, Suzette AU - Riddle S AD - Cardiovascular Pulmonary Research Laboratories, Departments of Pediatrics and Medicine, School of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. FAU - Brown, R Dale AU - Brown RD AD - Cardiovascular Pulmonary Research Laboratories, Departments of Pediatrics and Medicine, School of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. FAU - Yadav, Subhash Chandra AU - Yadav SC AD - Department of Anatomy, All India Institute of Medical Sciences, New Delhi, India. FAU - Roy, Micaela K AU - Roy MK AD - Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. FAU - Dzieciatkowska, Monika E AU - Dzieciatkowska ME AD - Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. FAU - D'Alessandro, Angelo AU - D'Alessandro A AD - Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. FAU - Hansen, Kirk C AU - Hansen KC AD - Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. FAU - Stenmark, Kurt R AU - Stenmark KR AD - Cardiovascular Pulmonary Research Laboratories, Departments of Pediatrics and Medicine, School of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA. LA - eng GR - P01 HL014985/HL/NHLBI NIH HHS/United States GR - P01 HL152961/HL/NHLBI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20211108 PL - United States TA - JCI Insight JT - JCI insight JID - 101676073 SB - IM MH - Animals MH - Cellular Reprogramming/*genetics MH - Disease Models, Animal MH - Extracellular Vesicles/*genetics MH - Fibroblasts/*metabolism MH - Humans MH - Hypertension, Pulmonary/*physiopathology MH - Macrophages/*metabolism MH - Mice PMC - PMC8663554 OTO - NOTNLM OT - Cardiovascular disease OT - Hypertension OT - Inflammation OT - Macrophages OT - Pulmonology COIS- Conflict of interest: AD is the founder and chief scientific officer of Omix Technologies Inc. and Altis Bioscience LLC. KRS is a consultant for Actelion Pharmaceuticals US Inc. EDAT- 2021/09/10 06:00 MHDA- 2022/03/12 06:00 PMCR- 2021/11/08 CRDT- 2021/09/09 17:17 PHST- 2021/02/05 00:00 [received] PHST- 2021/09/08 00:00 [accepted] PHST- 2021/09/10 06:00 [pubmed] PHST- 2022/03/12 06:00 [medline] PHST- 2021/09/09 17:17 [entrez] PHST- 2021/11/08 00:00 [pmc-release] AID - e148382 [pii] AID - 148382 [pii] AID - 10.1172/jci.insight.148382 [doi] PST - epublish SO - JCI Insight. 2021 Nov 8;6(21):e148382. doi: 10.1172/jci.insight.148382.