TY - JOUR
T1 - 3’UTR shortening of HAS2 promotes hyaluronan hyper-synthesis and bioenergetic dysfunction in pulmonary hypertension
AU - Tseng, Victor
AU - Collum, Scott D.
AU - Allawzi, Ayed
AU - Crotty, Kathryn
AU - Yeligar, Samantha
AU - Trammell, Aaron
AU - Ryan Smith, M.
AU - Kang, Bum Yong
AU - Sutliff, Roy L.
AU - Ingram, Jennifer L.
AU - Jyothula, Soma S.S.K.
AU - Thandavarayan, Rajarajan A.
AU - Huang, Howard J.
AU - Nozik, Eva S.
AU - Wagner, Eric J.
AU - Michael Hart, C.
AU - Karmouty-Quintana, Harry
N1 - Funding Information:
We thank Dr. Dean Jones for helpful commentary and discussion; Drs. Viranuj Sueblinvong and Nicholas Maurice for assistance with fibroblast and epithelial cell culture experiments; Dr. Clare Prohaska, Jennifer Kleinhenz, Tammy Murphy, Sarah Chang, Jing Ma, David Michael, and Cory Sylber for valuable technical assistance; and Ivy McDermott for assistance with flow cytometry. VT, SC, CMH, and HKQ designed the experiments, analyzed the data, and interpreted the findings. VT and SC executed the experiments and collected data. VT composed the manuscript. SC, CMH, and HKQ made major contributions and revised the manuscript. AA, KC, TM, and SY performed and analyzed flow cytometry experiments for cell-specific HAS expression and apoptosis analysis. VT and RLS performed vascular contractility studies. ABYK assisted with RT-qPCR quantitation of HA-regulating genes in the SU-HYP mouse model. MRS and AWT assisted with bioenergetics studies and scientific discussions. JLI provided analysis and scientific discussions pertaining to murine HAS2 overexpression. SSJ, RAT, and HJH assisted with procurement and analysis of human IPAH tissues. ENG, RLS, CMH, and HKQ contributed to scientific discussions, experimental design, manuscript editing, and extensive data interpretation.
Funding Information:
This work was supported by NHLBI 5T32HL116271 and the Actelion Pharmaceuticals Young Investigator Award to VT; VA Merit Review Award 1I01BX004263 to CMH; NHLBI 5R01HL102167 to CMH and RLS; NHLBI 5R01HL133053 to BYK, NHLBI 5R01HL138510 to HKQ; NHLBI 5R01HL086680 and 1R35HL139726-01 to ENG. Portions of this work were previously presented at scientific symposia.
Publisher Copyright:
© 2022 The Authors
PY - 2022/8
Y1 - 2022/8
N2 - Pulmonary hypertension (PH) comprises a diverse group of disorders that share a common pathway of pulmonary vascular remodeling leading to right ventricular failure. Development of anti-remodeling strategies is an emerging frontier in PH therapeutics that requires a greater understanding of the interactions between vascular wall cells and their extracellular matrices. The ubiquitous matrix glycan, hyaluronan (HA), is markedly elevated in lungs from patients and experimental models with PH. Herein, we identified HA synthase-2 (HAS2) in the pulmonary artery smooth muscle cell (PASMC) layer as a predominant locus of HA dysregulation. HA upregulation involves depletion of NUDT21, a master regulator of alternative polyadenylation, resulting in 3’UTR shortening and hyper-expression of HAS2. The ensuing increase of HAS2 and hyper-synthesis of HA promoted bioenergetic dysfunction of PASMC characterized by impaired mitochondrial oxidative capacity and a glycolytic shift. The resulting HA accumulation stimulated pro-remodeling phenotypes such as cell proliferation, migration, apoptosis-resistance, and stimulated pulmonary artery contractility. Transgenic mice, mimicking HAS2 hyper-synthesis in smooth muscle cells, developed spontaneous PH, whereas targeted deletion of HAS2 prevented experimental PH. Pharmacological blockade of HAS2 restored normal bioenergetics in PASMC, ameliorated cell remodeling phenotypes, and reversed experimental PH in vivo. In summary, our results uncover a novel mechanism of HA hyper-synthesis and downstream effects on pulmonary vascular cell metabolism and remodeling.
AB - Pulmonary hypertension (PH) comprises a diverse group of disorders that share a common pathway of pulmonary vascular remodeling leading to right ventricular failure. Development of anti-remodeling strategies is an emerging frontier in PH therapeutics that requires a greater understanding of the interactions between vascular wall cells and their extracellular matrices. The ubiquitous matrix glycan, hyaluronan (HA), is markedly elevated in lungs from patients and experimental models with PH. Herein, we identified HA synthase-2 (HAS2) in the pulmonary artery smooth muscle cell (PASMC) layer as a predominant locus of HA dysregulation. HA upregulation involves depletion of NUDT21, a master regulator of alternative polyadenylation, resulting in 3’UTR shortening and hyper-expression of HAS2. The ensuing increase of HAS2 and hyper-synthesis of HA promoted bioenergetic dysfunction of PASMC characterized by impaired mitochondrial oxidative capacity and a glycolytic shift. The resulting HA accumulation stimulated pro-remodeling phenotypes such as cell proliferation, migration, apoptosis-resistance, and stimulated pulmonary artery contractility. Transgenic mice, mimicking HAS2 hyper-synthesis in smooth muscle cells, developed spontaneous PH, whereas targeted deletion of HAS2 prevented experimental PH. Pharmacological blockade of HAS2 restored normal bioenergetics in PASMC, ameliorated cell remodeling phenotypes, and reversed experimental PH in vivo. In summary, our results uncover a novel mechanism of HA hyper-synthesis and downstream effects on pulmonary vascular cell metabolism and remodeling.
KW - 3' Untranslated Regions/genetics
KW - Animals
KW - Cell Proliferation
KW - Energy Metabolism/genetics
KW - Humans
KW - Hyaluronan Synthases/genetics
KW - Hyaluronic Acid/biosynthesis
KW - Hypertension, Pulmonary/enzymology
KW - Mice
KW - Mice, Transgenic
KW - Myocytes, Smooth Muscle/enzymology
KW - hypoxia
KW - smooth muscle cell
KW - RNA processing deficiency
KW - Pulmonary hypertension
KW - Vascular biology
KW - Hyaluronan
KW - metabolism
KW - extracellular matrix
UR - http://www.scopus.com/inward/record.url?scp=85132439660&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85132439660&partnerID=8YFLogxK
U2 - 10.1016/j.matbio.2022.06.001
DO - 10.1016/j.matbio.2022.06.001
M3 - Article
C2 - 35671866
AN - SCOPUS:85132439660
SN - 0945-053X
VL - 111
SP - 53
EP - 75
JO - Matrix Biology
JF - Matrix Biology
ER -