TY - JOUR
T1 - MicroRNA-21/PDCD4 Proapoptotic Signaling from Circulating CD341 Cells to Vascular Endothelial Cells
T2 - A Potential Contributor to Adverse Cardiovascular Outcomes in Patients with Critical Limb Ischemia
AU - Spinetti, Gaia
AU - Sangalli, Elena
AU - Tagliabue, Elena
AU - Maselli, Davide
AU - Colpani, Ornella
AU - Ferland-Mccollough, David
AU - Carnelli, Franco
AU - Orlando, Patrizia
AU - Paccagnella, Agostino
AU - Furlan, Anna
AU - Stefani, Piero Maria
AU - Sambado, Luisa
AU - Sambataro, Maria
AU - Madeddu, Paolo
N1 - Funding Information:
Acknowledgments. The authors thank Dr. Simona Rodighiero, Imaging Unit of the Department of Experimental Oncology, European Institute of Oncology, Milan, Italy, for the support in the generation of the microscopy data and immunofluorescence data presented herein. Funding. Funding and financial support was obtained from the Italian Ministry of Health, Ricerca Corrente to the IRCCS MultiMedica, Foundation Cariplo (2016-0922), and a British Heart Foundation program grant. Study 1 was also supported by Diabetic ONLUS Association, section of Treviso. Duality of Interest. No potential conflicts of interest relevant to this article were reported. Author Contributions. G.S. and P.M. acquired funding, analyzed data, and wrote the manuscript. E.S. researched data relative to both study 1 and study 2. E.T. performed the statistical analyses for study 1. D.M. isolated cells and performed the in vitro molecular analyses of study 2. O.C. conducted tissue and cells immune characterization. D.F.-M. isolated cells and provided biological samples. F.C. and P.O. provided human BM samples and contributed to the discussion. A.P. acquired funding and contributed to the discussion. A.F., P.M.S., and L.S. were involved in collection of the human BM samples and recording the clinical data. M.S. acquired funding, provided human BM samples, and corrected the manuscript. G.S. and P.M. are the guarantors of this work and, as such, had full access to all the data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis. Prior Presentation. Parts of this study were presented in abstract form at the Keystone Symposia: Novel Aspects of Bone Biology (E3), Snowbird, UT, 13–16 June 2018 and at the European Society of Cardiology Congress, Paris, France, 31 August to 4 September 2019.
Publisher Copyright:
© 2020 American Diabetes Association Inc.. All rights reserved.
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/7/1
Y1 - 2020/7/1
N2 - OBJECTIVE In patients with type 2 diabetes (T2D) and critical limb ischemia (CLI), migration of circulating CD341 cells predicted cardiovascular mortality at 18 months after revascularization. This study aimed to provide long-term validation and mechanistic understanding of the biomarker. RESEARCH DESIGN AND METHODS The association between CD341 cell migration and cardiovascular mortality was reassessed at 6 years after revascularization. In a new series of T2D-CLI and control subjects, immuno-sorted bone marrow CD341 cells were profiled for miRNA expression and assessed for apoptosis and angiogenesis activity. The differentially regulatedmiRNA-21and its proapoptotic target, PDCD4,weretitrated to verify their contribution in transferring damaging signals from CD341 cells to endothelial cells. RESULTS Multivariable regression analysis confirmed that CD341 cell migration forecasts long-term cardiovascular mortality. CD341 cells from T2D-CLI patients were more apoptotic and less proangiogenic than those from control subjects and featured miRNA-21 downregulation, modulation of several long noncoding RNAs acting as miRNA-21 sponges, and upregulation of the miRNA-21 proapoptotic target PDCD4. Silencing miR-21 in control CD341 cells phenocopied the T2D-CLI cell behavior. In coculture, T2D-CLI CD341 cells imprinted naive endothelial cells, increasing apoptosis, reducing network formation, and modulating the TUG1 sponge/miRNA-21/PDCD4 axis. Silencing PDCD4 or scavenging reactive oxygen species protected endothelial cells from the negative influence of T2D-CLI CD341 cells. CONCLUSIONS Migration of CD341 cells predicts long-term cardiovascular mortality in T2D-CLI patients. An altered paracrine signaling conveys antiangiogenic and proapoptotic features from CD341 cells to the endothelium. This damaging interaction may increase the risk for life-threatening complications.
AB - OBJECTIVE In patients with type 2 diabetes (T2D) and critical limb ischemia (CLI), migration of circulating CD341 cells predicted cardiovascular mortality at 18 months after revascularization. This study aimed to provide long-term validation and mechanistic understanding of the biomarker. RESEARCH DESIGN AND METHODS The association between CD341 cell migration and cardiovascular mortality was reassessed at 6 years after revascularization. In a new series of T2D-CLI and control subjects, immuno-sorted bone marrow CD341 cells were profiled for miRNA expression and assessed for apoptosis and angiogenesis activity. The differentially regulatedmiRNA-21and its proapoptotic target, PDCD4,weretitrated to verify their contribution in transferring damaging signals from CD341 cells to endothelial cells. RESULTS Multivariable regression analysis confirmed that CD341 cell migration forecasts long-term cardiovascular mortality. CD341 cells from T2D-CLI patients were more apoptotic and less proangiogenic than those from control subjects and featured miRNA-21 downregulation, modulation of several long noncoding RNAs acting as miRNA-21 sponges, and upregulation of the miRNA-21 proapoptotic target PDCD4. Silencing miR-21 in control CD341 cells phenocopied the T2D-CLI cell behavior. In coculture, T2D-CLI CD341 cells imprinted naive endothelial cells, increasing apoptosis, reducing network formation, and modulating the TUG1 sponge/miRNA-21/PDCD4 axis. Silencing PDCD4 or scavenging reactive oxygen species protected endothelial cells from the negative influence of T2D-CLI CD341 cells. CONCLUSIONS Migration of CD341 cells predicts long-term cardiovascular mortality in T2D-CLI patients. An altered paracrine signaling conveys antiangiogenic and proapoptotic features from CD341 cells to the endothelium. This damaging interaction may increase the risk for life-threatening complications.
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U2 - 10.2337/dc19-2227
DO - 10.2337/dc19-2227
M3 - Article
C2 - 32358022
AN - SCOPUS:85086793270
VL - 43
SP - 1520
EP - 1529
JO - Diabetes Care
JF - Diabetes Care
SN - 1935-5548
IS - 7
ER -