Amro Elgheznawy1, Lei Shi1, Jiong Hu1, Ilka Wittig1, Hebatullah Laban1, Joachim Pircher1, Alexander Mann1, Patrick Provost1, Voahanginirina Randriamboavonjy1, Ingrid Fleming2. 1. From the Institute for Vascular Signaling, Centre for Molecular Medicine, and DZHK (German Centre for Cardiovascular Research) partner site Rhine-Main, Frankfurt, Germany (A.E., L.S., J.H., H.L., V.R., I.F.); Functional Proteomics, SFB 815 Core Unit, Goethe-University, Frankfurt, Germany (I.W.); Walter-Brendel-Centre of Experimental Medicine and DZHK partner site Munich Heart Alliance, Ludwig-Maximilians-Universität, Munich, Germany (J.P.); Endokrinologikum Frankfurt, Frankfurt, Germany (A.M.); and Centre Hospitalier Universitaire de Québec Research Center, and Faculty of Medicine, Université Laval, Quebec, Canada (P.P.). 2. From the Institute for Vascular Signaling, Centre for Molecular Medicine, and DZHK (German Centre for Cardiovascular Research) partner site Rhine-Main, Frankfurt, Germany (A.E., L.S., J.H., H.L., V.R., I.F.); Functional Proteomics, SFB 815 Core Unit, Goethe-University, Frankfurt, Germany (I.W.); Walter-Brendel-Centre of Experimental Medicine and DZHK partner site Munich Heart Alliance, Ludwig-Maximilians-Universität, Munich, Germany (J.P.); Endokrinologikum Frankfurt, Frankfurt, Germany (A.M.); and Centre Hospitalier Universitaire de Québec Research Center, and Faculty of Medicine, Université Laval, Quebec, Canada (P.P.). fleming@em.uni-frankfurt.de.
Abstract
RATIONALE: MicroRNAs (miRNAs) are short noncoding RNA species generated by the processing of longer precursors by the ribonucleases Drosha and Dicer. Platelets contain large amounts of miRNA that are altered by disease, in particular diabetes mellitus. OBJECTIVE: This study determined why platelet miRNA levels are attenuated in diabetic individuals and how decreased levels of the platelet-enriched miRNA, miR-223, affect platelet function. METHODS AND RESULTS: Dicer levels were altered in platelets from diabetic mice and patients, a change that could be attributed to the cleavage of the enzyme by calpain, resulting in loss of function. Diabetes mellitus in human subjects as well as in mice resulted in decreased levels of platelet miR-142, miR-143, miR-155, and miR-223. Focusing on only 1 of these miRNAs, miR-223 deletion in mice resulted in modestly enhanced platelet aggregation, the formation of large thrombi and delayed clot retraction compared with wild-type littermates. A similar dysregulation was detected in platelets from diabetic patients. Proteomic analysis of platelets from miR-223 knockout mice revealed increased levels of several proteins, including kindlin-3 and coagulation factor XIII-A. Whereas, kindlin-3 was indirectly regulated by miR-223, factor XIII was a direct target and both proteins were also altered in diabetic platelets. Treating diabetic mice with a calpain inhibitor prevented loss of platelet dicer as well as the diabetes mellitus-induced decrease in platelet miRNA levels and the upregulation of miR-223 target proteins. CONCLUSIONS: Thus, calpain inhibition may be one means of normalizing platelet miRNA processing as well as platelet function in diabetes mellitus.
RATIONALE: MicroRNAs (miRNAs) are short noncoding RNA species generated by the processing of longer precursors by the ribonucleases Drosha and Dicer. Platelets contain large amounts of miRNA that are altered by disease, in particular diabetes mellitus. OBJECTIVE: This study determined why platelet miRNA levels are attenuated in diabetic individuals and how decreased levels of the platelet-enriched miRNA, miR-223, affect platelet function. METHODS AND RESULTS: Dicer levels were altered in platelets from diabeticmice and patients, a change that could be attributed to the cleavage of the enzyme by calpain, resulting in loss of function. Diabetes mellitus in human subjects as well as in mice resulted in decreased levels of platelet miR-142, miR-143, miR-155, and miR-223. Focusing on only 1 of these miRNAs, miR-223 deletion in mice resulted in modestly enhanced platelet aggregation, the formation of large thrombi and delayed clot retraction compared with wild-type littermates. A similar dysregulation was detected in platelets from diabeticpatients. Proteomic analysis of platelets from miR-223 knockout mice revealed increased levels of several proteins, including kindlin-3 and coagulation factor XIII-A. Whereas, kindlin-3 was indirectly regulated by miR-223, factor XIII was a direct target and both proteins were also altered in diabetic platelets. Treating diabeticmice with a calpain inhibitor prevented loss of platelet dicer as well as the diabetes mellitus-induced decrease in platelet miRNA levels and the upregulation of miR-223 target proteins. CONCLUSIONS: Thus, calpain inhibition may be one means of normalizing platelet miRNA processing as well as platelet function in diabetes mellitus.
Authors: Jesse W Rowley; Stéphane Chappaz; Aurélie Corduan; Mark M W Chong; Robert Campbell; Amanda Khoury; Bhanu Kanth Manne; Jeremy G T Wurtzel; James V Michael; Lawrence E Goldfinger; Michele M Mumaw; Marvin T Nieman; Benjamin T Kile; Patrick Provost; Andrew S Weyrich Journal: Blood Date: 2016-01-14 Impact factor: 22.113
Authors: Dorothee Kaudewitz; Philipp Skroblin; Lukas H Bender; Temo Barwari; Peter Willeit; Raimund Pechlaner; Nicholas P Sunderland; Karin Willeit; Allison C Morton; Paul C Armstrong; Melissa V Chan; Ruifang Lu; Xiaoke Yin; Filipe Gracio; Katarzyna Dudek; Sarah R Langley; Anna Zampetaki; Emanuele de Rinaldis; Shu Ye; Timothy D Warner; Alka Saxena; Stefan Kiechl; Robert F Storey; Manuel Mayr Journal: Circ Res Date: 2015-12-08 Impact factor: 17.367
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