title: Dicer cleavage by calpain determines platelet microRNA levels and function in diabetes.
abstract: BACKGROUND
  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.
  
  
  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.
authors.affiliation: 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.).
authors.forename: Amro
authors.initials: A
authors.lastname: Elgheznawy
authors.affiliation: 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.).
authors.forename: Lei
authors.initials: L
authors.lastname: Shi
authors.affiliation: 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.).
authors.forename: Jiong
authors.initials: J
authors.lastname: Hu
authors.affiliation: 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.).
authors.forename: Ilka
authors.initials: I
authors.lastname: Wittig
authors.affiliation: 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.).
authors.forename: Hebatullah
authors.initials: H
authors.lastname: Laban
authors.affiliation: 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.).
authors.forename: Joachim
authors.initials: J
authors.lastname: Pircher
authors.affiliation: 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.).
authors.forename: Alexander
authors.initials: A
authors.lastname: Mann
authors.affiliation: 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.).
authors.forename: Patrick
authors.initials: P
authors.lastname: Provost
authors.affiliation: 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.).
authors.forename: Voahanginirina
authors.initials: V
authors.lastname: Randriamboavonjy
authors.affiliation: 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.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Fleming
mesh_terms: D000328:Adult; D000818:Animals; D001792:Blood Platelets / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism / Q000502:physiology; D002118:Calcium / Q000494:pharmacology; D002154:Calpain / Q000097:blood* / Q000172:deficiency; D003598:Cytoskeletal Proteins / Q000097:blood; D053487:DEAD-box RNA Helicases / Q000097:blood*; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000201:enzymology; D003924:Diabetes Mellitus, Type 2 / Q000097:blood* / Q000201:enzymology; D005176:Factor XIII / Q000378:metabolism; D005260:Female; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D008297:Male; D008565:Membrane Proteins / Q000097:blood; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D035683:MicroRNAs / Q000097:blood* / Q000235:genetics; D018895:Microsatellite Repeats; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000097:blood; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology*; D020543:Proteome; D043244:Ribonuclease III / Q000097:blood*
chemical_list: D003598:Cytoskeletal Proteins; C477322:FERMT3 protein, human; D008565:Membrane Proteins; D035683:MicroRNAs; D009363:Neoplasm Proteins; D020543:Proteome; C527818:kindlin-3 protein, mouse; D015759:Ionomycin; D005176:Factor XIII; C408087:DICER1 protein, human; C509781:Dicer1 protein, mouse; D043244:Ribonuclease III; D002154:Calpain; C112682:Capn1 protein, mouse; D053487:DEAD-box RNA Helicases; D002118:Calcium
keywords: diabetes mellitus; microRNA; platelets; signal transduction