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Novel Peptide Micellar Nanoformulation for Treatment of Thrombotic Complications of Sepsis
Author(s) -
Bae Misuk,
Rusu Luiza,
Castellon Maricela,
Du Xiaoping,
Onyuksel Hayat,
Minshall Richard D
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.674.8
Subject(s) - myr , in vivo , sepsis , von willebrand factor , medicine , peptide , in vitro , ex vivo , pharmacology , chemistry , platelet , biochemistry , immunology , biology , gene , microbiology and biotechnology , genome
Sepsis is one of the leading causes of death in hospital intensive care units in the U.S. Despite supportive care, microvascular thrombosis and disseminated intravascular coagulation in septic patients leads to ~40% mortality. Since there are no anti‐thrombotic drugs for sepsis on the market, new drugs are desperately needed. Interestingly, the plasma level of von Willebrand factor (vWF), the glue to which platelets adhere to form a clot, is elevated in septic patients and high levels predict poor outcome. We recently showed Gα12 is required for αSNAP‐dependent vWF secretion from Weibel Palade bodies. We identified a 6 aa sequence in the N‐terminus of Gα12, generated a cell permeable myristoylated alpha‐ S NAP B inding D omain peptide, Myr‐ SBD , and showed that this blocked thrombin and shear‐stress induced vWF secretion from human endothelial cells in vitro . In addition, intravenous delivery of Myr‐SBD to mice at the time of cecal ligation and puncture (CLP) surgery reduced vWF levels, decreased microvascular thrombosis in the kidney, and increased survival. Here, we extend these initial studies using a well‐characterized sterically‐stabilized simple micelle (SSM) system to generate a nanoformulation of Myr‐SBD peptide for in vivo delivery. Pilot pharmacokinetic and efficacy parameters of Myr‐SBD SSM were investigated by fluorescence spectrometry in vitro and in vivo . Fluorescent probe‐labeled Myr‐SBD (Myr‐SBD‐PDAM), prepared by modifying the carboxylic acid group of Myr‐SBD via PDAM conjugation was confirmed by fluorescence spectrometry. Myr‐SBD‐PDAM SSM was prepared by incubating the peptide with DSPE‐PEG2000 using a film‐rehydration method. Following SSM preparation, micelle size and morphology assessed by electron microscopy (TEM) and light scattering (DLS) revealed uniform micelles of 14.1 ± 2.7 nm in diameter. Uptake and intracellular accumulation of Myr‐SBD‐PDAM SSM was confirmed by fluorescence and confocal microscopy. Intravenous delivery of Myr‐SBD SSM to mice after CLP surgery improved survival compared to the untreated CLP group. Thus, nanoformulation of Myr‐SBD peptide may enable in vivo delivery for treatment of thrombotic complications of severe inflammatory response syndrome. Support or Funding Information This work was supported by MPI R01 HL125356 (RDM, XD), the Michael Reese Research and Education Foundation (RDM), and AHA (LR).