Targeting a cell-specific microRNA repressor of CXCR4 ameliorates atherosclerosis in mice

The CXC chemokine receptor 4 (CXCR4) in endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) is crucial for vascular integrity. The atheroprotective functions of CXCR4 in vascular cells may be counteracted by atherogenic functions in other nonvascular cell types. Thus, strategies for cell-specifically augmenting CXCR4 function in vascular cells are crucial if this receptor is to be useful as a therapeutic target in treating atherosclerosis and other vascular disorders. Here, we identified miR-206-3p as a vascular-specific CXCR4 repressor and exploited a target-site blocker (CXCR4-TSB) that disrupted the interaction of miR-206-3p with CXCR4 in vitro and in vivo. In vitro, CXCR4-TSB enhanced CXCR4 expression in human and murine ECs and VSMCs to modulate cell viability, proliferation, and migration. Systemic administration of CXCR4-TSB in Apoe-deficient mice enhanced Cxcr4 expression in ECs and VSMCs in the walls of blood vessels, reduced vascular permeability and monocyte adhesion to endothelium, and attenuated the development of diet-induced atherosclerosis. CXCR4-TSB also increased CXCR4 expression in B cells, corroborating its atheroprotective role in this cell type. Analyses of human atherosclerotic plaque specimens revealed a decrease in CXCR4 and an increase in miR-206-3p expression in advanced compared with early lesions, supporting a role for the miR-206-3p–CXCR4 interaction in human disease. Disrupting the miR-206-3p–CXCR4 interaction in a cell-specific manner with target-site blockers is a potential therapeutic approach that could be used to treat atherosclerosis and other vascular diseases. Description Disrupting the interaction of miR-206-3p with CXCR4 boosts CXCR4 expression in vascular cells and offers a therapeutic opportunity in atherosclerosis. Editor’s summary The C-X-C motif chemokine receptor 4 (CXCR4) is important for vascular integrity, but boosting CXCR4 to treat atherosclerosis is limited by on-target effects in nonvascular cells. Here Cimen et al. showed that the microRNA miR-206-3p is a vascular-specific repressor of CXCR4 and that disrupting this repression with an mRNA target-site blocker increased CXCR4 in vascular cells in vitro. In a mouse model of atherosclerosis, treatment with the target-site blocker after disease onset reversed pathology and improved CXCR4 function. Human atherosclerotic plaques also had decreased CXCR4 and increased miR-206-3p, supporting the preclinical findings. These results suggest that increasing CXCR4 in vascular cells might be a potential therapeutic strategy for atherosclerosis in humans. —Brandon Berry

[1]  A. Habenicht,et al.  Novel mechanisms and therapeutic targets in atherosclerosis: inflammation and beyond. , 2023, European heart journal.

[2]  P. Willeit,et al.  High immunoglobulin-M levels to oxidation-specific epitopes are associated with lower risk of acute myocardial infarction , 2023, Journal of lipid research.

[3]  D. Santovito,et al.  Non-canonical features of microRNAs: paradigms emerging from cardiovascular disease , 2022, Nature Reviews Cardiology.

[4]  G. Rabinovich,et al.  Galectin-1 prevents pathological vascular remodeling in atherosclerosis and abdominal aortic aneurysm , 2022, Science advances.

[5]  R. Rad,et al.  Endothelial Retargeting of AAV9 In Vivo , 2022, Advanced science.

[6]  Hsi-Yuan Huang,et al.  miRTarBase update 2022: an informative resource for experimentally validated miRNA–target interactions , 2021, Nucleic Acids Res..

[7]  Lucas E. Wange,et al.  Prime-seq, efficient and powerful bulk RNA sequencing , 2021, bioRxiv.

[8]  Z. Mallat,et al.  Humoral immunity in atherosclerosis and myocardial infarction: from B cells to antibodies. , 2021, Cardiovascular research.

[9]  Jacob C. Ulirsch,et al.  Discovery and systematic characterization of risk variants and genes for coronary artery disease in over a million participants , 2021, Nature Genetics.

[10]  S. Ylä-Herttuala,et al.  Profiling of Primary and Mature miRNA Expression in Atherosclerosis-Associated Cell Types , 2021, Arteriosclerosis, thrombosis, and vascular biology.

[11]  S. V. Bontha,et al.  Chemokine Receptor-6 Promotes B-1 Cell Trafficking to Perivascular Adipose Tissue, Local IgM Production and Atheroprotection , 2021, Frontiers in Immunology.

[12]  M. Cybulsky,et al.  Radiation Impacts Early Atherosclerosis by Suppressing Intimal LDL Accumulation. , 2021, Circulation research.

[13]  Sathish Kumar Jayapal,et al.  Global Burden of Cardiovascular Diseases and Risk Factors, 1990–2019 , 2020, Journal of the American College of Cardiology.

[14]  S. Chi,et al.  Position-specific oxidation of miR-1 encodes cardiac hypertrophy , 2020, Nature.

[15]  C. Weber,et al.  Small Things Matter: Relevance of MicroRNAs in Cardiovascular Disease , 2020, Frontiers in Physiology.

[16]  M. Daemen,et al.  Noncanonical inhibition of caspase-3 by a nuclear microRNA confers endothelial protection by autophagy in atherosclerosis , 2020, Science Translational Medicine.

[17]  Lawrence A Leiter,et al.  Two Phase 3 Trials of Inclisiran in Patients with Elevated LDL Cholesterol. , 2020, The New England journal of medicine.

[18]  R. Samulski,et al.  Engineering adeno-associated virus vectors for gene therapy , 2020, Nature Reviews Genetics.

[19]  C. Weber,et al.  B-Cell Specific CXCR4 Protects Against Atherosclerosis Development and Increases Plasma IgM Levels. , 2020, Circulation research.

[20]  C. Weber,et al.  Immunotherapy for cardiovascular disease. , 2019, European heart journal.

[21]  J. Witztum,et al.  Diversification and CXCR4-Dependent Establishment of the Bone Marrow B-1a Cell Pool Governs Atheroprotective IgM Production Linked To Human Coronary Atherosclerosis. , 2019, Circulation research.

[22]  E. Boersma,et al.  Anti-Oxidized LDL Antibodies and Coronary Artery Disease: A Systematic Review , 2019, Antioxidants.

[23]  C. Weber,et al.  Double bond configuration of palmitoleate is critical for atheroprotection , 2019, Molecular metabolism.

[24]  D. Rader,et al.  CXCL12 Derived From Endothelial Cells Promotes Atherosclerosis to Drive Coronary Artery Disease. , 2019, Circulation.

[25]  R. Zimmer,et al.  Dicer in Macrophages Prevents Atherosclerosis by Promoting Mitochondrial Oxidative Metabolism , 2018, Circulation.

[26]  David P. Bartel,et al.  The biochemical basis of microRNA targeting efficacy , 2018, Science.

[27]  R. Zimmer,et al.  miR-103 promotes endothelial maladaptation by targeting lncWDR59 , 2018, Nature Communications.

[28]  M. Sperandio,et al.  Chrono-pharmacological Targeting of the CCL2-CCR2 Axis Ameliorates Atherosclerosis. , 2018, Cell metabolism.

[29]  D. Bartel.  Metazoan MicroRNAs , 2018, Cell.

[30]  Diana Domanska,et al.  MirGeneDB 2.0: the metazoan microRNA complement , 2018, bioRxiv.

[31]  J. Joung,et al.  Gene therapy comes of age , 2018, Science.

[32]  Jay W. Shin,et al.  An integrated expression atlas of miRNAs and their promoters in human and mouse , 2017, Nature Biotechnology.

[33]  Christoph Ziegenhain,et al.  zUMIs - A fast and flexible pipeline to process RNA sequencing data with UMIs , 2017, bioRxiv.

[34]  C. Weber,et al.  Functional ex-vivo Imaging of Arterial Cellular Recruitment and Lipid Extravasation. , 2017, Bio-protocol.

[35]  Christopher J. Mitchell,et al.  Toward the human cellular microRNAome , 2017, bioRxiv.

[36]  S. Watkins,et al.  Prevention of atherosclerosis by bioactive palmitoleate through suppression of organelle stress and inflammasome activation , 2016, Science Translational Medicine.

[37]  Arndt F. Siekmann,et al.  MicroRNA 139-5p coordinates APLNR-CXCR4 crosstalk during vascular maturation , 2016, Nature Communications.

[38]  A. Schober,et al.  Endothelial Dicer promotes atherosclerosis and vascular inflammation by miRNA-103-mediated suppression of KLF4 , 2016, Nature Communications.

[39]  E. Hovig,et al.  A Uniform System for the Annotation of Vertebrate microRNA Genes and the Evolution of the Human microRNAome. , 2015, Annual review of genetics.

[40]  C. Glass,et al.  The selection and function of cell type-specific enhancers , 2015, Nature Reviews Molecular Cell Biology.

[41]  Daehyun Baek,et al.  mRNA destabilization is the dominant effect of mammalian microRNAs by the time substantial repression ensues. , 2014, Molecular cell.

[42]  Y. Pi,et al.  TRPV1 activation impedes foam cell formation by inducing autophagy in oxLDL-treated vascular smooth muscle cells , 2014, Cell Death and Disease.

[43]  F. Kiessling,et al.  MicroRNA-126-5p promotes endothelial proliferation and limits atherosclerosis by suppressing Dlk1 , 2014, Nature Medicine.

[44]  Erling Falk,et al.  Update on acute coronary syndromes: the pathologists' view. , 2013, European heart journal.

[45]  J. Danesh,et al.  Large-scale association analysis identifies new risk loci for coronary artery disease , 2012, Nature Genetics.

[46]  Rongkun Shen,et al.  Capturing microRNA targets using an RNA-induced silencing complex (RISC)-trap approach , 2012, Proceedings of the National Academy of Sciences.

[47]  Gabriele Sales,et al.  MAGIA2: from miRNA and genes expression data integrative analysis to microRNA–transcription factor mixed regulatory circuits (2012 update) , 2012, Nucleic Acids Res..

[48]  E. Braunwald,et al.  A tale of coronary artery disease and myocardial infarction. , 2012, The New England journal of medicine.

[49]  C. Weber,et al.  Atherosclerosis: current pathogenesis and therapeutic options , 2011, Nature Medicine.

[50]  Marcel Martin.  Cutadapt removes adapter sequences from high-throughput sequencing reads , 2011 .

[51]  Ciro Indolfi,et al.  The knockout of miR-143 and -145 alters smooth muscle cell maintenance and vascular homeostasis in mice: correlates with human disease , 2009, Cell Death and Differentiation.

[52]  M. Cybulsky,et al.  GM-CSF regulates intimal cell proliferation in nascent atherosclerotic lesions , 2009, The Journal of experimental medicine.

[53]  Deepak Srivastava,et al.  miR-145 and miR-143 Regulate Smooth Muscle Cell Fate Decisions , 2009, Nature.

[54]  A. Tall,et al.  Macrophage deficiency of p38alpha MAPK promotes apoptosis and plaque necrosis in advanced atherosclerotic lesions in mice. , 2009, The Journal of clinical investigation.

[55]  C. Burge,et al.  Most mammalian mRNAs are conserved targets of microRNAs. , 2008, Genome research.

[56]  M. Biffoni,et al.  A three-step pathway comprising PLZF/miR-146a/CXCR4 controls megakaryopoiesis , 2008, Nature Cell Biology.

[57]  F. Tacke,et al.  Protective Role of CXC Receptor 4/CXC Ligand 12 Unveils the Importance of Neutrophils in Atherosclerosis , 2008, Circulation research.

[58]  D. Link,et al.  Suppression of CXCL12 production by bone marrow osteoblasts is a common and critical pathway for cytokine-induced mobilization. , 2007, Blood.

[59]  D. Duan,et al.  Systemic AAV-9 transduction in mice is influenced by animal age but not by the route of administration , 2007, Gene Therapy.

[60]  Michael T. McManus,et al.  Dysregulation of Cardiogenesis, Cardiac Conduction, and Cell Cycle in Mice Lacking miRNA-1-2 , 2007, Cell.

[61]  T. Nagasawa,et al.  Maintenance of the hematopoietic stem cell pool by CXCL12-CXCR4 chemokine signaling in bone marrow stromal cell niches. , 2006, Immunity.

[62]  N. Rajewsky,et al.  Cell-type-specific signatures of microRNAs on target mRNA expression. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[63]  A. Zernecke,et al.  SDF-1&agr;/CXCR4 Axis Is Instrumental in Neointimal Hyperplasia and Recruitment of Smooth Muscle Progenitor Cells , 2005, Circulation research.

[64]  H. Horiguchi,et al.  Oxidized LDL in Carotid Plaques and Plasma Associates With Plaque Instability , 2002, Arteriosclerosis, thrombosis, and vascular biology.

[65]  Kouji Matsushima,et al.  The chemokine receptor CXCR4 is essential for vascularization of the gastrointestinal tract , 1998, Nature.

[66]  R. Evans,et al.  Oxidized LDL Regulates Macrophage Gene Expression through Ligand Activation of PPARγ , 1998, Cell.

[67]  J. Witztum,et al.  Cloning of monoclonal autoantibodies to epitopes of oxidized lipoproteins from apolipoprotein E-deficient mice. Demonstration of epitopes of oxidized low density lipoprotein in human plasma. , 1996, The Journal of clinical investigation.

[68]  Klaus Rajewsky,et al.  A B cell-deficient mouse by targeted disruption of the membrane exon of the immunoglobulin μ chain gene , 1991, Nature.

[69]  OUP accepted manuscript , 2021, Nucleic Acids Research.

[70]  A. Schober,et al.  Dicer generates a regulatory microRNA network in smooth muscle cells that limits neointima formation during vascular repair , 2016, Cellular and Molecular Life Sciences.

[71]  Piero Carninci,et al.  Detecting expressed genes using CAGE. , 2014, Methods in molecular biology.

[72]  H. Soifer,et al.  Silencing of gene expression by gymnotic delivery of antisense oligonucleotides. , 2012, Methods in molecular biology.

[73]  D. Gagnon,et al.  A R T I C L E , 2022 .