Adoptive transfer of CX3CR1 transduced-T regulatory cells improves homing to the atherosclerotic plaques and dampens atherosclerosis progression.

AIM Loss of immunosuppressive response supports inflammation during atherosclerosis. We tested whether adoptive cell therapy (ACT) with Tregulatory cells (Tregs) engineered to selectively migrate in the atherosclerotic plaque would dampen the immune-inflammatory response in the arterial wall in animal models of Familial Hypercholesterolemia (FH). METHODS AND RESULTS FH patients presented a decreased Tregs suppressive function associated to an increased inflammatory burden. A similar phenotype was observed in Ldlr -/- mice accompanied by a selective increased expression of the chemokine CX3CL1 in the aorta but not in other districts (lymph nodes, spleen and liver). Treg overexpressing CX3CR1 were thus generated (CX3CR1+-Treg) to drive Treg selectively to the plaque. CX3CR1+-Treg were injected (i.v.) in Ldlr -/- fed high-cholesterol diet (WTD) for 8 weeks. CX3CR1+-Treg were detected in the aorta, but not in other tissues, of Ldlr -/- mice 24h after ACT, corroborating the efficacy of this approach. After 4 additional weeks of WTD, ACT with CX3CR1+-Treg resulted in reduced plaque progression and lipid deposition, ameliorated plaque stability by increasing collagen and smooth muscle cells content, while decreasing the number of pro-inflammatory macrophages. Shotgun proteomics of the aorta showed a metabolic rewiring in CX3CR1+-Treg treated Ldlr -/- mice compared to controls that was associated with the improvement of inflammation-resolving pathways and disease progression. CONCLUSION ACT with vasculotropic Treg appears as a promising strategy to selectively target immune activation in the atherosclerotic plaque. TRANSLATIONAL RELEVANCE Improving pro-resolutive inflammatory response represents a promising therapeutic approach to control atherosclerosis progression. Meanwhile, selective immunosuppression at the atherosclerotic plaque looks critical to limit unspecific inhibition of inflammation in other tissues. Our work demonstrates that engineering of immunosuppressive T regulatory cells to be hijacked in the atherosclerotic plaque limits atherosclerosis progression by targeting local inflammation.

[1]  J. Todd,et al.  Chronic Immune Activation in Systemic Lupus Erythematosus and the Autoimmune PTPN22 Trp620 Risk Allele Drive the Expansion of FOXP3+ Regulatory T Cells and PD-1 Expression , 2019, Front. Immunol..

[2]  L. Joosten,et al.  Treatment with Statins Does Not Revert Trained Immunity in Patients with Familial Hypercholesterolemia. , 2019, Cell metabolism.

[3]  G. Norata,et al.  Immunometabolic function of cholesterol in cardiovascular disease and beyond. , 2019, Cardiovascular research.

[4]  Ira Tabas,et al.  Inflammation and its resolution in atherosclerosis: mediators and therapeutic opportunities , 2019, Nature Reviews Cardiology.

[5]  G. Norata,et al.  Lysosomal Acid Lipase: From Cellular Lipid Handler to Immunometabolic Target. , 2019, Trends in pharmacological sciences.

[6]  G. Lombardi,et al.  Past, Present, and Future of Regulatory T Cell Therapy in Transplantation and Autoimmunity , 2019, Front. Immunol..

[7]  Hamideh Amirfakhryan.  Vaccination against atherosclerosis: An overview. , 2019, Hellenic journal of cardiology : HJC = Hellenike kardiologike epitheorese.

[8]  G. Gusarova,et al.  Regulatory T Cells Promote Macrophage Efferocytosis during Inflammation Resolution , 2018, Immunity.

[9]  G. Biolo,et al.  Evaluation of the performance of Dutch Lipid Clinic Network score in an Italian FH population: The LIPIGEN study. , 2018, Atherosclerosis.

[10]  A. Annoni,et al.  Myeloid apolipoprotein E controls dendritic cell antigen presentation and T cell activation , 2018, Nature Communications.

[11]  H. Randeva,et al.  Familial Hypercholesterolemia: New Horizons for Diagnosis and Effective Management , 2018, Front. Pharmacol..

[12]  D. Hafler,et al.  Regulatory T cells in autoimmune disease , 2018, Nature Immunology.

[13]  Zhen Yang,et al.  Regulatory T cells as a new therapeutic target for atherosclerosis , 2018, Acta Pharmacologica Sinica.

[14]  G. Hansson,et al.  Hypercholesterolemia Enhances T Cell Receptor Signaling and Increases the Regulatory T Cell Population , 2017, Scientific Reports.

[15]  G. Norata,et al.  Anti-PCSK9 antibodies for the treatment of heterozygous familial hypercholesterolemia: patient selection and perspectives , 2017, Vascular health and risk management.

[16]  M. Kallikourdis,et al.  Improving homing in T cell therapy. , 2017, Cytokine & growth factor reviews.

[17]  E. Stroes,et al.  PCSK9 monoclonal antibodies reverse the pro-inflammatory profile of monocytes in familial hypercholesterolaemia , 2017, European heart journal.

[18]  P. Shah,et al.  Vaccine against arteriosclerosis: an update , 2017, Therapeutic advances in vaccines.

[19]  G. Roselli,et al.  Tailored chemokine receptor modification improves homing of adoptive therapy T cells in a spontaneous tumor model , 2016, Oncotarget.

[20]  C. Weber,et al.  Regulatory T cells in atherosclerosis: critical immune regulatory function and therapeutic potential , 2015, Cellular and Molecular Life Sciences.

[21]  A. G. Betz,et al.  Peak inflammation in atherosclerosis, primary biliary cirrhosis and autoimmune arthritis is counter-intuitively associated with regulatory T cell enrichment , 2015, Immunobiology.

[22]  B. Bernstein,et al.  Th17 cells transdifferentiate into regulatory T cells during resolution of inflammation , 2015, Nature.

[23]  Ying Sun,et al.  Chemokine CX3CL1 and its receptor CX3CR1 are associated with human atherosclerotic lesion volnerability. , 2015, Thrombosis research.

[24]  A. Lichtman,et al.  Modulation of Atherosclerotic Lesions by Circulating Cells : The Translational Spectrum Series , 2014 .

[25]  D. Greaves,et al.  Fractalkine Promotes Human Monocyte Survival via a Reduction in Oxidative Stress , 2014, Arteriosclerosis, thrombosis, and vascular biology.

[26]  L. Qi,et al.  Impaired Circulating CD4+LAP+ Regulatory T Cells in Patients with Acute Coronary Syndrome and Its Mechanistic Study , 2014, PloS one.

[27]  Constance Auvynet,et al.  Pharmacological Inhibition of the Chemokine Receptor, CX3CR1, Reduces Atherosclerosis in Mice , 2013, Arteriosclerosis, thrombosis, and vascular biology.

[28]  S. Achenbach,et al.  Decreased numbers of regulatory T cells are associated with human atherosclerotic lesion vulnerability and inversely correlate with infiltrated mature dendritic cells. , 2013, Atherosclerosis.

[29]  Catherine Boileau,et al.  Familial hypercholesterolaemia is underdiagnosed and undertreated in the general population: guidance for clinicians to prevent coronary heart disease , 2013, European heart journal.

[30]  C. Riccardi,et al.  Characterization of a new regulatory CD4+ T cell subset in primary Sjögren's syndrome. , 2013, Rheumatology.

[31]  C. Ballantyne,et al.  New therapeutic principles in dyslipidaemia: focus on LDL and Lp(a) lowering drugs. , 2013, European heart journal.

[32]  T. Lüscher,et al.  Depletion of FOXP3+ regulatory T cells promotes hypercholesterolemia and atherosclerosis. , 2013, The Journal of clinical investigation.

[33]  P. Tipping,et al.  Cytokine Therapy With Interleukin-2/Anti–Interleukin-2 Monoclonal Antibody Complexes Expands CD4+CD25+Foxp3+ Regulatory T Cells and Attenuates Development and Progression of Atherosclerosis , 2012, Circulation.

[34]  J. Bauersachs,et al.  Fractalkine Is Expressed in Early and Advanced Atherosclerotic Lesions and Supports Monocyte Recruitment via CX3CR1 , 2012, PloS one.

[35]  A. Maseri,et al.  Effector Memory T cells Are Associated With Atherosclerosis in Humans and Animal Models , 2012, Journal of the American Heart Association.

[36]  E. Abraham,et al.  Intracellular HMGB1 Negatively Regulates Efferocytosis , 2011, The Journal of Immunology.

[37]  Hong Liu,et al.  Fractalkine/CX3CR1 and atherosclerosis. , 2011, Clinica chimica acta; international journal of clinical chemistry.

[38]  G. Hansson,et al.  Immunotherapy With Tolerogenic Apolipoprotein B-100–Loaded Dendritic Cells Attenuates Atherosclerosis in Hypercholesterolemic Mice , 2011, Circulation.

[39]  K. Garlaschelli,et al.  Circulating CD4+CD25hiCD127lo Regulatory T-Cell Levels Do Not Reflect the Extent or Severity of Carotid and Coronary Atherosclerosis , 2010, Arteriosclerosis, thrombosis, and vascular biology.

[40]  R. Klingenberg,et al.  Intranasal Immunization With an Apolipoprotein B-100 Fusion Protein Induces Antigen-Specific Regulatory T Cells and Reduces Atherosclerosis , 2010, Arteriosclerosis, thrombosis, and vascular biology.

[41]  M. Linton,et al.  Macrophage LRP-1 Controls Plaque Cellularity by Regulating Efferocytosis and Akt Activation , 2010, Arteriosclerosis, thrombosis, and vascular biology.

[42]  R. Toes,et al.  Vaccination using oxidized low-density lipoprotein-pulsed dendritic cells reduces atherosclerosis in LDL receptor-deficient mice. , 2010, Cardiovascular research.

[43]  T. Akasaka,et al.  Upregulation of fractalkine and its receptor, CX3CR1, is associated with coronary plaque rupture in patients with unstable angina pectoris. , 2010, Circulation journal : official journal of the Japanese Circulation Society.

[44]  D. Greaves,et al.  Fractalkine has anti-apoptotic and proliferative effects on human vascular smooth muscle cells via epidermal growth factor receptor signalling , 2009, Cardiovascular research.

[45]  T. Mrkvan,et al.  In Vivo Expansion of Activated Naive CD8+ T Cells and NK Cells Driven by Complexes of IL-2 and Anti-IL-2 Monoclonal Antibody As Novel Approach of Cancer Immunotherapy1 , 2009, The Journal of Immunology.

[46]  D. Dichek,et al.  TGF-&bgr;1 Limits Plaque Growth, Stabilizes Plaque Structure, and Prevents Aortic Dilation in Apolipoprotein E–Null Mice , 2009, Arteriosclerosis, thrombosis, and vascular biology.

[47]  M. Daemen,et al.  Atherosclerosis in Mice Lactadherin Deficiency Leads to Apoptotic Cell Accumulation and Accelerated , 2007 .

[48]  G. Keren,et al.  Role of Naturally Occurring CD4+CD25+ Regulatory T Cells in Experimental Atherosclerosis , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[49]  A. G. Betz,et al.  Alloantigen-enhanced accumulation of CCR5+ ‘effector’ regulatory T cells in the gravid uterus , 2007, Proceedings of the National Academy of Sciences.

[50]  G. Norata,et al.  Effects of Fractalkine Receptor Variants on Common Carotid Artery Intima-Media Thickness , 2006, Stroke.

[51]  O. Quehenberger,et al.  Expression of Fractalkine (CX3CL1) and its Receptor, CX3CR1, Is Elevated in Coronary Artery Disease and Is Reduced During Statin Therapy , 2005, Arteriosclerosis, thrombosis, and vascular biology.

[52]  W. Janssen,et al.  Cell-Surface Calreticulin Initiates Clearance of Viable or Apoptotic Cells through trans-Activation of LRP on the Phagocyte , 2005, Cell.

[53]  A. Tedgui,et al.  Induction of a Regulatory T Cell Type 1 Response Reduces the Development of Atherosclerosis in Apolipoprotein E–Knockout Mice , 2003, Circulation.

[54]  P. Debré,et al.  Decreased Atherosclerotic Lesion Formation in CX3CR1/Apolipoprotein E Double Knockout Mice , 2003, Circulation.

[55]  B. McManus,et al.  Characterization of fractalkine (CX3CL1) and CX3CR1 in human coronary arteries with native atherosclerosis, diabetes mellitus, and transplant vascular disease. , 2002, Cardiovascular pathology : the official journal of the Society for Cardiovascular Pathology.

[56]  T. Imai,et al.  Fractalkine and vascular injury. , 2001, Trends in immunology.

[57]  S. Fazio,et al.  Familial Hypercholesterolemia: Genes and Beyond , 2000 .

[58]  B. Rollins,et al.  Monocyte chemoattractant protein-1 accelerates atherosclerosis in apolipoprotein E-deficient mice. , 1999, Arteriosclerosis, thrombosis, and vascular biology.