Single systemic transfer of a human gene associated with exceptional longevity halts the progression of atherosclerosis and inflammation in ApoE knockout mice through a CXCR4-mediated mechanism

• Published in: European heart journal Vol. 41; no. 26; pp. 2487 - 2497
• Main Authors: Puca, Annibale Alessandro, Carrizzo, Albino, Spinelli, Chiara, Damato, Antonio, Ambrosio, Mariateresa, Villa, Francesco, Ferrario, Anna, Maciag, Anna, Fornai, Francesco, Lenzi, Paola, Valenti, Valentina, di Nonno, Flavio, Accarino, Giulio, Madonna, Michele, Forte, Maurizio, Calì, Gaetano, Baragetti, Andrea, Norata, Giuseppe Danilo, Catapano, Alberico Luigi, Cattaneo, Monica, Izzo, Raffaele, Trimarco, Valentina, Montella, Francesco, Versaci, Francesco, Auricchio, Alberto, Frati, Giacomo, Sciarretta, Sebastiano, Madeddu, Paolo, Ciaglia, Elena, Vecchione, Carmine
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 07-07-2020
• Subjects: Aged; Animals; Apolipoproteins E; Atherosclerosis - genetics; Basic Science; Carotid Intima-Media Thickness; Female; Humans; Inflammation; Intercellular Signaling Peptides and Proteins; Longevity; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Knockout, ApoE; Middle Aged; Phosphoproteins; Plaque, Atherosclerotic; Receptors, CXCR4; Low-density lipoprotein; Vascular function; Atherosclerosis; Immune system
• ISSN: 0195-668X; 1522-9645
• DOI: 10.1093/eurheartj/ehz459

Abstract
Aims
Here, we aimed to determine the therapeutic effect of longevity-associated variant (LAV)-BPIFB4 gene therapy on atherosclerosis.
Methods and results
ApoE knockout mice (ApoE−/−) fed a high-fat diet were randomly allocated to receive LAV-BPIFB4, wild-type (WT)-BPIFB4, or empty vector via adeno-associated viral vector injection. The primary endpoints of the study were to assess (i) vascular reactivity and (ii) atherosclerotic disease severity, by Echo-Doppler imaging, histology and ultrastructural analysis. Moreover, we assessed the capacity of the LAV-BPIFB4 protein to shift monocyte-derived macrophages of atherosclerotic mice and patients towards an anti-inflammatory phenotype. LAV-BPIFB4 gene therapy rescued endothelial function of mesenteric and femoral arteries from ApoE−/− mice; this effect was blunted by AMD3100, a CXC chemokine receptor type 4 (CXCR4) inhibitor. LAV-BPIFB4-treated mice showed a CXCR4-mediated shift in the balance between Ly6Chigh/Ly6Clow monocytes and M2/M1 macrophages, along with decreased T cell proliferation and elevated circulating levels of interleukins IL-23 and IL-27. In vitro conditioning with LAV-BPIFB4 protein of macrophages from atherosclerotic patients resulted in a CXCR4-dependent M2 polarization phenotype. Furthermore, LAV-BPIFB4 treatment of arteries explanted from atherosclerotic patients increased the release of atheroprotective IL-33, while inhibiting the release of pro-inflammatory IL-1β, inducing endothelial nitric oxide synthase phosphorylation and restoring endothelial function. Finally, significantly lower plasma BPIFB4 was detected in patients with pathological carotid stenosis (>25%) and intima media thickness >2 mm.
Conclusion
Transfer of the LAV of BPIFB4 reduces the atherogenic process and skews macrophages towards an M2-resolving phenotype through modulation of CXCR4, thus opening up novel therapeutic possibilities in cardiovascular disease.