VCAM-1 Targeted Lipopolyplexes as Vehicles for Efficient Delivery of shRNA-Runx2 to Osteoblast-Differentiated Valvular Interstitial Cells; Implications in Calcific Valve Disease Treatment.
Geanina VoicuDaniela RebleanuCristina Ana MocanuGabriela TankoIonel DrocCristina-Mariana UrituMariana PintealaIleana ManduteanuMaya SimionescuManuela CalinPublished in: International journal of molecular sciences (2022)
Calcific aortic valve disease (CAVD) is a progressive inflammatory disorder characterized by extracellular matrix remodeling and valvular interstitial cells (VIC) osteodifferentiation leading to valve leaflets calcification and impairment movement. Runx2, the master transcription factor involved in VIC osteodifferentiation, modulates the expression of other osteogenic molecules. Previously, we have demonstrated that the osteoblastic phenotypic shift of cultured VIC is impeded by Runx2 silencing using fullerene (C60)-polyethyleneimine (PEI)/short hairpin (sh)RNA-Runx2 (shRunx2) polyplexes. Since the use of polyplexes for in vivo delivery is limited by their instability in the plasma and the non-specific tissue interactions, we designed and obtained targeted, lipid-enveloped polyplexes (lipopolyplexes) suitable for (1) systemic administration and (2) targeted delivery of shRunx2 to osteoblast-differentiated VIC (oVIC). Vascular cell adhesion molecule (VCAM)-1 expressed on the surface of oVIC was used as a target, and a peptide with high affinity for VCAM-1 was coupled to the surface of lipopolyplexes encapsulating C60-PEI/shRunx2 (V-LPP/shRunx2). We report here that V-LPP/shRunx2 lipopolyplexes are cyto- and hemo-compatible and specifically taken up by oVIC. These lipopolyplexes are functional as they downregulate the Runx2 gene and protein expression, and their uptake leads to a significant decrease in the expression of osteogenic molecules (OSP, BSP, BMP-2). These results identify V-LPP/shRunx2 as a new, appropriately directed vehicle that could be instrumental in developing novel strategies for blocking the progression of CAVD using a targeted nanomedicine approach.
Keyphrases
- aortic valve
- transcription factor
- cell adhesion
- transcatheter aortic valve replacement
- aortic stenosis
- extracellular matrix
- transcatheter aortic valve implantation
- aortic valve replacement
- induced apoptosis
- mesenchymal stem cells
- cancer therapy
- poor prognosis
- cell cycle arrest
- genome wide identification
- bone marrow
- dna binding
- mitral valve
- oxidative stress
- multiple sclerosis
- signaling pathway
- bone regeneration
- binding protein
- drug delivery
- endothelial cells
- coronary artery disease
- ejection fraction
- endoplasmic reticulum stress
- long non coding rna
- gene expression
- heart failure
- cell proliferation
- combination therapy
- chronic kidney disease
- fatty acid
- pi k akt