Ganglioside functionalized lipid-coated nanoparticles: fabrication, characterization and application
Embargo Date
2027-02-05
OA Version
Citation
Abstract
Siglecs (sialic acid binding immunoglobulin-like lectins) are a family of lectins characterized by an amino-terminal V-set immunoglobulin domain that mediates sialic acid binding and variable numbers of C2-set immunoglobulin domains. Sialic acids are a family of nine-carbon acidic monosaccharides that occur naturally at the end of glycoconjugates, such as glycoproteins and glycolipids. Mammalian glycome contains numerous sialylated glycans that can be potentially recognized as ligands by Siglecs. Among them, it was discovered that binding of the monosialodihexosylgangliosid (GM3) to Siglec‐1 (Sialoadhesin, CD169) plays an important role in the glycoprotein‐independent sequestration of HIV‐1 particles in non-lysosomal virus‐containing compartments (VCCs) in CD169+ myeloid cells. It has been shown previously that GM3-functionalized gold nanoparticles recapitulate key aspects of CD169-dependent HIV-1 uptake and trafficking in CD169-expressing dendritic cells and macrophages. Therefore, CD169 becomes a potential target utilizing the GM3-NPs for multiple therapeutic applications. In this dissertation, we first studied the GM3 incorporated lipid-coated mesoporous silica nanoparticles (LMSN) as antiretroviral drugs delivery carrier. GM3 is incorporated into the membrane to facilitate LMSN binding to CD169-expressing THP-1 cells. The lipid layer around the silica core provides stealth properties that dampen the inflammatory cytokine response in human monocyte-derived macrophages. Quantification of antiretrovirals release from nanoparticles, and assessment of antiviral potency to HIV-1 infection in vitro reveals that antiretrovirals co-formulated into LMSN retain optimal antiviral potency for 90 days when stored at room temperature. These findings suggest that GM3-NP equip the MSN core with lipid-derived properties for surface passivation and lipid-mediated binding are of high interest for achieving an effective delivery of antiretrovirals to tissue reservoirs of HIV-1 replication.
Next, we studied the Chimeric Antigen Receptor (CAR) T cell activation by the GM3-NPs. Activation of CAR T cells at the immunological synapse (IS) formed between antigen presenting cell (APC) and T cell is thought to promote strong, persistent antigen-specific T cell-mediated immune responses but requires integration of CAR ligands into the APC/T cell interface. We demonstrated that CAR ligand functionalized, lipid-coated, biodegradable polymer NPs that contain the ganglioside GM3 bind to CD169-expressing APCs and localize to the cell contact site between APCs and CAR T cells upon initiation of cell conjugates. The CD169+ APC/CAR T cell interface is characterized by a strong optical colocalization of GM3-NPs and CARs, enrichment of F-actin, and recruitment of ZAP-70, indicative of integration of GM3-NPs into a functional IS. Overall, it identifies GM3-NPs as a potential antigen delivery platform for active targeting of CD169 expressing APCs and enhancement of CAR T cell activation at the NP-containing IS.
Lastly, we investigated the GM3-NP as T cell modulator by conjugating anti-CD3 (Signal 1) and anti-CD28 (Signal 2) antibodies on the NP. The conjugation of stimulatory molecules on the GM3-NP was quantified. The stimulatory effect on T cells was tested in vitro. The APC-mediated activation resulted in a higher level of T cell when loading antibodies conjugated GM3-NPs on to CD169+ cells than by using antibodies conjugated GM3-NP alone. The in vivo delivery and accumulation of GM3-NPs were explored by two administration routes.
Description
2023
License
Attribution-NonCommercial-ShareAlike 4.0 International