Remyelination-Promoting DNA Aptamer Conjugate Myaptavin-3064 Binds to Adult Oligodendrocytes In Vitro

Abstract

We previously applied Systematic Evolution of Ligands by EXponential enrichment (SELEX) technology to identify myelin-specific DNA aptamers, using crude mouse central nervous system myelin as bait. This selection identified a 40-nucleotide aptamer (LJM-3064). Multiple biotinylated LJM-3064 molecules were conjugated to a streptavidin core to mimic a multimeric immunoglobulin M (IgM) antibody, generating 3064-BS-streptavidin (Myaptavin-3064). We previously showed that Myaptavin-3064 induces remyelination in the Theiler’s murine encephalomyelitis virus (TMEV) model of chronic spinal cord demyelination. While details of target binding and the mechanism of action remain unclear, we hypothesized that Myaptavin-3064 induces remyelination by binding to oligodendrocytes (OLs). We now report the results of binding assays using the human oligodendroglioma (HOG) cell line, applying both flow cytometry and immunocytochemistry (IC) to assay aptamer conjugate binding to cells. IC assays were applied to compare aptamer conjugate binding to primary embryonic mouse mixed cortical cultures and primary adult rat mixed glial cultures. We show that Myaptavin-3064 binds to HOG cells, with increased binding upon differentiation. In contrast, a negative control aptamer conjugate, 3060-BS, which did not promote central nervous system (CNS) remyelination, does not bind to HOG cells. Myaptavin-3064 did not bind to lung (L2) or kidney (BHK) cell lines. Total internal reflection fluorescence (TIRF) imaging indicates that Myaptavin-3064 binds at the cell membrane of live cells. In addition to HOG cells, Myaptavin-3064 binds to adult rat OLs, but not to embryonic mouse mixed cortical cultures. These data support the hypothesis that Myaptavin-3064 binds to a surface molecule on both rodent and human OLs in a manner that triggers a remyelination signal pathway.

Karakterisasi Mikrorna-15a Terenkapsulasi Liposom Dengan Konjugasi Dna Aptamer Myaptavin-3064 Sebagai Novel Terapi Remielinisasi Sel Oligodendrosit Pada Skizofrenia

“…Gambar 3. Gambar menunjukan ilustrasi dari hipotesis terkait proses remielinisasi sel oligodendrosit yang diinduksi oleh miaptavin-3064 (31).…”

Application of aptamers in regenerative medicine

“…Their further study found that a streptavidin tetramer could mediate the coupling of four aptamers. This formed the tetramer aptamer complex Myaptavin-3064, which mimics the activity of polyvalent antibodies ( Fereidan-Esfahani et al, 2020 ). The biological stability of the aptamer complex was enhanced compared with that of the aptamer monomer.…”

“…In 2020, Fereidan-Esfahani et al (2020) used flow cytometry and immunocytochemistry to determine the cell-binding properties of Myaptavin-3064 in vitro . Myaptavin-3064 shows relatively specific binding to the human oligodendroglioma (HOG) cell line.…”

“…Two synergistic biological effects of myelin regeneration and antiinflammation were achieved in the multiple sclerosis mouse model induced by a myelin oligodendrocyte glycoprotein, which expanded the application of the LJM-3064 aptamer (Hosseini et al, 2019). In 2020, Fereidan-Esfahani et al (2020 used flow cytometry and immunocytochemistry to determine the cell-binding properties of Myaptavin-3064 in vitro. Myaptavin-3064 shows relatively specific binding to the human oligodendroglioma (HOG) cell line.…”

Aptamers’ Potential to Fill Therapeutic and Diagnostic Gaps

“…Finally, Dr. Maher III’s group aimed to understand the biological mechanism of a previously described DNA aptamer conjugate with streptavidine, also capable of adopting G-quadruplex structure, shown to promote remyelination in a mouse model of chronic spinal cord demyelination [ 90 ]. In their most recent paper, the authors provide evidence that this macromolecular aptamer-based tool binds to the cell membrane of human and adult rat oligodendrocytes, suggesting that binding to a membrane molecule triggers the remyelination pathway [ 91 ].…”