Remyelination-Promoting DNA Aptamer Conjugate Myaptavin-3064 Binds to Adult Oligodendrocytes In Vitro.
Remyelination-Promoting DNA Aptamer Conjugate Myaptavin-3064 Binds to Adult Oligodendrocytes In Vitro.
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DOI：
10.3390/ph13110403
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发表时间：
2020-11-19
期刊：
影响因子：
--
通讯作者：
Maher LJ 3rd
中科院分区：
文献类型：
--
作者：
Fereidan-Esfahani M;Yue WY;Wilbanks B;Johnson AJ;Warrington AE;Howe CL;Rodriguez M;Maher LJ 3rd
我们以前应用系统进化配体指数富集（SELEX）技术，以确定髓鞘特异性DNA适体，使用粗小鼠中枢神经系统髓鞘作为诱饵。该选择鉴定了40个核苷酸的适体（LJM-3064）。将多个生物素化的LJM-3064分子缀合至链霉亲和素核心以模拟多聚体免疫球蛋白M（IgM）抗体，产生3064-BS-链霉亲和素（Myaptavin-3064）。我们先前显示Myaptavin-3064在慢性脊髓脱髓鞘的Theiler小鼠脑脊髓炎病毒（TMEV）模型中诱导髓鞘再生。虽然靶结合的细节和作用机制仍不清楚，但我们假设Myaptavin-3064通过结合少突胶质细胞（OL）诱导髓鞘再生。我们现在报告的结果，使用人少突胶质细胞瘤（HOG）细胞系的结合试验，应用流式细胞术和免疫细胞化学（IC）来测定适体结合细胞。应用IC测定来比较适体缀合物与原代胚胎小鼠混合皮质培养物和原代成年大鼠混合神经胶质培养物的结合。我们表明Myaptavin-3064与HOG细胞结合，在分化后结合增加。相反，不促进中枢神经系统（CNS）髓鞘再生的阴性对照适体缀合物3060-BS不结合HOG细胞。Myaptavin-3064不与肺（L2）或肾（BHK）细胞系结合。全内反射荧光（TIRF）成像表明Myaptavin-3064结合在活细胞的细胞膜上。除了HOG细胞，Myaptavin-3064结合成年大鼠OL，但不结合胚胎小鼠混合皮质培养物。这些数据支持Myaptavin-3064以触发髓鞘再生信号途径的方式结合啮齿动物和人OL两者上的表面分子的假设。
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.
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影响因子：
12.7
作者：
Bradl M;Lassmann H
通讯作者：
Lassmann H
DOI：
10.1073/pnas.1808064115
发表时间：
2018-12-11
影响因子：
11.1
作者：
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通讯作者：
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影响因子：
6.2
作者：
KAHN, MA;DEVELLIS, J
通讯作者：
DEVELLIS, J
影响因子：
3.7
作者：
Nastasijevic B;Wright BR;Smestad J;Warrington AE;Rodriguez M;Maher LJ 3rd
通讯作者：
Maher LJ 3rd
DOI：
10.1073/pnas.0812500106
发表时间：
2009-04-21
影响因子：
11.1
作者：
Duncan, I. D.;Brower, A.;Schultz, R. D.
通讯作者：
Schultz, R. D.