Targeting Human Osteoarthritic Chondrocytes with Ligand Directed Bacteriophage-Based Particles.
Targeting Human Osteoarthritic Chondrocytes with Ligand Directed Bacteriophage-Based Particles.
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以配体导向的噬菌体颗粒为靶点的人骨关节炎软骨细胞。
DOI：
10.3390/v13122343
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发表时间：
2021-11-23
期刊：
影响因子：
--
通讯作者：
Pothacharoen P
中科院分区：
文献类型：
--
作者：
Chongchai A;Waramit S;Wongwichai T;Kampangtip J;Phitak T;Kongtawelert P;Hajitou A;Suwan K;Pothacharoen P
骨关节炎（OA）是一种退行性关节疾病，其特征是关节软骨进行性恶化和丧失。目前尚无治疗方法可以逆转 OA 的发作。因此，我们开发了一种靶向递送策略，将基因转移到原代人类软骨细胞中，作为概念验证研究。我们在基于 M13 丝状噬菌体（噬菌体）的颗粒的 pIII 小外壳蛋白上展示了软骨细胞亲和肽（CAP），该颗粒在巨细胞病毒 CMV 启动子和腺相关病毒血清型 2（AAV-2）的反向末端重复序列（ITR）顺式元件下携带哺乳动物转基因盒。使用原代人关节软骨细胞（HAC）作为体外模型，并表征了 CAP 配体与 HAC 致病条件相关的选择性和结合特性。我们发现 CAP 配体对致病性 HAC 具有高度选择性。此外，还评估了 CAP 展示噬菌体 (CAP.Phage) 的稳定性、细胞毒性和基因传递效率。我们发现噬菌体颗粒在较宽的温度和 pH 值范围内稳定，同时对 HAC 没有细胞毒性。重要的是，携带分泌型荧光素酶 (Lucia) 报告基因的 CAP.Phage 颗粒可以高效、选择性地将转基因表达传递至 HAC。总之，发现CAP配体优先结合致病性软骨细胞，并且CAP.噬菌体颗粒成功地将转基因靶向并递送至HAC。
Osteoarthritis (OA) is a degenerative joint disease characterized by progressive deterioration and loss of articular cartilage. There is currently no treatment to reverse the onset of OA. Thus, we developed a targeted delivery strategy to transfer genes into primary human chondrocytes as a proof-of-concept study. We displayed a chondrocyte-affinity peptide (CAP) on the pIII minor coat protein of the M13 filamentous bacteriophage (phage)-based particle carrying a mammalian transgene cassette under cytomegalovirus CMV promoter and inverted terminal repeats (ITRs) cis elements of adeno-associated virus serotype 2 (AAV-2). Primary human articular chondrocytes (HACs) were used as an in vitro model, and the selectivity and binding properties of the CAP ligand in relation to the pathogenic conditions of HACs were characterized. We found that the CAP ligand is highly selective toward pathogenic HACs. Furthermore, the stability, cytotoxicity, and gene delivery efficacy of the CAP-displaying phage (CAP.Phage) were evaluated. We found that the phage particle is stable under a wide range of temperatures and pH values, while showing no cytotoxicity to HACs. Importantly, the CAP.Phage particle, carrying a secreted luciferase (Lucia) reporter gene, efficiently and selectively delivered transgene expression to HACs. In summary, it was found that the CAP ligand preferably binds to pathogenic chondrocytes, and the CAP.Phage particle successfully targets and delivers transgene to HACs.
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