Dabrowski Alexandra C, Logan Alexandria R, Rayaji Rameshwari, Rodriguez Brianna, Cai Li, Vazquez Maribel
Department of Biomedical Engineering, Rutgers, The State University of New Jersey-New Brunswick, Piscataway, NJ 08854, US.
Explor Biomat X. 2025;2. doi: 10.37349/ebmx.2025.101335. Epub 2025 Apr 14.
This study evaluated the impact of retinal extracellular matrix (ECM) and key biomaterial substrates on the motility of transplantable retinal cells with genomic manipulation, using the therapeutic molecule, Topoisomerase II beta (Top2b), as a model.
Tests first applied in ovo electroporation to examine the effects of a pharmacological Top2b inhibitor (ICRF-193) on progenitor motility and development of embryonic retina. Complementary qRT-PCR tests measured changes in select cadherin molecules in response to treatment. In vitro transfection produced cultured retinal progenitor cell groups with Top2b overexpression and Top2b knockdown. Differences in the adhesion and motility of Top2b altered groups, compared to wildtype cells, were measured upon biomaterial substrates used in emerging transplantation matrixes.
Data illustrated significant differences in the number and spacing of retinal ganglion cells when retina was treated with ICRF-193, as well as downregulation of several key cadherin molecules. Cultured retinal progenitors with Top2b knockdown and Top2b overexpression exhibited different expression of chemotactic receptors, adhesion parameters, and modalities of migration upon substrates of laminin, poly-L-lysine, and collagen IV. Significant changes in cell morphology and surface area were also measured compared to wildtype cells.
Corroborating in vivo and in vitro data support Top2b as a therapeutic target for retinal progenitor motility but indicate significant differences in the migration of Top2b altered cells upon substrates used in transplantation. These data highlight the therapeutic advantages of bioinspired materials developed to aid the motility of replacement cells with modified genetic expression to improve transplantation outcomes across the nervous system.
本研究以治疗性分子拓扑异构酶IIβ（Top2b）为模型，评估视网膜细胞外基质（ECM）和关键生物材料底物对经基因组操作的可移植视网膜细胞运动性的影响。
首先在鸡胚电穿孔中进行试验，以检查一种Top2b药理学抑制剂（ICRF-193）对胚胎视网膜祖细胞运动性和发育的影响。互补定量逆转录聚合酶链反应（qRT-PCR）试验测量了治疗后选定钙黏蛋白分子的变化。体外转染产生了Top2b过表达和Top2b敲低的培养视网膜祖细胞群。在新兴移植基质中使用的生物材料底物上，测量Top2b改变组与野生型细胞相比在黏附性和运动性方面的差异。
数据表明，用ICRF-193处理视网膜时，视网膜神经节细胞的数量和间距存在显著差异，同时几种关键钙黏蛋白分子下调。在层粘连蛋白、聚-L-赖氨酸和IV型胶原底物上，Top2b敲低和Top2b过表达的培养视网膜祖细胞表现出趋化受体表达、黏附参数和迁移方式的不同。与野生型细胞相比，细胞形态和表面积也有显著变化。
体内和体外数据相互印证，支持Top2b作为视网膜祖细胞运动性的治疗靶点，但表明Top2b改变的细胞在移植所用底物上的迁移存在显著差异。这些数据突出了生物启发材料的治疗优势，这些材料旨在帮助具有修饰基因表达的替代细胞运动，以改善整个神经系统的移植效果。
