Voznesenskaya Anna, Berggren Per-Olof, Ilegems Erwin
The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Stockholm, Sweden.
Front Bioeng Biotechnol. 2023 Jul 19;11:1147244. doi: 10.3389/fbioe.2023.1147244. eCollection 2023.
Genetic modification of pancreatic islet organoids, assembled prior to transplantation is an emerging alternative to direct genetic manipulations for a number of clinical and research applications. We have previously shown that dispersion of islet cells followed by re-aggregation into islet organoids, or pseudoislets, allows for efficient transduction with viral vectors, while maintaining physiological functions of native islets. Among viruses currently used for genetic manipulations, adeno-associated viruses (AAVs) have the most attractive safety profile making them suitable for gene therapy applications. Studies reporting on pseudoislet transduction with AAVs are, however, lacking. Here, we have characterized in detail the performance of AAV serotype 8 in transduction of islet cells during pseudoislet formation in comparison with human adenovirus type 5 (AdV5). We have assessed such parameters as transduction efficiency, expression kinetics, and endocrine cell tropism of AAV8 alone or in combination with AdV5. Data provided within our study may serve as a reference point for future functional studies using AAVs for gene transfer to islet cell organoids and will facilitate further development of engineered pseudoislets of superior quality suitable for clinical transplantation.
在移植前组装的胰岛类器官的基因改造，对于许多临床和研究应用来说，是直接基因操作的一种新兴替代方法。我们之前已经表明，将胰岛细胞分散后再重新聚集形成胰岛类器官或假胰岛，能够在保持天然胰岛生理功能的同时，通过病毒载体进行高效转导。在目前用于基因操作的病毒中，腺相关病毒（AAV）具有最具吸引力的安全特性，使其适用于基因治疗应用。然而，关于AAV对假胰岛转导的研究报道却很缺乏。在这里，我们详细比较了AAV血清型8与人5型腺病毒（AdV5）在假胰岛形成过程中对胰岛细胞的转导性能。我们评估了单独使用AAV8或与AdV5联合使用时的转导效率、表达动力学和内分泌细胞嗜性等参数。我们研究中提供的数据可为未来使用AAV将基因转移到胰岛细胞类器官的功能研究提供参考，并将促进适合临床移植的高质量工程化假胰岛的进一步发展。
