CD90型
遗传增强
干细胞
离体
造血
造血干细胞
生物
病毒载体
祖细胞
川地34
移植
病毒学
细胞生物学
体内
重组DNA
基因
医学
遗传学
外科
作者
Kurt Berckmueller,Justin Thomas,Eman A. Taha,Seunga Choo,Ravishankar Madhu,Greta Kanestrom,Peter B. Rupert,Roland K. Strong,Hans‐Peter Kiem,Stefan Radtke
标识
DOI:10.1016/j.ymthe.2023.08.003
摘要
Hematopoietic stem cell (HSC) gene therapy is currently performed on CD34+ hematopoietic stem and progenitor cells containing less than 1% true HSCs and requiring a highly specialized infrastructure for cell manufacturing and transplantation. We have previously identified the CD34+CD90+ subset to be exclusively responsible for short- and long-term engraftment. However, purification and enrichment of this subset is laborious and expensive. HSC-specific delivery agents for the direct modification of rare HSCs are currently lacking. Here, we developed novel targeted viral vectors to specifically transduce CD90-expressing HSCs. Anti-CD90 single chain variable fragments (scFvs) were engineered onto measles- and VSV-G-pseudotyped lentiviral vectors that were knocked out for native targeting. We further developed a custom hydrodynamic titration methodology to assess the loading of surface-engineered capsids, measure antigen recognition of the scFv, and predict the performance on cells. Engineered vectors formed with minimal impairment in the functional titer, maintained their ability to fuse with the target cells, and showed highly specific recognition of CD90 on cells ex vivo. Most important, targeted vectors selectively transduced human HSCs with secondary colony-forming potential. Our novel HSC-targeted viral vectors have the potential to significantly enhance the feasibility of ex vivo gene therapy and pave the way for future in vivo applications.
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