San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy.
San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy.
Mol Ther. 2023 Jan 4;31(1):230-248. doi: 10.1016/j.ymthe.2022.08.011. Epub 2022 Aug 17.
Mesenchymal stromal cells (MSCs) have been employed in vitro to support hematopoietic stem and progenitor cell (HSPC) expansion and in vivo to promote HSPC engraftment. Based on these studies, we developed an MSC-based co-culture system to optimize the transplantation outcome of clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 gene-edited (GE) human HSPCs. We show that bone marrow (BM)-MSCs produce several hematopoietic supportive and anti-inflammatory factors capable of alleviating the proliferation arrest and mitigating the apoptotic and inflammatory programs activated in GE-HSPCs, improving their expansion and clonogenic potential in vitro. The use of BM-MSCs resulted in superior human engraftment and increased clonal output of GE-HSPCs contributing to the early phase of hematological reconstitution in the peripheral blood of transplanted mice. In conclusion, our work poses the biological bases for a novel clinical use of BM-MSCs to promote engraftment of GE-HSPCs and improve their transplantation outcome.
间充质基质细胞（MSCs）已被用于体外支持造血干细胞和祖细胞（HSPC）的扩增，并在体内促进 HSPC 的植入。基于这些研究，我们开发了一种基于 MSC 的共培养系统，以优化簇状规则间隔短回文重复（CRISPR）-Cas9 基因编辑（GE）人类 HSPC 的移植结果。我们表明，骨髓（BM）-MSCs 产生几种造血支持和抗炎因子，能够缓解 GE-HSPC 中增殖停滞，并减轻激活的细胞凋亡和炎症程序，从而提高其在体外的扩增和集落形成能力。使用 BM-MSCs 可实现更好的人植入，并增加 GE-HSPC 的克隆输出，有助于移植小鼠外周血中血液学重建的早期阶段。总之，我们的工作为 BM-MSCs 促进 GE-HSPC 植入和改善其移植结果的新的临床应用提供了生物学基础。
