Immunomodulation of the donor lung with CRISPR-mediated activation of IL-10 expression.
Immunomodulation of the donor lung with CRISPR-mediated activation of IL-10 expression.
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DOI：
10.1016/j.healun.2023.06.001
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发表时间：
2023-10
影响因子：
8.9
通讯作者：
Keshavjee, Shaf
中科院分区：
文献类型：
--
作者：
Mesaki, Kumi;Juvet, Stephen;Yeung, Jonathan;Guan, Zehong;Wilson, Gavin W.;Hu, Jim;Davidson, Alan R.;Kleinstiver, Benjamin P.;Cypel, Marcelo;Liu, Mingyao;Keshavjee, Shaf
供体肺的炎性损伤仍然是肺移植中的一个持续挑战，限制了供体器官的使用和移植后的结果。在供体器官中诱导免疫调节能力可以解决这个未解决的临床问题。我们试图将CRISPR-Cas技术应用于供体肺以微调免疫调节基因表达，首次探索CRISPR介导的转录激活在整个供体肺中的治疗用途。我们在体外和体内探索了CRISPR介导的IL-10（一种关键的免疫调节细胞因子）转录上调的可行性。我们首先评估了大鼠和人细胞系中基因激活的效力、可滴定性和多样性。接下来，在大鼠肺中表征体内CRISPR介导的IL-10活化。最后，将IL-10活化的供体肺移植到受体大鼠中以评估移植环境中的可行性。靶向转录激活在体外诱导稳健的和可滴定的IL-10上调。向导RNA的组合还促进多重基因调节，即IL-10和IL 1受体拮抗剂的同时活化。体内分析表明，使用免疫抑制将基于Cas9的激活剂腺病毒递送至肺是可行的，免疫抑制常规应用于器官移植受体。转录调节的供体肺在同基因和同种异体受体中保持IL-10上调。我们的研究结果强调了CRISPR表观基因组编辑通过在供体器官中创造更有利的免疫调节环境来改善肺移植结果的潜力，这一范例可能扩展到其他器官移植。
Inflammatory injury in the donor lung remains a persistent challenge in lung transplantation that limits donor organ usage and post-transplant outcomes. Inducing immunomodulatory capacity in donor organs could address this unsolved clinical problem. We sought to apply CRISPR-Cas technologies to the donor lung to fine-tune immunomodulatory gene expression, exploring for the first time the therapeutic use of CRISPR-mediated transcriptional activation in the whole donor lung. We explored the feasibility of CRISPR-mediated transcriptional upregulation of IL-10, a key immunomodulatory cytokine, in vitro and in vivo. We first evaluated the potency, titratability, and multiplexibility of the gene activation in rat and human cell lines. Next, in vivo CRISPR-mediated IL-10 activation was characterized in rat lungs. Finally, the IL-10-activated donor lungs were transplanted into recipient rats to assess the feasibility in a transplant setting. The targeted transcriptional activation induced robust and titrable IL-10 upregulation in vitro. The combination of guide RNAs also facilitated multiplex gene modulation, i.e. simultaneous activation of IL-10 and IL1 receptor antagonist. In vivo profiling demonstrated that adenoviral delivery of Cas9-based activators to the lung was feasible with the use of immunosuppression, which is routinely applied to organ transplant recipients. The transcriptionally modulated donor lungs retained IL-10 upregulation in isogeneic and allogeneic recipients. Our findings highlight the potential of CRISPR epigenome editing to improve lung transplant outcomes by creating a more favorable immunomodulatory environment in the donor organ, a paradigm that may be extendable to other organ transplants.
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影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
76.2
作者：
Loor, Gabriel;Warnecke, Gregor;Van Raemdonck, Dirk
通讯作者：
Van Raemdonck, Dirk
影响因子：
82.9
作者：
Maeder, Morgan L.;Stefanidakis, Michael;Jiang, Haiyan
通讯作者：
Jiang, Haiyan
影响因子：
5.1
作者：
Martins, S;de Perrot, M;Keshavjee, S
通讯作者：
Keshavjee, S
DOI：
10.1016/s0022-5223(03)00114-4
发表时间：
2003-10-01
影响因子：
6
作者：
Fischer, S;de Perrot, M;Keshavjee, S
通讯作者：
Keshavjee, S