Cas9-mediated knockout of Ndrg2 enhances the regenerative potential of dendritic cells for wound healing
Cas9-mediated knockout of Ndrg2 enhances the regenerative potential of dendritic cells for wound healing
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DOI：
10.1101/2022.03.14.484360
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发表时间：
2022-03
影响因子：
16.6
通讯作者：
Dominic Henn;Dehua Zhao;Dharshan Sivaraj;Artem A. Trotsyuk;C. Bonham;Katharina S. Fischer;Tim Kehl;Tobias Fehlmann;Autumn H. Greco;Hudson C. Kussie;Sylvia E Moortgat Illouz;Jagannath Padmanabhan;Janos A. Barrera;U. Kneser;H. Lenhof;Michael Januszyk;B. Levi;A. Keller;M. Longaker;Kellen Chen;Lei S. Qi;G. Gurtner
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作者：
Dominic Henn;Dehua Zhao;Dharshan Sivaraj;Artem A. Trotsyuk;C. Bonham;Katharina S. Fischer;Tim Kehl;Tobias Fehlmann;Autumn H. Greco;Hudson C. Kussie;Sylvia E Moortgat Illouz;Jagannath Padmanabhan;Janos A. Barrera;U. Kneser;H. Lenhof;Michael Januszyk;B. Levi;A. Keller;M. Longaker;Kellen Chen;Lei S. Qi;G. Gurtner
慢性伤口给广大患者群体带来了沉重的医疗负担。基于细胞的疗法虽然已显示出治疗慢性伤口的益处，但尚未广泛应用于临床实践。在这里，我们开发了一种新型 CRISPR/Cas9 方法来精确编辑树突状细胞 (DC)，以增强其治愈慢性伤口的治疗潜力。通过对耐受性 DC 进行单细胞 RNA 测序 (scRNA-seq)，我们发现 N-myc 下调基因 2 (Ndrg2) 标记了 DC 祖细胞的特定群体，是 CRISPR 敲除 (KO) 的有希望的靶标。 Ndrg2-KO 改变 DC 的转录组谱，并保留具有强大的促血管生成和再生能力的未成熟细胞状态。然后，我们将基于 CRISPR 的细胞工程融入水凝胶技术中，用于体内细胞递送，并开发了一种基于 DC 的免疫疗法的高效转化方法，可加速非糖尿病和糖尿病小鼠模型中全层伤口的愈合。这些发现可能为未来在树突状细胞中使用安全基因编辑来治疗各种类型的慢性伤口的临床试验打开大门。
Chronic wounds impose a significant healthcare burden to a broad patient population. Cell based therapies, while having shown benefits for the treatment of chronic wounds, have not achieved widespread adoption into clinical practice. Here, we developed a novel CRISPR/Cas9 approach to precisely edit dendritic cells (DCs) to enhance their therapeutic potential for healing chronic wounds. Using single-cell RNA sequencing (scRNA-seq) of tolerogenic DCs, we discover N-myc downregulated gene 2 (Ndrg2), which marks a specific population of DC progenitors, as a promising target for CRISPR knockout (KO). Ndrg2-KO alters the transcriptomic profile of DCs and preserves an immature cell state with a strong, pro-angiogenic and regenerative capacity. We then incorporated our CRISPR-based cell engineering within a hydrogel technology for in vivo cell delivery and developed a highly effective translational approach for DC based immunotherapy that accelerated healing of full-thickness wounds in both non-diabetic and diabetic mouse models. These findings could open the door to future clinical trials using safe gene editing in DCs for treating various types of chronic wounds.