Development of a protein therapeutics to counteract mustard-induced skin injury

项目总结 硫磺芥末(SM)和氮芥末(NM)是会导致皮肤起泡的发泡剂。AS 烷化剂，SM/NM通过复杂的细胞事件导致皮肤损伤，涉及DNA损伤，免费 自由基的形成和脂质过氧化。针对多细胞的治疗方法的发展 组织损伤修复过程可能为对抗发泡剂引起的损伤提供有效的对策 皮肤损伤。我们的研究小组先前发现MG53是细胞膜修复的重要组成部分。 与野生型小鼠相比，MG53-/-小鼠对NM引起的皮肤损伤的敏感性增加， 而过表达MG53的转基因小鼠对NM的皮肤暴露具有抵抗力。使用活细胞 在成像方面，我们有了一个意想不到的潜在令人兴奋的发现，在人类卵泡干细胞(Hfscs)中。 经NM处理后，MG53细胞固有的膜修复功能受损。然而，应用程序 外源重组人MG53(RhMG53)蛋白可恢复NM诱导的细胞膜完整性 角质形成细胞和hfSCs的损伤。我们已经生成了初步数据，表明局部给药 重组人MG53对NM致小鼠皮肤损伤有一定的保护作用。这项探索性的研究和 发展中的R21项目专注于了解MG53在皮肤伤口修复中的机制作用 在NM暴露后(目标1)，建立rhMG53的有效剂量和有效时间窗 应用于NM致小鼠皮肤损伤的治疗(目标2)。在等待R21项目的积极结果之前， 我们计划提交一个更大的中和U01申请，以进一步建立大型动物的概念验证数据 SM诱导的皮肤损伤模型，并寻求与FDA和BARDA合作推进rhMG53 作为一种新的蛋白质疗法，可作为治疗皮肤(和其他器官)的药物储备 化学战中的伤害。

PROJECT SUMMARY Sulfur mustard (SM) and nitrogen mustard (NM) are vesicant agents that cause blistering of the skin. As alkylating agents, SM/NM causes skin injuries through complicated cellular events, involving DNA damage, free radical formation, and lipid peroxidation. Development of therapeutic approaches that target the multi-cellular process of tissue injury-repair can potentially provide effective countermeasures to combat vesicant-induced dermal lesions. Our group previously identified MG53 as an essential component of cell membrane repair. Compared with the wild type mice, the mg53-/- mice show increased susceptibility to NM-induced dermal injuries, whereas transgenic mice with overexpression of MG53 are resistant to dermal exposure of NM. Using live-cell imaging, we made an unexpected and potentially exciting finding that in human follicle stem cells (HFSCs) treated with NM, the intrinsic membrane repair function of MG53 was compromised. However, the application of exogenous recombinant human MG53 (rhMG53) protein could restore cell membrane integrity from NM-induced injury in both keratinocytes and HFSCs. We have generated preliminary data to show that topical administration of rhMG53 can mitigate NM-induced dermal injury in mice. Studies proposed in this Exploratory and Developmental R21 project are focused on understanding the mechanistic action of MG53 in skin wound repair following NM exposure (Aim 1), and establishing the efficacious dose and effective time window for rhMG53 application to treat NM-induced skin injuries in mice (Aim 2). Pending on positive findings from this R21 project, we plan to submit a larger CounterACT U01 application to further build the proof-of-concept data in large animal models of SM-induced cutaneous injury, and to seek partnership with FDA and BARDA for advancing rhMG53 as a novel protein therapeutic that can be stockpiled as a medical reserve to treat cutaneous (and other organs) injuries in the event of chemical warfare.