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

项目摘要 硫芥（SM）和氮芥（NM）是引起皮肤起泡的起泡剂。作为 烷化剂SM/NM通过复杂的细胞事件引起皮肤损伤，涉及DNA损伤，游离 自由基形成和脂质过氧化。靶向多细胞肿瘤的治疗方法的发展 组织损伤修复过程可以潜在地提供有效的对策，以对抗水疱剂引起的 皮肤损伤我们的小组先前确定MG 53是细胞膜修复的重要组成部分。 与野生型小鼠相比，mg 53-/-小鼠对NM诱导的皮肤损伤表现出增加的易感性， 而MG 53过表达的转基因小鼠对NM的皮肤暴露具有抗性。使用活细胞 通过成像，我们发现了一个意想不到的和潜在的令人兴奋的发现，在人类卵泡干细胞（HFSC）中， 经NM处理后，MG 53的内在膜修复功能受损。然而，应用 外源性重组人MG 53蛋白（rhMG 53）可以恢复NM诱导的细胞膜完整性， 角质形成细胞和HFSC两者的损伤。我们已经生成了初步数据，表明局部给药 rhMG 53能减轻NM诱导的小鼠皮肤损伤。本探索性研究中提出的研究， 开发R21项目的重点是了解MG 53在皮肤伤口修复中的机制作用 NM暴露后（目标1），并确定rhMG 53的有效剂量和有效时间窗 应用于治疗小鼠中NM诱导的皮肤损伤（目的2）。在R21项目取得积极成果之前， 我们计划提交更大的CounterACT U 01申请，以进一步构建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和巴尔达的合作，以推进rhMG 53 作为一种新的蛋白质治疗剂，可以储存作为医疗储备，以治疗皮肤（和其他器官） 在化学战中受伤。

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.