Gadde Rajitha, Shah Shrey, Böhlke Mark, Kim Jonghan, Betharia Swati
Department of Pharmaceutical Sciences, MCPHS University, School of Pharmacy, Boston, MA, USA.
Department of Biomedical and Nutritional Sciences, Zuckerberg College of Health Sciences, University of Massachusetts Lowell, MA, USA.
Free Radic Biol Med. 2025 May;232:421-436. doi: 10.1016/j.freeradbiomed.2025.02.046. Epub 2025 Mar 1.
Wilson's Disease (WD) is a rare autosomal recessive disorder caused by mutations in the ATP7B gene. These mutations lead to defective copper (Cu) transport and to accumulation of Cu in tissues, primarily in the liver and brain. Current treatment options such as D-penicillamine, trientine, and zinc salts focus on increasing Cu excretion or reducing Cu absorption, but often cause debilitating side effects. N,N'-bis(2-mercaptoethyl)isophthalamide (NBMI) is a lipophilic thiol-based compound originally developed for environmental decontamination. It has been shown to chelate toxic metals such as mercury, lead, and cadmium. This study was designed to evaluate the efficacy of NBMI to mitigate Cu overload using both in vitro and in vivo models of WD. HepG2 cells with the ATP7B gene knocked down had increased sensitivity to copper sulfate (CuSO) compared to wild-type (WT) cells, validating the cell model for WD. Pretreatment with NBMI (2.5-50 μM) improved cell viability, reduced Cu-induced oxidative stress, decreased metallothionein levels, mitigated resulting DNA damage, and reduced overall levels of free intracellular Cu. In an established toxic milk mouse (tx-J) model of WD, 1% dietary NBMI effectively lowered hepatic, cerebral, and renal Cu levels. Treatment with 1% NBMI also improved liver function, as evidenced by reduced ALT levels and normalized hepatocyte morphology. Tx-J mice displayed higher liver-to-body weight ratios compared to WT mice, and treatment with 1% NBMI effectively reduced this ratio. While NBMI did not impact the elevated white blood cell counts and low platelet levels characteristic of tx-J mice, it also did not cause any detrimental effects on red blood cell, hemoglobin, and hematocrit levels. This dose of NBMI also restored homeostasis of other dysregulated essential metal ions in tx-J mice. These findings suggest that dietary administration of NBMI effectively chelates excess free Cu, ameliorates WD symptoms and offers a promising alternative to existing chelators.
威尔逊病（WD）是一种由ATP7B基因突变引起的罕见常染色体隐性疾病。这些突变导致铜（Cu）转运缺陷以及Cu在组织中蓄积，主要是在肝脏和大脑中。目前的治疗方法，如D-青霉胺、曲恩汀和锌盐，主要侧重于增加Cu排泄或减少Cu吸收，但往往会引起使人衰弱的副作用。N，N'-双（2-巯基乙基）间苯二甲酰胺（NBMI）是一种基于硫醇的亲脂性化合物，最初是为环境净化而开发的。它已被证明能螯合汞、铅和镉等有毒金属。本研究旨在使用WD的体外和体内模型评估NBMI减轻Cu过载的疗效。与野生型（WT）细胞相比，ATP7B基因敲低的HepG2细胞对硫酸铜（CuSO）的敏感性增加，验证了WD的细胞模型。用NBMI（2.5-50 μM）预处理可提高细胞活力，降低Cu诱导的氧化应激，降低金属硫蛋白水平，减轻由此产生的DNA损伤，并降低细胞内游离Cu的总体水平。在已建立的WD毒性乳鼠（tx-J）模型中，1%的饮食NBMI有效地降低了肝脏、大脑和肾脏中的Cu水平。用1%的NBMI治疗也改善了肝功能，这可通过降低ALT水平和使肝细胞形态正常化来证明。与WT小鼠相比，tx-J小鼠的肝体重比更高，用1%的NBMI治疗可有效降低该比例。虽然NBMI对tx-J小鼠特有的白细胞计数升高和血小板水平降低没有影响，但它也没有对红细胞、血红蛋白和血细胞比容水平造成任何有害影响。该剂量的NBMI还恢复了tx-J小鼠中其他失调的必需金属离子的稳态。这些发现表明，饮食中给予NBMI可有效螯合过量的游离Cu，改善WD症状，并为现有螯合剂提供了一种有前景的替代方案。
