Poole Brittney, Oshins Regina, Huo Zhiguang, Aranyos Alek, West Jesse, Duarte Sergio, Clark Virginia C, Beduschi Thiago, Zarrinpar Ali, Brantly Mark, Khodayari Nazli
Department of Medicine, Division of Pulmonary, Critical Care and Sleep Medicine, University of Florida, Gainesville, Florida, USA.
Department of Biostatistics, College of Public Health, University of Florida, Gainesville, Florida, USA.
Hepatol Commun. 2024 Jan 29;8(2). doi: 10.1097/HC9.0000000000000370. eCollection 2024 Feb 1.
Alpha-1 antitrypsin deficiency (AATD) is a genetic disease caused by misfolding and accumulation of mutant alpha-1 antitrypsin (ZAAT) in the endoplasmic reticulum of hepatocytes. Hepatic ZAAT aggregates acquire a toxic gain-of-function that impacts the endoplasmic reticulum which is theorized to cause liver disease in individuals with AATD who present asymptomatic until late-stage cirrhosis. Currently, there is no treatment for AATD-mediated liver disease except liver transplantation. In our study of mitochondrial RNA, we identified that Sirtuin3 (SIRT3) plays a role in the hepatic phenotype of AATD.
Utilizing RNA and protein analysis in an in vitro AATD model, we investigated the role of SIRT3 in the pathophysiology of AATD-mediated liver disease while also characterizing our novel, transgenic AATD mouse model.
We show lower expression of SIRT3 in ZAAT-expressing hepatocytes. In contrast, the overexpression of SIRT3 increases hepatic ZAAT degradation. ZAAT degradation mediated by SIRT3 appeared independent of proteasomal degradation and regular autophagy pathways. We observed that ZAAT-expressing hepatocytes have aberrant accumulation of lipid droplets, with ZAAT polymers localizing on the lipid droplet surface in a direct interaction with Perilipin2, which coats intracellular lipid droplets. SIRT3 overexpression also induced the degradation of lipid droplets in ZAAT-expressing hepatocytes. We observed that SIRT3 overexpression induces lipophagy by enhancing the interaction of Perilipin2 with HSC70. ZAAT polymers then degrade as a consequence of the mobilization of lipids through this process.
In this context, SIRT3 activation may eliminate the hepatic toxic gain-of-function associated with the polymerization of ZAAT, providing a rationale for a potential novel therapeutic approach to the treatment of AATD-mediated liver disease.
α-1抗胰蛋白酶缺乏症(AATD)是一种遗传性疾病,由突变的α-1抗胰蛋白酶(ZAAT)在肝细胞内质网中错误折叠和积累所致。肝脏中的ZAAT聚集体获得了一种有毒的功能增益,影响内质网,理论上这会导致AATD患者出现肝脏疾病,这些患者在晚期肝硬化之前通常没有症状。目前,除了肝移植外,尚无针对AATD介导的肝病的治疗方法。在我们对线粒体RNA的研究中,我们发现沉默调节蛋白3(SIRT3)在AATD的肝脏表型中起作用。
利用体外AATD模型中的RNA和蛋白质分析,我们研究了SIRT3在AATD介导的肝病病理生理学中的作用,同时还对我们新的转基因AATD小鼠模型进行了表征。
我们发现表达ZAAT的肝细胞中SIRT3的表达较低。相反,SIRT3的过表达增加了肝脏中ZAAT的降解。SIRT3介导的ZAAT降解似乎独立于蛋白酶体降解和常规自噬途径。我们观察到,表达ZAAT的肝细胞中脂滴异常积累,ZAAT聚合物定位于脂滴表面,并与包裹细胞内脂滴的外周脂膜蛋白2直接相互作用。SIRT3的过表达还诱导了表达ZAAT的肝细胞中脂滴的降解。我们观察到,SIRT3的过表达通过增强外周脂膜蛋白2与热休克蛋白70(HSC70)的相互作用来诱导脂噬。ZAAT聚合物随后因通过该过程的脂质动员而降解。
在这种情况下,SIRT3激活可能消除与ZAAT聚合相关的肝脏毒性功能增益,为治疗AATD介导的肝病提供了一种潜在新治疗方法的理论依据。