Targeting the site encoded by SERPINA1*E342K for treating alpha‐1 antitrypsin deficiency‐associated liver diseases

Zhang, Xiaojuan; Pham, Kien; Li, Danmeng; Schutte, Ryan J.; Brantly, Mark; Liu, Chen; Ostrov, David A.

doi:10.1002/1873-3468.13452

Cited by 3 publications

(6 citation statements)

References 15 publications

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“…The ratio of intracellular and extracellular hAAT levels in this cell line is consistent with clinical data [9]. In contrast, the human hepatoma cell line HepG2 was used as a negative control since it lacks PASD resistant staining and Z-hAAT polymers, and the majority of the hAAT in HepG2 cells is secreted, which suggests HepG2 cells express wild-type human AAT [9,20,22]. PASD, hAAT, or hAAT polymer were not detected in livers from C57BL/6J mice, confirming the specificity of the assays for Z-hAAT [9].…”

Section: Discussionsupporting

confidence: 88%

“…This cell line resembled disease characteristics demonstrated by PASD resistant staining and positive staining by an antibody raised against Z-hAAT polymer in immunohistochemistry stain. The ratio of intracellular and extracellular hAAT levels in this cell line is consistent with clinical data [9]. In contrast, the human hepatoma cell line HepG2 was used as a negative control since it lacks PASD resistant staining and Z-hAAT polymers, and the majority of the hAAT in HepG2 cells is secreted, which suggests HepG2 cells express wild-type human AAT [9,20,22].…”

Section: Discussionsupporting

confidence: 80%

“…We previously developed a cell line comprised of malignant hepatocytes that stably expresses Z-hAAT generated from a PiZ mouse liver for testing candidate compounds for effects on intracellular Z-hAAT aggregation [9]. This cell line resembled disease characteristics demonstrated by PASD resistant staining and positive staining by an antibody raised against Z-hAAT polymer in immunohistochemistry stain.…”

Section: Discussionmentioning

confidence: 99%

“…Z-hAAT mouse malignant hepatocytes [9] were cultured in advanced DMEM/F12, 10% fetal bovine serum, 6 mM l-glutamine, 1% penicillin/streptomycin, and 40 ng/mL dexamethasone in 5% CO 2 at 37 • C. Cultures of the human HepG2 hepatoma cell line (ATCC, Manassas, VA, USA) were similarly maintained. Z-hAAT hepatocytes (1-2 × 10 4 ) and HepG2 cells (6-8 × 10 4 ) were seeded on 96-well tissue culture plates.…”

Section: Cell Culture and Compound Treatmentmentioning

confidence: 99%

“…In addition, PiZ mice develop hepatocyte pathology that resembles diseased liver in humans with AATD [7,8]. In our previous study, a novel PiZ hepatocyte culture system was developed to study the kinetics of Z-hAAT production, polymerization, retention, and cellular response [9]. This novel system also serves as a platform to identify and characterize novel therapeutics capable of mitigating proteotoxicity [9].…”

Section: Introductionmentioning

confidence: 99%

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A Novel Small Molecule Inhibits Intrahepatocellular Accumulation of Z-Variant Alpha 1-Antitrypsin In Vitro and In Vivo

Zhang

Pham

et al. 2019

Cells

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Alpha 1-antitrypsin deficiency (AATD) is the most common genetic cause of liver disease in children and is associated with early-onset chronic liver disease in adults. AATD associated liver injury is caused by hepatotoxic retention of polymerized mutant alpha 1-antitrypsin molecules within the endoplasmic reticulum. Currently, there is no curative therapy for AATD. In this study, we selected small molecules with the potential to bind mutant alpha 1-antitrypsin (Z-variant) to inhibit its accumulation in hepatocytes. We used molecular docking to select candidate compounds that were validated in cell and animal models of disease. A crystal structure of polymerized alpha 1-antitrypsin molecule was used as the basis for docking 139,735 compounds. Effects of the top scoring compounds were investigated in a cell model that stably expresses Z-variant alpha 1-antitrypsin and in PiZ mice expressing Z-variant human alpha 1-antitrypsin (Z-hAAT), encoded by SERPINA1*E342K. 4′,′5-(Methylenedioxy)-2-nitrocinnamic acid was predicted to bind cleaved alpha 1-antitrypsin at the polymerization interface, and observed to co-localize with Z-hAAT, increase Z-hAAT degradation, inhibit intracellular accumulation of Z-hAAT, and alleviate liver fibrosis.

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Section: Discussionsupporting

confidence: 88%

Section: Discussionsupporting

confidence: 80%

Section: Discussionmentioning

confidence: 99%

Section: Cell Culture and Compound Treatmentmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Novel Small Molecule Inhibits Intrahepatocellular Accumulation of Z-Variant Alpha 1-Antitrypsin In Vitro and In Vivo

Zhang

Pham

et al. 2019

Cells

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Identification of small molecules by screening a mixture-based scaffold compound library for treatment of alpha-1 antitrypsin deficiency

Zhang

Santos

Debevec

et al. 2020

Biochemical and Biophysical Research Communications

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Human placental proteomics and exon variant studies link AAT/SERPINA1 with spontaneous preterm birth

Tiensuu

Haapalainen

Tissarinen

et al. 2022

BMC Med

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Background Preterm birth is defined as live birth before 37 completed weeks of pregnancy, and it is a major problem worldwide. The molecular mechanisms that lead to onset of spontaneous preterm birth are incompletely understood. Prediction and evaluation of the risk of preterm birth is challenging as there is a lack of accurate biomarkers. In this study, our aim was to identify placental proteins that associate with spontaneous preterm birth. Methods We analyzed the proteomes from placentas to identify proteins that associate with both gestational age and spontaneous labor. Next, rare and potentially damaging gene variants of the identified protein candidates were sought for from our whole exome sequencing data. Further experiments we performed on placental samples and placenta-associated cells to explore the location and function of the spontaneous preterm labor-associated proteins in placentas. Results Exome sequencing data revealed rare damaging variants in SERPINA1 in families with recurrent spontaneous preterm deliveries. Protein and mRNA levels of alpha-1 antitrypsin/SERPINA1 from the maternal side of the placenta were downregulated in spontaneous preterm births. Alpha-1 antitrypsin was expressed by villous trophoblasts in the placenta, and immunoelectron microscopy showed localization in decidual fibrinoid deposits in association with specific extracellular proteins. siRNA knockdown in trophoblast-derived HTR8/SVneo cells revealed that SERPINA1 had a marked effect on regulation of the actin cytoskeleton pathway, Slit–Robo signaling, and extracellular matrix organization. Conclusions Alpha-1 antitrypsin is a protease inhibitor. We propose that loss of the protease inhibition effects of alpha-1 antitrypsin renders structures critical to maintaining pregnancy susceptible to proteases and inflammatory activation. This may lead to spontaneous premature birth.

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Targeting the site encoded by SERPINA1*E342K for treating alpha‐1 antitrypsin deficiency‐associated liver diseases

Cited by 3 publications

References 15 publications

A Novel Small Molecule Inhibits Intrahepatocellular Accumulation of Z-Variant Alpha 1-Antitrypsin In Vitro and In Vivo

A Novel Small Molecule Inhibits Intrahepatocellular Accumulation of Z-Variant Alpha 1-Antitrypsin In Vitro and In Vivo

Identification of small molecules by screening a mixture-based scaffold compound library for treatment of alpha-1 antitrypsin deficiency

Human placental proteomics and exon variant studies link AAT/SERPINA1 with spontaneous preterm birth

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