Search bioRxiv⌕ Search

Biology subjects

Aranyos, A. M.

Publications and source records attributed to Aranyos, A. M..

2 recordsLinked to original sources

Novel Transgenic Humanized Alpha-1 Antitrypsin Deficiency Mouse Model on Murine SERPINA1 Null Background

Alpha-1 antitrypsin deficiency (AATD) is a rare genetic disorder caused by accumulation of misfolded -1 antitrypsin within hepatocytes. AATD patients are prone to develop liver disease that remains undiagnosed until the late stages of the disease. Due to challenges in manipulating the -1 antitrypsin genes in mice, determining a true loss of function of -1 antitrypsin in previous AATD mouse models has been challenging. Here, we report generation and liver characterization of a new humanized transgenic mouse model for AATD with a background of a CRISPR-Cas9 generated SERPINA1-null mouse. Male and female transgenic mice for normal (Pi*M) and mutant (Pi*Z) variants of human -1 antitrypsin at 4-6 months of age were subjected to this study. The accumulation of human -1 antitrypsin in the hepatocytes and fibrotic features of the liver were monitored by performing an in vivo study. We demonstrate a strong liver phenotype satisfying clinically relevant manifestations of liver pathology associated with AATD, including hepatic accumulation of human -1 antitrypsin globules, liver deposition of extracellular matrix proteins, hepatic ER stress, and liver fibrosis in Pi*Z mice, in addition to mild systemic inflammation. In addition to major phenotypic criteria of AATD-associated liver fibrosis, accompanying single-nucleus RNA-seq data demonstrate activation of pathways associated with liver metabolic changes, inflammation, and regeneration. Data from this study suggest our humanized transgenic AATD mouse model could provide a suitable model to study -1 antitrypsin loss of function, replicate the pathophysiology of AATD associated liver disease, and evaluate therapeutic reagents against this disease. NEW & NOTEWORTHYWe have characterized a new humanized transgenic mouse model of -1 antitrypsin deficiency with a SERPINA1-null background that shows strong manifestations of liver disease. Our data explores the altered phenotype of -1 antitrypsin deficient hepatocytes and suggests a relationship between liver cell types during disease progression. This model may become a useful tool for investigating -1 antitrypsin loss of function, pathogenic mechanisms, and for drug discovery aimed at both prevention and treatment of the disease.

molecular biology↗

Development of an on-chip detection of Zika virus and antibodies simultaneously using array of nanowells

Zika virus (ZIKV) infections are an emerging health pandemic of significant medical importance. ZIKV appeared recently in the Americas from Africa via the South Pacific. The current outbreak has garnered attention by exhibiting unique characteristics of devastating neurodevelopmental defects in newborns of infected pregnant women. Current guidelines for ZIKV diagnostics developed by the Center of Diseases Control and Prevention (CDC) consist of nucleic acid testing, plaque reduction neutralization test (PRNT), and a serologic test for IgM detection. To better accommodate and comply with these guidelines, we developed a simultaneous on-chip detection of ZIKV and anti-ZIKV antibodies using an array of nanowells. Using on-chip microengraving, we were able to detect anti-ZIKV antibodies and their immunoglobulin isotypes. In parallel, applying on-chip real-time PCR with epifluorescence microscopy, we were able to quantify ZIKV viral load as low as one copy. To test clinical samples of patients at the postconvalescent stage, we analyzed samples from 8 patients. The on-chip nanowells could effectively identify antibodies that reacted against ZIKV envelope protein and their isotypes with high sensitivity and specificity. The small sample requirement with high specificity and sensitivity and combined molecular and serological tests could potentially be very advantageous and beneficial in accurate detection of Zika infection for better disease monitoring and management.

bioengineering↗