Lung epithelial protein disulfide isomerase A3 (PDIA3) plays an important role in influenza infection, inflammation, and airway mechanics

© 2019 Protein disulfide isomerases (PDI) are a family of redox chaperones that catalyze formation or isomerization of disulfide bonds in proteins. Previous studies have shown that one member, PDIA3, interacts with influenza A virus (IAV) hemagglutinin (HA), and this interaction is required for efficient oxidative folding of HA in vitro. However, it is unknown whether these host-viral protein interactions occur during active infection and whether such interactions represent a putative target for the treatment of influenza infection. Here we show that PDIA3 is specifically upregulated in IAV-infected mouse or human lung epithelial cells and PDIA3 directly interacts with IAV-HA. Treatment with a PDI inhibitor, LOC14 inhibited PDIA3 activity in lung epithelial cells, decreased intramolecular disulfide bonds and subsequent oligomerization (maturation) of HA in both H1N1 (A/PR8/34) and H3N2 (X31, A/Aichi/68) infected lung epithelial cells. These reduced disulfide bond formation significantly decreased viral burden, and also pro-inflammatory responses from lung epithelial cells. Lung epithelial-specific deletion of PDIA3 in mice resulted in a significant decrease in viral burden and lung inflammatory-immune markers upon IAV infection, as well as significantly improved airway mechanics. Taken together, these results indicate that PDIA3 is required for effective influenza pathogenesis in vivo, and pharmacological inhibition of PDIs represents a promising new anti-influenza therapeutic strategy during pandemic and severe influenza seasons

Conjugated bile acids attenuate allergen-induced airway inflammation and hyperresposiveness by inhibiting UPR transducers

© 2019 American Society for Clinical Investigation. Conjugated bile acids (CBAs), such as tauroursodeoxycholic acid (TUDCA), are known to resolve the inflammatory and unfolded protein response (UPR) in inflammatory diseases, such as asthma. Whether CBAs exert their beneficial effects on allergic airway responses via 1 arm or several arms of the UPR, or alternatively through the signaling pathways for conserved bile acid receptor, remains largely unknown. We used a house dust mite-induced (HDM-induced) murine model of asthma to evaluate and compare the effects of 5 CBAs and 1 unconjugated bile acid in attenuating allergen-induced UPR and airway responses. Expression of UPRassociated transcripts was assessed in airway brushings from human patients with asthma and healthy subjects. Here we show that CBAs, such as alanyl β-muricholic acid (AβM) and TUDCA, significantly decreased inflammatory, immune, and cytokine responses; mucus metaplasia; and airway hyperresponsiveness, as compared with other CBAs in a model of allergic airway disease. CBAs predominantly bind to activating transcription factor 6α (ATF6α) compared with the other canonical transducers of the UPR, subsequently decreasing allergen-induced UPR activation and resolving allergic airway disease, without significant activation of the bile acid receptors. TUDCA and AβM also attenuated other HDM-induced ER stress markers in the lungs of allergic mice. Quantitative mRNA analysis of airway epithelial brushings from human subjects demonstrated that several ATF6α-related transcripts were significantly upregulated in patients with asthma compared with healthy subjects. Collectively, these results demonstrate that CBA-based therapy potently inhibits the allergen-induced UPR and allergic airway disease in mice via preferential binding of the canonical transducer of the UPR, ATF6α. These results potentially suggest a novel avenue to treat allergic asthma using select CBAs

Glutathione <em>S</em>-transferase P1 (<em>GSTP1</em>) directly influences platinum drug chemosensitivity in ovarian tumour cell lines

BACKGROUND: Chemotherapy response in ovarian cancer patients is frequently compromised by drug resistance, possibly due to altered drug metabolism. Platinum drugs are metabolised by glutathione S-transferase P1 (GSTP1), which is abundantly, but variably expressed in ovarian tumours. We have created novel ovarian tumour cell line models to investigate the extent to which differential GSTP1 expression influences chemosensitivity. METHODS: Glutathione S-transferase P1 was stably deleted in A2780 and expression significantly reduced in cisplatin-resistant A2780DPP cells using Mission shRNA constructs, and MTT assays used to compare chemosensitivity to chemotherapy drugs used to treat ovarian cancer. Differentially expressed genes in GSTP1 knockdown cells were identified by Illumina HT-12 expression arrays and qRT–PCR analysis, and altered pathways predicted by MetaCore (GeneGo) analysis. Cell cycle changes were assessed by FACS analysis of PI-labelled cells and invasion and migration compared in quantitative Boyden chamber-based assays. RESULTS: Glutathione S-transferase P1 knockdown selectively influenced cisplatin and carboplatin chemosensitivity (2.3- and 4.83-fold change in IC(50), respectively). Cell cycle progression was unaffected, but cell invasion and migration was significantly reduced. We identified several novel GSTP1 target genes and candidate platinum chemotherapy response biomarkers. CONCLUSIONS: Glutathione S-transferase P1 has an important role in cisplatin and carboplatin metabolism in ovarian cancer cells. Inter-tumour differences in GSTP1 expression may therefore influence response to platinum-based chemotherapy in ovarian cancer patients

Data for: Lung epithelial protein disulfide isomerase A3 (PDIA3) plays an important role in influenza infection, inflammation, and airway mechanics

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Data for: Lung epithelial protein disulfide isomerase A3 (PDIA3) plays an important role in influenza infection, inflammation, and airway mechanics

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Hormonal induction of supermale golden rosy barb and isolation of Y-chromosome specific markers

Viable supermales of the golden rosy barb Puntius conchonius are generated through hormonal sex reversal and progeny testing. Discrete immersion (3 h/day on the second, fourth, and sixth day after hatching) of the fry in Estradiol-17&#946; (E<SUB>2</SUB>) at doses of 400-600 &#956;g/L ensures greater than 40% survival and production of more than 98% F<SUB>1</SUB> females; of these 50% are heterogametic females, that when crossed with normal males, sire 25% YY supermales. Supermales sire female progenies at the frequencies between 0 and 8%. Reproductive performance of hormonally sex-reversed females and androgenetic females is inferior to the normal ones. Conversely, the performance of androgenetic males is superior but suffers from low fertilizability. The relative performance of supermale produced by breeding sex-reversed parents is superior to those produced by androgenesis. Using the SRY-specific primers, the PCR analysis of the genomic DNA of the male golden rosy barb produces three products of 588, 333, and 200 bp length. However only the 200 bp product is amplified in the female genome. Hence it is possible to use the first two products as molecular markers to rapidly identify fish possessing a Y chromosome. The presence of 333 and 588 bp fragments in normal (X<SUP>1</SUP>Y<SUP>2</SUP>), hormonally induced (Y<SUP>1</SUP>Y<SUP>2</SUP>) and androgenetic (Y<SUP>2</SUP>Y<SUP>2</SUP>) males and the absence of relation between the 200 bp fragment and the X-chromosome indicates that the male specific markers are specific to Y-chromosome. For the first time, a Y-chromosome specific molecular marker for a cyprinid has been identified, isolated, and characterized

Erratum to Molecular cloning of growth hormone-encoding cDNA of an Indian major carp, Labeo rohita, and its expression in Escherichia coli and zebrafish [Gen. Comp. Endocrinol. 125, (2002), 236-247]

10.1016/j.ygcen.2010.01.013General and Comparative Endocrinology1671180-GCEN

Enhanced expression of β-thymosin mRNA in the ovary of GnRH analog or estradiol-17 β-treated paradise fish, Macropodus opercularis

cDNA clones were isolated as expressed sequence tags (ESTs) from the ovarian cDNA library of Macropodus opercularis. The EST sequences showed similarity with many housekeeping genes and ribosomal proteins. One of the ESTs showed similarity to &#946;-thymosin, a 5-kDa polypeptide expressed under different physiological conditions. The cDNA corresponding to &#946;-thymosin of M. opercularis is 368 bp in length and codes for a putative polypeptide of 42 amino acids. Multiple alignment of the deduced amino acid sequence showed 61% similarity with piscine &#946;-thymosins and 56% similarity with mammalian &#946;-thymosins. Administration of a gonadotropin releasing hormone analog or estradiol-17&#946; induced an increase in the gonadosomatic index, oocyte diameter and also enhanced expression of &#946;-thymosin m-RNA in the recrudizing ovary. This report indicates that both GnRH analog and E2 might induce similar pathways for the differentiation of ovarian cells for the maturation of oocytes

Erratum: Cloning, sequencing and expression of cDNA encoding growth hormone from Indian catfish (Heteropneustes fossilis) (Journal of Biosciences (2001) 26:3 (315-324))

10.1007/s12038-012-9292-5Journal of Biosciences381179-JOBS

Molecular cloning of growth hormone-encoding cDNA of an Indian major carp, Labeo rohita, and its expression in Escherichia coli and zebrafish

10.1006/gcen.2001.7759General and Comparative Endocrinology1252236-247GCEN