HECT, UBA and WWE domain containing 1 represses cholesterol efflux during CD4+ T cell activation in Sjögren’s syndrome

Introduction: Sjögren’s syndrome (SS) is a chronic autoimmune disorder characterized by exocrine gland dysfunction, leading to loss of salivary function. Histological analysis of salivary glands from SS patients reveals a high infiltration of immune cells, particularly activated CD4+ T cells. Thus, interventions targeting abnormal activation of CD4+ T cells may provide promising therapeutic strategies for SS. Here, we demonstrate that Hect, uba, and wwe domain containing 1 (HUWE1), a member of the eukaryotic Hect E3 ubiquitin ligase family, plays a critical role in CD4+ T-cell activation and SS pathophysiology.Methods: In the context of HUWE1 inhibition, we investigated the impact of the HUWE1 inhibitor BI8626 and sh-Huwe1 on CD4+ T cells in mice, focusing on the assessment of activation levels, proliferation capacity, and cholesterol abundance. Furthermore, we examined the therapeutic potential of BI8626 in NOD/ShiLtj mice and evaluated its efficacy as a treatment strategy.Results: Inhibition of HUWE1 reduces ABCA1 ubiquitination and promotes cholesterol efflux, decreasing intracellular cholesterol and reducing the expression of phosphorylated ZAP-70, CD25, and other activation markers, culminating in the suppressed proliferation of CD4+ T cells. Moreover, pharmacological inhibition of HUWE1 significantly reduces CD4+ T-cell infiltration in the submandibular glands and improves salivary flow rate in NOD/ShiLtj mice.Conclusion: These findings suggest that HUWE1 may regulate CD4+ T-cell activation and SS development by modulating ABCA1-mediated cholesterol efflux and presents a promising target for SS treatment

RNA-Seq Analysis Identifies Transcription Factors Involved in Anthocyanin Biosynthesis of ‘Red Zaosu’ Pear Peel and Functional Study of <i>PpPIF8</i>

Red-skinned pears are favored by people for their attractive appearance and abundance of anthocyanins. However, the molecular basis of anthocyanin biosynthesis in red pears remains elusive. Here, a comprehensive transcriptome analysis was conducted to explore the potential regulatory mechanism of anthocyanin biosynthesis in ‘Red Zaosu’ pear (Pyrus pyrifolia × Pyrus communis). Gene co-expression analysis and transcription factor mining identified 263 transcription factors, which accounted for 6.59% of the total number of transcription factors in the pear genome in two gene modules that are highly correlated with anthocyanin biosynthesis. Clustering, gene network modeling with STRING-DB, and local motif enrichment analysis (CentriMo) analysis suggested that PpPIF8 may play a role in anthocyanin biosynthesis. Furthermore, eight PIFs were identified in the pear genome, of which only PpPIF8 was rapidly induced by light. Functional studies showed that PpPIF8 localizes in the nucleus and is preferentially expressed in the tissue of higher levels of anthocyanin. The overexpression of PpPIF8 in pear peel and pear calli promotes anthocyanin biosynthesis and upregulates the expression of anthocyanin biosynthesis genes. Yeast-one hybrid and transgenic analyses indicated that PpPIF8 binds to the PpCHS promoter to induce PpCHS expression. The positive effect of PpPIF8 on anthocyanin biosynthesis is different from previously identified negative regulators of PyPIF5 and MdPIF7 in pear and apple. Taken together, our data not only provide a comprehensive view of transcription events during the coloration of pear peel, but also resolved the regulatory role of PpPIF8 in the anthocyanin biosynthesis pathway

Influence of Bagging on Fruit Quality, Incidence of Peel Browning Spots, and Lignin Content of ‘Huangguan’ Pears

The ‘Huangguan’ pear is one of the high-quality pear cultivars produced in China. However, the bagged fruit of the ‘Huangguan’ pear often suffers from peel browning spots after rain during their mature period. In this study, in an effort to discover the impact of bagging treatments on the occurrence of peel browning spots and fruit quality, fruits were covered by single-layer, two-layer, or triple-layer paper bags six weeks after reaching full bloom. The results showed that the bagged fruits were characterized by smooth surfaces and reduced lenticels compared with the unbagged ones. The unbagged and the two-layer bagged fruits had yellow/green peels, while the single- and triple-layer bagged ones had yellow/white peels. Compared with the unbagged fruits, the bagged fruits had higher vitamin C (Vc) contents and values of peel color indexes L and a and lower soluble solid contents (SSCs), titratable acid (TA) contents, absorbance index differences (IAD), and b values. Additionally, the triple-layer bagged group was superior to other groups in terms of fruit quality, but it also had the maximum incidence of peel browning spots. Before and after the appearance of peel browning spots, the bagged fruits had smoother and thinner cuticles compared with the unbagged ones. Furthermore, the triple-layer bagged fruits had minimum lignin contents and maximum phenolic contents in their peels, with minimum activity of lignin synthesis-related enzymes such as phenylalanine ammonia lyase (PAL), peroxidase (POD), and polyphenol oxidase (PPO), as well as minimum expressions of relevant genes such as cinnamyl alcohol dehydrogenase (CAD), cinnamoyl CoA reductase (CCR), 4-coumarate: coenzyme A ligase (4CL6), and cinnamate 4-hydroxylase (C4H1). It was deduced that POD activity and the relative expressions of CAD9, CCR3, CCR4, and CCR5 may play key roles in the occurrence of peel browning spots. In summary, lignin synthesis affected the incidence of peel browning spots in bagged ‘Huangguan’ pears. This study provides a theoretical basis for understanding the incidence of peel browning spots in ‘Huangguan’ pears