Comprehensive Transcriptome Analysis Research Articles

352 Evaluation of allergenic properties and uptake of cobalt nanoparticles in human skin; a comprehensive transcriptomic analysis and skin permeation assessment

352 Evaluation of allergenic properties and uptake of cobalt nanoparticles in human skin; a comprehensive transcriptomic analysis and skin permeation assessment

Genome-wide analysis of the R2R3-MYB gene family and identification of candidate genes that regulate isoflavone biosynthesis in red clover (Trifolium pratense)

Red clover (Trifolium pratense) is a perennial legume with high feeding and medicinal value attributed to its abundant isoflavone content. Previous studies reported that R2R3-MYB transcription factors are involved in the biosynthesis of isoflavones; however, their specific role in red clover remains poorly understood. Through comprehensive genome-wide and transcriptome analyses, a total of 138 TpR2R3-MYB genes were identified and classified into 30 distinct subgroups within a phylogenetic tree. Importantly, six of these subgroups showed associations with isoflavone biosynthesis in red clover. The majority of segmental duplication events (Ka/Ks < 1) indicated that the TpR2R3-MYB gene underwent strong purifying selection during evolution. The qRT-PCR analysis demonstrated high expression levels of TpMYB79 and TpMYB53 in Minshan red clover at full flowering stage, consistent with the trend for isoflavone content determination, suggesting that TpMYB79 and TpMYB53 might be important regulators of isoflavone biosynthesis in red clover. Additionally, we observed nucleus and vacuole membrane localization of TpMYB53 and TpMYB79, with TpMYB53 primarily exerting transcriptional activation through its C-terminal activation motifs while TpMYB79 exhibited no transcriptional activity. These findings provided a foundation for the study of the biological function of R2R3-MYB transcription factors in red clover.

Integrated omics profiles for exploring the potential mechanism underlying aroma formation in the terpenoid-rich aromatic plant Opisthopappus taihangensis and the bioactivity of its leaf essential oil

Opisthopappus taihangensis (Anthemideae, Asteraceae), which is rich in bioactive components, produces flowers and leaves with robust fragrances. In this study, we conducted comprehensive metabolomic and transcriptomic analyses to identify changes in terpenoid metabolites and associated gene expression across various O. taihangensis tissues (leaves, buds, and inflorescences at the exposure, initial-bloom, and full-bloom stages). We identified 1,370 metabolites using headspace solid-phase micro-extraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS), 308 of which were terpenoid metabolites. The expression of terpenoid synthesis-related genes was relatively consistent with the synthesis of terpenoid metabolites at each examined developmental stage. An analysis of gene networks governing terpenoid synthesis revealed that MCT genes (OtMCT1, OtMCT2, and OtMCT3), TPS genes (OtTPS5, OtTPS9, and OtTPS10), and OtISPS1 may be crucial for synthesizing specific metabolites in different tissues. Additionally, the essential oil extracted from leaves by water distillation showed thatthujone and camphor were the predominant components. The considerable antioxidant activity of the leaf essential oil was comparable to that of vitamin C (16 μg/mL). Notably, its antimicrobial effects against Staphylococcus aureus and Escherichia coli growth were greater than those of ampicillin and vancomycin at the same concentrations. Scanning electron microscopy images indicated that the leaf essential oil significantly disrupted bacterial cell structures. This study thoroughly analyzed the network of genes regulating terpenoid metabolites in different O. taihangensis tissues, and elucidated the antioxidant and antibacterial potential of the leaf essential oil, thus providing valuable insights for breeding, molecular characterization, and the potential application of O. taihangensis in developing useful essential oil-based natural products.

Proteomics and transcriptomic analyses provide new insights into the pectin polysaccharide biosynthesis in Premna puberula Pamp

Premna puberula Pamp. is a plant with similarities to both food and medicine, which has attracted the attention of researchers because of the rich pectin in its leaves. However, the mechanism of efficient pectin accumulation remains unclear. Through transcriptomic and proteomic analyses, this study investigated the differentially expressed genes and proteins associated with pectin during various developmental stages of P. puberula. The combined omics approach revealed that the majority of differential genes are mainly involved in the anabolic metabolism of primary and secondary metabolites. Notable differential pathways include starch and sucrose metabolism, amino acid sugar metabolism, and nucleotide sugar metabolism. The SWEET gene family was identified and analyzed, leading to the cloning of PpSWEET15 and subcellular localization. Overexpression of PpSWEET15 resulted in a significant decrease in sucrose content in leaves from 134.44 μg·g−1 to <10 μg·g−1. Glucose content decreased from 407.75 μg·g−1 to 318.38 μg·g−1, while fructose levels showed no significant difference between the two groups of leaves. This comprehensive transcriptomic and proteomic analysis provides valuable insights into the molecular mechanisms underlying the efficient accumulation of pectin in P. puberula.

New direction: identification of immunoinflammatory subtypes and potential therapeutic targets for cholangiocarcinoma

BackgroundCholangiocarcinoma (CHOL) is a rare cancer with low survival rates. Despite advances in precision medicine targeting molecular subtypes, the immune subtypes of CHOL remain poorly understood. This study aimed to identify immune subtypes of CHOL and investigate their implications in the metabolic regulation of macrophage functions in inflammation.MethodsWe conducted a comprehensive analysis of transcriptome and single-cell sequencing data from multiple databases to classify the immune subtypes of CHOL. Immune cell infiltration within the tumor microenvironment (TME) and the metabolic pathways involved in macrophage activation and polarization were also analyzed.ResultsTwo distinct immune subtypes, immune-infiltrated CS1 and immune-depleted CS2, were identified in CHOL. CS1 exhibited a highly active TME with substantial immune cell infiltration, including macrophages, and activation of immune-related signaling pathways, such as inflammatory and interferon pathways. In contrast, CS2 was characterized by a deficiency in immune components and poorer prognosis. Metabolic regulation of macrophages, particularly the downregulation of oxidative phosphorylation in CS1, suggested a shift towards glycolysis as an energy source for activated macrophages, contributing to the immune-responsive phenotype observed in CS1. Additionally, the oncogenic role of DLX5 in CHOL was revealed, with potential impacts on macrophage functions in inflammation.ConclusionThis study provides insights into immune subtype classification, novel biomarker identification, and the metabolic regulation of macrophage functions in CHOL. Understanding macrophage metabolic reprogramming within immune subtypes may contribute to the development of targeted immunotherapies for CHOL.

Comprehensive transcriptomic, proteomic, and intestinal microbiota analyses of largemouth bass (Micropterus salmoides) intestines reveal new insights into immune responses to Aeromonas hydrophila infection

This study investigates the immune responses of largemouth bass (Micropterus salmoides) to infection by Aeromonas hydrophila, a pathogen responsible for significant economic losses in freshwater aquaculture due to bacterial enteritis. We employed transcriptomic sequencing, label-free LC-MS/MS quantitative proteomics, and 16S RNA sequencing to evaluate the transcriptomic, proteomic, and intestinal microbiota changes in infected fish compared to healthycontrols. Each fish (approximately 60 g) in the infection group was injected with 0.5 ml of an A. hydrophila suspension (1.0 × 108 CFU/mL), while the control group received 0.5 ml of phosphate buffer. Samples were collected 72 h post-injection, with three biological replicates made from an equal mix of tissue samples from six fish each. A total of 1666 differentially expressed genes (DEGs) and 2477 differentially expressed proteins (DEPs) were identified. Integrated analysis revealed that the up-regulated DEGs/DEPs in the intestines of infected largemouth bass were primarily associated with immune-related pathways including "antigen processing and presentation", "MAPK signaling pathway", "ECM-receptor interaction", and "leukocyte transendothelial migration". Notable upregulated immune-related proteins included complement or antigen-presenting proteins like complement C1, complement C3, complement C6, ɑ-2-macroglobulin, laminin, serotransferrin, leukocyte cell-derived chemotaxin-2, immunoglobulin C1-set domain-containing protein, and the MHC class I alpha antigen. 16S RNA sequencing indicated a significant increase in Proteobacteria and a decrease in Fusobacteria in the intestines of infected fish compared to controls. Collectively, these findings demonstrate that A. hydrophila infection significantly alters gene and protein expression as well as intestinal microbiota in largemouth bass, providing insights into their immune defense mechanisms against infection.

Integrative transcriptomic analysis reveals the molecular responses of tobacco to magnesium deficiency

IntroductionMagnesium (Mg) is a crucial macronutrient for plants. Understanding the molecular responses of plants to different levels of Mg supply is important for improving cultivation practices and breeding new varieties with efficient Mg utilization.MethodsIn this study, we conducted a comprehensive transcriptome analysis on tobacco (Nicotiana tabacum L.) seedling leaves to investigate changes in gene expression in response to different levels of Mg supply, including Mg-deficient, 1/4-normal Mg, normal Mg, and 4×-normal Mg, with a particular focus on Mg deficiency at 5, 15 and 25 days after treatment (DAT), respectively.ResultsA total of 11,267 differentially expressed genes (DEGs) were identified in the Mg-deficient, 1/4-normal Mg, and/or 4×-normal Mg seedlings compared to the normal Mg seedlings. The global gene expression profiles revealed potential mechanisms involved in the response to Mg deficiency in tobacco leaves, including down-regulation of genes–two DEGs encoding mitochondria-localized NtMGT7 and NtMGT9 homologs, and one DEG encoding a tonoplast-localized NtMHX1 homolog–associated with Mg trafficking from the cytosol to mitochondria and vacuoles, decreased expression of genes linked to photosynthesis and carbon fixation at later stages, and up-regulation of genes related to antioxidant defenses, such as NtPODs, NtPrxs, and NtGSTs.DiscussionOur findings provide new insights into the molecular mechanisms underlying how tobacco responds to Mg deficiency.

A Comprehensive Transcriptomic and Proteomics Analysis of Candidate Secretory Proteins in Rose Grain Aphid, Metopolophium dirhodum (Walker)

The Rose grain aphid, a notable agricultural pest, releases saliva while feeding. Yet, there is a need for a comprehensive understanding of the specific identity and role of secretory proteins released during probing and feeding. Therefore, a combined transcriptomic and proteomic approach was employed in this study to identify putative secretory proteins. The transcriptomic sequencing result led to the assembly of 18,030 unigenes out of 31,344 transcripts. Among these, 705 potential secretory proteins were predicted and functionally annotated against publicly accessible protein databases. Notably, a substantial proportion of secretory genes (71.5%, 69.08%, and 60.85%) were predicted to encode known proteins in Nr, Pfam, and Swiss-Prot databases, respectively. Conversely, 27.37% and 0.99% of gene transcripts were predicted to encode known proteins with unspecified functions in the Nr and Swiss-Prot databases, respectively. Meanwhile, the proteomic analysis result identified, 15 salivary proteins. Interestingly, most salivary proteins (i.e., 60% of the proteins) showed close similarity to A. craccivora, while 46.67% showed close similarity to A. glycines, M. sacchari and S. flava. However, to verify the expression of these secretory genes and characterize the biological function of salivary proteins further investigation should be geared towards gene expression and functional analysis.

Protein lactylation within the nucleus independently predicts the prognosis of non‑specific triple‑negative breast cancer.

Protein lactylation represents a pervasive post-translational modification prevalent in histones and diverse proteins, fostering tumor initiation and progression. Nonetheless, the impact of protein lactylation on the prognosis of non-specific triple-negative breast cancer (TNBC) remains uncertain. In the present study, the pan-lysine lactylation (panKlac) levels in cytoplasmic and nuclear compartments were semi-quantitatively examined using a tissue microarray encompassing 77 non-specific TNBC tissues. The association of the prognosis of patients with the panKlac levels in the cytoplasmic and nuclear compartments or other tumor attributes was assessed using Kaplan-Meier and Cox regression analyses. Furthermore, the molecular pathways involved in the promotional effect of lactylation on cell proliferation were determined through a transcriptomic analysis. The results indicated that the panKlac levels were markedly higher in tumor tissues than in para-tumor mammary regions and showed no significant correlations with various clinicopathological parameters, such as tumor dimension, lymph node involvement or histological grading. Notably, high panKlac levels within the nucleus served as an independent predictor of recurrence-free survival, whereas high cytoplasmic panKlac levels were a protective factor for patient survival. The panKlac levels were also markedly elevated in the TNBC cell line, MDA-MB-231. Additionally, glycolysis inhibition significantly reduced the global panKlac levels and concurrently diminished cell proliferation. According to the comprehensive transcriptomic analysis results, pathways related to ribosomal subunit biosynthesis/assembly and aminoacyl-tRNA biosynthesis were involved in the tumor-promoting mechanisms of lactylation. Further results revealed the oncogenic propensity of tyrosyl-tRNA synthetase 1 (YARS1) and its association with lactate production. Overall, Klac levels within the nucleus are an independent prognostic indicator for patients with non-specific TNBC. It is imperative to delve deeper into the roles and mechanisms of nuclear protein lactylation and YARS1 in non-specific TNBC.

Deciphering the Genetic and Biochemical Drivers of Fruit Cracking in Akebia trifoliata.

This study investigates the molecular mechanisms underlying fruit cracking in Akebia trifoliata, a phenomenon that significantly impacts fruit quality and marketability. Through comprehensive physiological, biochemical, and transcriptomic analyses, we identified key changes in cell wall components and enzymatic activities during fruit ripening. Our results revealed that ventral suture tissues exhibit significantly elevated activities of polygalacturonase (PG) and β-galactosidase compared to dorsoventral line tissues, indicating their crucial roles in cell wall degradation and structural weakening. The cellulose content in VS tissues peaked early and declined during ripening, while DL tissues maintained relatively stable cellulose levels, highlighting the importance of cellulose dynamics in fruit cracking susceptibility. Transcriptomic analysis revealed differentially expressed genes (DEGs) associated with pectin biosynthesis and catabolism, cell wall organization, and oxidoreductase activities, indicating significant transcriptional regulation. Key genes like AKT032945 (pectinesterase) and AKT045678 (polygalacturonase) were identified as crucial for cell wall loosening and pericarp dehiscence. Additionally, expansin-related genes AKT017642, AKT017643, and AKT021517 were expressed during critical stages, promoting cell wall loosening. Genes involved in auxin-activated signaling and oxidoreductase activities, such as AKT022903 (auxin response factor) and AKT054321 (peroxidase), were also differentially expressed, suggesting roles in regulating cell wall rigidity. Moreover, weighted gene co-expression network analysis (WGCNA) identified key gene modules correlated with traits like pectin lyase activity and soluble pectin content, pinpointing potential targets for genetic manipulation. Our findings offer valuable insights into the molecular basis of fruit cracking in A. trifoliata, laying a foundation for breeding programs aimed at developing crack-resistant varieties to enhance fruit quality and commercial viability.

Genomic and transcriptomic analyses identify distinctive features of triple-negative inflammatory breast cancer

Triple-negative inflammatory breast cancer (TN-IBC) is the most aggressive type of breast cancer, yet its defining genomic, molecular, and immunological features remain largely unknown. In this study, we performed the largest and most comprehensive genomic and transcriptomic analyses of prospectively collected TN-IBC patient samples from a phase II clinical trial (ClinicalTrials.gov, NCT02876107, registered on August 22, 2016) and compared them to similarly analyzed stage III TN-non-IBC patient samples (ClinicalTrials.gov, NCT02276443, registered on October 21, 2014). We found that TN-IBC tumors have distinctive genomic, molecular, and immunological characteristics, including a lower tumor mutation load than TN-non-IBC, and an association of immunosuppressive tumor-infiltrating immune components with an unfavorable response to neoadjuvant chemotherapy. To our knowledge, this is the only study in which TN-IBC and TN-non-IBC samples were collected prospectively. Our analysis improves the understanding of the molecular landscape of the most aggressive subtype of breast cancer. Further studies are needed to discover novel prognostic biomarkers and druggable targets for TN-IBC.

CTNI-78. PNOC008: A PILOT TRIAL TESTING THE CLINICAL BENEFIT OF USING MOLECULAR PROFILING TO DETERMINE AN INDIVIDUALIZED TREATMENT PLAN IN CHILDREN AND YOUNG ADULTS WITH NEWLY DIAGNOSED HIGH-GRADE GLIOMA (EXCLUDING DIFFUSE INTRINSIC PONTINE GLIOMA)

Abstract BACKGROUND Despite multiple clinical trials in young patients with newly diagnosed high grade gliomas (HGG), survival remains poor. PNOC008 is a single-arm, multi-center pilot trial, investigating the feasibility, toxicity, and efficacy of a molecularly guided individualized treatment approach following radiotherapy. METHODS Patients aged ≤21 years with newly diagnosed, localized, hemispheric HGG (Stratum A) or non DIPG, diffuse midline glioma (DMG) (Stratum B) were eligible. Comprehensive molecular profiling (targeted gene panel, whole exome, and whole transcriptome sequencing) was performed on primary tumor tissue. The molecular data was reviewed by a dedicated tumor board that recommended an individualized treatment plan combining up to four FDA approved drugs. Patients were followed for toxicity and efficacy. Circulating tumor DNA (ctDNA), imaging, and quality of life (QOL) measures were collected at multiple timepoints. RESULTS Fifty-five HGG patients enrolled between 2018 and 2023 (median age 11 years [range 2-20], n=31 female, n=29 Stratum A), including H3K27-altered (n=17), H3/IDH-wildtype diffuse pediatric-type (n=16), and H3G34-mutant diffuse hemispheric glioma (n=12). In 44 patients that followed the recommended treatment, median overall survival (OS) from time of study enrollment was 26.5 months in Stratum A (lower 95% CI: 18.5) and 23.6 months in Stratum B (lower 95% CI: 16.8), and 30 months in H3G34-mutant patients (n=10, lower 95% CI:24.6) with median follow-up of 34.5 months for all patients (lower 95% CI: 32.2). The treatment recommendations most commonly included alkylator with targeted therapy combinations. The often novel drug combinations were generally well tolerated with grade 3 or 4 treatment-related adverse events being mostly hematologic. Central imaging, ctDNA, and QOL analyses are underway. CONCLUSIONS A personalized treatment approach using comprehensive transcriptomic and genomic analysis is feasible and well tolerated with encouraging survival data in children and young adults with HGG. Further analyses of molecular subgroups and correlatives are ongoing.

A combined analysis of transcriptome and proteome reveals the regulation mechanism of alginate oligosaccharides on alleviating energy deficit in postharvest strawberry

Strawberry exhibits a very short shelf life due to its rapid softening and decay, resulting in postharvest loss. Our previous study showed that AOS, an eco-friendly nontoxic natural product, plays a role in preserving the freshness of strawberries by regulating ABA content and its signaling pathway. Considering the critical role that energy deficit plays in the senescence process of fruits, coupled with the potential influence of plant hormones on energy metabolism. In this study, the molecular mechanism of strawberry postharvest senescence in response to AOS treatment was further explored from the perspective of energy metabolism regulation. This was achieved through a two-layered approach, integrating the direct determination of energy-related substances with a comprehensive joint analysis of transcriptome and proteome data. AOS treatment significantly delayed the loss of ATP, ADP, and AMP contents compared to control fruit, implying that maintaining cell energy is a crucial factor in AOS postponing senescence in strawberry. Subsequently, the mechanisms were further revealed by transcriptome and proteome analysis. Overall, 255 DEGs were screened from the AOS group, mainly related to carbohydrate and energy metabolism, defense, plant hormone, and secondary metabolism. Additionally, A total of 227 proteins were identified that were differentially expressed in the AOS group, and the majority were related to carbohydrate and energy metabolism (85 proteins). Two omics data implied that AOS could effectively regulate the energy metabolism system to maintain the cellular energy level of harvested strawberry during storage, ultimately leading to longer shelf life. Therefore, our findings provide comprehensive information regarding the molecular mechanism underlying the postharvest storage of strawberry treated with AOS. From our results, it can be concluded that AOS postharvest treatment is very useful for keeping fruit quality and extending shelf life by maintaining a higher energy level.

A comprehensive analysis of metabolomics and transcriptomics revealed the mTOR-autophagy pathway is involved in ammonia metabolism of yellow catfish Pelteobagrus fulvidraco

Nonionic ammonia (NH3) in water is toxic, ammonia intoxication increases susceptibility to pathogens, and threaten the survival and development of fish. The study endeavors to propose a novel mechanism for ammonia detoxification by observing alterations in biomolecules and metabolites in liver of yellow catfish exposed to ammonia stress (AS) for 96 h. This study conducted an in-depth analysis of the serum and liver of fish subjected to 0.72 mg/L NH3 for 96 h. Results revealed elevated serum ammonia levels, compromised serum antioxidant system, disrupted biochemical indexes, and severe liver damage. Liver transcriptomic analysis revealed 156 down-regulated and 55 genes up-regulated differentially expressed genes (DEGs) after AS. KEGG and GO enrichment analysis found that DEGs mainly concentrated in autophagy and mTOR signaling pathways. We examined the autophagy under AS at the molecular, protein and cellular levels by qRT-PCR, Western blot, and ultrastructural observations, respectively. The results showed that the autophagy mediated by mTOR under AS can be divided into two stages: the inhibition stage from 0 to 72 h and the autophagy activation stage from 72 to 96 h. Additionally, we demonstrate that activating autophagy through rapamycin enhances the survival rate when exposed to 3.6 mg/L NH3. Liver metabolomics revealed 278 significantly down-regulated and 31 significantly up-regulated differential metabolites (DEMs) after 96 h AS. Enrichment analysis showed that these DEMs were mainly concentrated in “arginine biosynthesis” and “valine leucine isoleucine biosynthesis”. We also found that the expression of genes related to urea and glutamine synthesis (UGS) in the liver increased significantly after AS, indicating that the detoxification of ammonia necessitates the utilization of substrates and intermediate metabolites essential for UGS.

Co-Colorectal cancer stem cells employ the FADS1/DDA axis to evade NK cell-mediated immunosuppression after co-cultured with NK cells under hypoxia

Colorectal cancer (CRC) ranks as China’s second most common cancer and fifth top cancer death cause. The study highlights the role of Natural Killer (NK) cells in targeting cancer stem cells (CSCs) that evade immune responses in CRC. Colorectal cancer stem cells (CCSCs) were stem from HT-29 cells and co-cultured with NK cells under normoxic or hypoxic conditions. The impact of this co-culture was evaluated using CCK8 assays for NK cell viability, ELISA for cytokine level changes, and flow cytometry for assessing NK cell apoptosis and activation. Comprehensive metabolomic and transcriptomic analyses were also performed to identify key genes and metabolites involved in the interaction between CCSCs and NK cells Co-culture of CCSCs with NK cells under hypoxia reduced NK cytotoxicity, increased NK apoptosis, and altered cytokine secretion by decreasing IFN-γ and TNF-α levels while increasing IL-6. Transcriptomic and metabolomic analysis identified 4 genes (FADS1, ALDH3A2, GCSH, MTCL1) and 3 metabolites (glyoxylic acid, spermine, DDA) as significant. Interfering with FADS1 counteracted the suppression of IFN-γ and TNF-α induced by CSC cells. Curiously, this inhibition caused by si-FADS1 could be neutralized by the addition of exogenous DDA. Co-culturing with NK cells notably increased spermine levels. Exogenous spermine resulted in a significant reduction in HT-29 cell death rates at 32 µM, 64 µM, and 128 µM, compared to NK cells without spermine. Our research explored CCSCs employed the FADS1/DDA axis to evade NK cell-mediated immunosuppression after co-cultured with NK cells under hypoxia.

Inflammasome-targeted therapy might prevent adverse perinatal outcomes of recurrent chronic intervillositis of unknown etiology

Chronic histiocytic intervillositis of unknown origin (CHI) is a rare placental disorder associated with adverse pregnancy outcomes, frequent recurrence, and a lack of effective preventive strategies. Recent insights indicate a potential link between CHI-associated inflammatory lesions and the inflammasome pathway, suggesting innovative therapeutic avenues. Here we show a potential role of the inflammasome pathway in CHI through comprehensive transcriptomic analysis of grade 2 or 3 histopathologic CHI samples, paired with placental controls. Additionally, we present case studies of three individuals with recurrent CHI, who have undergone treatment with anakinra and colchicine throughout pregnancy, resulting in improved perinatal outcomes. Notably, all cases are characterized by the birth of healthy, full-term infants, with reduced or absent intervillositis recurrence. Placental assessment unveils heightened activation of the NLRP3-PYCARD inflammasome pathway and IL-1β processing in CHI samples, with downregulation observed in treated pregnancy samples, devoid of intervillositis. Collectively, these findings suggest a potential therapeutic role for targeting the inflammasome pathway in preventing recurrent CHI in pregnant individuals.

The Molecular Classification of Pheochromocytomas and Paragangliomas: Discovering the Genomic and Immune Landscape of Metastatic Disease.

Pheochromocytomas (PCCs) and paragangliomas (PGLs, together PPGLs) are the most hereditary tumors known. PPGLs were considered benign, but the fourth edition of the World Health Organisation (WHO) classification redefined all PPGLs as malignant neoplasms with variable metastatic potential. The metastatic rate differs based on histopathology, genetic background, size, and location of the tumor. The challenge in predicting metastatic disease lies in the absence of a clear genotype-phenotype correlation among the more than 20 identified genetic driver variants. Recent advances in molecular clustering based on underlying genetic alterations have paved the way for improved cluster-specific personalized treatments. However, despite some clusters demonstrating a higher propensity for metastatic disease, cluster-specific therapies have not yet been widely adopted in clinical practice. Comprehensive genomic profiling and transcriptomic analyses of large PPGL cohorts have identified potential new biomarkers that may influence metastatic potential. It appears that no single biomarker alone can reliably predict metastatic risk; instead, a combination of these biomarkers may be necessary to develop an effective prediction model for metastatic disease. This review evaluates current guidelines and recent genomic and transcriptomic findings, with the aim of accurately identifying novel biomarkers that could contribute to a predictive model for mPPGLs, thereby enhancing patient care and outcomes.

Fungal Methane Production Under High Hydrostatic Pressure in Deep Subseafloor Sediments.

Fungi inhabiting deep subseafloor sediments have been shown to possess anaerobic methane (CH4) production capabilities under atmospheric conditions. However, their ability to produce CH4 under in situ conditions with high hydrostatic pressure (HHP) remains unclear. Here, Schizophyllum commune 20R-7-F01, isolated from ~2 km below the seafloor, was cultured in Seawater Medium (SM) in culture bottles fitted with sterile syringes for pressure equilibration. Subsequently, these culture bottles were transferred into 1 L stainless steel pressure vessels at 30 °C for 5 days to simulate in situ HHP and anaerobic environments. Our comprehensive analysis of bioactivity, biomass, and transcriptomics revealed that the S. commune not only survived but significantly enhanced CH4 production, reaching approximately 2.5 times higher levels under 35 MPa HHP compared to 0.1 MPa standard atmospheric pressure. Pathways associated with carbohydrate metabolism, methylation, hydrolase activity, cysteine and methionine metabolism, and oxidoreductase activity were notably activated under HHP. Specifically, key genes involved in fungal anaerobic CH4 synthesis, including methyltransferase mct1 and dehalogenase dh3, were upregulated 7.9- and 12.5-fold, respectively, under HHP. Enhanced CH4 production under HHP was primarily attributed to oxidative stress induced by pressure, supported by intracellular reactive oxygen species (ROS) levels and comparative treatments with cadmium chloride and hydrogen peroxide. These results may provide a strong theoretical basis and practical guidance for future studies on the contribution of fungi to global CH4 flux.

Characterization of Superoxide Dismutase (SOD) Gene Family and Their Responses to Salinity Stress and Fruit Development in Octoploid Strawberry

Superoxide dismutases (SODs), as the first line of defense against reactive oxygen species (ROS), play an essential role in protecting plants from adverse elicitors during plant growth and development. However, little is known about the SOD gene family and their response to salinity stress and fruit development in cultivated strawberries (Fragaria × ananassa). In this study, 32 SOD genes consisting of 16 Cu/ZnSODs, 11 FeSODs, and 5 MnSOD were identified, which presented three well-resolved clades in the phylogenetic tree. Each clade had similar motifs, and exon–intron structures, which in turn supported the evolutionary classification. Cis-acting element analysis suggested that FaSOD genes might be involved in the plant response to abiotic and biotic stresses, hormones, and light. The analysis of previously published transcriptome data revealed that FaSOD genes are expressed variably under salt stress. Among these SODs, FaMSD5 was expressed at relatively high levels in strawberry root and leaf, and its transcript abundance significantly increased after salt treatment. Some transcription factors related to photomorphogenesis, hormone signaling pathways, and hyperosmotic salinity response were predicted to bind to the FaMSD5 promoter. These outcomes implied that FaMSD5 might play an important role in protection against salt stress. In addition, the comprehensive transcriptome analysis of FaSOD genes in strawberry fruit showed that almost all FaCSDs and FaMSDs were more highly expressed than FaFSDs at different developmental stages, and the expression patterns of FaCSD1, FaCSD2, FaCSD7, FaCSD8, and FaCSD10 suggested that they were likely to be involved in fruit development and ripening. This study provides a basis for further exploration of the function of the FaSOD gene family in strawberry and provides candidate FaSOD genes for enhancing salinity tolerance and regulating fruit development and ripening.

The full transcription map of cottontail rabbit papillomavirus in tumor tissues.

Cottontail rabbit papillomavirus (CRPV), the first papillomavirus associated with tumor development, has been used as a powerful model to study papillomavirus pathogenesis for more than 90 years. However, lack of a comprehensive analysis of the CRPV transcriptome has impeded the understanding of CRPV biology and molecular pathogenesis. Here, we report the construction of a complete CRPV transcription map from Hershey CRPV-induced skin tumor tissues. By using RNA-seq in combination with long-reads PacBio Iso-seq, 5' and 3' RACE, primer-walking RT-PCR, Northern blot, and RNA in situ hybridization, we demonstrated that the CRPV genome transcribes its early and late RNA transcripts unidirectionally from at least five distinct major promoters (P) and polyadenylates its transcripts at two major polyadenylation (pA) sites. The viral early transcripts are primarily transcribed from three "early" promoters, P90, P156, and P907 and polyadenylated at nt 4368 by using an early polyadenylation signal (PAS) at nt 4351. Like other low-risk human papillomaviruses and animal papillomaviruses, CRPV E6 and E7 transcripts are transcribed from three separate early promoters. Transcripts from two "late" promoters, P7525, and P1225, utilize either an early PAS for E1^E4 or a late PAS at 7399 for L2 and L1 RNA polyadenylation at nt 7415 to express capsid L2 and L1 proteins respectively. By using the mapped four 5' splice sites and three 3' splice sites, CRPV RNA transcripts undergo extensive alternative splicing to produce more than 33 viral RNA isoforms for production of at least 12 viral proteins, some of which without codon optimization are expressible in rabbit RK13 and human HEK293T cells. The constructed full CRPV transcription map in this study for the first time will enhance our understanding of the structures and expressions of CRPV genes and their contribution to molecular pathogenesis and tumorigenesis.