Project description:Purpose: To identify the gene expression change under hypoxia on HBE cells. Methods: bulk RNA-seq was performed on primary human bronchial epithelial cells (HBE) that were cultured on air-liquid interface (ALI) condition and treated under normoxia or hypoxia (1% O2) for 6hr, 24hr, and 5days. Results: hypoxia-related genes were significantly upregulated under hypoxia including EGLN3.
Project description:Purpose: To identify the cell types change and gene expression change under hypoxia on HBE cells. Methods: The primary human bronchial epithelial cells (HBE) were cultured on air-liquid interface (ALI) conditions. After 4 weeks, the cells were cultured under normoxic or hypoxic (1% O2) conditions for 6h or 5days. The cells were dissociated with Accutase solution and proceeded to scRNA-seq. Results: Hypoxia did not induce any hypoxia-specific cell types, however, all cell types upregulated hypoxia-related, and senescence-related genes and downregulated cell proliferation genes.
Project description:Gestational hypoxia presents a significant stress to an unborn fetus that can lead to significant complications related to fetal growth restriction and resulting in diseases in the newborn as well as those manifesting later in life. Recent evidence indicates that inflammation and oxidative stress are contributing factors to hypoxia-related diseases. The Center for Perinatal Biology at Loma Linda University has studied gestational chronic hypoxia in a sheep model for over 20 years to study dysfunction of vascular and nonvascular tissues derived from mothers, fetuses and offspring. In this project we are attempting to use metabolomics to assess metabolic dysregulation in vascular tissues along with markers of oxidative stress and inflammation in the mother and offspring to determine the extent of dysregulation due to chronic hypoxia. Untargeted metabolomics analysis focused on sheep plasma and arteries from the lung, resistance arteries in the brain, uterine arteries, and cultured human myocytes will be used to explore markers of glucose and lipid metabolism disruption. Targeted analyses of oxylipins and endocannabinoids will be used on the same samples to explore markers of oxidative stress and inflammation, which should be increased during hypoxia. This study should delineate pathways and biomarkers that help explain how hypoxia leads to the development of neonatal as well as adult-onset diseases associated with chronic hypoxia that are inter-related with fetal growth restriction.
Project description:Gestational hypoxia presents a significant stress to an unborn fetus that can lead to significant complications related to fetal growth restriction and resulting in diseases in the newborn as well as those manifesting later in life. Recent evidence indicates that inflammation and oxidative stress are contributing factors to hypoxia-related diseases. The Center for Perinatal Biology at Loma Linda University has studied gestational chronic hypoxia in a sheep model for over 20 years to study dysfunction of vascular and nonvascular tissues derived from mothers, fetuses and offspring. In this project we are attempting to use metabolomics to assess metabolic dysregulation in vascular tissues along with markers of oxidative stress and inflammation in the mother and offspring to determine the extent of dysregulation due to chronic hypoxia. Untargeted metabolomics analysis focused on sheep plasma and arteries from the lung, resistance arteries in the brain, uterine arteries, and cultured human myocytes will be used to explore markers of glucose and lipid metabolism disruption. Targeted analyses of oxylipins and endocannabinoids will be used on the same samples to explore markers of oxidative stress and inflammation, which should be increased during hypoxia. This study should delineate pathways and biomarkers that help explain how hypoxia leads to the development of neonatal as well as adult-onset diseases associated with chronic hypoxia that are inter-related with fetal growth restriction.
Project description:DEAD-box RNA-binding proteins (RBPs) play a significant role in RNA metabolism to achieve cellular homeostasis, including miRNA biogenesis and transcription. Hypoxia induces stemness cell-like characteristics in cancer cells and promotes malignant progression. Despite the fact that hypoxia can induce the changes in protein and RNA modification, thereby regulating downstream gene expressions, how modifications at different molecular layers interplay with each other are poorly understood. Here we show that hypoxia induces HectH9-mediated K63-linked polyubiquitination of the DEAD-box protein DDX17 as well as reduces N6-methyladenosine (m6A) marks in pri-miRNAs. While m6A potentiates DDX17 binding to pri-miRNAs, decreased m6A modifications of pri-miRNAs and increased polyubiquitination of DDX17 under hypoxia lead to decreased DDX17 binding to pri-miRNAs binding. These events enhance the association of DDX17 with the ubiquitin receptor p300 and lead to a decrease in miRNA biogenesis, especially for miRNAs regulating stemness and stemness-related genes. In addition, polyubiquitinated DDX17 together with p300 upregulates H3K56Ac levels on the stemness and stemness-related genes, resulting in enhancement of tumor initiating ability. Post-transcriptionally, decreased miRNA production, including those targeting stemness genes or stemness-related genes, also facilitates tumor initiation. Together, hypoxia triggers DDX17 poly-ubiquitination, which orchestrates dual mechanisms to increase tumor initiating ability and promote tumor progression.
Project description:We investigated the effect of low oxygen culture on the proliferation and hair inductive capacity of human dermal papilla cells (DPCs) and dermal sheath cells (DSCs). DPCs and DSCs were cultured in atmospheric/hyperoxia (20% O2), physiological/normoxia (6% O2), or hypoxia (1% O2) conditions, respectively. Proliferation of DPCs and DSCs was highest under normoxia. Hypoxia inhibited proliferation of DPCs but enhanced proliferation of DSCs. In DPCs, hypoxia down-regulated expression of hair inductive capacity-related genes, including BMP4, LEF1, SOX2, and VCAN, and normoxia up-regulated expression of ALP. In DSCs, both normoxia and hypoxia up-regulated SOX2 expression, and hypoxia down-regulated BMP4 expression. Microarray analysis revealed increased expression of pluripotency-related genes, including SPRY, NR0B1, MSX2, IFITM1, and DAZL, under hypoxia. In an in vivo hair follicle reconstitution assay, cultured DPCs and DSCs were transplanted with newborn mouse epidermal keratinocytes into nude mice using a chamber method. In DPCs, normoxia allowed the most efficient induction of hair follicles. In DSCs, hypoxia allowed the most efficient induction and maturation of hair follicles. These results suggest that low oxygen culture enhances the proliferation and maintains functions of human DPCs and DSCs and could be used for skin engineering and clinical applications.
Project description:We investigated the effect of low oxygen culture on the proliferation and hair inductive capacity of human dermal papilla cells (DPCs) and dermal sheath cells (DSCs). DPCs and DSCs were cultured in atmospheric/hyperoxia (20% O2), physiological/normoxia (6% O2), or hypoxia (1% O2) conditions, respectively. Proliferation of DPCs and DSCs was highest under normoxia. Hypoxia inhibited proliferation of DPCs but enhanced proliferation of DSCs. In DPCs, hypoxia down-regulated expression of hair inductive capacity-related genes, including BMP4, LEF1, SOX2, and VCAN, and normoxia up-regulated expression of ALP. In DSCs, both normoxia and hypoxia up-regulated SOX2 expression, and hypoxia down-regulated BMP4 expression. Microarray analysis revealed increased expression of pluripotency-related genes, including SPRY, NR0B1, MSX2, IFITM1, and DAZL, under hypoxia. In an in vivo hair follicle reconstitution assay, cultured DPCs and DSCs were transplanted with newborn mouse epidermal keratinocytes into nude mice using a chamber method. In DPCs, normoxia allowed the most efficient induction of hair follicles. In DSCs, hypoxia allowed the most efficient induction and maturation of hair follicles. These results suggest that low oxygen culture enhances the proliferation and maintains functions of human DPCs and DSCs and could be used for skin engineering and clinical applications.
Project description:The aim of this study was to perform the multiscale correlation between quantitative texture features phenotype of pre-biopsy biparametric MRI (bpMRI) and targeted sequence-based RNA expression for hypoxia-related genes. Images from pre-biopsy 3T bpMRI scans in clinically localised prostate cancer (PCa) patients of various risk categories (n=15) were used to extract textural features. The genomic landscape of hypoxia-related genes expression was obtained using post-radical prostatectomy tissue for targeted RNA expression profiling using the TempO-sequence method. The nonparametric Games Howell test was used to correlate the differential expression of the three important hypoxia-related genes with 28 radiomic texture features. Following this, cBioportal was accessed and a gene-oriented query was conducted to extract Oncoprint genomic output graph of the selected hypoxia-related genes from The Cancer Genome Atlas (TCGA). Correlation analysis using Pearson's coefficients calculated against each selected gene profile; survival analysis using Kaplan-Meier estimators were carried out. We found the quantitative bpMR imaging textural features, including histogram and grey level co-occurrence matrix (GLCM), correlated with hypoxia related genes (ANGPTL4, VEGFA, and P4HA1) seen on RNA sequencing using TempO-Seq method. Further radiogenomic analysis, including data accessed on cBioportal genomic database, confirmed that overexpressed hypoxia-related genes significantly correlated with a poor survival outcome, with a median survival of 81.11: 133.00 months in those with and without alterations of genes respectively. In summary, radiomic texture features of bpMRI in localised PCa correlate with the expression of hypoxia-related genes expression in prostate cancer. The expression data analysis showed that hypoxia-related genes are associated with poor survival.