Project description:Purpose: There is a need for point-of-care diagnostics for future mass casualty events involving radiation exposure. The development of radiation exposure and dose prediction algorithms for biodosimetry is needed for screening of large populations during these scenarios, and exploration of the potential effects which sex, age, genetic heterogeneity, and physiological comorbidities may have on the utility of biodosimetry diagnostics is needed. In the current study, proteomic profiling was used to examine sex specific differences in age matched C57BL6 mice on the blood proteome following radiation exposure and the usefulness of development and application of biodosimetry algorithms using both male and female samples. Methods: C57BL6 male and female mice between 9-11 weeks of age received a single total body radiation exposure of either 2, 4 or 8 Gy with plasma collection at days 1, 3 and 7 post-irradiation. Plasma was then screened using the SomaScan v4.1 assay for ~7000 protein analytes. A subset panel of protein biomarkers demonstrated significant (FDR<0.05 and |logFC|>0.2) changes in expression following radiation exposure. All proteins were used for feature selection to build predictive models of radiation exposure using different sample and sex specific cohorts. Both binary (prediction of any radiation exposure) and multidose (prediction of specific radiation dose) model series were developed using either female and male samples combined or only female or only male samples. The binary series (Models 1, 2 and 3) and multidose series (Models 4, 5 and 6) series included female/male combined, female only and male only respectively. Results: Detectable values were obtained for all ~7000 proteins included in the SomaScan assay for all samples. Each model algorithm built using a unique sample cohort was validated with a training set of samples and tested with a separate new sample series. Overall predictive accuracies in the binary model series was ~100% at the model training level and when tested with fresh samples 97.9% for Model 1(Female and Male) and 100% for Model 2 (Female only) and Model 3 (Male only). When sex specific Models’ 2 and 3 were tested with the opposite sex, the overall predictive accuracy rate dropped to 62.5% for Model 2 and remained 100% for Model 3. The overall predictive accuracy rate in the multidose model series was 100% for all models at the model training level and when tested with fresh samples 83.3%, 75% and 83.3% for Multidose Models 4-6 respectively. When sex specific Models’ 5 (Female only) and Model 6 (Male only) were tested with the opposite sex the overall predictive accuracy rate dropped to 52.1% and 68.8% respectively. Conclusion: These models represent novel predictive panels of radiation responsive proteomic biomarkers and illustrate the utility and necessity of considering sex specific differences in development of radiation biodosimetry prediction algorithms. As sex specific differences were observed in this study, and as use of point-of-care radiation diagnostics in future mass casualty settings will necessarily include persons of both sexes, consideration of sex specific variation is essential to ensure these diagnostic tools have practical utility in the field.
Project description:Background/Aims: Tibetan chickens, a unique plateau breed, have good performances to adapt to high-altitude hypoxic environments. A number of positively selected genes have been reported in Tibetan chickens; however, the mechanisms of gene expression for hypoxia adaptation are not fully understood. Methods: Eggs from Tibetan (TC) and Chahua (CH) chickens were incubated under hypoxic and normoxic conditions, and vessel density index (VDI) in the chorioallantoic membrane (CAM) of embryos was measured. Meanwhile, Transcriptomic and proteomic analyses of CAM tissues were performed in TC and CH embryos under hypoxic incubation using RNA-seq and iTRAQ. Results: We found that the vessel density index (VDI) in CAM of TCs was lower than in CHs under hypoxia incubation. In the transcriptomic and proteomic analyses, 160 differentially expressed genes (DEGs) and 387 differentially expressed proteins (DEPs) that were mainly enriched in angiogenesis, vasculature development, blood vessel morphogenesis, blood circulation, renin-angiotensin system, and HIF-1 and VEGF signaling pathways. Twenty-six genes involved in angiogenesis and blood circulation, two genes involved in ion transport, and six genes that regulated energy metabolism were identified as candidate functional genes in regulating hypoxic adaption of chicken embryos. Conclusion: Combination of transcriptomic and proteomic data revealed several key candidate regulators and pathways that might play high-priority roles in the hypoxic adaptation of Tibetan chickens by regulating angiogenesis and promoting blood circulation, thus explaining the blunt responses to hypoxic conditions on CAM angiogenesis in Tibetan chicken embryos. This research provided insights into the molecular mechanism of hypoxia adaptation in Tibetan chickens.
Project description:This SuperSeries is composed of the following subset Series: GSE23672: COMPARATIVE TRANSCRIPTOMIC AND PROTEOMIC ANALYSIS OF LGR5+ve STEM CELLS AND THEIR DAUGHTERS (AGILENT ARRAYS) GSE33948: COMPARATIVE TRANSCRIPTOMIC AND PROTEOMIC ANALYSIS OF LGR5+ve STEM CELLS AND THEIR DAUGHTERS (AFFYMETRIX ARRAYS) Refer to individual Series
Project description:Understanding mammalian preimplantation development, particularly in humans, at the proteomic level remains limited. Here, we applied our Comprehensive Solution of Ultrasensitive Proteome Technology to measure the proteomic profiles of oocytes and early embryos, and identified nearly 8,000 proteins in humans and over 6,300 proteins in mice. We observed distinct proteomic dynamics before and around zygotic genome activation (ZGA) between the two species. Integrative analysis with translatome data revealed extensive divergence between translation activation and protein accumulation. Multi-omic analysis indicated that ZGA transcripts contribute to protein accumulation in blastocysts. Consequently, we identified several transcriptional regulators critical for early development in mice, thereby linking ZGA to the first lineage specification. Furthermore, single-embryo proteomics of poor quality embryos from over 100 patient-couples provided insights into preimplantation development failure. Our study may contribute to reshaping the framework of mammalian preimplantation development and opening new avenues for addressing human infertility.
Project description:Sulforaphane (SFN), an isothiocyanate found in cruciferous vegetables, is a potent inhibitor of experimental mammary carcinogenesis and may be an effective, safe chemopreventive agent for use in humans. SFN acts in part on the Keap1/Nrf2 pathway to regulate a battery of cytoprotective genes. In this study transcriptomic and proteomic changes in the estrogen receptor negative, non tumorigenic human breast epithelial MCF10A cell line were analyzed following SFN treatment or KEAP1 knockdown with siRNA using microarray and stable isotopic labeling with amino acids in culture (SILAC), respectively. Changes in selected transcripts and proteins were confirmed by PCR and Western blot in MCF10A and MCF12A cells. There was strong correlation between the transcriptomic and proteomic responses in both the SFN treatment (R=0.679, P<0.05) and KEAP1 knockdown (R=0.853, P<0.05) experiments. Common pathways for SFN treatment and KEAP1 knockdown were xenobiotic metabolism and antioxidants, glutathione metabolism, carbohydrate metabolism and NADH/NADPH regeneration. Moreover, these pathways were most prominent in both the transcriptomic and proteomic analyses. The aldo-keto reductase family members, AKR1B10, AKR1C1, AKR1C2 and AKR1C3, as well as NQO1 and ALDH3A1, were highly upregulated at both the transcriptomic and proteomic level. Collectively, these studies served to identify potential biomarkers that can be used in clinical trials to investigate the initial pharmacodynamic action of SFN in the breast. MCF10A cells were treated with SFN or had KEAP1 knocked down by siRNA.
Project description:We performed both transcriptomic and proteomic analyses on gastric cancer cells infected by H. pylori isolates from GML patients and the gastric ulcer strain 26695 to investigate the differentially expressed molecular signatures that were induced by GML isolates. Transcriptomicd analysis revealed fifteen pathways, including the Wnt signaling pathway, the mTOR signaling pathway, the NOD-like receptor signaling pathway and the Hippo signaling pathway, were revealed to be related to GML isolates.
Project description:Heat stress is one of the primary abiotic stresses that limit crop production . Grape is a popular cultivated fruit with high economic value throughout the world, and whose growth and development is often influenced by high temperature. Alternative splicing (AS) is a widespread mechanism increasing transcriptome complexity and proteome diversity. We conducted high temperature treatments (35oC, 40oC and 45oC) on grapevines (Vitis vinifera), and assessed proteomic and transcriptomic (especially AS)changes in leaves. We found that nearly 70% of the genes were alternatively spliced under high temperature. Intron retention (IR), exon skipping (ES) and alternative donor/acceptor sites were markedly induced under different high temperatures. IR was the most abundant up- and down-regulated AS event; moreover, IR events at 40 and 45oC were far higher than those at 35oC. These results indicated AS, especially IR, is an important posttranscriptional regulatory during grape leaf responses to high temperature. Proteomic analysis showed that protein levels of the RNA binding proteins SR45, SR30, and SR34, and the nuclear ribonucleic protein U1A in grape leaves gradually rose as ambient temperature increased. The results also revealed why AS events occurred more frequently under high temperature in grape leaves. After integrating transcriptomic and proteomic data, we found that HSPs and some important transcript factors such as MBF1c and HSFA2 were mainly involved in heat tolerance in grape through up-regulating transcriptional and translational levels, and were especially modulated by AS. The results provide the first simultaneous evidence for grape leaf responses to high temperature at transcriptional, posttranscriptional and translational levels.
Project description:An increasing number of studies, including mutant expression profiling and comparative transcriptomic analyses, require reference RNA-seq data collections in mice. Particularly, to complement previous profiling data sets based on arrays, a full RNA-seq developmental series will be required for whole embryos. E10.5 is a key reference stage as it represents the early organogenesis stage. Here, we have performed high-throughput sequencing of total RNA form whole mice embryos at embryonic stage E10.5.
Project description:Comparative transcriptomic and proteomic analyses provide insights into functional genes for hypoxic adaptation in embryos of Tibetan chickens
Project description:Objective The purpose of this study was to find the key regulatory genes and proteins in condylar ossification of temporomandibular joint (TMJ) by transcriptomics and proteomics in porcine embryos. Method A total of 12 miniature pig embryos (6 in each group) at E45 and E85 days were used in this study. Six embryos for tissue sections (3 in each group) . The remaining six embryos were cut down the condyle along the parallel line between the lowest point of the sigmoid notch and the highest point of the condyle, and each condylar tissue sample was divided into two parts averagely. After RNA extraction, library construction and sequencing, one part tissue was analyzed by mapping to compare the differential expression of mRNA before and after ossification of embryonic condylar tissue. Another tissue was analyzed by protein extraction, enzymolysis and mass spectrometry to compare the differential expression of protein before and after ossification of embryonic condylar tissue. Finally, the differential genes and proteins were analyzed by GO and KEGG enrichment analysis. The differentially expressed genes in transcriptome and proteomic analysis were verified by QPCR. Result A total of 25322 genes were detected by transcriptome analysis, and there were 19584 differential genes between E85 days and E45 days. There are 1592 differential genes were screened to meet the Fold change≥2 or ≤0.5 of which 1086 genes were up-regulated and 506 genes were down-regulated in E85 days compared with E45 days. A total of 4613 proteins were detected by proteomic analysis, there were 419 differential proteins between E85 and E45 days, including 313 up-regulated proteins and 106 down-regulated proteins. A total of 37 differentially expressed genes/proteins were found by cross transcriptome and proteomic analysis. QPCR analysis showed that 13 of 15 genes were consistent with transcriptome analysis. Conclusion Condylar transcriptome and proteomic analysis during the development of temporomandibular joint in miniature pigs revealed the regulatory genes/proteins of condylar ossification. These regulatory genes/proteins can provide basic data for human TMJ development research and disease treatment.