Project description:We performed shallow whole genome sequencing (WGS) on circulating free (cf)DNA extracted from plasma or cerebrospinal fluid (CSF), and shallow WGS on the tissue DNA extracted from the biopsy in order to evaluate the correlation between the two biomaterials. After library construction and sequencing (Hiseq3000 or Ion Proton), copy number variations were called with WisecondorX.
Project description:Whole genome sequencing (WGS) of tongue cancer samples and cell line was performed to identify the fusion gene translocation breakpoint. WGS raw data was aligned to human reference genome (GRCh38.p12) using BWA-MEM (v0.7.17). The BAM files generated were further analysed using SvABA (v1.1.3) tool to identify translocation breakpoints. The translocation breakpoints were annotated using custom scripts, using the reference GENCODE GTF (v30). The fusion breakpoints identified in the SvABA analysis were additionally confirmed using MANTA tool (v1.6.0).
Project description:Nanostring miRNA Array on PGP human bipolar spindle neuron For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODE_Data_Use_Policy_for_External_Users_03-07-14.pdf
Project description:Breast Cancer Resistance Protein (Bcrp) and P-glycoprotein (Pgp) are xenobiotic efflux transporters that limit permeability of their substrates in the gastrointestinal system and brain, and increase drug clearance in liver and kidney. Bcrp and Pgp are also known to transport different endogenous metabolites such as riboflavin, urate, sulfated steroids and glucocorticoids. However, a systemic approach to understand the endogenous substrates and function of these transporters is still lacking. We hypothesized that Bcrp and Pgp have important endogenous functions and play a crucial role in maintaining metabolomic homeostasis in the biological system. To understand the impact of Bcrp and Pgp on endogenous metabolome, we performed untargeted metabolomics of CSF and plasma of male Sprague Dawley (SD) rats (WT) and SD rats lacking both Bcrp and Pgp (double knock-out, dKO rats) (n= 8 per genotype). In addition, we performed genome-wide microarray analysis of brain frontal cortex, kidney and liver from the same animals to analyze changes in gene expression (n=5 per genotype). Our results indicate distinct differences in metabolites and gene expression between WT and dKO rats. Rank order based on both fold change (>1.5 or <0.67) and significance (p <0.05) identified 29 metabolites in CSF and 47 metabolites in plasma as most significantly altered between WT and dKO rats, which includes, among other biochemicals, known substrates of Bcrp e.g., riboflavin, urate, daidzein and its metabolites. Significantly altered metabolites in plasma as well as CSF showed signatures of altered fatty acid metabolism and oxidative stress, which was further supported by the pathway analysis of significantly altered genes. These findings may lead to new insights in understanding the endogenous function of Bcrp and Pgp transporters and identification of functional biomarkers of these transporters
Project description:A drug currently efficient for cerebral stroke therapy is Semax, a synthetic peptide bearing a fragment of ACTH (4â7) and the C-terminal tripeptide Pro-Gly-Pro (PGP) was included to ensure resistance to peptidases.The genome-wide expression changes induced by Semax and PGP in rat brain cortex tissues damaged by focal ischemia were studied using the genome-wide RatRef-12 Expression BeadChip (Illumina, USA), which contains 22,226 genes, according to NCBI. We compared the biochip data obtained at 3 h and 24 h after permanent middle cerebral artery occlusion (pMCAO) in each of the three groups (âischemia,â âischemia + Semax,â and âischemia + PGPâ). The transcriptome profiles were examined at 24 h vs. 3 h after pMCAO in rats that produced ischemic cortical injury and in rats with the same injury treated with Semax or PGP.
Project description:In principle, whole-genome sequencing (WGS) of the human genome even at low coverage offers higher resolution for genomic copy number variation (CNV) detection compared to array-based technologies, which is currently the first-tier approach in clinical cytogenetics. There are, however, obstacles in replacing array-based CNV detection with that of low-coverage WGS such as cost, turnaround time, and lack of systematic performance comparisons. With technological advances in WGS in terms of library preparation, instrument platforms, and data analysis algorithms, obstacles imposed by cost and turnaround time are fading. However, a systematic performance comparison between array and low-coverage WGS-based CNV detection has yet to be performed. Here, we compared the CNV detection capabilities between WGS (short-insert, 3kb-, and 5kb-mate-pair libraries) at 1X, 3X, and 5X coverages and standardly used high-resolution arrays in the genome of 1000-Genomes-Project CEU genome NA12878. CNV detection was performed using standard analysis methods, and the results were then compared to a list of Gold Standard NA12878 CNVs distilled from the 1000-Genomes Project. Overall, low-coverage WGS is able to detect drastically more (approximately 5 fold more on average) Gold Standard CNVs compared to arrays and is accompanied with fewer CNV calls without secondary validation. Furthermore, we also show that WGS (at ≥1X coverage) is able to detect all seven validated deletions larger than 100 kb in the NA12878 genome whereas only one of such deletions is detected in most arrays. Finally, we show that the much larger 15 Mbp Cri-du-chat deletion can be clearly seen at even 1X coverage from short-insert WGS.
Project description:We evaluated linked-read whole genome sequencing (WGS) for detection of structural chromosomal rearrangements in primary samples of varying DNA quality from 12 patients diagnosed with ALL. Linked-read WGS enabled precise, allele-specific, digital karyotyping at a base-pair resolution for a wide range of structural variants including complex rearrangements, aneuploidy assessment and gene deletions. Additional RNA-sequencing and copy number aberrations (CNA) data from Illumina Infinium arrays were also generated and assessed against the linked-read WGS data. RNA-sequencing data was used to support structural chromosomal rearrangements detected in the linked-read WGS data by detecting expressed fusion genes as a consequence of the rearrangements. Illumina Infinium arrays (450k array and/or SNP array) were used to assess CNA status to further support the findings in the linked-read WGS data. The processed CNA data from the primary ALL patient samples has been deposited to GEO. RNA-sequencing, linked-read WGS data, and raw SNP array data from the primary ALL patient samples will not be deposited because the patient/parent consent does not cover depositing data that may be used for large-scale determination of germline variants in a repository. The ALL samples were collected 10-20 years ago from pediatric patients aged 2-15 years, some whom have deceased. The linked-read WGS data and the RNA-sequencing data sets generated in the study are available upon reasonable request from the corresponding author Jessica.Nordlund@medsci.uu.se.
Project description:We evaluated linked-read whole genome sequencing (WGS) for detection of structural chromosomal rearrangements in primary samples of varying DNA quality from 12 patients diagnosed with ALL. Linked-read WGS enabled precise, allele-specific, digital karyotyping at a base-pair resolution for a wide range of structural variants including complex rearrangements, aneuploidy assessment and gene deletions. Additional RNA-sequencing and copy number aberrations (CNA) data from Illumina Infinium arrays were also generated and assessed against the linked-read WGS data. RNA-sequencing data was used to support structural chromosomal rearrangements detected in the linked-read WGS data by detecting expressed fusion genes as a consequence of the rearrangements. Illumina Infinium arrays (450k array and/or SNP array) were used to assess CNA status to further support the findings in the linked-read WGS data. The processed CNA data from the primary ALL patient samples has been deposited to GEO. RNA-sequencing, linked-read WGS data, and raw SNP array data from the primary ALL patient samples will not be deposited because the patient/parent consent does not cover depositing data that may be used for large-scale determination of germline variants in a repository. The ALL samples were collected 10-20 years ago from pediatric patients aged 2-15 years, some whom have deceased. The linked-read WGS data and the RNA-sequencing data sets generated in the study are available upon reasonable request from the corresponding author Jessica.Nordlund@medsci.uu.se.