ABSTRACT: A DIA workflow, which is constructed on a quadrupole-Orbitrap LC-MS platform, is used to reveal perturbed proteins between ccRCC and para-carcinoma tissues.
Project description:This is a supplementary study to the CPTAC CCRCC Discovery Study - Proteome. Unlabeled, digested peptide material from individual tissue samples (ccRCC and NAT) was spiked with index Retention Time (iRT) peptides (Biognosys) and subjected to data-independent acquisition (DIA) analysis.
Kidney cancer is among the 10 most common cancers in both men and women and each year there are approximately 60,000 new cases with over 14,000 deaths (<a href="https://seer.cancer.gov/statfacts/html/kidrp.html" target="_blank">NCI, Surveillance, Epidemiology and End Results (SEER) Program</a>). Several histological and molecular subtypes have been identified and clear cell renal cell carcinoma (CCRCC) is the most prevalent (<a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5936048/" target="_blank">Hsieh el al., 2017 Nat Rev Dis Primers</a>). To advance the proteogenomic understanding of CCRCC, the CPTAC program has investigated 110 tumors (CPTAC discovery cohort) and subjected these samples to global proteome and phosphoproteome analysis. An optimized workflow for mass spectrometry of tissues using isobaric tags (TMT (tandem mass tags)-10) was used (<a href="https://www.nature.com/articles/s41596-018-0006-9" target="_blank">Mertins et al., Nature Protocols 2018</a>). Proteome and phosphoproteome data from the CCRCC tumors is available below along with peptide spectrum analyses (PSMs) and protein summary reports from the CPTAC common data analysis pipeline (CDAP).
Clinical data is provided. Additional attributes along with genotypes will be available as cohort characterization proceeds.
Genomic data will be available from the NCI Genomic Data Commons.
<em>Note: Sample-wise assessment of genomic profiles in this cohort identified seven tumor samples with molecular aberrations atypical for ccRCC. While these seven non-ccRCC samples (C3L-00359-01, C3N-00313-03, C3N-00435-05, C3N-00492-04, C3N-00832-01, C3N-01175-01, C3N-01180-01) and their corresponding NATs (C3N-00435-06, C3N-00492-05, C3N-01175-05) were excluded from the ccRCC cohort in all downstream analyses, the non-ccRCC samples served as useful controls to highlight ccRCC-specific features. These seven samples were therefore annotated as non-ccRCC samples.</em>
<ul><li>Dataset imported into MassIVE from <a href="https://pdc.cancer.gov/pdc/study/PDC000200" target="_blank">https://pdc.cancer.gov/pdc/study/PDC000200</a> on 10/04/23</li></ul>
Project description:To illustrate the molecular mechanism of the overexpression of RELB in esophageal squamous cell carcinoma compared with para-carcinoma tissues We then calculated bismark analysis using data obtained from bisulfite target sequencing data of six paires of esophageal squamous cell carcinoma and para-carcinoma tissues
Project description:Clear cell renal cell carcinoma (ccRCC) is the dominant subtype of renal cancer. With currently available therapies, cure of advanced and metastatic ccRCC is achieved only in rare cases. Here, we developed a workflow integrating different -omics technologies to identify ccRCC-specific HLA-presented peptides as potential drug targets for ccRCC immunotherapy. We analyzed frequent ccRCC-specific peptides by MS-based HLA ligandomics of 55 ccRCC tumors (cohort 1), paired non-tumor renal tissues and 158 benign tissues from other organs. Pathways enriched in ccRCC compared to its cell type of origin were identified by transcriptome and gene set enrichment analyses in 51 tumor tissues of the same cohort. To retrieve a list of candidate target genes with involvement in ccRCC pathogenesis, ccRCC-specific pathway genes were intersected with the source genes of tumor-exclusive peptides. The candidates were validated in an independent cohort from the Cancer Genome Atlas (TCGA KIRC, n=452), yielding 113 candidate genes. DNA methylation (TCGA KIRC, n=273), and somatic mutations (TCGA KIRC, n=392), as well as correlations with tumor metabolites (cohort 1, n=30) and immune-oncological markers (cohort 1, n=37) were analyzed to refine regulatory and functional involvements of candidates. Immunogenicity analysis identified candidate epitopes able to activate native CD8+ T cells. Functional analysis of EGLN3, a candidate with frequent ccRCC-specific immunogenic peptides, revealed possible tumor-promoting functions. Integration of HLA ligandomics, transcriptomics, genetic and epigenetic data leads to the identification of novel functionally relevant therapeutic targets for ccRCC immunotherapy. Validation of the identified targets is now mandatory to expand the treatment landscape of ccRCC.
Project description:To screen the related miRNAs in the occurence and development of sebaceous gland carcinoma(SGC) of the eyelid , we have employed Human Agilient microarray expression profiling as a discovery platform to identify differentially expressed miRNAs for SGC and TP53 mtation related miRNAs. Human formalin-fixed paraffin-embedded (FFPE) SGC tissues and para-carcinoma sebaceous gland(SG) FFPE samples from SGC patients were screened using microarray assay. Human FFPE SGC tissues containing TP53 mutation and human FFPE SGC tissues without TP53 mutation were screened using microarray. The overlap of two sets of microarray differentially upregulated miRNAs were selected as our research object.
Project description:<p>A discovery-based lipid profiling study of serum samples from a cohort that included patients with clear cell Renal Cell Carcinoma (ccRCC) stage I, II, III, and IV (n=112) and controls (n=52) was performed using ultraperformance liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry and machine learning techniques. Multivariate models based on support vector machines and the LASSO variable selection method yielded two discriminant lipid panels for ccRCC detection and early diagnosis. A 16-lipid panel allowed discriminating ccRCC patients from controls with 95.7% accuracy in a training set under cross- validation, and 77.1% accuracy in an independent test set. A second model trained to discriminate early (I and II) from late (III and IV) stage ccRCC yielded a panel of 26 compounds that classified stage I patients from an independent test set with 82.1% accuracy. Thirteen species, including cholic acid, undecylenic acid, lauric acid, LPC(16:0/0:0), and PC(18:2/18:2) identified with level 1 exhibited significant lower levels in samples from ccRCC patients compared to controls. Moreover, 3α-hydroxy-5α-androstan-17-one 3-sulfate, cis-5-dodecenoic acid, arachidonic acid, cis-13-docosenoic acid, PI(16:0/18:1), PC(16:0/18:2), and PC(O-16:0/20:4) contributed to discriminate early from late ccRCC stage patients. The results are auspicious for early ccRCC diagnosis after validation of the panels in larger and different cohorts.</p>
Project description:Clear cell renal cell carcinoma (ccRCC) is a disease with a potentially high level of tumour heterogeneity and inherited predisposition. Although many studies performed comprehensive molecular characterisation of ccRCC, the DNA regulatory mechanism is still unknown, especially at the single-cell level. Single-cell RNA sequencing (scRNA-seq) and single-cell assay for transposase-accessible chromatin using sequencing (scATAC-seq) were performed on 19 ccRCC samples. A total of 102,723 high-quality single-cell transcriptome information, 61,693 high-quality nuclei and 190,916 unique peaks were captured. We used whole-exome sequencing to understand the heterogeneity between individuals, validate the function of long noncoding RNAs (lncRNAs) in vitro and identify protein interactions by protein mass spectrometry. Single-cell transcriptome and chromatin accessibility maps of ccRCC were constructed. We found the gene expression characteristics of different tumour cell subtypes in ccRCC and predicted their sensitivity to targeted and conventional drugs. Interestingly, we discovered that two lncRNAs (RP11-661C8.2 and CTB-164N12.1) promoted the invasion and migration of ccRCC, which were verified by in vitro experiments. Taken together, this study demonstrated the comprehensive gene expression and DNA regulation information of ccRCC and discovered and validated lncRNAs that promote tumour cell invasion and migration, which will provide new insights into the treatment of ccRCC.
Project description:TGF-β-SMAD signaling pathway plays an important role in progression of various tumors. However, posttranscriptional regulation such as N6-methyladenosine (m6A) of TGF-β-SMAD signaling axis remains incompletely understood. Here, we reveal that Insulin like growth factor 2 mRNA binding protein 2 (IGF2BP2) is low expression as well as associated with poor prognosis in clear cell renal cell carcinoma (ccRCC) patients and inhibits proliferation as well as promotes metastasis of ccRCC cells. To explore the underlying mechanism of IGF2BP2 in ccRCC, RNA-seq, RIP-seq and LACE-seq were performed to investigate the downstream targets of IGF2BP2.
Project description:TGF-β-SMAD signaling pathway plays an important role in progression of various tumors. However, posttranscriptional regulation such as N6-methyladenosine (m6A) of TGF-β-SMAD signaling axis remains incompletely understood. Here, we reveal that Insulin like growth factor 2 mRNA binding protein 2 (IGF2BP2) is low expression as well as associated with poor prognosis in clear cell renal cell carcinoma (ccRCC) patients and inhibits proliferation as well as promotes metastasis of ccRCC cells. To explore the underlying mechanism of IGF2BP2 in ccRCC, RNA-seq, RIP-seq and LACE-seq were performed to investigate the downstream targets of IGF2BP2.
Project description:TGF-β-SMAD signaling pathway plays an important role in progression of various tumors. However, posttranscriptional regulation such as N6-methyladenosine (m6A) of TGF-β-SMAD signaling axis remains incompletely understood. Here, we reveal that Insulin like growth factor 2 mRNA binding protein 2 (IGF2BP2) is low expression as well as associated with poor prognosis in clear cell renal cell carcinoma (ccRCC) patients and inhibits proliferation as well as promotes metastasis of ccRCC cells. To explore the underlying mechanism of IGF2BP2 in ccRCC, RNA-seq, RIP-seq and LACE-seq were performed to investigate the downstream targets of IGF2BP2.
Project description:Inactivation of the von Hippel-Lindau (VHL) E3 ubiquitin ligase protein is a hallmark of clear cell renal cell carcinoma (ccRCC). Identifying how pathways affected by VHL loss contribute to ccRCC remains challenging. We used a genome-wide in vitro expression strategy to identify proteins that bound VHL only when hydroxylated. Zinc fingers and homeoboxes 2 (ZHX2) was found as a VHL target and its hydroxylation allowed VHL to regulate its protein stability. Tumor cells from ccRCC patients with VHL loss-of-function mutations usually had increased ZHX2 amount and nuclear localization. Functionally, depletion of ZHX2 inhibited VHL-deficient ccRCC cell growth in vitro and in vivo. Mechanistically, integrated ChIP-Seq and microarray analysis showed that ZHX2 promoted NF-kB activation. These studies reveal ZHX2 as a potential therapeutic target for ccRCC.