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ABSTRACT: Bulk 3-prime RNA-sequencing of macrodissected neuroendocrine and exocrine tumor components of FFPE-embedded mouse gastric neuroendocrine carcinomas

PROVIDER: PRJEB61968 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR11422919_1.fastq.gz	Fastqsanger.gz
	ERR11422919_2.fastq.gz	Fastqsanger.gz
	ERR11422920_1.fastq.gz	Fastqsanger.gz
	ERR11422920_2.fastq.gz	Fastqsanger.gz
	ERR11422921_1.fastq.gz	Fastqsanger.gz

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mRNA profiles in formalin-fixed and paraffin embedded (FFPE) breast cancer tissues

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Pulmonary Carcinoids and Low-Grade Gastrointestinal Neuroendocrine Tumors Show Common MicroRNA Expression Profiles, Different from Adenocarcinomas and Small Cell Carcinomas

Project description:Background: It is still uncertain whether carcinoids of the lung and gastrointestinal (GI) tract have a common origin or whether they are closer in origin to carcinomas of the same organs. MicroRNA (miRNA) expression may clarify their nature and origin. Methods: First, to verify whether formalin-fixed paraffin-embedded (FFPE) samples retain the expression signature of the tissue, miRNA expression was compared between FFPE and frozen samples. Second, we selected surgically resected FFPE samples of pulmonary and GI carcinoids, as well as other types of tumors and normal tissues from each organ, and we compared the comprehensive expression patterns of miRNAs by microarray. These data were analyzed by hierarchical clustering and consensus clustering with non-negative matrix factorization (NMF). Results: The miRNA expression profiles of FFPE and frozen samples correlated quite well. In the first hierarchical clustering, most of the carcinoids formed one major cluster with loose subpartitioning into each organ type, while the second major cluster mainly comprised adenocarcinomas and normal tissues. The NMF approach largely supported hierarchical clustering. In the additional cluster analysis comparing carcinoids to small-cell lung carcinomas (SCLCs), carcinoids formed a distinct cluster, while SCLCs grouped together with pulmonary adenocarcinomas and normal lung tissues in another major cluster. Furthermore, we found some miRNAs that exhibited significant expression in carcinoids. Conclusion: Carcinoids had a characteristic pattern of miRNA expression, suggesting a common origin for pulmonary and GI carcinoids. The expression profiles were different in carcinoids and SCLCs, indicating distinct histogenesis of these neuroendocrine tumors.

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Identification of HER2 peptides for quantification in formalin-fixed paraffin-embedded breast cancer tissues

Project description:The ability to accurately quantify proteins in formalin-fixed paraffin-embedded (FFPE) tissues using targeted mass spectrometry opens exciting perspectives for biomarker discovery. We have developed and evaluated a selected reaction monitoring (SRM) assay for the human receptor tyrosine-protein kinase erbB-2 (HER2) in FFPE breast tumors. Peptide candidates were identified using an untargeted mass spectrometry approach in relevant cell lines. A selected reaction monitoring (SRM) assay was developed for the six best candidate peptides and evaluated for linearity, precision and lower limit of quantification. The six HER2 peptides of interest were then quantified by SRM in a cohort of 40 archival formalin-fixed paraffin-embedded tumor tissues from women with invasive breast carcinomas, showing different levels of HER2 gene amplification. Finally, the agreement was tested between data generated by SRM and data generated using standard clinical pathology methods, namely immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). The high agreement observed indicates that SRM is a suitable method to quantify peptides from FFPE material, and therefore represents a powerful approach for biomarker discovery studies.

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