Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Validation of a laboratory assay for prediction of relapse in stage I non-small cell lung cancer (NSCLC) using miRNA profiling of formalin-fixed paraffin-embedded (FFPE) tissue specimens.

ABSTRACT: Technical validation of miRNA assay for prediction of relapse in NSCLC.

ORGANISM(S): Homo sapiens

SUBMITTER: Jesper Dahlgaard

PROVIDER: E-MTAB-618 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Analytical variables influencing the performance of a miRNA based laboratory assay for prediction of relapse in stage I non-small cell lung cancer (NSCLC).

Dahlgaard Jesper J Mazin Wiktor W Jensen Thomas T Pøhl Mette M Bshara Wiam W Hansen Anker A Kanisto Eric E Hamilton-Dutoit Stephen Jacques SJ Hansen Olfred O Hager Henrik H Ditzel Henrik J HJ Yendamuri Sai S Knudsen Steen S

BMC research notes 20111019

<h4>Background</h4>Laboratory assays are needed for early stage non-small lung cancer (NSCLC) that can link molecular and clinical heterogeneity to predict relapse after surgical resection. We technically validated two miRNA assays for prediction of relapse in NSCLC. Total RNA from seventy-five formalin-fixed and paraffin-embedded (FFPE) specimens was extracted, labeled and hybridized to Affymetrix miRNA arrays using different RNA input amounts, ATP-mix dilutions, array lots and RNA extraction- ...[more]

PMID: 22011393

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Project description:BACKGROUND: The ability of gene profiling to predict treatment response and prognosis in breast cancers has been demonstrated in many studies using DNA microarray analyses on RNA from fresh frozen tumor specimens. In certain clinical and research situations, performing such analyses on archival formalin fixed paraffin-embedded (FFPE) surgical specimens would be advantageous as large libraries of such specimens with long-term follow-up data are widely available. However, FFPE tissue processing can cause fragmentation and chemical modifications of the RNA. A number of recent technical advances have been reported to overcome these issues. Our current study evaluates whether or not the technology is ready for clinical applications. METHODS: A modified RNA extraction method and a recent DNA microarray technique, cDNA-mediated annealing, selection, extension and ligation (DASL, Illumina Inc) were evaluated. The gene profiles generated from FFPE specimens were compared to those obtained from paired fresh fine needle aspiration biopsies (FNAB) of 25 breast cancers of different clinical subtypes (based on ER and Her2/neu status). Selected RNA levels were validated using RT-qPCR, and two public databases were used to demonstrate the prognostic significance of the gene profiles generated from FFPE specimens. RESULTS: Compared to FNAB, RNA isolated from FFPE samples was relatively more degraded, nonetheless, over 80% of the RNA samples were deemed suitable for subsequent DASL assay. Despite a higher noise level, a set of genes from FFPE specimens correlated very well with the gene profiles obtained from FNAB, and could differentiate breast cancer subtypes. Expression levels of these genes were validated using RT-qPCR. Finally, for the first time we correlated gene expression profiles from FFPE samples to survival using two independent microarray databases. Specifically, over-expression of ANLN and KIF2C, and under-expression of MAPT strongly correlated with poor outcomes in breast cancer patients. CONCLUSION: We demonstrated that FFPE specimens retained important prognostic information that could be identified using a recent gene profiling technology. Our study supports the use of FFPE specimens for the development and refinement of prognostic gene signatures for breast cancer. Clinical applications of such prognostic gene profiles await future large-scale validation studies.

Dataset Information

Validation of a laboratory assay for prediction of relapse in stage I non-small cell lung cancer (NSCLC) using miRNA profiling of formalin-fixed paraffin-embedded (FFPE) tissue specimens.

Publications

Analytical variables influencing the performance of a miRNA based laboratory assay for prediction of relapse in stage I non-small cell lung cancer (NSCLC).

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