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2015 ; 10
(7
): e0132831
Nephropedia Template TP
gab.com Text
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English Wikipedia
A Multidisciplinary Biospecimen Bank of Renal Cell Carcinomas Compatible with
Discovery Platforms at Mayo Clinic, Scottsdale, Arizona
#MMPMID26181416
Ho TH
; Nateras RN
; Yan H
; Park JG
; Jensen S
; Borges C
; Lee JH
; Champion MD
; Tibes R
; Bryce AH
; Carballido EM
; Todd MA
; Joseph RW
; Wong WW
; Parker AS
; Stanton ML
; Castle EP
PLoS One
2015[]; 10
(7
): e0132831
PMID26181416
show ga
To address the need to study frozen clinical specimens using next-generation RNA,
DNA, chromatin immunoprecipitation (ChIP) sequencing and protein analyses, we
developed a biobank work flow to prospectively collect biospecimens from patients
with renal cell carcinoma (RCC). We describe our standard operating procedures
and work flow to annotate pathologic results and clinical outcomes. We report
quality control outcomes and nucleic acid yields of our RCC submissions (N=16) to
The Cancer Genome Atlas (TCGA) project, as well as newer discovery platforms, by
describing mass spectrometry analysis of albumin oxidation in plasma and 6 ChIP
sequencing libraries generated from nephrectomy specimens after histone H3 lysine
36 trimethylation (H3K36me3) immunoprecipitation. From June 1, 2010, through
January 1, 2013, we enrolled 328 patients with RCC. Our mean (SD) TCGA RNA
integrity numbers (RINs) were 8.1 (0.8) for papillary RCC, with a 12.5% overall
rate of sample disqualification for RIN <7. Banked plasma had significantly less
albumin oxidation (by mass spectrometry analysis) than plasma kept at 25 °C
(P<.001). For ChIP sequencing, the FastQC score for average read quality was at
least 30 for 91% to 95% of paired-end reads. In parallel, we analyzed frozen
tissue by RNA sequencing; after genome alignment, only 0.2% to 0.4% of total
reads failed the default quality check steps of Bowtie2, which was comparable to
the disqualification ratio (0.1%) of the 786-O RCC cell line that was prepared
under optimal RNA isolation conditions. The overall correlation coefficients for
gene expression between Mayo Clinic vs TCGA tissues ranged from 0.75 to 0.82.
These data support the generation of high-quality nucleic acids for genomic
analyses from banked RCC. Importantly, the protocol does not interfere with
routine clinical care. Collections over defined time points during disease
treatment further enhance collaborative efforts to integrate genomic information
with outcomes.