Page 241 - index.html

P06.11

WHOLE-EXOME SEQUENCING IN BRAZILIAN PATIENTS
WITH HEPATOCELLULAR CARCINOMA

1
1
Andreza Correa Texeira , Wilson Silva-Junior , Rodrigo Tocantins Calado ,
* 1
Kamila Peronni , Jessica Plaça , Daniela Tirapelli , Alice Barros , Guilherme
1
1
1
1
1
1
1
1
Rosa , Enio Mente , Jorge Elias-Junior , Valdair Muglia , Lucas Monsignore , Ana
1
LC Martinelli 1
1 Medical School of Ribeirao Preto-USP, Ribeirao Preto, Brazil
Corresponding author’s email: andrezacteixeira@gmail.com
Introduction: Genetic alterations in hepatocarcinogenesis are important events in the
development and progression of hepatocellular carcinoma (HCC). Emerging technologies
to perform genomic analyses have been important tools to improve tumor characterization.
Aims: The aim of this study was to evaluate the whole-exome sequencing in subjects with
chronic hepatitis C and hepatocellular carcinoma.
Material and Methods: We evaluated the whole-exome sequencing in six patients with ePOSTER ABSTRACTS
hepatocelullar carcinoma (HCC) and cirrhosis due to chronic hepatitis C (CHC), and in
two subjects with CHC, but without liver cancer (one with mild fibrosis and other with
cirrhosis). The exome capture was performed using the Nextera Exome Rapid Capture kit
(Illumina Inc., San Diego, CA, USA). Then, the captured DNA was sequenced on Illumina
Genome Analyzer IIx (GAIIx), based on the Solexa or SBS technology (Sequencing-by-
Synthesis) using the TruSeq SBS kit v5 (Illumina Inc., San Diego, CA, USA), configured to
2x75bp paired-end. The reads were analyzed for quality, and those with phred-score higher
than 30 were aligned to the human reference genome (GRCh37) using the Burrows–
Wheeler Aligner (BWA, version 0.7.12). The variants were called with the Genome Analysis
Toolkit (GATK, version 3.5) and annotated with Spneff (version 4.2).

Results: The whole-exome sequencing of all samples generated 135,699 variants (one
variant for each 22,388 bases). We found 15,449 somatic mutations, of which 6,806
were in the 3’untranslated region (UTR); 2,929 were missense mutations; 2,803 silent
mutations; and 1,128 were in the 5’UTR. The most mutated genes were MUC4 (n=76);
ZNF717 (n=65); HLA-DRB1 (n=38); and PDE4DIP (n=36). When evaluating the HCC
samples, the most mutated genes were RSPH3 (n=3; one missense mutation and 2 silent);
and NOTCH4 (n=3; one missense mutation and 2 silent). The variants MORN1 (intron),
DUSP28 (5’UTR) and TP63 (3’UTR) were observed only in HCC patients.

EASL HCC Summit • Geneva, Switzerland • 2-5 February, 2017 241