Genome editing to map the role of 7p14.3 locus in prostate cancer

[puxi]

B. Stringa, S. Garritano, A. Romanel, P. Gasperini, G. Petris, A.

Cereseto, F. Demichelis

University of Trento, Trento, Italy

Prostate cancer (PCa), the second most common cancer

among men, is a highly heritable molecularly and clinically

heterogeneous disease. In this work we focused on the study

of germline variants as putative responsible for early

somatic events in PCa. In silico analysis by our group

identified a non-coding polymorphic regulatory element at

the 7p14.3 locus that is associated with DNA repair and

hormone regulated transcript levels and with a prostate

cancer specific subclass with high genomic instability. In

vitro studies confirmed the enhancer activity of the locus

and the binding affinity for two transcription factors (TFs);

androgen receptor (AR) and CCAAT/Enhancer Binding

Protein (C/EBP) beta (CEBPB). To first proof a functional

role of the 7p14.3 locus I deleted 731bp of the genomic area

of interest in PC-3 cells by using CRISPR-Cas9. RNA-seq

analyses performed upon AR overexpression and/or

CEBPB silencing revealed significant deregulation of the

transcriptome in edited versus non-edited cells (control

cells). To well-characterize TFs binding motifs flaking the

locus, I am performing the disruption of AR and CEBPB

motifs via CRISPR-Cas9 in prostate cell lines. Moreover, to

better study the role of minor allele of the polymorphism on

genomic instability, I am currently working on the establishment of isogenic PC-3 cells via CRISPR-Cas9. Once

clones with the three possible genotypes are selected, I plan

on testing DNA damage response under relevant conditions.

This work is a proof of concept of germline predisposition

to molecularly distinct cancer subclasses and has the

potential to nominate new mechanisms of cancer

development.

Abstracts from the 51st European Society of Human Genetics Conference: Posters 459

B. Stringa: None. S. Garritano: None. A. Romanel:

None. P. Gasperini: None. G. Petris: None. A. Cereseto:

None. F. Demichelis: None.

[j-o-h-n]

Ciao....

Good Luck, Good Health and Good Humor.

j-o-h-n Friday 07/12/2019 6:36 PM DST