eprintid: 35580
rev_number: 22
eprint_status: archive
userid: 8536
dir: disk0/00/03/55/80
datestamp: 2024-11-18 11:03:55
lastmod: 2024-11-21 12:35:07
status_changed: 2024-11-18 11:03:55
type: doctoralThesis
metadata_visibility: show
creators_name: Jin, Fukang
title: ARID1A mutations in colorectal cancer: Biological and therapeutic implications
subjects: ddc-610
divisions: i-61800
adv_faculty: af-06
abstract: Introduction: The AT-interacting domain-rich protein 1A (ARID1A) gene undergoes mutations in approximately 10% of human colorectal cancer (CRC). The loss of function (LOF) associated with these mutations is intricately linked to the onset and advancement of CRC, and it serves as a prognostic indicator for unfavorable outcomes in CRC patients. Despite this, the exploration of the clinical implications and applications of ARID1A mutations remains relatively constrained.

Objectives: To delve deeper into the connection between ARID1A mutations and CRC, as well as to unravel the associated signaling pathways. This investigation seeks to identify and screen clinical drugs that specifically target ARID1A mutations.

Methods: In this investigation, the Cre-loxP system was employed to create Arid1a mutant mouse models. We produced CRC organoids, namely Arid1a flox/flox;Apc flox/flox;Kras ki/wt;Trp53 flox/flox (ArAKPf) and Apc flox/flox;Kras ki/wt;Trp53 flox/flox (AKPf), as well as human CRC cell line models featuring ARID1A knockout (KO) through CRISPR technology. These models were subsequently utilized in functional experiments.

Results: In comparison to the AKPf cell line, the ArAKPf cell line exhibited a significant augmentation in colony formation. This proliferative enhancement was substantiated by the cell counting kit-8 (CCK-8) assay, affirming heightened proliferative capabilities in the ArAKPf and HCT116 ARID1A KO cell line. Wound healing assays manifested a substantial increase in migratory distance for the ARID1A mutant cell line, and Boyden chamber assays unveiled a pronounced escalation in both migratory and invasive capacities of the ArAKPf and HCT116 ARID1A KO cell line. Subsequent bioinformatics analyses implicated Arid1a in the modulation of molecules integral to the phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway. This proposition was validated through quantitative reverse transcription PCR (RT-qPCR) assessments. Furthermore, drug screening delineated an extensive resistance profile in Arid1a-mutated mouse CRC cells. However, these cells demonstrated susceptibility to B-cell lymphoma 2 (BCL-2) inhibitors.

Conclusion: The findings suggest that ARID1A mutation confers heightened proliferative, migratory, and invasive capacities upon mouse and human CRC cells, concurrently resulting in a broad spectrum of drug resistance. Notably, BCL-2 inhibitors emerge as potential therapeutic alternatives in addressing these phenotypic changes.
date: 2024
id_scheme: DOI
id_number: 10.11588/heidok.00035580
ppn_swb: 1909314560
own_urn: urn:nbn:de:bsz:16-heidok-355800
date_accepted: 2024-07-11
advisor: HASH(0x55e83b063488)
language: eng
bibsort: JINFUKANGARID1AMUTA
full_text_status: public
place_of_pub: Heidelberg
citation:   Jin, Fukang  (2024) ARID1A mutations in colorectal cancer: Biological and therapeutic implications.  [Dissertation]     
document_url: https://archiv.ub.uni-heidelberg.de/volltextserver/35580/1/Fukang%20Jin_dissertation.pdf