Secretariat, Alumni Association, IDAC
Date	Wednesday, 11 May 2022, 17:00～
Room	7th Floor, Seminar Room 1,IDAC Center for Basic Aging Research
Title	Functional Validation of Somatic Copy Number Alteration (SCNA) in KRAS driver mutation Lung Adenocarcinoma
Speaker	Dr. Nagano, Ai
Affiliation	Cancer Institute, University College London
Organizer	Ogasawara, Koetsu (Department of Immunobiology・ext 8579)
Abstract	Introduction: The somatic copy number deletions on chr9p21 and chr3p21 are prevalent in KRAS driver mutation lung adenocarcinoma (LUAD). While recent genomic studies catalogued the significance of somatic copy number alterations (SCNA) in cancer cohorts, the functions of each copy number segment are still unknown. Material and Method: Here, we developed a new clone tracking system, COBALTseq (CRISPR-mediated copy number editing with barcoded clone tracking by sequencing) to quantify one of the functions, the cell growth advancement. We designed the target vectors of chr9p21 and chr3p21 using lentivirus to introduce two DNA double-strand breaks to the junctions of each segment. Using this approach, we measured the cell growth advantage on four KRAS driver LUAD cell lines, NCI-H358 and A549. Result: We tracked on average 200 clones edited with SCNA target vectors.  The significant increase of cell growth was observed on chr9p21 deletion on NCI-H358 and on chr3p21 on A549 cell line. Conclusion: In conclusion, a novel approach, COBALTseq demonstrated the quantitative cell growth advancement from initiating the cancer driving genomic alterations.  Despite the low genome-editing efficiency of 2-3% on SCNA region, the main strength of this approach is to detect the cell growth advantage by clone tracking which allows us to statistically focus on a small proportion of successfully edited clones within a population.