Comprehensive genetic evaluation of ductal carcinoma in situ

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The aim of this project (part 1) was to identify the genetic signature that predict the progression of ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) of the breast in clinical specimens. From the interim analysis of 236 specimens, we confirmed that the major genetic mutations such as PIK3CA, AKT1, and TP53 were consistently detected as previously reported in the TCGA dataset. In 2020, we collected more data from clinical sequencing and the cohort now includes total 313 specimens (252 from IDC and 61 from DCIS). Thus, we are updating the analysis of the entire cohort and attempt to identify the candidate genes responsible for progression from DCIS to IDC even in less frequently mutated genes.The aim of this project (part 2) was to describe the molecular mechanism of the breast cancer progression in the model. We obtained cell lines such as MCF10A and MCF10DCIS as models of normal epithelia and DCIS tumor in the breast, respectively. We optimized the conditions for CRISPR/Cas9 screen for both of the cell lines. The optimization included the identification of accurate doubling time of the cell lines, spinfection conditions such as cell density and polybrene concentration and appropriate puromycin/blasticidin concentrations to selected the infected cells.