Effects of a humanized CD47 antibody and SIRPa on triple negative breast carcinoma stem cells
Signal regulatory protein alpha (SIRPa, designated CD172a), also called SHPS-1 (SHP substrate 1) binds to CD47, a receptor for Thromobospondin-1 (TSP1). To block CD47-SIRP interaction, several CD47 blocking agents are in clinical trials. The purpose of this project is to explore the effects of SIRPFc signaling on bCSCs (breast cancer stem cells) derived from MDA-MB- 231 cells, when bCSCs were treated with either SIRPFc or CD47 blocking antibody CC90002 from Celgene Corporation. Overall design: MDA-MB-231 were cultured routinely as described earlier(PMID: 26840086).  MDA-MB-231 cells were cultured 72h using Aggrewell media from stem cell technologies according to manufacture instructions. The cells were treated with SIRPa-Fc or CD47 blocking antibody CC90002 (Celgene Corporation) for 72hrs. The Total RNA was extracted using ISOLATE II RNA Mini Kit from BIOLINE and RNA integrity and quantification was measured using RNA-Bio analyzer.