Single cell RNA-sequencing of normal breast tissue samples before and after 12-week anti-progestin (ulipristal acetate) treatment

Vacuum-assisted biopsy (VAB) from normal breast tissue were obtained from women participating in the Breast Cancer - Anti-Progestin Prevention Study 1 (BC-APPS1). A baseline VAB was performed in the luteal phase of the menstrual cycle and a second VAB was performed after a 12-week anti-progestin (ulipristal acetate, daily 5mg oral tablet) treatment. Breast tissue minced into small fragments was digested overnight with collagenase/hyaluronidase. Following further digestion with trypsin and dispase, a dissociated breast single cell suspension was obtained and single-cell RNA-sequencing was performed. Standard workflow for the 10X Genomics Single Cell 3' RNA Kit was used in two batches of 6 samples (3 participants per run with pre and post-ulipristal acetate treatment).

本研究的正常乳腺组织真空辅助活检（Vacuum-assisted biopsy, VAB）标本均来自参与乳腺癌抗孕激素预防研究1（Breast Cancer - Anti-Progestin Prevention Study 1, BC-APPS1）的女性受试者。受试者分别于月经周期黄体期接受基线真空辅助活检，并在接受12周抗孕激素治疗（醋酸乌利司他，每日口服5mg片剂）后再次接受真空辅助活检。将乳腺组织剪切成细小碎片后，使用胶原酶/透明质酸酶进行过夜消化。经胰蛋白酶和分散酶进一步消化后，获得解离的乳腺单细胞悬液，并开展单细胞RNA测序（single-cell RNA-sequencing）。实验采用10X Genomics单细胞3'端RNA建库试剂盒的标准流程，分两批处理共6份样本（每轮实验包含3名受试者的治疗前及治疗后样本）。