The temporal progression of lung immune remodeling during breast cancer metastasis

The dynamics of immune reprogramming in the breast cancer lung metastastic niche remains incomplete. We used MULTI-seq (McGinnis, Patterson, et al., Nature Methods, 2019) to perform multiplexed single-cell RNA-sequencing analysis of cryopreserved lung immune cells isolated from PyMT mice, freshly-isolated lung immune cells isolated from PyMT mice, and cryopreserved lung immune cells isolated from 4T1 tumor-bearing mice from the onset of primary tumorigenesis through the formation of macrometastatic lesions. Cryopreserved PyMT samples were collected from 6-14 weeks of age, fresh PyMT samples were collected at 6, 10 and 13 weeks of age (with age-matched wild-type controls) MULTI-seq barcoded CD45+ CD31- Ter119- mouse immune cells were isolated using FACS and analyzed using 3' scRNA-seq (10x Genomics)