Periostin+ stromal cells guide lymphovascular invasion by cancer cells [DSP]
Cancer cell dissemination to sentinel lymph nodes associates with poor patient outcomes, particularly in breast cancer. The process by which cancer cells egress from the primary tumor upon interfacing with the lymphatic vasculature is complex and driven by dynamic interactions between cancer cells and stromal cells, including cancer associated fibroblasts (CAFs). The matricellular protein periostin can distinguish CAF subtypes in breast cancer and is associated with increased desmoplasia and disease recurrence in patients. However, since periostin is secreted, periostin-expressing CAFs are difficult to characterize in situ, limiting our understanding of their specific contribution to cancer progression. Here, we used in vivo genetic labeling and ablation to lineage trace periostin+ cells and characterize their functions during tumor growth and metastasis. Periostin-expressing CAFs were spatially found at periductal and perivascular margins, were enriched at lymphatic vessel peripheries, and were differentially activated by highly-metastatic cancer cells versus poorly-metastatic counterparts. Surprisingly, genetically depleting periostin+ CAFs slightly accelerated primary tumor growth but impaired intratumoral collagen organization and inhibited lymphatic, but not lung, metastases. Periostin ablation in CAFs impaired their ability to deposit aligned collagen matrices and inhibited cancer cell invasion through collagen and across lymphatic endothelial cell monolayers. Thus, highly-metastatic cancer cells mobilize periostin-expressing CAFs in the primary tumor site that promote collagen remodeling and collective cell invasion within lymphatic vessels and ultimately to sentinel lymph nodes. Overall design: We used digital spatial profiling (DSP) technology to quantitate transcript in spatially distinct populations of periostin-expressing cells located proximal to lymphatic vessels and near blood vessels and ducts in the naive mammary gland of periostin lineage tracing mice. To quantitate gene expression, we used GeoMx mouse Whole Transcriptome Atlas (WTA) panel from NanoString with fluorescence labelled morphology markers; green fluorescent protein (ZSGreen) for periostin-expressing cells, PDPN for lymphatic vessel marker and SMA for ducts and blood vessels to visualize morphology of tissues on GeoMx DSP platform.