Deciphering the temporal heterogeneity of cancer-associated fibroblast subpopulations in breast cancer

Journal of Experimental & Clinical Cancer Research

Freja Albjerg Venning¹  Kamilla Westarp Zornhagen¹  Janine Terra Erler¹  Lena Wullkopf¹  Chris Denis Madsen²  Morteza Chalabi Hajkarim³  Kyoung Jae Won³  Carmen Rodriguez-Cupello⁴  Mikkel Morsing⁴  Pontus Kjellman⁴  Jonas Sjölund⁴ 

[1] Biotech Research and Innovation Centre (BRIC), University of Copenhagen (UCPH), Ole Maaløes Vej 5, 2200, Copenhagen N, Denmark;Biotech Research and Innovation Centre (BRIC), University of Copenhagen (UCPH), Ole Maaløes Vej 5, 2200, Copenhagen N, Denmark;Department of Laboratory Medicine, Division of Translational Cancer Research, Lund University, Scheelevägen 2, 22381, Lund, Sweden;Biotech Research and Innovation Centre (BRIC), University of Copenhagen (UCPH), Ole Maaløes Vej 5, 2200, Copenhagen N, Denmark;Novo Nordisk Foundation Center for Stem Cell Biology, DanStem, Faculty of Health and Medical Sciences, University of Copenhagen, 2200, Copenhagen N, Denmark;Department of Laboratory Medicine, Division of Translational Cancer Research, Lund University, Scheelevägen 2, 22381, Lund, Sweden;

关键词: Cancer-associated fibroblast (CAF);    CAF heterogeneity;    CAF subpopulations;    Flow cytometry analysis;    Breast cancer progression;   

DOI  :  10.1186/s13046-021-01944-4

来源: Springer

【 摘 要 】

BackgroundCancer-associated fibroblasts (CAFs) comprise a heterogeneous population of stromal cells within the tumour microenvironment. CAFs exhibit both tumour-promoting and tumour-suppressing functions, making them exciting targets for improving cancer treatments. Careful isolation, identification, and characterisation of CAF heterogeneity is thus necessary for ex vivo validation and future implementation of CAF-targeted strategies in cancer.MethodsMurine 4T1 (metastatic) and 4T07 (poorly/non-metastatic) orthotopic triple negative breast cancer tumours were collected after 7, 14, or 21 days. The tumours were analysed via flow cytometry for the simultaneous expression of six CAF markers: alpha smooth muscle actin (αSMA), fibroblast activation protein alpha (FAPα), platelet derived growth factor receptor alpha and beta (PDGFRα and PDGFRβ), CD26/DPP4 and podoplanin (PDPN). All non-CAFs were excluded from the analysis using a lineage marker cocktail (CD24, CD31, CD45, CD49f, EpCAM, LYVE-1, and TER-119). In total 128 murine tumours and 12 healthy mammary fat pads were analysed.ResultsWe have developed a multicolour flow cytometry strategy based on exclusion of non-CAFs and successfully employed this to explore the temporal heterogeneity of freshly isolated CAFs in the 4T1 and 4T07 mouse models of triple-negative breast cancer. Analysing 128 murine tumours, we identified 5–6 main CAF populations and numerous minor ones based on the analysis of αSMA, FAPα, PDGFRα, PDGFRβ, CD26, and PDPN. All markers showed temporal changes with a distinct switch from primarily PDGFRα+ fibroblasts in healthy mammary tissue to predominantly PDGFRβ+ CAFs in tumours. CD26+ CAFs emerged as a large novel subpopulation, only matched by FAPα+ CAFs in abundance.ConclusionWe demonstrate that multiple subpopulations of CAFs co-exist in murine triple negative breast cancer, and that the abundance and dynamics for each marker differ depending on tumour type and time. Our results form the foundation needed to isolate and characterise specific CAF populations, and ultimately provide an opportunity to therapeutically target specific CAF subpopulations.

【 授权许可】

CC BY