Pro-cathepsin D interacts with the extracellular domain of the beta chain of LRP1 and promotes LRP1-dependent fibroblast outgrowth.
Interactions between cancer cells and fibroblasts are crucial in cancer progression. We have previously shown that the aspartic protease cathepsin D (cath-D), a marker of poor prognosis in breast cancer that is overexpressed and highly secreted by breast cancer cells, triggers mouse embryonic fibroblast outgrowth via a paracrine loop. Here, we show the requirement of secreted cath-D for human mammary fibroblast outgrowth using a three-dimensional co-culture assay with breast cancer cells that do or do not secrete pro-cath-D. Interestingly, proteolytically-inactive pro-cath-D remains mitogenic, indicating a mechanism involving protein-protein interaction. We identify the low-density lipoprotein (LDL) receptor-related protein-1, LRP1, as a novel binding partner for pro-cath-D in fibroblasts. Pro-cath-D binds to residues 349-394 of the β chain of LRP1, and is the first ligand of the extracellular domain of LRP1β to be identified. We show that pro-cath-D interacts with LRP1β in cellulo. Interaction occurs at the cell surface, and overexpressed LRP1β directs pro-cath-D to the lipid rafts. Our results reveal that the ability of secreted pro-cath-D to promote human mammary fibroblast outgrowth depends on LRP1 expression, suggesting that pro-cath-D-LRP1β interaction plays a functional role in the outgrowth of fibroblasts. Overall, our findings strongly suggest that pro-cath-D secreted by epithelial cancer cells promotes fibroblast outgrowth in a paracrine LRP1-dependent manner in the breast tumor microenvironment.
Beaujouin
Mélanie
M
IRCM, Institut de Recherche en Cancérologie de Montpellier, Montpellier, F-34298, France.
Prébois
Christine
C
Derocq
Danielle
D
Laurent-Matha
Valérie
V
Masson
Olivier
O
Pattingre
Sophie
S
Coopman
Peter
P
Bettache
Nadir
N
Grossfield
Jami
J
Hollingsworth
Robert E
RE
Zhang
Hongyu
H
Yao
Zemin
Z
Hyman
Bradley T
BT
van der Geer
Peter
P
Smith
Gary K
GK
Liaudet-Coopman
Emmanuelle
E
D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D002277:Carcinoma / Q000378:metabolism* / Q000473:pathology; D002402:Cathepsin D / Q000378:metabolism*; D048708:Cell Growth Processes; D002461:Cell Line, Transformed; D018920:Coculture Techniques; D004792:Enzyme Precursors / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D026503:Low Density Lipoprotein Receptor-Related Protein-1 / Q000235:genetics / Q000378:metabolism*; D021962:Membrane Microdomains / Q000235:genetics; D051379:Mice; D019899:Paracrine Communication; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics
D015703:Antigens, CD; D004792:Enzyme Precursors; C475476:LRP1 protein, human; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D034741:RNA, Small Interfering; C058345:procathepsin D; D002402:Cathepsin D
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