% IMPORTANT: The following is UTF-8 encoded.  This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.

@ARTICLE{Schubert:300227,
      author       = {A. Schubert$^*$ and A. Mongkolsittisilp and A. Kobitski and
                      M. Schulz and O. Voloshanenko$^*$ and M. Schaffrinski and N.
                      Winkler$^*$ and M. Nessling$^*$ and K. Richter$^*$ and D.
                      Kranz$^*$ and K. Nienhaus and D. Jäger and L. Trümper and
                      J. Büntzel and C. Binder and G. U. Nienhaus and M.
                      Boutros$^*$},
      title        = {{WNT}5a export onto extracellular vesicles studied at
                      single-molecule and single-vesicle resolution.},
      journal      = {The FEBS journal},
      volume       = {nn},
      issn         = {0014-2956},
      address      = {Oxford [u.a.]},
      publisher    = {Wiley-Blackwell},
      reportid     = {DKFZ-2025-00689},
      pages        = {nn},
      year         = {2025},
      note         = {#EA:B110#LA:B110# / epub},
      abstract     = {WNT signaling governs development, homeostasis, and aging
                      of cells and tissues, and is frequently dysregulated in
                      pathophysiological processes such as cancer. WNT proteins
                      are hydrophobic and traverse the intercellular space between
                      the secreting and receiving cells on various carriers,
                      including extracellular vesicles (EVs). Here, we address the
                      relevance of different EV fractions and other vehicles for
                      WNT5a protein, a non-canonical WNT ligand that signals
                      independently of beta-catenin. Its highly context-dependent
                      roles in cancer (either tumor-suppressive or
                      tumor-promoting) have been attributed to two distinct
                      isoforms, WNT5a Short (WNT5aS) and WNT5a Long (WNT5aL),
                      resulting from different signal peptide cleavage sites. To
                      explore possible differences in secretion and extracellular
                      transport, we developed fusion constructs with the
                      fluorescent proteins (FPs) mScarlet and mOxNeonGreen.
                      Functional reporter assays revealed that both WNT5a isoforms
                      inhibit canonical WNT signaling, and EVs produced by
                      WNT5a-bearing tumor cells, carrying either of the WNT5a
                      isoforms, induced invasiveness of the luminal A breast
                      cancer cell line MCF7. We used fluorescence intensity
                      distribution analysis (FIDA) and fluorescence correlation
                      spectroscopy (FCS) to characterize at single-molecule
                      sensitivity WNT5aL-bearing entities secreted by HEK293T
                      cells. Importantly, we found that most WNT5aL proteins
                      remained monomeric in the supernatant after
                      ultracentrifugation; only a minor fraction was EV-bound. We
                      further determined the average sizes of the EV fractions and
                      the average number of WNT5aL proteins per EV. Our detailed
                      biophysical analysis of the physical nature of the EV
                      populations is an important step toward understanding
                      context-dependent WNT cargo loading and signaling in future
                      studies.},
      keywords     = {WNT signaling (Other) / WNT transport (Other) /
                      extracellular vesicles (Other) / fluorescence correlation
                      spectroscopy (Other) / number and brightness analysis
                      (Other)},
      cin          = {B110 / W230},
      ddc          = {610},
      cid          = {I:(DE-He78)B110-20160331 / I:(DE-He78)W230-20160331},
      pnm          = {312 - Funktionelle und strukturelle Genomforschung
                      (POF4-312)},
      pid          = {G:(DE-HGF)POF4-312},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:40165582},
      doi          = {10.1111/febs.70074},
      url          = {https://inrepo02.dkfz.de/record/300227},
}