PIK3CA mutations are specifically localized to lymphatic endothelial cells of lymphatic malformations.

Blesinger, Hannah; Kaulfuß, Silke; Aung, Thiha; Schwoch, Sonja; Prantl, Lukas; Rössler, Jochen Karl; Wilting, Jörg; Becker, Jürgen (2018). PIK3CA mutations are specifically localized to lymphatic endothelial cells of lymphatic malformations. PLoS ONE, 13(7), e0200343. Public Library of Science 10.1371/journal.pone.0200343

[img]
Preview
Text
journal.pone.0200343.pdf - Published Version
Available under License Creative Commons: Attribution (CC-BY).

Download (36MB) | Preview

Lymphatic malformations (LM) are characterized by the overgrowth of lymphatic vessels during pre- and postnatal development. Macrocystic, microcystic and combined forms of LM are known. The cysts are lined by lymphatic endothelial cells (LECs). Resection and sclerotherapy are the most common treatment methods. Recent studies performed on LM specimens in the United States of America have identified activating mutations in the phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) gene in LM. However, whole tissue but not isolated cell types were studied. Here, we studied LM tissues resected at the University Hospitals Freiburg and Regensburg, Germany. We isolated LECs and fibroblasts separately, and sequenced the commonly affected exons 8, 10, and 21 of the PIK3CA gene. We confirm typical monoallelic mutations in 4 out of 6 LM-derived LEC lines, and describe two new mutations i.) in exon 10 (c.1636C>A; p.Gln546Lys), and ii.) a 3bp in-frame deletion of GAA (Glu109del). LM-derived fibroblasts did not possess such mutations, showing cell-type specificity of the gene defect. High activity of the PIK3CA-AKT- mTOR pathway was demonstrated by hyperphosphorylation of AKT-Ser473 in all LM-derived LECs (including the ones with newly identified mutations), as compared to normal LECs. Additionally, hyperphosphorylation of ERK was seen in all LM-derived LECs, except for the one with Glu109del. In vitro, the small molecule kinase inhibitors Buparlisib/BKM-120, Wortmannin, and Ly294002, (all inhibitors of PIK3CA), CAL-101 (inhibitor of PIK3CD), MK-2206 (AKT inhibitor), Sorafenib (multiple kinases inhibitor), and rapamycin (mTOR inhibitor) significantly blocked proliferation of LM-derived LECs in a concentration-dependent manner, but also blocked proliferation of normal LECs. However, MK-2206 appeared to be more specific for mutated LECs, except in case of Glu109 deletion. In sum, children that are, or will be, treated with kinase inhibitors must be monitored closely.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Gynaecology, Paediatrics and Endocrinology (DFKE) > Clinic of Paediatric Medicine

UniBE Contributor:

Rössler, Jochen Karl

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1932-6203

Publisher:

Public Library of Science

Language:

English

Submitter:

Karen Lidzba

Date Deposited:

05 Feb 2019 14:10

Last Modified:

26 Nov 2019 23:28

Publisher DOI:

10.1371/journal.pone.0200343

PubMed ID:

29985963

BORIS DOI:

10.7892/boris.124288

URI:

https://boris.unibe.ch/id/eprint/124288

Actions (login required)

Edit item Edit item
Provide Feedback