RT Journal Article
T1 Pathogenic variant in EPHB4 results in central conducting lymphatic anomaly
A1 Li, Dong
A1 Wenger, Tara
A1 Seiler, Christoph
A1 March, Michael
A1 Gutiérrez Uzquiza, Álvaro
A1 Kao, Charlly
A1 Bhoj, Elizabeth
A1 Tian, Lifeng
A1 Rosenbach, Misha
A1 Liu, Yichuan
A1 Robinson, Nora
A1 Behr, Mechenzie
A1 Chiavacci, Rosetta
A1 Hou, Cuiping
A1 Wang, Tiancheng
A1 Bakay, Marina
A1 Pellegrino da Silva, Renata
A1 Perkins, Jonathan
A1 Sleiman, Patrick
A1 Levine, Michael
A1 Hicks, Patricia
A1 Itkin, Maxim
A1 Dori, Yoav
A1 Hakonarson, Hakon
AB Central conducting lymphatic anomaly (CCLA) is one of the complex lymphatic anomalies characterized by dilated lymphatic channels, lymphatic channel dysmotility and distal obstruction affecting lymphatic drainage. We performed whole exome sequencing (WES) of DNA from a four-generation pedigree and examined the consequences of the variant by transfection of mammalian cells and morpholino and rescue studies in zebrafish. WES revealed a heterozygous mutation in EPHB4 (RefSeq NM_004444.4; c.2334 + 1G>C) and RNA-Seq demonstrated that the EPHB4 mutation destroys the normal donor site, which leads to the use of a cryptic splice donor that results in retention of the intervening 12-bp intron sequence. Transient co-expression of the wild-type and mutant EPHB4 proteins showed reduced phosphorylation of tyrosine, consistent with a loss-of-function effect. Zebrafish ephb4a morpholino resulted in vessel misbranching and deformities in the lymphatic vessel development, indicative of possible differentiation defects in lymphatic vessels, mimicking the lymphatic presentations of the patients. Immunoblot analysis using zebrafish lysates demonstrated over-activation of mTORC1 as a consequence of reduced EPHB4 signaling. Strikingly, drugs that inhibit mTOR signaling or RAS-MAPK signaling effectively rescued the misbranching phenotype in a comparable manner. Moreover, knock-in of EPHB4 mutation in HEK293T cells also induced mTORC1 activity. Our data demonstrate the pathogenicity of the identified EPHB4 mutation as a novel cause of CCLA and suggesting that ERK inhibitors may have therapeutic benefits in such patients with complex lymphatic anomalies.
PB Oxford University Press
SN 0964-6906
SN 1460-2083
YR 2018
FD 2018
LK https://hdl.handle.net/20.500.14352/115929
UL https://hdl.handle.net/20.500.14352/115929
LA eng
NO Dong Li, Tara L Wenger, Christoph Seiler, Michael E March, Alvaro Gutierrez-Uzquiza, Charlly Kao, Elizabeth Bhoj, Lifeng Tian, Misha Rosenbach, Yichuan Liu, Nora Robinson, Mechenzie Behr, Rosetta Chiavacci, Cuiping Hou, Tiancheng Wang, Marina Bakay, Renata Pellegrino da Silva, Jonathan A Perkins, Patrick Sleiman, Michael A Levine, Patricia J Hicks, Maxim Itkin, Yoav Dori, Hakon Hakonarson, Pathogenic variant in EPHB4 results in central conducting lymphatic anomaly, Human Molecular Genetics, Volume 27, Issue 18, September 2018, Pages 3233–3245,
DS Docta Complutense
RD 16 abr 2025