ErbB signalling is a potential therapeutic target for vascular lesions with fibrous component
Abstract
Background: Sporadic venous malformation (VM) and angiomatosis of soft tissue (AST) are benign, congenital vascular anomalies affecting venous vasculature. Depending on the size and location of the lesion, symptoms vary from motility disturbances to pain and disfigurement. Due to high recurrence of the lesions more effective therapies are needed.
Methods: As targeting stromal cells has been an emerging concept in anti-angiogenic therapies, here, by using VM/AST patient samples, RNA-sequencing, cell culture techniques and a xenograft mouse model, we investigated the crosstalk of endothelial cells (EC) and fibroblasts and its effect on vascular lesion growth.
Results: We report, for the first time, expression and secretion of transforming growth factor A (TGFA) in ECs or intervascular stromal cells in AST and VM lesions. TGFA induced secretion of VEGF-A paracrinally, and regulated EC proliferation. Oncogenic PIK3CA variant in p.H1047R, a common somatic mutation found in these lesions, increased TGFA expression, enrichment of hallmark hypoxia, and in a mouse xenograft model, lesion size and vascularization. Treatment with afatinib, a pan-ErbB tyrosine-kinase inhibitor, decreased vascularization and lesion size in mouse xenograft model with ECs expressing oncogenic PIK3CA p.H1047R variant and fibroblasts.
Conclusions: Based on the data, we suggest that targeting of both intervascular stromal cells and ECs is a potential treatment strategy for vascular lesions having a fibrous component.
Funding: Academy of Finland, Ella and Georg Ehnrooth foundation, the ERC grants, Sigrid Jusélius Foundation, Finnish Foundation for Cardiovascular Research, Jane and Aatos Erkko Foundation, and Department of Musculosceletal and Plastic Surgery, Helsinki University Hospital.
Data availability
RNA-seq data has been submitted to NCBI Gene Expression Omnibus under accession numbers GSE130807 and GSE196311 (GEO reviewer access tokens; wbivkayaxhojdqp and mbehiikgvtmfryh, respectively).
-
Activation of Epidermal Growth Factor Receptor Pathway in Slow-Flow Vascular MalformationsNCBI Gene Expression Omnibus, GSE130807.
-
Gene expression profiling of HUVEC-s expressing PIK3CA with H1047R point mutationNCBI Gene Expression Omnibus, GSE196311.
Article and author information
Author details
Funding
Academy of Finland (328835)
- Johanna P Laakkonen
ERC grant (GA670951)
- Seppo Ylä-Herttuala
ERC grant (802825)
- Minna U Kaikkonen
Sigrid Jusélius Foundation
- Seppo Ylä-Herttuala
Sigrid Jusélius Foundation
- Minna U Kaikkonen
Finnish Foundation for Cardiovascular Research
- Johanna P Laakkonen
Finnish Foundation for Cardiovascular Research
- Seppo Ylä-Herttuala
Finnish Foundation for Cardiovascular Research
- Minna U Kaikkonen
Jane and Aatos Erkko Foundation
- Minna U Kaikkonen
Department of Musculosceletal and Plastic Surgery, Helsinki University Hospital
- Pia Vuola
Academy of Finland (321535)
- Johanna P Laakkonen
Academy of Finland (353376)
- Johanna P Laakkonen
Academy of Finland (287478)
- Minna U Kaikkonen
Academy of Finland (294073)
- Minna U Kaikkonen
Ella and Georg Ehnrooth foundation
- Johanna P Laakkonen
CoE of Cardiovascular and Metabolic Disease (307402)
- Seppo Ylä-Herttuala
GeneCellNano Flagship Program (337120)
- Johanna P Laakkonen
GeneCellNano Flagship Program (337120)
- Seppo Ylä-Herttuala
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: Animal experiments were approved by National Experimental Animal Board of Finland (Decision No Esavi-2019-004672) and carried out in accordance with guidelines of the Finnish Act on Animal Experimentation.
Human subjects: Patient sample collection was approved by the Ethical Committee of the Helsinki University hospital, Helsinki, Finland (Decision No 127/13/03/02/2010 and No 1394/2020). Control sample collection was approved by the Research Ethics Committee of the Northern Savo Hospital District, Kuopio, Finland (Decision No 139/2015). Umbilical cord collection for HUVEC isolation was performed with approval from the Research Ethics Committee of the Northern Savo Hospital District, Kuopio, Finland (Decision No 341/2015). Informed consent, and consent to publish, was obtained from all patients included in the study.
Copyright
© 2023, Jauhiainen et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 956
- views
-
- 128
- downloads
-
- 3
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Further reading
-
- Cell Biology
- Evolutionary Biology
Maintenance of rod-shape in bacterial cells depends on the actin-like protein MreB. Deletion of mreB from Pseudomonas fluorescens SBW25 results in viable spherical cells of variable volume and reduced fitness. Using a combination of time-resolved microscopy and biochemical assay of peptidoglycan synthesis, we show that reduced fitness is a consequence of perturbed cell size homeostasis that arises primarily from differential growth of daughter cells. A 1000-generation selection experiment resulted in rapid restoration of fitness with derived cells retaining spherical shape. Mutations in the peptidoglycan synthesis protein Pbp1A were identified as the main route for evolutionary rescue with genetic reconstructions demonstrating causality. Compensatory pbp1A mutations that targeted transpeptidase activity enhanced homogeneity of cell wall synthesis on lateral surfaces and restored cell size homeostasis. Mechanistic explanations require enhanced understanding of why deletion of mreB causes heterogeneity in cell wall synthesis. We conclude by presenting two testable hypotheses, one of which posits that heterogeneity stems from non-functional cell wall synthesis machinery, while the second posits that the machinery is functional, albeit stalled. Overall, our data provide support for the second hypothesis and draw attention to the importance of balance between transpeptidase and glycosyltransferase functions of peptidoglycan building enzymes for cell shape determination.
-
- Cell Biology
Polynucleotide kinase phosphatase (PNKP) has enzymatic activities as 3′-phosphatase and 5′-kinase of DNA ends to promote DNA ligation and repair. Here, we show that cyclin-dependent kinases (CDKs) regulate the phosphorylation of threonine 118 (T118) in PNKP. This phosphorylation allows recruitment to the gapped DNA structure found in single-strand DNA (ssDNA) nicks and/or gaps between Okazaki fragments (OFs) during DNA replication. T118A (alanine)-substituted PNKP-expressing cells exhibited an accumulation of ssDNA gaps in S phase and accelerated replication fork progression. Furthermore, PNKP is involved in poly (ADP-ribose) polymerase 1 (PARP1)-dependent replication gap filling as part of a backup pathway in the absence of OFs ligation. Altogether, our data suggest that CDK-mediated PNKP phosphorylation at T118 is important for its recruitment to ssDNA gaps to proceed with OFs ligation and its backup repairs via the gap-filling pathway to maintain genome stability.