ProfessorBeata Wojciak-Stothard

Research Focus
The research focus of my group is to understand the mechanism of endothelial dysfunction in pulmonary hypertension. The objective of this work is to develop new therapeutic strategies for the treatment of this disease. To achieve this we use data from proteomic, genomic and metabolomic screening of pulmonary hypertensive patient samples available in our Centre. This allows us to identify signalling mediators linked to endothelial dysfunction. We also manipulate the expression levels and activity of selected protein targets in cultured pulmonary vascular cells in order to verify their role in pulmonary vascular remodelling.

CURRENT PROJECTS
Organ-on-a-chip (OOaC) modelling of vascular dysfunction in PH. To address the problem of significant drug failure in clinical trials and to reduce animal experimentation, my group have produced a novel device, a three dimensional model of human pulmonary artery in a microchip. The device was used to create the first inducible model of PAH, utilising patient-derived blood outgrowth endothelial cells. This pioneering work attracted industrial funding (Astra Zeneca) and resulted in numerous collaborations within Imperial College and abroad and won Provost’s Award For Excellence in Animal Research (3R’s).
Other OOac projects involve: (i) an alveoli-vascular model (BHF studentship and BHF CRE Intermediate Fellowship (ii) a model of vascular geometry in CTEPH (President’s PhD scholarship) and (iii) a model of sex-dependent signalling in PAH. Another novel platform utilising human pulmonary artery smooth muscle cells to measure vasoreactivity in response to drug treatment (EPATs) is being developed by Dr Adam Fellows, an NHLI Intermediate Fellow in my lab.

Targeting of endothelial proteins involved in pro-proliferative, pro-inflammatory and pro-angiogenic responses of PAH endothelium – translational studies to address an unmet clinical need of identifying effective treatment strategy in PAH. The group of proteins of interest includes Ras, Rho and Arf GTPases, intracellular chloride channel (CLIC) proteins and Kruppel like transcription factor family , KLF2, 4 and 6. My group utilise patient cells in vitro and rodent PAH models to establish effects of manipulation of expression and activity of these proteins on endothelial dysfunction and pulmonary vascular remodelling. Targeting approaches involve modification of protein prenylation as well as design of new inhibitors of Ras family regulatory proteins, guanosine exchange factors (GEFs).
Other recent projects focused on flow-induced changes in micro-RNAs:
- exosomal delivery of flow-induced therapeutic micro-RNAs. We identified a subset of exosomal microRNAs that are induced by homeostatic transcription factor KLF-2 in the lung endothelial cell but are reduced in PAH patients and animals and characterised the effects of liposomal delivery of these microRNAs on vascular remodelling in animal PAH models and endothelial function in vitro and in vivo.

- liposomal delivery of miR-150, known to be reduced in PAH and correlate with poor survival. Collaboration with Silence Therapeutics allowed us to use a liposomal preparation (DACC) that facilitates delivery of miRs into the lung endothelium.

PAST PROJECTS
Earlier projects involved studying the role of neutrophils and neutrophil extracellular traps (NETs) in pulmonary vascular remodelling in pulmonary hypertension and therapeutic targeting of enzymes increasing nitric oxide bioavailability, DDAH.

Our group were first to demonstrate that deficiency of oxygen and nitric oxide is a key factor in the disease process, inducing pulmonary endothelial barrier dysfunction and abnormal angiogenic responses as a result of an imbalance in the activation of actin cytoskeleton regulators, Rho GTPases: RhoA and Rac1.

We have identified another Rho GTPase, RhoB which is involved in pulmonary vascular remodelling. RhoB regulates actin polymerisation and the intracellular trafficking of growth factor receptors in pulmonary endothelial and smooth muscle cells. Prenylation (the addition of hydrophobic molecules, farnesyl or a geranyl-geranyl moiety to the C-terminal cysteine(s) of the target protein) of RhoB is required for its membrane localization and activation. We have identified an effective strategy of pharmacological targeting of RhoB by drugs used in the treatment of cancer, farnesyltransferase inhibitors. We show that these inhibitors may exert better control over disease-related increases in Rho proteins expression and activity than general prenylation inhibitors, statins.

Fig. 1. Overexpression of geranylgeranylated RhoB induces stress fibre formation in human pulmonary artery endothelial cells (HPAECs). GG-RhoB (blue), F-actin (red). Bar=10µm.

We have recently identified a novel pathway regulating membrane trafficking and the stability of bone morphogenetic protein receptor II (BMPRII) in pulmonary endothelium involving Arf family of proteins. We show that activation of one member of this family, Arf6 inhibits membrane recycling of BMPRII and prompts proteolytic degradation of the receptor. Inhibition of Arf6 activity with cytohesin inhibitors, such as SecinH3, restores BMPRII expression and reduces inflammatory responses in cultured endothelial cells and the lungs of animal models of pulmonary hypertension and prevents development of the disease in preclinical models of PAH. This work identifies a new class of pharmacological targets in PAH. It also provides an important insight into basic biological mechanisms responsible for the regulation of localization and stability of membrane receptors.
Figure 2. Trajectories of YFP-GGA1 (yellow fluorescent protein–labelled golgi-associated, gamma adaptin ear-containing, ARF-binding protein 1) vesicles in CLIC4-overexpressing human pulmonary artery endothelial cells (HPAECs). Bar=2µm. (Circ Res 2019)

Research Group
Pulmonary Vascular Biology Group

PhD Students:

Salina Nicoleau; salina.nicoleau14@imperial.ac.uk

Maike Haensel; maike.haensel18@imperial.ac.uk

Rehab Alharbi; r.alharbi20@imperial.ac.uk

Alexander Ainscough; alex.ainscough14@imperial.ac.uk

Hebah Sindi; h.sindi14@imperial.ac.uk

Mai Alzaydi; m.alzaydi15@imperial.ac.uk

Tim Smith; tahjsmartguy@gmail.com

Lucio Iannone; l.iannone@imperial.ac.uk

Lulwah Aldabbous; l.aldabbous12@imperial.ac.uk

Lisa Storck; lisa.storck@imperial.ac.uk
Post-Docs:

Dr Adam Fellows; a.fellows@imperial.ac.uk

Dr Sandro Satta

Dr Giusy Russomanno; g.russomanno@imperial.ac.uk

Dr Vahitha Abdul-Salam; vahitha.abdul-salam02@imperial.ac.uk

Dr Lucie Duluc; l.duluc@imperial.ac.uk

Dr Hilda Tsang
If you are interested in joining my research group, please contact me by email (b.wojciak-stothard@imperial.ac.uk ), enclosing a CV, short description of your research experience to date and proposed source of funding for your work (if applicable).
MSc student supervision

Rachel Hayley MSc in Molecular Medicine (Feb 2020-August2020)

Salina Nicoleau MSc in Molecular Medicine (Feb 2020-August 2020)

Pei Haoyu MRes in Translational Medicine (Oct 2018-Sept 2019)

Angelos Glynos MRes in Translational Medicine (Oct 2017-September 2018)

Kyeong Beom Jo MSc in Molecular Medicine (Feb 2018-August 2018)

Ahmed Osman MRes in Translational Medicine (Oct 2016-September 2017)

Abdul Mahomed MRes in Translational Medicine (Oct 15-Aug 16)

Sara Fonseca MSc in Molecular Medicine (Feb-August 2015)

Daria Belik MSc in Molecular Medicine (February – August 2013)

Dogukan Dalgalan MSc in Molecular Medicine (February–August 2012)

Wu Yixing MSc in Molecular Medicine (February-August 2011)

Andia Redpath MRes in Biomedical research, BMR stream October 2010 – May 2011
BSc student supervision

Miu Shing Hung BSc in Biomedical Sciences (Nov 2020-April 2021)

Nayana Iyler BSc in Biomedical Sciences (Nov 2019-April 2020)

Aditya Vijay BSc project (Haematology) (Feb –May 2019)

Jasraj Panesar BSc project (Haematology) (Feb-May 2015)

Veidika Chohan BSc project (Haematology) (February – May 2014)

Kim Dain BSc project (Haematology)(February – May 2014)

Dexter Valencia BSc project (Haematology) (February –May 2012)

Kirupananthan Thanighan BSc project (Haematology) (February –May 2012)

Wing kin Liu BSc project (Haematology) (February –May 2012)

Ahmed Syed BSc project (Haematology) (March- May 2010)