PhD Studentship: Transgenic mesenchymal stem cell production of eppin as a novel therapeutic agent

Location
United Kingdom
Posted
Jan 29, 2018
Closes
Apr 06, 2018
Organization Type
University and College
Hours
Full Time
Details

Main research theme:

Respiratory diseases acute respiratory distress syndrome (ARDS) and chronic obstructive pulmonary disease (COPD) have many causative factors such as trauma, environmental exposure, sepsis or genetic predisposition, yet this broad range of disease phenotypes are all characterised by an unbalanced protease-antiprotease shield and excessive inflammation in the lung.

The most well described condition caused by loss of antiprotease is emphysema due to α-1 antitrypsin deficiency, the most abundant neutrophil elastase inhibitor in the lung. This results in significant remodelling of the airways, leading to emphysema- a key pathologic feature of COPD1. Serine antiproteases, such as SLPI and elafin, have also been implicated in the pathology of disease when this balance is disturbed or overwhelmed, such as in cystic fibrosis2 and ALI3.

SLPI and elafin are the most prominent members of the WFDC family of proteins which also includes human epididymal protease inhibitor (eppin) and human epididymis protein 4. A total of 18 WFDC proteins have been identified to date, yet many remain uncharacterised.

SLPI and elafin are multifactorial proteins, known to possess anti-bacterial, anti-viral and immunomodulatory properties4,5. and are therefore considered to play key roles in innate immunity at mucosal surfaces. Both proteins have been investigated as potential therapeutics through aerosolised administration6. Recombinant protein therapies rely on production of active protein yet the WFDC motifs which characterise this family of proteins include four disulphide bonds, making refolding of these proteins to an active conformation a difficult process. Concerns over both the delivery systems and their susceptibility to cleavage in a protease rich environment have also hampered their development.

The importance of eppin was overlooked until recently as it was thought to be exclusively expressed in the epididymis, however, recent work has shown eppin expression a variety of myeloid cell types, including monocytes and neutrophils7. In addition to antibacterial activity, we have demonstrated the immunomodulatory properties of eppin7, yet, the mechanism of action remains to be fully elucidated. In a model of lung inflammation (IT-LPS) recombinant eppin decreased neutrophil influx to the lung and significantly attenuated serum chemokine production7.

The inherent anti-inflammatory activity of mesenchymal stem cells (MSC) has been shown in vitro and in several animal models8. Based on this data, MSC therapies are currently being investigated to treat acute9 and chronic10 respiratory inflammation, yet no significant reduction in disease severity or patient mortality has yet been achieved. To obtain sufficient cells for therapy, MSCs must be passaged through several generations, yet successive passage is known to reduce their immunomodulatory potency. We propose the production of transgenic MSC (tMSC) which over express eppin, as a novel therapeutic agent. When introduced intravenously, over 70% of these cells will home to the lung where, in combination with the inherent anti-inflammatory effects of MSCs, the production of eppin in its native conformation will help to address both the protease burden and excessive inflammatory response.

In this project we aim to:

  1. Assess the status and function of eppin in BAL and serum in patients acute and chronic inflammatory lung disease.
  2. Production and validation of tMSC overexpressing eppin.
  3. Further explore the mechanism of immunomodulatory activity in vitro using LPS challenged alveolar macrophages.
  4. Investigation of the utility of these transgenic cells as a novel intervention strategy using human lung tissue (precision cut lung slice) and in a murine models of lung injury.

 

Person Specification

Applicants should have a strong background in biomedical sciences and ideally in molecular biology including qPCR and cloning. Applicant will be expected to show both willingness and ability to undertake in vivo studies and ideally have undertaken the A(SP)A personal licence course and possess experience in small animal handling.

They should have a commitment to research in Respiratory disease and have at least an Upper Second Class Honours Degree in a relevant Biomedical sciences degree. Applicants must also hold a UK driving licence.

 

How to apply

Informal enquiries:

Dr.Scott, e-mail: a.scott@bham.ac.uk

 

Funding Notes

Only eligible to UK/Ireland and EU applicants

 

References

  1.  Stone et al Respir Med. 2014, PMID: 24176989
  2. Weldon et al J Immunol. 2009 PMID: 20007580
  3. Kerrin et al Thorax. 2013 PMID: 23242946
  4. Scott et al Biochem Soc Trans. 2011 PMID: 21936829
  5. Williams et al Clin Sci (Lond). 2006 PMID: 16336202
  6. Small et al Biol Chem. 2017 PMID: 27930359
  7. Scott et al Eur Respir J. 2017 PMID: 28705940
  8. Matthay et al Chest 2010 PMCID: PMC2951759
  9. Johnson et al Stem Cells Transl Med. 2017 PMID: 28186706
  10. Wecht and Rojas Respirology. 2016 PMID: 27688156