Transendothelial migration of mesenchymal stem cells

Lead Research Organisation: Keele University
Department Name: Unlisted

Abstract

When tissue becomes injured or damaged it undergoes a repair process with the aim of bringing the tissue back to its normal functioning. In order to enhance this repair, which sometimes does not occur adequately, there is much interest in targeting stem cells to tissues to enhance repair. In adults there is a type of stem cell called mesenchymal stem cells (MSC) that can form a variety of tissues and have been administered to patients to try and enhance tissue regeneration. When these cells are injected into the blood they can migrate into tissues to some extent. Nothing is known about how these cells migrate across the wall of the blood vessel into the injured tissue. In this process the MSC have to cross endothelial cells which cover the inner lining of the blood vessels. This grant proposal aims to understand this mechanism. An experimental system will be set up to observe the migration of MSC across endothelial cells. The MSC will be flowed across endothelial cells to mimic blood flow and the molecules involved in transendothelial migration analysed. Transendothelial migration of white blood cells (leukocytes) is well characterised where specific molecules called adhesion molecules and chemokines are involved. Adhesion molecules are known to 'stick' leukocytes to endothelial cells so that migration of leukocytes across these cells can occur. Chemokines are molecules that attract cells and cause them to migrate and are involved in directing leukocytes out the blood stream and into the tissue. Some of these adhesion molecules and chemokines have been detected on MSCs but little or nothing is known about which are functional in migration. This will be investigated in the project. It will also be examined if it is possible to label MSC with microscopically small magnetic particles and to use magnets to capture and immobilise MSC on endothelial cells. It will then be investigated if chemokines can further enhance the migration of the magnetic MSC across the endothelium. The successful outcome the project would identify which adhesion molecules and chemokines are involved in directing MSC across endothelial cells and if magnetic particles can enhance this. Once this is known one may be able to increase the recruitment of MSC into damaged or injured tissues to help regenerate them.

Technical Summary

Mesenchymal stem cells (MSC) are multipotent cells capable of differentiating into a variety of tissues. There is currently much interest in these cells for tissue engineering purposes, to enhance the regeneration of injured or damaged tissue. There is evidence that when MSC are administered in the circulation they show some degree of extravasation and engraftment into damaged tissue. Nothing is known regarding the mechanism that these cells use to transmigrate across the endothelium. This is in contrast to leukocytes whose recruitment mechanisms into tissues are comparatively well-described. MSC have been shown to express adhesion molecules and chemokine receptors, but it is not known which of these are functional in transendothelial migration. Furthermore it may be possible to enhance transendothelial migration using magnetic nanoparticles to capture circulating MSC in a selected region of the vasculature. The objectives of the research are to: 1) Establish an in vitro model of MSC adhesion and migration across endothelial cells under conditions of flow; 2) To use this model to identify adhesion molecules, chemokines and chemokine receptors involved in the adhesion and transendothelial migration of MSCs; 3) To establish if magnetic nanoparticles in combination with chemokines can be used to enhance the capture and transendothelial migration of MSCs. MSCs and human umbilical vein endothelial cells (HUVEC) will be treated with blocking antibodies to adhesion molecules, chemokines and chemokine receptors. The effects of these antibodies on MSC rolling, stable adhesion and transmigration will be examined under conditions of flow. As a positive control results will be compared to those of neutrophils whose adhesive and migratory characteristics are well-described in this model. The adhesion and migration of MSCs across endothelial cells from skin and heart origin will be compared with those of HUVEC. MSC will be loaded with magnetic nanoparticles and flowed over endothelial cells. It will be investigated if magnets can capture these MSC on the endothelium, and in the presence of chemokines lead to enhanced transendothelial migration. The successful outcome of the project would identify functional adhesion molecules, chemokines and their receptors in MSC migration and if this migration can be augmented by magnetic nanoparticles. The research could reveal factors that stimulate stem cells to relocate to sites of tissue injury, leading to therapeutic strategies to improve homing of stem cells and tissue repair.

Publications

10 25 50
 
Description How stem cells migrate into inflamed tissues. The therapeutic effects of stem cells in animal model of rheumatoid arthrhtis
Exploitation Route How to improve the targeting of stem cells for repair of tissues . Therapeutics.
Sectors Chemicals,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

 
Description They have been used as a therapy in animals with rheumatoid arthrhtis
First Year Of Impact 2014
Sector Healthcare,Pharmaceuticals and Medical Biotechnology
Impact Types Economic

 
Description Eleanor Peel Trust
Amount £98,735 (GBP)
Organisation The Dowager Countess Eleanor Peel Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2013 
End 10/2016
 
Description Institute of Orthopaedics at Oswestry, Project grant
Amount £113,000 (GBP)
Organisation Institute of Orthopaedics 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2009 
End 01/2011
 
Title Bidirectional migration of stem cells 
Description We found a method for examing how stem cells migrate across endothelail cells of blood vessels, in two different directions. This was published. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
 
Description Ed Rainger 
Organisation University of Birmingham
Country United Kingdom 
Sector Academic/University 
PI Contribution Allowed the study of the migration of mesenchymal stem cells
Collaborator Contribution Use of a flow assay
Impact numerous papers, see publications,
Start Year 2007
 
Description Transendothelail migration of stem cells 
Organisation Novartis
Country Global 
Sector Private 
PI Contribution collaboration in the use of methods
Collaborator Contribution Chemotaxis methods
Impact papers published. See publications
Start Year 2006
 
Title Bidirectional migration of stem cells 
Description We found a method for examing how stem cells migrate across endothelail cells of blood vessels, in two different directions. This was published. 
Type Of Technology New/Improved Technique/Technology 
Impact No actual Impacts realised to date 
 
Description 11 invitations as speaker and chair at national and international conferences/seminars 
Form Of Engagement Activity Scientific meeting (conference/symposium etc.)
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Participants in your research or patient groups
Results and Impact 11 invitations as speaker and chair at national and international conferences/seminars I was invited to speak/chair based on the work of the grant

no actual impacts realised to date
Year(s) Of Engagement Activity 2006
 
Description Charity talk 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Talk about our research to a patient and fund raising group

Interest in funding the research
Year(s) Of Engagement Activity 2010
 
Description Open Day 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Talk about or research, practical demonstrations at a hospital open day


Increase in public awarness
Year(s) Of Engagement Activity 2007
 
Description Open day 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Talk about or research, practical demonstrations at a hospital open day

Increase in public awarness
Year(s) Of Engagement Activity 2009
 
Description Open day 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Talk about or research, practical demonstrations at a hospital open day

Increase in public awarness
Year(s) Of Engagement Activity 2009
 
Description Open day 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Talk about or research, practical demonstrations at a hospital open day



Increase in public awarness
Year(s) Of Engagement Activity 2007
 
Description Open day 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Talk about or research, practical demonstrations at a hospital open day


Increase in public awarness
Year(s) Of Engagement Activity 2008
 
Description Research Day 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? Yes
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact A day when members of the public visit our labs and see the research

The public had an increased understanding of our research
Year(s) Of Engagement Activity 2007,2008,2009
 
Description Research Day 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact 50 local members of the public and profesional scienstists attended talks and poster sessions, and were shown around the labs.

Local fund raisers donated funds for our research.
Year(s) Of Engagement Activity 2010
 
Description School visit - Oswestry 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? Yes
Geographic Reach Regional
Primary Audience Schools
Results and Impact They heard about the research and performed some hands-on experiments

They generated further interest in science and pathology
Year(s) Of Engagement Activity 2009