Regulation of lung stem cells in homeostasis and tumorigenesis. Role of Stress-Activated Protein Kinases

Lead Research Organisation: University of Cambridge
Department Name: Wellcome Trust - MRC Cam Stem Cell Inst

Abstract

Lung diseases and specially lung cancer, are among the more prevalent and one of the first causes of mortality in humans. Dysruption of lung homeostasis is underlying these pathologies. The alveolar epithelium is the area where gas exchange takes place. Thus the maintenance of the proper morphology and cellular composition is a key event for lung function. The turnover of lung epithelia is responsibility of a small group of cells named lung stem/ progenitor cells that have the potentiality to differentiate into any of the cell types of the alveolar epithelia. However, very little is known about lung stem cells and only recently have been isolated. I have been able to isolate lung stem/progenitor cells and culture them. Using this system I plan to study the mechanisms involved in the regulation of lung stem cell homeostasis. I will also search for markers that may be used for the detection and spotting of these cells in vitro and in vivo.
In-depth characterisation of the mechanisms regulating lung stem cells will make possible controlling their fate to self-renewal or production of alveolar epithelia by manipulation of some specific proteins. Transferring of these findings to humans will open new ways in the study and therapy of lung diseases.

Technical Summary

In the lung there is a population of stem/progenitor cells able to produce Alveolar type-II and Clara cells that constitute lung epithelia. I have developed a modified protocol to culture these lung stem/progenitor cells and induce their differentiation in vitro. I have demonstrated that a member of the Stress-Activated Protein Kinases (SAPKs), p38a, is essential in the regulation of the lung stem cell homeostasis.
I aim to study and characterise the mechanisms underlying the control of lung stem cells by p38a and the crosstalk with other pathways that may be also involved in lung stem cell homeostasis. I will use a double approach using tissue cultured lung stem cells and examined their in vivo properties with animal experimentation.
Lung cancer is among the most prevalent and fatal in western countries. Currently, a new body of research focuses on stem cells as the origin of cancer. It remains controversial the existence of cancer stem cells and the connection of these cells to normal stem cells. I have recently found the regulation of lung stem cell homeostasis by the stress-activated protein kinase (SAPK) p38a and the consequences of losing this control to the lung tumorigenesis induced by K-RasV12.

To that purpose I will
1. delineate the phenotypic and molecular attributes of lung stem/progenitor cells,
2. dissect the signaling networks that collaborate with p38a to regulate these cells in vitro and in vivo.
3. study the lung stem cell contribution to tumorigenesis and the role of p38a in this process.
I will examine these working hypotheses:

(i) There is a specific population of lung stem/progenitor cells with the potential to differentiate into all alveolar cell types. As I have isolated cells that act in vitro as potential alveolar stem cells. We need to assay that potentiality in vivo.
(ii) SAPKs are essential regulators of the intracellular mechanisms involved in lung stem cell regulation. Molecular and phenotypical characterisation of lung stem cells and their regulation. Use of a double promoter system to trace lung stem cells searching for specific markers that allow differential localization and isolation.
(iii) Lung stem cells are the origin for cancer stem cells. Using tissue culture and animal injections I will characterise genetically, biochemically and functionally putative cancer stem cells isolated from lung tumors (K-Ras) using cancer (TTF-1) and stem cell markers (Side Population, CD133+, CD44, alpha2beta1 integrin). I will examine the potentiality of lung stem cells to be transformed and become cancer stem cells.
 
Description AICR grants
Geographic Reach Europe 
Policy Influence Type Participation in advisory committee
 
Description AICR grants
Geographic Reach Multiple continents/international 
Policy Influence Type Participation in advisory committee
 
Description AICR grants
Geographic Reach Multiple continents/international 
Policy Influence Type Participation in advisory committee
 
Description Juan de la Cierva programme
Geographic Reach National 
Policy Influence Type Participation in advisory committee
 
Description Ramon and Cajal Programme
Geographic Reach National 
Policy Influence Type Participation in advisory committee
 
Description Reviewer for Cancer Research UK grants
Geographic Reach National 
Policy Influence Type Participation in advisory committee
 
Description Reviewer for Cancer Research grants
Geographic Reach National 
Policy Influence Type Participation in advisory committee
 
Description Reviewer for Wellcome Trust Grants
Geographic Reach National 
Policy Influence Type Participation in advisory committee
 
Description Collaboration Grant
Amount £619,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 09/2010 
End 08/2013
 
Title An specific lung alveolar stem cell line 
Description Isolation of a discrete population of cells from the lung alveolar epithelium with the potential of in vitro and in vivo differentiation to the different distal lung cell types. 
Type Of Material Cell line 
Year Produced 2009 
Provided To Others? Yes  
Impact Genetic and mechanistic characterization of lung stem cell self-renewal and differentiation 
 
Title Lgr6 is a marker of cancer stem cell like cells selected during human lung adenocarcinoma progression 
Description Dystuption of a miR-17-92/p38a axis in lung adenocarcinoma cells selects Lgr6+ cells by enhancing Wnt signaling during cancer progression. Lgr6+ cells have more tumorigenic potential than Lgr6- and harbor stem cell potential. 
Type Of Material Model of mechanisms or symptoms - human 
Year Produced 2013 
Provided To Others? Yes  
Impact This could open a new way for detection and diagnosis of human lung adenocarcinoma and new targets for more effective anticancer drugs. 
 
Title Lgr6 labelles a new human lung stem cell population 
Description A new population of cells isolated from human lungs with potential to differentiate and regenerate lung bronchioalveolar epithelium and unlimited expansion 
Type Of Material Cell line 
Year Produced 2012 
Provided To Others? Yes  
Impact Potential use to regenerate damage lung epithelium in vivo or generate bronchioalveolar tissue in vitro. Use for drug testing and generate in vitro models of lung pathologies. 
 
Title Lung alveolar-specific p38a Tet-On conditional knockout 
Description Mice expressing a Cre under the control of SP-C promoter (only active in At2 and lung alveolar stem cells), timely activated after tetracyclin administration that may excise a p38a-lox site to generate lung alveolar cell specific p38a knockout 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Provided To Others? No  
Impact Alveolar cell-specific contribution of p38a to lung alveolar homeostasis and regeneration. 
 
Title Lung regeneration by human/mouse lung stem cells in a bleomycin-induced lung injury model 
Description Using a model of bleomycin induced lung injury we have been able to use human or mouse lung stem cells to partially regenerate the damaged tissue. 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Provided To Others? No  
Impact This will provide a final support to the existence and possibility of isolation of multipotent lung alveolar stem cells for future possible cellular lung regeneration therapies 
 
Title New Mechanism of lung stem cell regulation 
Description New crosstalk between the p38MAPK and miR-17-92 pathways regulate lung stem cell differentiation and self-renewal. New cell lines carrying shRNAs to knockdown p38 or mir-17-92 members 
Type Of Material Model of mechanisms or symptoms - human 
Year Produced 2012 
Provided To Others? Yes  
Impact New potential targets to direct drugs used in lung pathologies involving the dysregulation of these pathways. 
 
Title Optimization of lung alveolar stem cell isolation and culturing 
Description We have improved a way to isolate multipotent progenitors from the lung alveolar epithelium and their in vitro expansion. 
Type Of Material Technology assay or reagent 
Year Produced 2009 
Provided To Others? Yes  
Impact Facilitate to obtaining lung alveolar stem cells from different backgrounds for in vitro studies. 
 
Title Potential assessment of lung stem cells in kidney capsule engraftments 
Description We have been able to assess the differentiation potential of mouse and human lung stem cells using kidney capsule engraftments. 
Type Of Material Physiological assessment or outcome measure 
Year Produced 2010 
Provided To Others? Yes  
Impact It is the first time that individual isolated lung stem/progenitor cells of either mouse or human origin have been assess to produce lung alveolar epithelium in vivo. 
 
Title Use of ex-vivo lung explants to study regeneration and tumorigenesis 
Description We have developed a way to in-vitro culture of adult lung explants to use for models of lung injury and lung tumorigenesis. We can test the potential of our lung stem cells injecting them into these explants and further analyises of their differentiation or transformation potential. 
Type Of Material Technology assay or reagent 
Year Produced 2009 
Provided To Others? Yes  
Impact It will allow the assessment of lung stem cells prior to in vivo analysis minimizing the number of animals used in future studies. 
 
Description ''Role of RACO-1 in tissue homeostasis and cancer'' 
Organisation Cancer Research UK
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution We are going to provide our expertise and reagents, as animal models and lung stem cells to the development of the project.
Collaborator Contribution exchange of reagents and scientific advisory
Impact It just started but there is some data that may be included in a manuscript submitted to a scientific journal.
Start Year 2010
 
Description Are There Really Lung Stem Cells? 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "Are There Really Lung Stem Cells?" Ventura, JJ. (05/15/2009), Joint Retreat (A Behrens, S Marino, Nic Jones labs), Keynote speaker. Powys, Wales, UK.

Reinforce collaborations amongst groups in UK
Year(s) Of Engagement Activity 2009
 
Description Characterisation of a Human Lung Alveolar Multipotent Population 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "Characterisation of a Human Lung Alveolar Multipotent Population" Ventura, JJ. (23/03/2012), Imperial College, London, UK.

meeting with researchers in the field of lung to share findings and start collaborations
Year(s) Of Engagement Activity 2012
 
Description Connecting Lung Epithelial Stem Cells and Lung Cancer by Stress-Activated Protein Kinases 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "Connecting Lung Epithelial Stem Cells and Lung Cancer by Stress-Activated Protein Kinases" Ventura, JJ. (04/07/2009), Cambridge Seminars in Oncology Series, Hutchinson/MRC Research Centre, University of Cambridge (Cambridge).

New collaborations in the university
Year(s) Of Engagement Activity 2009
 
Description Defining the cellular hierarchy in bronchioalveolar regeneration 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "Defining the cellular hierarchy in bronchioalveolar regeneration" Ventura, JJ. (04/09/2013). Leibniz Institute for Age Research / Fritz Lipmann Institute, Jena, Germany.

new collaborations abroad
Year(s) Of Engagement Activity 2013
 
Description Dissection of the cellular and molecular hierarchy of the bronchioalveolar epithelium in homeostasis and disease 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact • "Dissection of the cellular and molecular hierarchy of the bronchioalveolar epithelium in homeostasis and disease" Ventura, JJ. (26/10/2012), Developmental Biology Series, Gurdon Institute, Cambridge, UK.

show our research to students and postdocs in our institution
Year(s) Of Engagement Activity 2012
 
Description In Search of Lung Stem Cells 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "In Search of Lung Stem Cells" Ventura, JJ. (03/19/2010), Institute for Stem Cell Research, Edinburgh, UK.

show our new findings around the UK
Year(s) Of Engagement Activity 2010
 
Description In Search of Lung Stem Cells 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "In Search of Lung Stem Cells" Ventura, JJ. (04/13/2010), Paterson Institute for Cancer Research, Manchester, UK.

show our research around the UK
Year(s) Of Engagement Activity 2010
 
Description In Search of Lung Stem Cells 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "In Search of Lung Stem Cells" Ventura, JJ. (03/17/2010), Cellular and Molecular Pathology Divisional Symposium, University of Cambridge, Cambridge, UK.

new collaborations and advisory from people in the department
Year(s) Of Engagement Activity 2010
 
Description In Search of Lung Stem Cells 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "In Search of Lung Stem Cells" Ventura, JJ. (02/16/2010), MRC National Institute for Medical Research (NIMR), London, UK.

New collaborators
Year(s) Of Engagement Activity 2010
 
Description Lung Stem Cells and Cancer 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Health professionals
Results and Impact • "Lung Stem Cells and Cancer" Ventura, JJ. (07/14/2010), Stem Cell Summer School for Clinicians, Clare College, University of Cambridge, Cambridge, UK.

Opened our research to clinicians
Year(s) Of Engagement Activity 2010
 
Description Regulation of Lung Cancer by p38alpha MAPK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "Regulation of Lung Cancer by p38alpha MAPK" Ventura, JJ. (07/25/2008), External Seminar, Cancer Research UK-LRI (London).

New collaborators
Year(s) Of Engagement Activity 2008
 
Description Regulation of Lung Epithelial Stem Cells by Stress-Activated Protein Kinases 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "Regulation of Lung Epithelial Stem Cells by Stress-Activated Protein Kinases" Ventura, JJ. (03/25/2009), Faculty of Veterinary Medicine, University of Glasgow (Glasgow).

New collaborations
Year(s) Of Engagement Activity 2009
 
Description SAPKs, new players in lung tumorigenesis 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "SAPKs, new players in lung tumorigenesis" Ventura, JJ. (06/12/2008) Mouse Modelers' meeting, Cancer Research UK-CRI (Cambridge).

introduced our research to new audiences
Year(s) Of Engagement Activity 2008
 
Description Signalling Networks Regulating Human Lung Stem Cells 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "Signalling Networks Regulating Human Lung Stem Cells" Ventura, JJ. (06/30/2011), School of Life Sciences, University of Manchester, Manchester, UK.

Started new collaborations
Year(s) Of Engagement Activity 2011
 
Description Signalling Networks Regulating Human Lung Stem Cells 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "Signalling Networks Regulating Human Lung Stem Cells" Ventura, JJ. (09/19/2011), London Research Institute, Cancer Research-UK, London, UK.

Exhibition of our new findings to scientist in different fields
Year(s) Of Engagement Activity 2011
 
Description Signalling Networks Regulating Human Lung Stem Cells 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact • "Signalling Networks Regulating Human Lung Stem Cells" Ventura, JJ. (04/07/2011), Stem Cell Initiative Seminars, LRM, University of Cambridge, Cambridge, UK.

Sharing our findings with researchers in our university
Year(s) Of Engagement Activity 2011
 
Description The nature and liaisons of Human lung bronchioalveolar stem cells 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact • "The nature and liaisons of Human lung bronchioalveolar stem cells" Ventura, JJ. (11/10/2012), Stem Cell Club, Cambridge, UK.

show our research to students and postdocs in the university
Year(s) Of Engagement Activity 2012
 
Description • Regulation of Lung Epithelial Stem Cells by Stress-Activated Protein Kinases 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact • Regulation of Lung Epithelial Stem Cells by Stress-Activated Protein Kinases" Ventura, JJ. (11/04/2008), CCCRR Vascular Biology Research Seminar, School of Clinical Medicine (Cambridge).

Collaborations with medics
Year(s) Of Engagement Activity 2008