Defining the cellular origin of chronic lymphocytic leukaemia

Lead Research Organisation: University of Glasgow
Department Name: College of Medical, Veterinary &Life Sci

Abstract

Leukaemia is a cancer that results from a disruption in the normal development of cells within the blood. This disruption can occur when regulatory elements responsible for controlling white blood cell development malfunction. We recently discovered that a disturbance in the function of a ubiquitously expressed element protein kinase C during white blood cell development, initiates a specific type of leukaemia, chronic lymphocytic leukaemia (CLL). CLL is the most common adult blood cancer in the Western world, for which there is currently no cure. While many research groups are engaged in investigating the mechanisms that can induce CLL cell death in patients as a mechanism to remove the leukaemic cells, little is known about the origin of CLL cells. Until now it has been almost impossible to address this question, as the factors responsible for this initiating this leukaemia have remained elusive. However, our novel mouse model provides us with a unique opportunity to gain fundamental knowledge about the cellular origin of CLL, and may assist in the generation of novel treatments to cure CLL. Within our university-based research laboratory, we will perform established cellular and molecular techniques to identify which cell is responsible initiating CLL. In parallel with these experiments, we will characterise the CLL-initiating cells in CLL-patient cell samples. In this way, we will uncover important information regarding the cell of origin for CLL.

Technical Summary

B-cell chronic lymphocytic leukaemia (CLL) is the most prevalent leukaemia in the Western world with an incidence of 3 per 100,000, for which there is currently no cure. CLL cells morphologically resemble mature small B lymphocytes with a characteristic immunophenotype (CD5+CD19hiCD23+mIgMlo), a resistance to apoptosis, with up-regulation of anti-apoptotic proteins such as Bcl-2 and Mcl-1, and cell cycle arrest at the G0/G1 phase of the cell cycle. Clinically, CLL is a heterogeneous disorder in which some patients survive over a decade with stable disease, while in others the leukaemia behaves aggressively with survival measured in months. Over the last decade, several markers of poor prognostic significance have been identified, however to date no single genetic event has been associated with the initiation of CLL. In addition, the cell of origin of CLL has not been formally identified, although it is thought to originate from a maure B cell population, due to phenotypic and gene expression profiles. In an effort to address this question, we developed a unique mouse CLL model. This was achieved by retrovirally-infecting haemopoietic stem cell (HSC)-enriched cells, derived from wild type mice, with a dominant negative protein kinase C alpha (PKCa-KR) construct and culturing the cells either in in vitro or in vivo B cell generation systems. This resulted in the spontaneous generation of B lineage cells that resemble human CLL cells both at the phenotypic and molecular level, being refractory to apoptosis - possibly due to an upregulation in Bcl-2 and Mcl-1 expression levels, and arrested in G0/G1 ex vivo (Cancer Res. 66:527). These studies indicated that subversion of PKCa activity served to initiate the development of CLL. In this application, we show preliminary data demonstrating that our mouse CLL cells can be generated from recombinase activating gene deficient mice, which exhibit a development block at an early stage of B cell development, thus establishing that CLL can be derived from either an HSC or an early B cell progenitor population. Therefore, this proposal will phenotypically identify and molecularly characterise the CLL cell of origin both in our novel CLL mouse model and importantly in samples derived from CLL patients. We will then functionally characterise the mouse and human leukaemia initiating cell, both in vitro and in vivo. Collectively, the proposed studies will provide a deeper understanding of the biology of CLL, which may in turn illuminate novel avenues for therapeutic intervention.
 
Description GlaxoSmithKline (GSK)
Amount £102,928 (GBP)
Organisation GlaxoSmithKline (GSK) 
Sector Private
Country Global
Start 04/2011 
End 11/2012
 
Description KKLF Project Grant
Amount £53,487 (GBP)
Organisation The Kay Kendall Leukaemia Fund 
Sector Academic/University
Country United Kingdom
Start 09/2010 
End 08/2013
 
Description LLR Project grant
Amount £249,000 (GBP)
Funding ID 13012 
Organisation Leukaemia and Lymphoma Research 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2013 
End 09/2016
 
Description Lady Tata Memorial Trust Award
Amount £15,000 (GBP)
Organisation Lady Tata Memorial Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2009 
End 09/2010
 
Description Leukaemia Research Project Grant
Amount £20,000 (GBP)
Organisation Leukaemia and Lymphoma Research 
Sector Charity/Non Profit
Country United Kingdom
Start 06/2007 
End 08/2008
 
Description Lister Bellahouston Travelling Fellowship
Amount £800 (GBP)
Organisation Bellahouston Bequest Fund 
Sector Charity/Non Profit
Country United Kingdom
Start 05/2009 
End 06/2009
 
Description MRC Research Grant
Amount £327,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 06/2009 
End 05/2012
 
Description MRC/AZD Project grant
Amount £485,000 (GBP)
Funding ID MR/K014854/1 
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 03/2013 
End 02/2016
 
Description Tenovus Scotland
Amount £5,000 (GBP)
Funding ID S12/21 
Organisation Tenovus Cancer Care 
Department Tenovus Scotland
Sector Charity/Non Profit
Country United Kingdom
Start 03/2013 
End 02/2014
 
Description University of Glasgow studentship
Amount £34,600 (GBP)
Organisation Government of Scotland 
Department Scottish Funding Council
Sector Public
Country United Kingdom
Start 10/2009 
End 09/2011
 
Title CLL cell bank 
Description Cryopreserved DNA/RNA/protein/cell samples from CLL patients with linked clinical data. 
Type Of Material Biological samples 
Year Produced 2009 
Provided To Others? Yes  
Impact To enable further studies to investigate the biology of CLL 
 
Title CLL model in vivo 
Description We have developed a novel CLL mouse model which we are developing to be used as an in vivo therapeutic model 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Provided To Others? No  
Impact Publications prior to the start of this grant (2) and recognition of the model - invited for talks within University departments and conferences 
 
Description EliLilly 
Organisation Eli Lilly & Company Ltd
Department PKCb inhibitors
Country United Kingdom 
Sector Private 
PI Contribution We have been carrying out research with one of their compounds, enzastaurin
Collaborator Contribution They have provided the drug free of charge
Impact Published a paper in Haematologica in 2015 (Nakagawa et al).
Start Year 2009
 
Description Regulation of B cell signalling during development 
Organisation University of Glasgow
Department Institute of Infection, Immunity and Inflammation
Country United Kingdom 
Sector Academic/University 
PI Contribution Assisting with experiments, troubleshooting between groups in common areas of research
Collaborator Contribution Co-supervision of post-graduate and undergraduate students. Sharing of equipment and reagents
Impact Enabled award of an MRC grant: G0800167
 
Description University of Dundee 
Organisation University of Dundee
Department Dundee Cancer Centre
Country United Kingdom 
Sector Academic/University 
PI Contribution Providing expertise on pre-clinical modeling for novel drugs. Carrying out initial in vitro experiments on cells with a novel compound
Collaborator Contribution Discussion of molecular mechanisms of compounds and provision of compound
Impact None as yet
Start Year 2014
 
Description Funding letter 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Described in laymans terms the research that I do in the lab

Create awareness of our research
Year(s) Of Engagement Activity 2009,2012
 
Description Jordanhill 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Local
Primary Audience Schools
Results and Impact The talk was part of the Jordanhill School secondary school Health Week, discussing the importance of diet, impact on cancer and where therapies come from. Atleast 200 pupils were present over two 1 hour talks.

Lab visits have been arranged since this talk.
Year(s) Of Engagement Activity 2011
 
Description Nexxus Horizons, UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact I gave a seminar about the work that we were doing in the lab

Networking
Year(s) Of Engagement Activity 2007
 
Description Open Day 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Showed members of the public around our research facilities

Understanding from the public on the research we carry out
Year(s) Of Engagement Activity 2008,2009,2010,2014
 
Description Open Day specifically for CLL 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Gave an informal presentation to the public with an interest in CLL

None yet, except that of interest.
Year(s) Of Engagement Activity 2010,2014