Tracing stem cell lineages in human epithelial tissues

Lead Research Organisation: Queen Mary, University of London
Department Name: Blizard Institute of Cell and Molecular

Abstract

Most people have become familiar with the concept of using stem cells for the healing of tissues ravaged by old age or disease. Most of the headlines have been made by embryonic stem cells, cells that can be extracted from early embryos, and which possess the versatility of being seemingly able to be coaxed into almost any type of tissue. Less well known are adult stem cells; these are thought to be present in almost all tissues and their continued multiplication throughout life is necessary to maintain tissues like the gut lining and skin where millions of cells are exfoliated every day. We know very little about the identity of such stem cells and thus are not able to extract them, perhaps with the notable exception of blood stem cells which have been used for many decades in life-saving transplants. We are going to use some very novel genetic technology to identify adult stem cells and their progeny in a number of human tissues, principally the large intestine, skin, liver and pancreas. In the large intestine and skin we are seeking to discover if cancers arise from these cells, while in the pancreas if we could locate stem cells, we may be able to expand them outside the body and make them into insulin-producing cells for subsequent transplantation for diabetes. Many people are now suffering from liver failure and cannot be treated because a shortage of livers for transplantation. An alternative strategy is cell transplantation into the failing liver using a healthy donor liver?s own cells called hepatocytes. If we could identify stem cells in the liver, these could be greatly expanded in number outside the body, converted to functional hepatocytes, and transplanted. Thus, our investigation centres on the role of stem cells in human diseases, principally bowel cancer, diabetes and liver failure, diseases that all have a dismal outlook and for which there are still no prospects of cure, despite many years of research.

Technical Summary

This project proposal sets out a programme of work designed to define stem cell behaviour in several human epithelial tissues namely the colon, liver, pancreas and skin, where our knowledge of their very nature, location and subsequent fate of their progeny is unknown. We aim to locate clonal proliferative units of tissue and infer the location of the stem cell niche. Following this we will deduce the role of stem/long-lived progenitor cells in the colon, pancreas and epidermis, and their role in the development of liver cirrhosis. We will exploit a technique we have been developing that can detect cells that are deficient in mitochondrial DNA (mtDNA) encoded cytochrome c oxidase (CCO), the terminal enzyme (component IV) in the electron transport chain. These are non-pathogenic mutations that mtDNA acquires; over time they dominate a cell that becomes histologically recognised as CCO-deficient. In the colon at least, this process takes many years, and so is believed to occur in stem cells. Initial observations can be made on routine paraffin wax-embedded tissue sections, but definitive analyses will be carried out on fresh frozen sections where CCO deficient cells can be detected by simple enzyme histochemistry. Subsequently, individual cells can be laser-capture microdissected from multiple sites within a ribbon/patch of similarly CCO deficient cells and their complete mtDNA genome sequenced. The finding of the same mutation in all cells of a group establishes monoclonality, since the odds of two cells acquiring the same mutation is 1:2.48 x 109.
We propose to examine normal and diseased human intestine, liver, pancreas and epidermis. Our preliminary results in the colon places the stem cell niche towards at the base of the crypt. We have also discovered clones originating outwith the conventional stem cell niche, which may be long-lived progenitors and we seek to establish their potentiality. We are developing other techniques based on the methylation sequence tags of non-expressed genes not only to identify clones, but as methylation status is a dynamic process, to study the kinetics of niche succession. We seek the identity of the stem cell niche in all tissues and the role of stem/progenitor cells in disease, e.g. cancer and liver regenerative nodules.

Publications

10 25 50
 
Description Belgian Science Policy consumables Grant
Amount € 55,000 (EUR)
Organisation Belgian Science Policy Office 
Sector Public
Country Belgium
Start 09/2012 
End 08/2017
 
Description Consumables
Amount £20,000 (GBP)
Organisation CORE Charity 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2010 
End 12/2010
 
Description Project Grant
Amount £188,000 (GBP)
Organisation Barts Charity 
Sector Charity/Non Profit
Country United Kingdom
Start 12/2012 
End 11/2015
 
Description Start-up Grant
Amount £13,000 (GBP)
Organisation Breast Cancer Campaign (BCC) 
Sector Charity/Non Profit
Country United Kingdom
Start 07/2012 
End 06/2013
 
Description Computer Modelling 
Organisation Northwick Park Hospital
Department Histopathology
Country United Kingdom 
Sector Hospitals 
PI Contribution Provided appropriately treated tissue for 3D modelling
Collaborator Contribution Designed appropriate computer programmes for 3D modelling of serially sectioned and labelled neoplastic gastrointestinal tissue.
Impact 2 papers so far, more to follow Fellous TG, Islam S, Tadrous PJ, Elia G, Kocher HM, Bhattacharya S, Mears L, Turnbull DM, Taylor RW, Greaves LC, Chinnery PF, Taylor G, McDonald SA, Wright NA, Alison MR. Locating the stem cell niche and tracing hepatocyte lineages in human liver. Hepatology. 2009; 49: 1655-1663. Fellous TG, McDonald SA, Burkert J, Humphries A, Islam S, De-Alwis NM, Gutierrez-Gonzalez L, Tadrous PJ, Elia G, Kocher HM, Bhattacharya S, Mears L, El-Bahrawy M, Turnbull DM, Taylor RW, Greaves LC, Chinnery PF, Day CP, Wright NA, Alison MR. A Methodological Approach To Tracing Cell Lineage In Human Epithelial Tissues. Stem Cells 2009; 27: 1410-1420.
Start Year 2008
 
Description Provision of Diseased Liver Tissue 
Organisation Queen Mary University of London
Department Barts and The London School of Medicine and Dentistry
Country United Kingdom 
Sector Academic/University 
PI Contribution We are using the liver tissue from our partners to further our research
Collaborator Contribution Provision of human liver tissue with specific diseases.
Impact 3 papers published Fellous TG, Islam S, Tadrous PJ, Elia G, Kocher HM, Bhattacharya S, Mears L, Turnbull DM, Taylor RW, Greaves LC, Chinnery PF, Taylor G, McDonald SA, Wright NA, Alison MR. Locating the stem cell niche and tracing hepatocyte lineages in human liver. Hepatology. 2009; 49: 1655-1663. Fellous TG, McDonald SA, Burkert J, Humphries A, Islam S, De-Alwis NM, Gutierrez-Gonzalez L, Tadrous PJ, Elia G, Kocher HM, Bhattacharya S, Mears L, El-Bahrawy M, Turnbull DM, Taylor RW, Greaves LC, Chinnery PF, Day CP, Wright NA, Alison MR. A Methodological Approach To Tracing Cell Lineage In Human Epithelial Tissues. Stem Cells 2009; 27: 1410-1420. Lin WR, Lim SA, McDonald SA, Graham T, Wright V, Peplow C, Humphries A, Kocher HM, Wright NA, Dhillon AP, Alison MR. The histogenesis of regenerative nodules in human liver cirrhosis. Hepatology 2010; 51: 1017-1026.
Start Year 2009
 
Description Provision of Diseased Liver Tissue 
Organisation University of Birmingham
Department University of Birmingham, Liver Tissue bank
Country United Kingdom 
Sector Academic/University 
PI Contribution We are using the liver tissue from our partners to further our research
Collaborator Contribution Provision of human liver tissue with specific diseases.
Impact 3 papers published Fellous TG, Islam S, Tadrous PJ, Elia G, Kocher HM, Bhattacharya S, Mears L, Turnbull DM, Taylor RW, Greaves LC, Chinnery PF, Taylor G, McDonald SA, Wright NA, Alison MR. Locating the stem cell niche and tracing hepatocyte lineages in human liver. Hepatology. 2009; 49: 1655-1663. Fellous TG, McDonald SA, Burkert J, Humphries A, Islam S, De-Alwis NM, Gutierrez-Gonzalez L, Tadrous PJ, Elia G, Kocher HM, Bhattacharya S, Mears L, El-Bahrawy M, Turnbull DM, Taylor RW, Greaves LC, Chinnery PF, Day CP, Wright NA, Alison MR. A Methodological Approach To Tracing Cell Lineage In Human Epithelial Tissues. Stem Cells 2009; 27: 1410-1420. Lin WR, Lim SA, McDonald SA, Graham T, Wright V, Peplow C, Humphries A, Kocher HM, Wright NA, Dhillon AP, Alison MR. The histogenesis of regenerative nodules in human liver cirrhosis. Hepatology 2010; 51: 1017-1026.
Start Year 2009
 
Description Provision of Diseased Liver Tissue 
Organisation University of Leuven
Department Department of Imaging and Pathology
Country Belgium 
Sector Academic/University 
PI Contribution We are using the liver tissue from our partners to further our research
Collaborator Contribution Provision of human liver tissue with specific diseases.
Impact 3 papers published Fellous TG, Islam S, Tadrous PJ, Elia G, Kocher HM, Bhattacharya S, Mears L, Turnbull DM, Taylor RW, Greaves LC, Chinnery PF, Taylor G, McDonald SA, Wright NA, Alison MR. Locating the stem cell niche and tracing hepatocyte lineages in human liver. Hepatology. 2009; 49: 1655-1663. Fellous TG, McDonald SA, Burkert J, Humphries A, Islam S, De-Alwis NM, Gutierrez-Gonzalez L, Tadrous PJ, Elia G, Kocher HM, Bhattacharya S, Mears L, El-Bahrawy M, Turnbull DM, Taylor RW, Greaves LC, Chinnery PF, Day CP, Wright NA, Alison MR. A Methodological Approach To Tracing Cell Lineage In Human Epithelial Tissues. Stem Cells 2009; 27: 1410-1420. Lin WR, Lim SA, McDonald SA, Graham T, Wright V, Peplow C, Humphries A, Kocher HM, Wright NA, Dhillon AP, Alison MR. The histogenesis of regenerative nodules in human liver cirrhosis. Hepatology 2010; 51: 1017-1026.
Start Year 2009
 
Description Provision of neoplastic gastrointestinal tissue 
Organisation Queen Mary University of London
Department Barts and The London School of Medicine and Dentistry
Country United Kingdom 
Sector Academic/University 
PI Contribution Using this tissue to further the aims of the Grant
Collaborator Contribution Provision of neoplastic tissue
Impact 5 recent publications Lineage tracing reveals dynamics of stem cell clone evolution within human colorectal adenomas Adam Humphries, Laura Gay, Biancastella Cereser, Daniel Miller, Bibek Das, Alice, Gutteridge, George Elia, Emma Nye, Marco Novelli, Manuel Rodriguez-Justo, Stuart AC McDonald, Nicholas A Wright, Trevor A Graham. In preparation Zeki SS, Graham TA & McDonald SAC. Utilizing DNA mutations to trace epithelial cell lineages in human tissues. Edited by Mace KA & Braun KM; Progenitor Cells: Methods and Protocols, Methods in Molecular Biology, vol. 916, DOI 10.1007/978-1-61779-980-8_22, Springer Science+Business Media, LLC 2012, pubmed 22914949 Nicholson AM, Graham TA, Simpson A, Humphries A, Burch N, Rodriguez-Justo M, Novelli MR, Harrison R, Wright NA, McDonald SAC* and Jankowski JA. Barrett¹s epithelium shares a common progenitor with the surrounding squamous epithelium and glands are clonal units containing multiple stem cells. Gut 2011 doi:10.1136/gutjnl-2011-301174 pubmed 22200839 Graham TA, Humphries A, Sanders T, Rodriguez-Justo M, Tadrous PJ, Nicholson AM, Novelli MR, Leedham SJ, McDonald SAC and Wright NA. Stem cell dynamics and epigenetic drift restrict the use of methylation patterns as markers of clonal expansion in the human colon. Gastroenterology 2011 140(4):1241-1250, pubmed 21192938 Gutierrez-Gonzalez L, Graham TA, Rodriguez-Justo M, Leedham SJ Novelli M, Gay LJ, Ventayol-Garcia T, Green A, Stoker DL, Bamba S, Yamada E, Kishi Y, Jankowski JA, Wright NA and McDonald SAC. The clonal origins of dysplasia from metaplasia in the human stomach. Gastroenterology 2011 140(4):1251-1260, pubmed 21223968
Start Year 2006
 
Description Provision of neoplastic gastrointestinal tissue 
Organisation University College London Hospital
Department Histopathology
Country United Kingdom 
Sector Academic/University 
PI Contribution Using this tissue to further the aims of the Grant
Collaborator Contribution Provision of neoplastic tissue
Impact 5 recent publications Lineage tracing reveals dynamics of stem cell clone evolution within human colorectal adenomas Adam Humphries, Laura Gay, Biancastella Cereser, Daniel Miller, Bibek Das, Alice, Gutteridge, George Elia, Emma Nye, Marco Novelli, Manuel Rodriguez-Justo, Stuart AC McDonald, Nicholas A Wright, Trevor A Graham. In preparation Zeki SS, Graham TA & McDonald SAC. Utilizing DNA mutations to trace epithelial cell lineages in human tissues. Edited by Mace KA & Braun KM; Progenitor Cells: Methods and Protocols, Methods in Molecular Biology, vol. 916, DOI 10.1007/978-1-61779-980-8_22, Springer Science+Business Media, LLC 2012, pubmed 22914949 Nicholson AM, Graham TA, Simpson A, Humphries A, Burch N, Rodriguez-Justo M, Novelli MR, Harrison R, Wright NA, McDonald SAC* and Jankowski JA. Barrett¹s epithelium shares a common progenitor with the surrounding squamous epithelium and glands are clonal units containing multiple stem cells. Gut 2011 doi:10.1136/gutjnl-2011-301174 pubmed 22200839 Graham TA, Humphries A, Sanders T, Rodriguez-Justo M, Tadrous PJ, Nicholson AM, Novelli MR, Leedham SJ, McDonald SAC and Wright NA. Stem cell dynamics and epigenetic drift restrict the use of methylation patterns as markers of clonal expansion in the human colon. Gastroenterology 2011 140(4):1241-1250, pubmed 21192938 Gutierrez-Gonzalez L, Graham TA, Rodriguez-Justo M, Leedham SJ Novelli M, Gay LJ, Ventayol-Garcia T, Green A, Stoker DL, Bamba S, Yamada E, Kishi Y, Jankowski JA, Wright NA and McDonald SAC. The clonal origins of dysplasia from metaplasia in the human stomach. Gastroenterology 2011 140(4):1251-1260, pubmed 21223968
Start Year 2006
 
Description School Visit 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Primary Audience Schools
Results and Impact Several. Inspiring school pupils with no expectation of going to University, particularly Medical School.

More applicants from local schools
Year(s) Of Engagement Activity 2008,2009,2010,2011