Programming and re-programming of haemopoietic cell fate
Lead Research Organisation:
University of Oxford
Abstract
The blood is composed of a variety of different types of cells that each have specialised and distinct functions. As these cells die they are replenished by a small pool of long-lived cells known as stem cells which have the capacity both to produce more stem cells (a process termed self-renewal) or give rise to all the specialized blood cell types of the blood (a process termed differentiation). Our research programme attempts to understand, at the molecular level, how stem cells decide whether to self-renew, or to differentiate and in the latter case, into which cell type (a process termed lineage-specification). These working decisions that stem cells make are ultimately effected at the level of differential gene activity or usage, and gaining an understanding of how this process works is an important problem in developmental biology, transplantation medicine, stem cell based gene therapy and haematological malignancy. We study this process by genetically altering the activities of candidate molecular regulators of these processes. We anticipate that these studies will illuminate how normal cell fate decisions are instigated and inform approaches aimed at manipulating the expansion, directed-differentiation or reprogramming of stem cell fate for therapeutic or commercial advantage.
Technical Summary
All the different specialised cell types that constitute the blood are derived from a multipotential haemopoietic stem cell (HSC). HSCs are long-lived and in addition to having the capacity to give rise, through differentiation, to all blood lineages, HSC are capable of extensive self-renewal. It remains unclear at the molecular level how the balance between self-renewal and differentiation is regulated and how the different blood lineages are specified. However we can presume that the process of cell type restriction involves the formation of stable transcription factor complexes that initiate and consolidate (or repress) exclusive programmes of gene expression. Transcription factors have therefore been intensively studied as candidate instigators of cell fate decisions. This capacity has been emphasised by recent studies that indicate that individual transcription factors can reprogramme the development fate of already committed cell types. In our own laboratory we have shown that the erythroid-affiliated transcription factor GATA-1 can re-specify the fate of freshly isolated, bone marrow-derived, committed neutrophil-monocyte progenitors. In response to exogenous GATA-1 activity these cells adopt erythroid or other GATA-1-affiliated cell fates. These observations provide a mechanism for reprogramming of committed haematopoietic progenitors and an experimental entry point for directly identifying the key loci involved. Since manipulation of GATA-1 activity can also alter cell fate in multipotential progenitors that express endogenous GATA-1 and retain both erythroid and neutrophil monocyte potential, the same mechanisms and loci are likely to also be directly involved in normal uni-lineage differentiation from multipotent progenitor cells. Thus our approach is to examine transcription factor-mediated alterations in cell fate primarily in the experimental context of reprogramming and apply our results to the more normal context of cell fate programming from the multipotential uncommitted state. We will ask the following questions: 1) Which cells can be reprogrammed (lineage/stage of differentiation) by a single transcription factor. 2) Which transcription factors can reprogram. 3) What is the mechanism of reprogramming, is it de-differentiation or trans-differentiation? 4) How complete is the reprogramming i.e. do the cells show residual features of their original lineage and are they fully functional? 5) To what extent is the mechanism the same regardless of the lineage/stage being reprogrammed and to what extent is it lineage/stage specific? 6) What is the detailed mechanism of reprogramming in respect of target genes, chromatin re-modelling etc. 7) Does reprogramming require cell division. 8) Can human cells be reprogrammed? 9) To what extent to the processes which mediate reprogramming also function to programme the cell fate output of multipotent cells. We anticipate that these studies will contribute to our understanding of how normal cell fate decisions are instigated and inform approaches aimed at manipulating the expansion, directed-differentiation or reprogramming of stem cell fate for therapeutic or commercial advantage.
People |
ORCID iD |
| Tariq Enver (Principal Investigator) |
Publications
Anderson K
(2011)
Genetic variegation of clonal architecture and propagating cells in leukaemia.
in Nature
Anguita E
(2016)
A somatic mutation of GFI1B identified in leukemia alters cell fate via a SPI1 (PU.1) centered genetic regulatory network.
in Developmental biology
Banerji CR
(2013)
Cellular network entropy as the energy potential in Waddington's differentiation landscape.
in Scientific reports
Bridge G
(2012)
The microRNA-30 family targets DLL4 to modulate endothelial cell behavior during angiogenesis.
in Blood
Carpenter L
(2011)
Human induced pluripotent stem cells are capable of B-cell lymphopoiesis.
in Blood
Chan A
(2007)
CD56bright human NK cells differentiate into CD56dim cells: role of contact with peripheral fibroblasts.
in Journal of immunology (Baltimore, Md. : 1950)
Chickarmane V
(2009)
Computational modeling of the hematopoietic erythroid-myeloid switch reveals insights into cooperativity, priming, and irreversibility.
in PLoS computational biology
Ciau-Uitz A
(2010)
Tel1/ETV6 specifies blood stem cells through the agency of VEGF signaling.
in Developmental cell
Das Neves RP
(2010)
Connecting variability in global transcription rate to mitochondrial variability.
in PLoS biology
De Gobbi M
(2011)
Generation of bivalent chromatin domains during cell fate decisions
in Epigenetics & Chromatin
Duan CW
(2014)
Leukemia propagating cells rebuild an evolving niche in response to therapy.
in Cancer cell
Dzierzak E
(2008)
Stem cell researchers find their niche
in Development
Enver T
(2009)
Developmental biology: Instructions writ in blood.
in Nature
Enver T
(2009)
Stem cell states, fates, and the rules of attraction.
in Cell stem cell
Fan D
(2015)
Stem cell programs are retained in human leukemic lymphoblasts.
in Oncogene
Ferreira R
(2020)
Advances and challenges in retinoid delivery systems in regenerative and therapeutic medicine.
in Nature communications
Ford AM
(2009)
The TEL-AML1 leukemia fusion gene dysregulates the TGF-beta pathway in early B lineage progenitor cells.
in The Journal of clinical investigation
Glasow A
(2008)
DNA methylation-independent loss of RARA gene expression in acute myeloid leukemia.
in Blood
Goardon N
(2011)
Coexistence of LMPP-like and GMP-like leukemia stem cells in acute myeloid leukemia.
in Cancer cell
Graf T
(2009)
Forcing cells to change lineages.
in Nature
Guitart AV
(2013)
Hif-2a is not essential for cell-autonomous hematopoietic stem cell maintenance.
in Blood
Gupta R
(2009)
Molecular targeting of cancer stem cells.
in Cell stem cell
Gupta R
(2007)
NOV (CCN3) functions as a regulator of human hematopoietic stem or progenitor cells.
in Science (New York, N.Y.)
Gupta R
(2020)
Nov/CCN3 Enhances Cord Blood Engraftment by Rapidly Recruiting Latent Human Stem Cell Activity.
in Cell stem cell
Hong D
(2008)
Initiating and cancer-propagating cells in TEL-AML1-associated childhood leukemia.
in Science (New York, N.Y.)
Huang S
(2007)
Bifurcation dynamics in lineage-commitment in bipotent progenitor cells.
in Developmental biology
Kosak ST
(2007)
Coordinate gene regulation during hematopoiesis is related to genomic organization.
in PLoS biology
Kranc KR
(2015)
Acute loss of Cited2 impairs Nanog expression and decreases self-renewal of mouse embryonic stem cells.
in Stem cells (Dayton, Ohio)
Kranc KR
(2009)
Cited2 is an essential regulator of adult hematopoietic stem cells.
in Cell stem cell
Lawrie CH
(2008)
MicroRNA expression in lymphocyte development and malignancy.
in Leukemia
Lawrie CH
(2007)
MicroRNA expression distinguishes between germinal center B cell-like and activated B cell-like subtypes of diffuse large B cell lymphoma.
in International journal of cancer
Lima AF
(2018)
Osmotic modulation of chromatin impacts on efficiency and kinetics of cell fate modulation.
in Scientific reports
Migueles RP
(2017)
Transcriptional regulation of Hhex in hematopoiesis and hematopoietic stem cell ontogeny.
in Developmental biology
Morrison GM
(2008)
Anterior definitive endoderm from ESCs reveals a role for FGF signaling.
in Cell stem cell
Nijnik A
(2007)
DNA repair is limiting for haematopoietic stem cells during ageing.
in Nature
Nimmo R
(2013)
MiR-142-3p controls the specification of definitive hemangioblasts during ontogeny.
in Developmental cell
Nimmo RA
(2015)
Primed and ready: understanding lineage commitment through single cell analysis.
in Trends in cell biology
Pina C
(2015)
Single-Cell Network Analysis Identifies DDIT3 as a Nodal Lineage Regulator in Hematopoiesis.
in Cell reports
Pina C
(2012)
Inferring rules of lineage commitment in haematopoiesis.
in Nature cell biology
Pina C
(2008)
MLLT3 regulates early human erythroid and megakaryocytic cell fate.
in Cell stem cell
Robert-Moreno A
(2008)
Impaired embryonic haematopoiesis yet normal arterial development in the absence of the Notch ligand Jagged1.
in The EMBO journal
Rodrigues NP
(2008)
GATA-2 regulates granulocyte-macrophage progenitor cell function.
in Blood
Soneji S
(2007)
Inference, validation, and dynamic modeling of transcription networks in multipotent hematopoietic cells.
in Annals of the New York Academy of Sciences
Stavish D
(2020)
Generation and trapping of a mesoderm biased state of human pluripotency
in Nature Communications
Stewart MH
(2015)
The histone demethylase Jarid1b is required for hematopoietic stem cell self-renewal in mice.
in Blood
| Description | Cord Blood |
| Geographic Reach | National |
| Policy Influence Type | Participation in a guidance/advisory committee |
| Description | ESTOOLs - EU Framework6 integrated Project to examine mechanisms of human ES cell self renewal and differentiation |
| Amount | € 205,854 (EUR) |
| Funding ID | 18739 |
| Organisation | Sixth Framework Programme (FP6) |
| Sector | Public |
| Country | European Union (EU) |
| Start | |
| Description | EUROCSC - "STREP" Consortium for the study of Cancer Stem Cells |
| Amount | £148,170 (GBP) |
| Funding ID | 37632 |
| Organisation | European Commission |
| Sector | Public |
| Country | European Union (EU) |
| Start | |
| Description | EuroSystem European Federation for Systematic Stem Cell Biology |
| Amount | £394,168 (GBP) |
| Funding ID | 200720 |
| Organisation | European Commission |
| Sector | Public |
| Country | European Union (EU) |
| Start | |
| Description | Functional enhancement of Human Cord Blood derived Haematopoietic Stem Cells through Nov. |
| Amount | 2,400,000 kr (SEK) |
| Funding ID | 2019-01752 |
| Organisation | Swedish Research Council |
| Sector | Public |
| Country | Sweden |
| Start | 01/2020 |
| End | 12/2022 |
| Description | STEMEXPAND - Stem Cell Expansion - Expansion and engraftment of haematopoietic and mesenchymal stem cells |
| Amount | £384,233 (GBP) |
| Funding ID | 222989 |
| Organisation | European Commission |
| Sector | Public |
| Country | European Union (EU) |
| Start | |
| Description | Transcriptional programming of pre-leukaemia and leukaemia stem and progenitor cells in childhood ALL |
| Amount | £1,451,548 (GBP) |
| Organisation | Leukaemia and Lymphoma Research |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | |
| Title | Network Database |
| Description | A database of gene expression profiles and transcription factor targets throughout blood lineage development. |
| Type Of Material | Biological samples |
| Year Produced | 2009 |
| Provided To Others? | Yes |
| Impact | This database is serving to foster collaborative interactions with European colleagues including Gerald de Haan - University of Gronigen, Netherlands, Carsten Petersen, University of Lund, Sweden, Ingo Roeder University of Leipzig, Germany and Ruud Delwel Erasmus University, The Netherlands |
| Description | Micro Array Development |
| Organisation | Oxford Gene Technology |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | We have provided consultation about stem cell biology. We have provided stem cells and cell lines and identified gene targets for technology validation. |
| Collaborator Contribution | OGT have provided my laboratory with state of the art micro array platforms and assisted in design of array based experiments. |
| Impact | Interactions with Oxford Gene Technology have lead to partnership in a new Eu framework 7 programme EuroSYSTEM. They have resulted in on going development of novel array based methods for analysis of gene expression in single cells. |
| Description | Network reconstruction |
| Organisation | Lund University |
| Country | Sweden |
| Sector | Academic/University |
| PI Contribution | We have provided data sets and in depth knowledge of the biology of blood based stem cell systems. |
| Collaborator Contribution | This partnership has provided a PhD student based in the department of theoretical physics in Lund University under the direction of Carsten Petersen. This student's project is solely based on mathematical analysis of data produced by our MRC programme. |
| Impact | Co-authored publication in PLOS Computational Biology PubMed ID 19165316 There has been an exchange of my MRC funded bioinformatician (Shamit Soneji) between sites. Additionally we have received bioinformatical support from other members of the theoretical physics department. |
| Description | Stem cell expansion |
| Organisation | Lund University |
| Country | Sweden |
| Sector | Academic/University |
| PI Contribution | We have provided candidate genes involved in self renewal and molecular biology expertise. |
| Collaborator Contribution | Stefan Karlsson's laboratory has assisted us with in-depth knowledge of short hair pin RNA vectors and gene therapy approaches for our studies of human blood stem cell self renewal. |
| Impact | We have achieved funding through the Haemato Linne Programme in Sweden to support a joint post doctoral fellow based in Sweden. |
| Start Year | 2007 |
| Description | Bryanstone School |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | Yes |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Prof Enver accompanied by Dr Emma Kemp from Public Outreach Division of EuroSystem (EU Framework 7) showed a movie (stem cell story) to students from the Bryanstone School Science Society - approx 70 children. Prof Enver then presented his own research in an accessible form and hosted a question and answer session on both topics. The event took approximately 2 hours. Several school children requested laboratory placements in Prof Enver's lab and they were directed to Emma Kemp at EuroSystem for further information. |
| Year(s) Of Engagement Activity | 2009 |
| Description | Geoff Thomas |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Policymakers/politicians |
| Results and Impact | Prof Enver attended House of Commons to brief parliamentarians and representatives of charitable and pharmasuitacle sectors on break throughs in cancer stem cell science. This presentation was in support of the activities of the Geoff Thomas Foundation.which is raising funds for Leukaemia Research. http://www.geoffthomasfoundation.org/site/index.php?option=com_content&view=article&id=20&Itemid=68 Raised awareness. |
| Year(s) Of Engagement Activity | 2008 |
| Description | ISSCR - Barcelona |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | Yes |
| Geographic Reach | International |
| Primary Audience | Public/other audiences |
| Results and Impact | Prof Enver assembled a panel of scientists, ethisists and health care professionals to assist in a panel discussion based around short movies describing stem cells. The event was entitled "Everything you wanted to know about Stem cells but were afraid to ask" and took place at Bosmo Caixa Museum, Barcelona. It was attended by a range of scientists, patient advocate, patients, policy makers, lay public and journalists. In addition through the offices of ESTOOLS we put on a photographic exhibit entitled "Smile of a Stem Cell" which was displayed at the Cosmo Caixa Museum and open to public and scientists alike. This is part of on-going public engagement and out reach by the ISSCR but most importantly this served as a platform for high lighting the activities of two major European networks focused on stem cell research namely EuroSystem and ESTOOLS. |
| Year(s) Of Engagement Activity | 2009 |
| Description | Leukaemia Research Fund Golf Charity Day, Ashridge |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | Prof Enver described his research to approx 100 members of the public who had participated in a charitable golf event. This event raised approx £10k for Leukaemia Research and enhanced the scientific understanding of patients and their families in respect of the diseases which afflict them. |
| Year(s) Of Engagement Activity | 2009 |
| Description | Lord Patel |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Policymakers/politicians |
| Results and Impact | Prof Enver described the state of the art in stem cell biology to an assembly and parliamentarians and policymakers at the invitation of Lord Narendra Patel. This was part of informing opinion around the issue of stem cells and there by helping to maintain progressive legislation and competitiveness in the stem cell area within the UK. |
| Year(s) Of Engagement Activity | 2006 |
| Description | Twins with Leukaemia |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Media (as a channel to the public) |
| Results and Impact | Prof Enver attended the Science Media Centre and presented findings recently published in Science Magazine, Hong et al 'Initiating and cancer-propagating cells in TEL-AML10 associated childhood leukaemia", which pertained to the origins of childhood leukaemia. This story was widely covered in national and international news media including the front page of The Independent and major tv coverage. |
| Year(s) Of Engagement Activity | 2008 |