About the Project
Transplantation of haematopoietic stem cells (HSCs) is a curative treatment for many malignant and non-malignant haematological disorders. Moreover, HSC transplantation offers a possibility to cure bone marrow failure resulting from intensive radio/chemotherapy in patients suffering from solid cancers. However, the availability of matched donors, the toxicity of the conditioning regimen and the risk of graft rejection are major hurdles in treating patients. Autologous HSCs generated via reprogramming of patient-specific somatic cells (via direct reprogramming) or stem cells (via directed differentiation) could be useful for treating cancer patients.
We have recently shown that ectopic expression of transcription factors SCL and LMO2 reprogram murine fibroblasts into haematopoietic stem and progenitor cells (HSPCs). Reprogrammed HSPCs stably expanded on stromal cells and exhibited multilineage clonogenic capacity in vivo when transplanted into recipient mice. However, it is not clear how these two transcription factors induce HSPC phenotype in a very distinct cell type such as fibroblasts. The overall goal of this project is to understand how these two transcription factors induce blood progenitor cell phenotype in fibroblasts. To this end, we have generated a mouse line (iSCL-LMO2) with an inducible expression of transcription factors SCL and LMO2. Initially, we will establish reprogramming methodologies using fibroblasts derived from day 14.5 mouse embryo (iSCL-LMO2) and characterize reprogrammed blood progenitor cells using standard haematopoietic functional assays such as colony formation (CFU), flow cytometry and long-term multilineage repopulation capacity. Further, we aim to understand the molecular mechanisms involved in fate conversion by investigating the genomic targets of SCL and LMO2, chromatin architecture (open chromatin regions/histone marks) and gene expression at different stages of reprogramming. Finally, we will validate genes/pathways, which are regulated by SCL and LMO2, using overexpression/silencing or small molecules targeting the pathways. Together, this study will have implications not only in generating autologous blood cells but also targeting pathways involved in leukaemia since altered expression of SCL and LMO2 are involved in leukaemias.
Entry Requirements:
Applications should be submitted online and candidates should make direct contact with the Manchester supervisor to discuss their application directly. Applicants must have obtained, or be about to obtain, at least an upper second class honours degree (or equivalent) in a relevant subject.
Funding Notes
This project is available to UK/EU candidates. Funding covers fees (UK/EU rate) and stipend for four years. Overseas candidates can apply providing they can pay the difference in fees and are from an eligible country. Candidates will be required to split their time between Manchester and Singapore, as outlined on www.manchester.ac.uk/singaporeastar.
As an equal opportunities institution we welcome applicants from all sections of the community regardless of gender, ethnicity, disability, sexual orientation and transgender status. All appointments are made on merit.
References
1. Batta K*, Florkowska M*, Kouskoff V, Lacaud G. Direct reprogramming of murine fibroblasts to hematopoietic progenitor cells. Cell Reports. 2014; 9(5):1871-84.
2. Cheng H, Ang HY, A El Farran C, Li P, Fang HT, Liu TM, Kong SL, Chin ML, Ling WY, Lim EK, Li H, Huber T, Loh KM, Loh YH*, Lim B. Reprogramming mouse fibroblasts into engraftable myeloerythroid and lymphoid progenitors. Nat Commun. 2016 Nov 21;7:13396. * Corresponding author
3. Fang HT, El Farran CA, Xing QR, Zhang LF, Li H, Lim B, Loh YH. Global H3.3 dynamic deposition defines its bimodal role in cell fate transition. Nat Commun. 2018 Apr 18;9:1537
4. Goode DK, Obier N, Vijayabaskar MS, Lie-A-Ling M, Lilly AJ, Hannah R, Lichtinger M, Batta K, Florkowska M, Assi SA, Cauchy P, Gilmour J, Westhead DR*, Lacaud G*, Kouskoff V*, Göttgens B* and Bonifer C*. Dynamic gene regulatory networks drive hematopoietic specification and differentiation. Developmental Cell. 2016; 36:1-16. * Co-senior authors
5. Thambyrajah R, Mazan M, Patel R, Moignard V, Stefanska M, Marinopoulou E, Li Y, Lancrin C, Clapes T, Möröy T, Robin C, Miller C, Cowley S, Göttgens B, Kouskoff V, Lacaud G. GFI1 proteins orchestrate the emergence of haematopoietic stem cells through recruitment of LSD1. Nat Cell Biol. 2016 Jan;18(1):21-32
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