A gene editing approach to identify tumor suppressor genes in acute myeloid leukemia and modeling of AML in human hematopoietic stem cells
Acute myeloid leukemia (AML) arises through the sequential acquisition of genetic and epigenetic aberrations, initially in hematopoietic stem cells (HSCs) and subsequently in committed myeloid progenitors or precursors, resulting in their transformation into leukemic stem cells (LSCs). Generation of accurate pre-clinical AML models is an essential tool, to both identify the critical changes that induce the disease and to develop targeted therapies which are less toxic than the current standard of chemotherapy. The aim of this project is to identify novel AML tumor suppressors, and to characterize their role in the development of resistance to both conventional and molecular therapies.
By comparing different LSC populations, transformed by distinct oncogenic mutations, to normal progenitors we have identified gene that consistently down-regulated upon leukemic transformation. These genes will be targeted by gene editing in order to identify those responsible for suppressing AML formation, initially in murine genetic AML models, and subsequently by modeling AML in human HSCs.
In this project a well-defined genetic AML model will be used to identify tumor suppressor genes. Using lentiviral CRISPR-Cas9 delivery into pre-leukemic murine HSCs the candidate genes will be individually targeted and those where deletion accelerates AML formation identified. The identified tumor suppressors will next be targeted in human HSCs along with co-operating mutations (IDH, TET, FLT3, NPM1) using CRISPR-Cas9–based lentiviral targeting, followed by xenografting of immunodeficient mice. The resulting leukemias will be characterized for their sensitivity to both standard chemotherapy and targeted inhibitors, in order to identify the effects of the novel tumor suppressors in the development of therapy resistance.
This project will be based in the MRC Molecular Hematology Unit at the MRC Weatherall Institute of Molecular Medicine, with access to state-of-the-art facilities. The project provides an opportunity for training in a broad range of different techniques, including stem cell biology as a research subject, as well as the use of transgenic mice, combining chromatin analysis with high throughput sequencing, bioinformatics, and the opportunity to translate findings to the human hematopoietic system, and human HSCs in particular.
As well as the specific training detailed above, students will have access to a wide-range of seminars and training opportunities through the many research institutes and centres based in Oxford. Students are also able to attend the Methods and Techniques course run by the MRC Weatherall Institute of Molecular Medicine. This course runs through the year, ensuring that students have the opportunity to build a broad-based understanding of differing research techniques. There are also courses on Immunology and Bioinformatics and others may be added. Institute Seminars are held on a weekly basis and regularly attract world-class scientists in haematopoiesis research. Informal exchange of ideas in the coffee area is encouraged and is an attractive feature of the MRC WIMM.
Generic skills training is offered through the Medical Sciences Division's Skills Training Programme. This programme offers a comprehensive range of courses covering many important areas of researcher development: knowledge and intellectual abilities, personal effectiveness, research governance and organisation, and engagement, influence and impact. Students are actively encouraged to take advantage of the training opportunities available to them.
The department has a successful mentoring scheme, open to graduate students, which provides an additional possible channel for personal and professional development outside the regular supervisory framework. We hold an Athena SWAN Silver Award in recognition of our efforts to support the careers of female students and staff.
Pharmacological targeting of the Wdr5-MLL interaction in C/EBPα N-terminal leukemia.
Grebien F, Vedadi M, Getlik M, Giambruno R, Grover A, Avellino R, Skucha A, Vittori S, Kuznetsova E, Smil D, Barsyte-Lovejoy D, Li F, Poda G, Schapira M, Wu H, Dong A, Senisterra G, Stukalov A, Huber KVM, Schönegger A, Marcellus R, Bilban M, Bock C, Brown PJ, Zuber J, Bennett KL, Al-Awar R, Delwel R, Nerlov C, Arrowsmith CH, Superti-Furga G.
Nat Chem Biol. 2015 Aug;11(8):571-578. doi: 10.1038/nchembio
Molecular and cellular effects of oncogene cooperation in a genetically accurate AML mouse model.
Reckzeh K, Bereshchenko O, Mead A, Rehn M, Kharazi S, Jacobsen SE, Nerlov C, Cammenga J.
Leukemia. 2012 Jul;26(7):1527-36. doi: 10.1038/leu.2012.37. Epub 2012 Feb 9
Hematopoietic stem cell expansion precedes the generation of committed myeloid leukemia-initiating cells in C/EBPalpha mutant AML.
Bereshchenko O, Mancini E, Moore S, Bilbao D, Månsson R, Luc S, Grover A, Jacobsen SE, Bryder D, Nerlov C.
Cancer Cell. 2009 Nov 6;16(5):390-400. doi: 10.1016/j.ccr.2009.09.036
|4||Co-existence of LMPP-like and GMP-like Leukemia Stem Cells in Acute Myeloid Leukemia. Goardon N, Marchi E, Atzberger A, Quek L, Schuh A, Woll P, Mead A, Alford KA, Rout R, Chaudhury S, Gilkes A, Knapper S, Soneji S, Beldjord K, Begum S, Rose S, Geddes N, Griffiths M, Standen G, Sternberg A, Cavenagh J, Hunter H, Bowen D, Killick S, Robinson L, Price A, Macintyre E, Virgo P, Burnett A, Craddock C, Enver T, Jacobsen SEW, Porcher C and Vyas P. Cancer Cell. (2011). Jan 18;19(1):138-52. PMID: 21251617|
|5||Functional and genetic heterogeneity of distinctive leukemic stem cell populations in CD34- human acute myeloid leukaemia. Quek L, Otto GW, Garnett C, Lhermitte L, Lau I, Karamitros D, Doondeea J, Usukhbayar B, Goardon N, Ivey A, Gu Y, Gale R, Davies B, Sternberg A, Killick S, Hunter H, Cahalin P, Price A, Carr A, Griffiths M, Virgo P, Mackinnon S, Hills R, Grimwade D, Freeman S, Burnett A, Russell N, Craddock C, Mead AJ, Peniket A, Porcher C & Vyas P (2016). JEM. (2016). Jul 25;213(8)1513-1535. PMID: 27377587|