Research
CRISPR Discovery & Translational Applications
Gene (and Genome) editing is revolutionizing molecular biology and translational research. For the first time, we have the ability to carry out wide-ranging genetic manipulation in mammalian systems. This gives us the ability to understand how genes function and interact in models of human disease with the potential to transform how we understand and treat patients. The best way to investigate the function of a gene is to (1) knockdown/overexpress it in cells or (2) modify its DNA sequence in the genome of a cell line to imitate the effect of known mutations associated with disease. My group has successfully established oncology related cell line models using CRISPR knockdown/knockout for a variety of cancer sub-types. Current studies are harnessing these models to better understand prognostic gene markers, and drug resistance mutations.
We are currently endeavoring to harness current gene-editing research beyond simple base editing and towards translational applications – (i) Crispr library screens, (ii) Synthetic-lethality studies, (iii) Gene delivery to primary patient cells, (iv) Editing in complex systems (e.g. organoids), (v) in vivo editing.
Immuno-Oncology
Innate Immune Modulation as a Cancer Therapy
Myeloid leukaemia is a type of blood cancer that originates in the bone marrow, classified into either acute or chronic myeloid leukaemia (AML/CML). Type-I IFN signaling appears to negatively impact the development of leukemia, in line with its downstream functions, such as increase in rate of apoptosis, increase in AML immunogenicity, and decrease in cell proliferation. Despite this, it is unclear as to why type-I IFN treatment for AML is not as robust an envisioned. Emerging evidence now suggests targeting innate immune sensing mechanisms that regulate type-I IFN activity in response to viral infections may be exploited and refined to deliver antileukemic effects in AML rather than systemic delivery of type-I IFN regimens.
Despite strong biological rationale for the use of type-I IFNs for the treatment of acute myeloid leukemia (AML), their usage is limited to few hematologic malignancies. We propose that innate immune sensing machinery, particularly the stimulator of IFN genes pathway, may be exploited to deliver antileukemic effects in AML.
Developing Antibody Drug Conjugates to treat Cancers
The focus of this research collaboration (BiVictrix) is to immunophenotype samples from acute myeloid leukemia (AML) patients and healthy bone marrow samples which will aid in the BiVictrix Therapeutics drug pipeline development, and to characterize these novel drugs in disease models. The preliminary results from this award established two highly promising methodologies (mass cytometry and live cell imaging) for investigating the action of bivalent ADCs in model cellular systems. Whilst these methodologies show much promise, they now require further development and crucially, validation with human samples. The data generated so far focusses on a subset of cell surface markers for AML currently under development by BiVictrix, but mass cytometry has the potential for screening many other cell markers (associated with a variety of other diseases also).
As part of this work we have successfully recruited an MRC DiMeN iCASE DTP PhD student (Mr. James Griffin; Oct. 2021-2025).
Cancer Signaling
Inpp4b as a prognostic marker in AML
Recently, we have identified a novel 'oncogene' in AML - Inpp4b, (with Dr. Lenny Salmena & Dr. Mark D. Minden) which was previously known as a tumour suppressor akin to PTEN (see Woolley et al 2015; Dzneladze et al. 2015). This work highlighted a paradoxical role for Inpp4b as a prognostic marker for poor outcome in AML across multiple patient databases and showed an in vitro association with malignant phenotypes and chemotherapy resistance.
Inpp4b as a stem cell gene
We have since continued with mechanistic studies of Inpp4b and have thus far identified roles for this gene in maintaining ‘stemness’, in both mouse and cell-line models (manuscript in preparation) . We have shown a requirement forInpp4b in the maintenance in the hematopoietic stem cell (HSC) pool in murine models. Following on from this observation we have demonstrated that Inpp4b-deficient HSCs are functionally impaired in vivo. We have since focused on mechanistic studies to determine the role Inpp4b plays in stemness. We have established that Inpp4b is not a requirement for leukemia development (using an MLL-AF9 model) and studies of Inpp4b in leukemia are ongoing.
Reactive oxygen species (ROS) and Antioxidant Signaling in Cancer
1) Hydrogen peroxide as a second messenger in FLT3-ITD AML (Woolley et al 2013; Woolley et al 2013). Identified signal transduction mediated by H2O2 in FLT3 mutated AML. From this work I identified a novel link in the FLT3-ITD/Jak/STAT signaling pathway mediated by p22phox-dependent Nadph oxidases (NOXs), ultimately facilitating Pim-1 expression and survival (Woolley et al 2012).
2) Bcr-Abl drives pro-survival signaling via ROS. I built on my earlier work to show that ROS-mediated signaling plays an essential role in PI3K and MAPK/ERK pathways downstream of Bcr-Abl in CML (Landry et al 2013Landry et al. Leukemia Research 2013 Feb;37(2):183-9).
3) NOX-generated H2O2 at the endoplasmic reticulum generates DNA damage in FLT3-ITD AML. We identified dichotomous roles for in H2O2 in FLT-ITD AML, generating DNA alongside its role in signal transduction (Stanicka et al 2015). This further establishes H2O2 as a therapeutic target in AML, as it drives pro-survival signaling and possibly tumor heterogeneity via DNA-damaging events.
4) In addition we showed that a tumorogenic inflammatory pathway in colon cancer is driven by NOX-derived H2O2 and plays a role in metastasis (O'Leary et al 2012)
Research grants
Translational Relevance of the Stimulator of Interferon Genes (STING) pathway and viral infection in Follicular Lymphoma
NORTH WEST CANCER RESEARCH INCORPORATING CLATTERBRIDGE CANCER RESEARCH (UK)
February 2023 - February 2026
Bench Fees for Fatma Mayouf Omir Rabia
LIBYAN EMBASSY IN LONDON🚩
January 2022 - January 2025
Bench Fees for Hamdah Eenizan M Alsaeedi 2024-25 (201457448)
ROYAL EMBASSY OF SAUDI ARABIA CULTURAL BUREAU IN LONDON (UK)
November 2021 - October 2025
Research Support Doctorate PHD Fees for Mr Mohammed Alsubhi
ROYAL EMBASSY OF SAUDI ARABIA CULTURAL BUREAU IN LONDON (UK)
September 2018 - August 2022
Bench Fees for Abdullah Meshal A Alsufyani
ROYAL EMBASSY OF SAUDI ARABIA CULTURAL BUREAU IN LONDON (UK)
March 2020 - August 2024
Research Support Fees for Rehab Nawaf Shayem Alanazi
ROYAL EMBASSY OF SAUDI ARABIA CULTURAL BUREAU IN LONDON (UK)
March 2020 - February 2021
Research collaborations
BiVictrix Therapeutics
Developing ADCs for Cancer Therapy
BiVictrix Therapeutics
Developing ADCs for Cancer Therapy
Dr. Meong Hi Son
CRISPR Screening in NBL
Samsung Medical Centre (Seoul)
Developing CRISPR-based screening technologies in pediatric Neuroblastoma, which will be used to model prognostic markers and drug screening platforms.
Dr. Leonardo Salmena
Targetting INPP4B in Leukemia
University of Toronto
Understanding the molecular functions of INPP4B, anovel biomarker for poor prognosisin in AML, to establish therapeutic interventions
Dr. Mark Minden
Targetting INPP4B in Leukemia
Ontario Cancer Center (Princess Margaret Hospital)
Understanding the molecular functions of INPP4B, anovel biomarker for poor prognosisin in AML, to establish therapeutic interventions