Periodic Reporting for period 1 - ProMeta (Non-histone protein acetylation targets of KAT2A in AML)
Reporting period: 2018-06-01 to 2020-05-31
Acute Myeloid Leukaemia (AML) has a dismal prognosis with less than 30% 5-year survival.
Mainstay therapy has remained essentially unchanged for the past three decades, with small advances in disease-free survival for the most part attributable to transplantation and improved supportive care.
The World Health Organization (WHO) classifies AML in seven categories based on morphology and immune phenotype criteria.
Genome sequencing studies have gone some way into defining the mutational spectrum of the disease, with identification of the most frequent recurrently mutated genes, and highlighting the presence of multiple mutations in most forms of the disease.
A significant number of the most commonly mutated targets in AML are histone modifiers, i.e. proteins or complexes that catalyse post-translational modifications in specific residues of the histone side chains.
Acetylation status of lysine residues in histone tails have been studied in some detail, and suggest evident epigenetic post-translational modifications that alter DNA-templated processes and facilitate malignant transformation.
An important acetyltransferase is KAT2A, the dominant mammalian orthologue of GCN5, in haematopoietic stem progenitor and leukaemic cells.
It has been shown that KAT2A regulates the activity of Peroxisome Proliferator Activated Receptor Gamma-Coactivator-1α, and through protein acetylation, directly regulating their transcriptional activity, key to glucose metabolism.
This proposal has had as main goal to position KAT2A as a critical regulator and therapeutic target in AML biology, crucially in the interaction with non-histone proteins and as a candidate regulator of AML metabolism.
I have performed quantitative proteomics and metabolomic analysis of Kat2a depletion in a primary mouse AML model, where we demonstrated a requirement for Kat2a in maintaining leukaemia stem cells (LSC).
I identified a defect in the usage of oxidative phosphorylation upon Kat2a loss, which did not correspond to a structural mitochondrial defect and was reversible upon cell loading with malate, a key metabolite in oxidative energy production.
Proteomics and metabolomics data can be articulated with my lab’s published datasets (where I have as well contributed ) on transcriptional and epigenetic targets of KAT2A, to define the network of events underlying KAT2A activity in AML and select potential druggable targets.
Mainstay therapy has remained essentially unchanged for the past three decades, with small advances in disease-free survival for the most part attributable to transplantation and improved supportive care.
The World Health Organization (WHO) classifies AML in seven categories based on morphology and immune phenotype criteria.
Genome sequencing studies have gone some way into defining the mutational spectrum of the disease, with identification of the most frequent recurrently mutated genes, and highlighting the presence of multiple mutations in most forms of the disease.
A significant number of the most commonly mutated targets in AML are histone modifiers, i.e. proteins or complexes that catalyse post-translational modifications in specific residues of the histone side chains.
Acetylation status of lysine residues in histone tails have been studied in some detail, and suggest evident epigenetic post-translational modifications that alter DNA-templated processes and facilitate malignant transformation.
An important acetyltransferase is KAT2A, the dominant mammalian orthologue of GCN5, in haematopoietic stem progenitor and leukaemic cells.
It has been shown that KAT2A regulates the activity of Peroxisome Proliferator Activated Receptor Gamma-Coactivator-1α, and through protein acetylation, directly regulating their transcriptional activity, key to glucose metabolism.
This proposal has had as main goal to position KAT2A as a critical regulator and therapeutic target in AML biology, crucially in the interaction with non-histone proteins and as a candidate regulator of AML metabolism.
I have performed quantitative proteomics and metabolomic analysis of Kat2a depletion in a primary mouse AML model, where we demonstrated a requirement for Kat2a in maintaining leukaemia stem cells (LSC).
I identified a defect in the usage of oxidative phosphorylation upon Kat2a loss, which did not correspond to a structural mitochondrial defect and was reversible upon cell loading with malate, a key metabolite in oxidative energy production.
Proteomics and metabolomics data can be articulated with my lab’s published datasets (where I have as well contributed ) on transcriptional and epigenetic targets of KAT2A, to define the network of events underlying KAT2A activity in AML and select potential druggable targets.
The goal of this project has been to explore the role of KAT2A in Acute Myeloid Leukaemia (AML) through investigation of its metabolic consequences and non-histone protein acetylation activity.
I have performed global proteomics and metabolomics to investigate the contribution of KAT2A-mediated acetylation events to AML maintenance.
I have combined the use of Tag Mass Tagging (TMT) Proteomics, liquid chromatography/mass spectrometry (LC/MS) and nuclear magnetic resonance (NMR) spectroscopy metabolomics with analysis of mitochondrial function in human AML cell lines and primary mouse leukaemia cells to integrate information about the effects of KAT2A on leukaemia metabolism.
In parallel, I have made some progress in developing and implementing protocols for acetyl-proteomic analysis of leukaemia cells, which will allow integration of epigenetic and non-histone protein consequences of KAT2A-mediated lysine acetylation in regulating and reading leukaemia metabolism.
The data obtained highlighted a specific vulnerability of the mitochondria to Kat2a depletion, which did not have structural consequences, as determined by transmission and scanning electron microscopy, and was reversible upon cell loading with the Krebs cycle intermediate malate.
Mitochondrial function was analysed by flow cytometry determination of mitochondrial mass and membrane potential, ROS production, and critically Seahorse XF analysis of oxidative phosphorylation and glycolytic activity.
In vitro leukaemia cell analysis suggests that reduced mitochondrial function is at least partially responsible for the observed loss of LSC self-renewal in Kat2a KO leukaemia, convincingly linking KAT2A vulnerability in AML with metabolic regulation.
These results are being written-up for publication and will constitute the core of a scientific manuscript which will be presented at scientific seminars and in the next MSCA conferences.
I have performed global proteomics and metabolomics to investigate the contribution of KAT2A-mediated acetylation events to AML maintenance.
I have combined the use of Tag Mass Tagging (TMT) Proteomics, liquid chromatography/mass spectrometry (LC/MS) and nuclear magnetic resonance (NMR) spectroscopy metabolomics with analysis of mitochondrial function in human AML cell lines and primary mouse leukaemia cells to integrate information about the effects of KAT2A on leukaemia metabolism.
In parallel, I have made some progress in developing and implementing protocols for acetyl-proteomic analysis of leukaemia cells, which will allow integration of epigenetic and non-histone protein consequences of KAT2A-mediated lysine acetylation in regulating and reading leukaemia metabolism.
The data obtained highlighted a specific vulnerability of the mitochondria to Kat2a depletion, which did not have structural consequences, as determined by transmission and scanning electron microscopy, and was reversible upon cell loading with the Krebs cycle intermediate malate.
Mitochondrial function was analysed by flow cytometry determination of mitochondrial mass and membrane potential, ROS production, and critically Seahorse XF analysis of oxidative phosphorylation and glycolytic activity.
In vitro leukaemia cell analysis suggests that reduced mitochondrial function is at least partially responsible for the observed loss of LSC self-renewal in Kat2a KO leukaemia, convincingly linking KAT2A vulnerability in AML with metabolic regulation.
These results are being written-up for publication and will constitute the core of a scientific manuscript which will be presented at scientific seminars and in the next MSCA conferences.
This project has positioned KAT2A as a key regulator of leukaemia cell metabolism and indicated specific targets for therapeutic exploration.
Identification of novel and personalised therapeutic targets is a key necessity in AML, which is currently treated by aggressive cytotoxic regimens and bone marrow transplantation which unfortunately cannot benefit elderly multi-pathology patients that are most frequently affected by the condition.
Precise dissection of the mechanisms of action of vulnerability genes, such as KAT2A, offers the possibility of sequentially or combinatorially target their downstream targets, with reduced potential for toxicity and acquisition of drug resistance.
This will be key to increasing the span and quality of life of AML patients and reducing the burden on health and social services, with positive consequences for the individual and for society.
Identification of novel and personalised therapeutic targets is a key necessity in AML, which is currently treated by aggressive cytotoxic regimens and bone marrow transplantation which unfortunately cannot benefit elderly multi-pathology patients that are most frequently affected by the condition.
Precise dissection of the mechanisms of action of vulnerability genes, such as KAT2A, offers the possibility of sequentially or combinatorially target their downstream targets, with reduced potential for toxicity and acquisition of drug resistance.
This will be key to increasing the span and quality of life of AML patients and reducing the burden on health and social services, with positive consequences for the individual and for society.