Gabriella M A Forte

The University of Manchester · Faculty of Life Sciences

AIMS Patients with glioblastoma have dismal outcomes and there is an urgent need for new treatment modalities. The POBIG trial is the first to evaluate the safety and feasibility of single-fraction preoperative radiotherapy for newly diagnosed glioblastoma. We report the results from our first dose arm. METHOD POBIG is a phase I radiotherapy dose...

Background Mucopolysaccharidosis IIIC (MPSIIIC) is one of four Sanfilippo diseases sharing clinical symptoms of severe cognitive decline and shortened lifespan. The missing enzyme, heparan sulfate acetyl-CoA: α-glucosaminide-N-acetyltransferase (HGSNAT), is bound to the lysosomal membrane, therefore cannot cross the blood-brain barrier or diffuse b...

Introduction: Through mechanisms that are poorly understood, sustained endurance exercise increases atrial fibrillation (AF) risk. Here we investigated involvement of electrophysiological pulmonary vein myocardium (PV) remodelling in a translational model of high-intensity endurance exercise.

Mucopolysaccharidosis type II (MPSII) is a lysosomal storage disease caused by a mutation in the IDS gene, resulting in deficiency of the enzyme iduronate-2-sulfatase (IDS) causing heparan sulfate (HS) and dermatan sulfate (DS) accumulation in all cells. This leads to skeletal and cardiorespiratory disease with severe neurodegeneration in two third...

Although, for many decades, the day–night rhythm in resting heart rate has been attributed to the parasympathetic branch of the autonomic nervous system (high vagal tone during sleep), recently we have shown that there is a circadian clock in the cardiac pacemaker, the sinus node, and the day–night rhythm in heart rate involves an intrinsic rhythmi...

Background: Glioblastoma is a high-grade aggressive neoplasm whose outcomes have not changed in decades. In the current treatment pathway, tumour growth continues and remains untreated for several weeks post-diagnosis. Intensified upfront therapy could target otherwise untreated tumour cells and improve the treatment outcome. POBIG will evaluate t...

Rationale: Athletes present with atrioventricular node (AV node) dysfunction manifesting as AV block. This can necessitate electronic pacemaker implantation, known to be more frequent in athletes with a long training history. Objective: AV block in athletes is attributed to high vagal tone. Here we investigated the alternative hypothesis that elect...

Background: Endurance athletes are prone to bradyarrhythmias, which in the long-term may underscore the increased incidence of pacemaker implantation reported in this population. Our previous work in rodent models has shown training-induced sinus bradycardia to be due to microRNA (miR)-mediated transcriptional remodeling of the HCN4 channel, leadin...

Mucopolysaccharidosis type II (MPSII or Hunter disease) is a lysosomal storage disease caused by a mutation in the IDS gene, resulting in heparan sulfate (HS) and dermatan sulfate (DS) accumulation. This leads to skeletal and cardiorespiratory disease with severe neurodegeneration in two thirds of suffers. Enzyme replacement therapy is ineffective...

Background: Glioblastoma (GBM) has been extensively researched over the last few decades, yet despite aggressive multi-modal treatment, recurrence is inevitable and second-line treatment options are limited. Here, we demonstrate how high throughput screening (HTS) in multicellular spheroids can generate physiologically relevant patient chemosensit...

Highly conserved signalling pathways controlled by mammalian target of rapamycin (mTOR) and AMP-activated protein kinase (AMPK) are central to cellular metabolism and cell proliferation1,2, and their dysregulation is implicated in the pathogenesis of major human diseases such as cancer and type 2 diabetes. AMPK pathways leading to reduced cell prol...

Glycolysis represents the fundamental metabolic pathway for glucose catabolism across biology, and glycolytic enzymes are amongst the most abundant proteins in cells. Their expression at such levels provides a particular challenge. Here we demonstrate that the glycolytic mRNAs are localized to granules in yeast and human cells. Detailed live cell a...

mRNA localization serves key functions in localized protein production, making it critical that the translation machinery itself is present at these locations. Here we show that translation factor mRNAs are localized to distinct granules within yeast cells. In contrast to many messenger RNP granules, such as processing bodies and stress granules, w...

AMP-activated kinase (AMPK) and target of rapamycin (TOR) signalling coordinate cell growth, proliferation, metabolism and cell survival with the nutrient environment of cells. The poor vasculature and nutritional stress experienced by cells in solid tumours raises the question: how do they assimilate sufficient nutrients to survive? Here, we show...

Background Chimeric mouse models generated via adoptive bone marrow transfer are the foundation for immune cell tracking in neuroinflammation. Chimeras that exhibit low chimerism levels, blood-brain barrier disruption and pro-inflammatory effects prior to the progression of the pathological phenotype, make it difficult to distinguish the role of im...

MPS IIIC is caused by mutations in the HGSNAT gene. HGSNAT deficiency affects lysosomal catabolism of heparan sulfate (HS), resulting in widespread CNS pathology in infants and children, leading to behavioural problems, cognitive decline and, finally, dementia and death before adulthood. Currently there are no existing treatments for MPS IIIC and t...

mRNA localization serves key functions in localized protein production making it critical that the translation machinery itself is present at these locations. Here we show that translation factor mRNAs are localized to distinct granules within yeast cells. In contrast to many mRNP granules, such as P-bodies and stress granules, which contain transl...

BACKGROUND Actinomycin-D (ACTD) is antineoplastic antibiotic and is used to treat a variety of childhood cancers, including neuroblastoma. Few studies have investigated the efficacy of ACTD in high grade glioma; however our 3D-high throughput assay system has identified ACTD to be highly cytotoxic over a panel of twelve patient-derived glioma stem-...

The pediatric lysosomal storage disorder mucopolysaccharidosis type II is caused by mutations in IDS, resulting in accumulation of heparan and dermatan sulfate, causing severe neurodegeneration, skeletal disease, and cardiorespiratory disease. Most patients manifest with cognitive symptoms, which cannot be treated with enzyme replacement therapy, a...

Cell proliferation, metabolism, migration and survival are coordinated through the tight control of two target of rapamycin (TOR) kinase complexes: TORC1 and TORC2. Here, we show that a novel phosphorylation of fission yeast Gad8 (AGC kinase) on the evolutionarily conserved threonine 6 (Thr6) prevents the physical association between Gad8 and TORC2...

Aicardi-Goutières syndrome is an inflammatory disease occurring due to mutations in any of TREX1, RNASEH2A, RNASEH2B, RNASEH2C, SAMHD1, ADAR or IFIH1. We report on 374 patients from 299 families with mutations in these seven genes. Most patients conformed to one of two fairly stereotyped clinical profiles; either exhibiting an in utero disease-onse...

Cell growth and cell-cycle progression are tightly coordinated to enable cells to adjust their size (timing of division) to the demands of proliferation in varying nutritional environments. In fission yeast, nitrogen stress results in sustained proliferation at a reduced size. Here, we show that cells can sense nitrogen stress to reduce target of r...

The type I interferon system is integral to human antiviral immunity. However, inappropriate stimulation or defective negative regulation of this system can lead to inflammatory disease. We sought to determine the molecular basis of genetically uncharacterized cases of the type I interferonopathy Aicardi-Goutières syndrome and of other undefined ne...

We recently observed mutations in ADAR1 to cause a phenotype of bilateral striatal necrosis (BSN) in a child with the type I interferonopathy Aicardi-Goutières syndrome (AGS). We therefore decided to screen patients with apparently non-syndromic BSN for ADAR1 mutations, and for an upregulation of interferon-stimulated genes (ISGs). We performed San...

Aicardi-Goutières syndrome (AGS) is an inflammatory disorder caused by mutations in any of six genes (TREX1, RNASEH2A, RNASEH2B, RNASEH2C, SAMHD1, and ADAR). The disease is severe and effective treatments are urgently needed. We investigated the status of interferon-related biomarkers in patients with AGS with a view to future use in diagnosis and...

ABSTRACT Aicardi-Goutières syndrome (AGS) is an inflammatory disorder resulting from mutations in TREX1, RNASEH2A/2B/2C, SAMHD1 or ADAR1. Here we provide molecular, biochemical and cellular evidence for the pathogenicity of two synonymous variants in RNASEH2A. Firstly, the c.69G>A (p.Val23Val) mutation causes the formation of a splice donor site wi...

Adenosine deaminases acting on RNA (ADARs) catalyze the hydrolytic deamination of adenosine to inosine in double-stranded RNA (dsRNA) and thereby potentially alter the information content and structure of cellular RNAs. Notably, although the overwhelming majority of such editing events occur in transcripts derived from Alu repeat elements, the biol...

Plasmids used in this study. (PDF)

Quantification of CPY translocation in the presence and absence of MetAP activity. Pulse-labelling of WT (MK) CPY and mutants with A, C, E, G, and S inserted at P2 was performed in wild-type (MAP1 Δprc1 Δpep4) and Δmap1 (Δprc1 Δpep4) yeast cells in the presence and absence of the Map2 inhibitor fumagillin. CPY was immunoprecipitated and analysed by...

N-acetylation of ppαF blocks translocation in vitro. Wild-type (MR), MSR, and MSRR ppαF were translated in vitro in rabbit reticulocyte lysate and then incubated with yeast microsomes (yRM). Position of non-translocated (ppαF) and signal-sequence cleaved, glycosylated (g-pαF) are indicated. (*) Ubiquitinylated ppαF generated in the absence of micro...

DHC-αF translocation is insensitive to a P2 residue that can promote N-acetylation. (A) DHC-αF comprises ppαF with the hydrophobic core of the signal sequence replaced with that of DPAP B, creating an SRP-dependent substrate. DHC-αF with the endogenous P2 residue (MR) or with a serine inserted at position 2 (MS) were translated in vitro in a yeast...

Peak hydrophobicity analysis of Yeast Signal Sequences. Mean peak hydrophobicity of yeast signal sequences group according to their predicted N-terminal processing. Peak hydrophobicity determined based on Kyte-Doolittle [57] with a window size of 11. The “acetylated,” “methionine cleaved not acetylated,” and “non-processed” groups had mean peak hyd...

Relative amino acid frequency at position 2 by compartment in yeast. (PDF)

Predicted relative frequency of N-terminal methionine cleavage. (PDF)

N-terminal sequence and predicted processing of yeast signal sequences. (PDF)

Relative P2 frequency of signal sequences from different organisms. (PDF)

Oligonucleotides used in this study. (PDF)

Supporting methods. (PDF)

MS-pPdi1p is Methionine-cleaved and N-acetylated in vivo. MS-pPdi1p-myc was affinity purified from yeast cells with anti-myc antiserum and analysed by SDS-PAGE and staining with Coomassie brilliant blue (Text S1). The MS-pPdi1p-myc precursor band was excised, digested with elastase, and analysed by LC-MS/MS (Text S1). Product ion spectra and associ...

N-terminal sequence and predicted processing of cytosolic proteins. (PDF)

Predicted frequency of N-terminal processing of signal sequences from different organisms. (PDF)

Yeast strains used in this study. (PDF)

Amino-terminal acetylation is probably the most common protein modification in eukaryotes with as many as 50%-80% of proteins reportedly altered in this way. Here we report a systematic analysis of the predicted N-terminal processing of cytosolic proteins versus those destined to be sorted to the secretory pathway. While cytosolic proteins were pro...

Misfolded proteins in the endoplasmic reticulum (ER) are exported to the cytosol for degradation by the proteasome in a process known as ER-associated degradation (ERAD). CPY* is a well characterized ERAD substrate whose degradation is dependent upon the Hrd1 complex. However, although the functions of some of the components of this complex are kno...

Misfolded proteins in the endoplasmic reticulum (ER) are exported to the cytosol for degradation by the proteasome in a process known as ER-associated degradation (ERAD). CPY* is a well characterized ERAD substrate whose degradation is dependent upon the Hrd1 complex. However, although the functions of some of the components of this complex are kno...