Kimberley Louise Alexander-Kaufman

Chris O’Brien Lifehouse · Neurosurgery

Extracellular vesicles (EVs) play key roles in glioblastoma (GBM) biology and represent novel sources of biomarkers that are detectable in the peripheral circulation. Despite this notionally non-invasive approach to assess GBM tumours in situ, a comprehensive GBM EV protein signature has not been described. Here, EVs secreted by six GBM cell lines...

Glioblastoma (GBM) tumour invasion is facilitated by cell migration and degradation of the extracellular matrix (ECM). Invadopodia are actin-rich structures that protrude from the plasma membrane in direct contact with the ECM and are proposed to participate in epithelial-mesenchymal-transition. We have characterised the invasiveness of nine establ...

Outcomes for melanoma patients with stage III disease, differ widely even within the same sub-category. Molecular signatures that more accurately predict prognosis are needed to stratify patients according to risk. Proteomic analyses were used to identify differentially abundant proteins in extracts of surgically excised samples from patients with...

One of the sequelae of chronic alcohol abuse is malnutrition. Importantly, a deficiency in thiamine (vitamin B(1)) can result in the acute, potentially reversible neurological disorder Wernicke encephalopathy (WE). When WE is recognized, thiamine treatment can elicit a rapid clinical recovery. If WE is left untreated, however, patients can develop...

Extracellular vesicles (EVs) play key roles in glioblastoma (GBM; astrocytoma grade IV) biology and are novel sources of biomarkers. EVs released from GBM tumors can cross the blood-brain-barrier into the periphery carrying GBM molecules, including small non-coding RNA (sncRNA). Biomarkers cargoed in circulating EVs have shown great promise for ass...

Improving outcomes for diffuse glioma patients requires methods that can accurately and sensitively monitor tumour activity and treatment response. Extracellular vesicles (EV) are membranous nanoparticles that can traverse the blood–brain-barrier, carrying oncogenic molecules into the circulation. Measuring clinically relevant glioma biomarkers car...

Cutaneous squamous cell carcinoma (cSCC) and its precursors, actinic keratosis (AK) and Bowen’s disease (BD), are the most common types of keratinocytic skin lesions (KSL) which account for the majority of non-melanoma skin cancer lethality. Currently, clinical and histopathological criteria are used for the diagnosis, classification and therapeuti...

There is a real need for biomarkers that can indicate glioma disease burden and inform clinical management, particularly in the recurrent glioblastoma (GBM; grade IV glioma) setting where treatment-associated brain changes can confound current and expensive tumour surveillance methods. In this regard, extracellular vesicles (EVs; 30-1000 nm membran...

Actinic keratosis, Bowen's disease and cutaneous squamous cell carcinoma (cSCC) are heterogeneous keratinocytic skin lesions. Biomarkers that can accurately stratify these lesion types are needed to support a new paradigm of personalized and precise management of skin neoplasia. In this paper, we used a data independent acquisition proteomics workf...

Diffuse gliomas (grades II–IV) are amongst the most frequent and devastating primary brain tumours of adults. Currently, patients are monitored by clinical examination and radiographic imaging, which can be challenging to interpret and insensitive to early signs of treatment failure and tumour relapse. While brain biopsy and histologic analysis can...

Exosomes are nano-sized extracellular vesicles released by many cells that contain molecules characteristic of their cell of origin, including microRNA. Exosomes released by glioblastoma cross the blood–brain barrier into the peripheral circulation and carry molecular cargo distinct to that of “free-circulating” miRNA. In this pilot study, serum ex...

Glioblastoma, WHO‐grade IV glioma, carries a dismal prognosis owing to its infiltrative growth and limited treatment options. Glioblastoma‐derived extracellular vesicles (EVs; 30–1000nm membranous particles) influence the microenvironment to mediate tumour aggressiveness and carry oncogenic cargo across the blood‐brain‐barrier into the circulation....

Therapeutic dendritic cell vaccination for GBM using primary blood dendritic cells (BDC) has the potential to improve response rates compared with more commonly used monocyte-derived dendritic cells due to their superior antigen-presentation and migration. Combination of vaccination with immune checkpoint inhibitors such as anti-PD-1 antibodies may...

Cancer vaccines represent a novel and promising approach for aggressive neoplasms such as glioblastoma multiforme (GBM) where other treatment modalities have not been effective. In this regard, cancer vaccines have been developed which exploit the key central immunoregulatory dendritic cell (DC) to maximize vaccine efficacy. Such “DC vaccine” strat...

The role of astrocytes is becoming increasingly important to understanding how glioblastoma (GBM) tumor cells diffusely invade the brain. Yet, little is known of the contribution of extracellular vesicle (EV) signaling in GBM/astrocyte interactions. We modeled GBM-EV signaling to normal astrocytes in vitro to assess whether this mode of intercellul...

PURPOSE : Actinic keratoses (AK) are premalignant tumors that can be clinically difficult to differentiate from skin cancer. An easy, quick and reliable non‐invasive alternative to biopsy is needed to definitively confirm the clinical diagnoses. This study evaluates Tape Stripping (TS) of stratum corneum (SC) for non‐invasive biomarker analysis of...

Background: Exosomes are nano-sized extracellular vesicles released by many cells that contain molecules characteristic of their cell-of-origin, including microRNA. Exosomes released by glioblastoma cross the blood-brain-barrier into the peripheral circulation, and carry molecular cargo distinct to that of free-circulating miRNA. Methods: Serum exo...

Background: The boundaries between actinic keratosis (AK), Bowen's disease (BD), and cutaneous squamous cell carcinoma (cSCC) are sometimes not clear. Large-scale proteomic profiling studies of these lesions are also non-existent. Objective: To evaluate proteomic changes between normal epidermis, AK, BD and cSCC that could support a molecular cl...

Background: Cutaneous squamous cell carcinoma (cSCC) is a common type of skin cancer but there are no comprehensive proteomic studies on this entity. Materials and methods: We employed liquid chromatography coupled with tandem mass spectrometry (MS/MS) using formalin-fixed paraffin-embedded (FFPE) cSCC material to study the tumor and normal skin...

An antibody microarray (DotScanTM) has been developed for profiling clinical melanoma specimens. Immobilized antibodies capture live cells expressing corresponding antigens to produce a dot pattern that represents the surface profile or immunophenotype. The unique signatures obtained may correlate with disease subtype, tumor progression, and clinic...

Clinical trials of heat shock protein 90 (Hsp90) inhibitors have been limited by high toxicity. We previously showed that the Hsp90 inhibitor, SNX-7081, synergizes with and restores sensitivity to fludarabine nucleoside (2-FaraA) in human chronic lymphocytic leukemia (CLL) cells with lesions in the p53 pathway (Best OG, et al., Leukemia Lymphoma 53...

Patients with a stable chronic lymphocytic leukemia (CLL) double their blood lymphocyte count in >5 years, but may develop progressive disease with lymphocytes doubling in <12 months. To identify a protein signature for progressive CLL, whole cell extracts of peripheral blood mononuclear cells from CLL patients (n = 27) were screened using iTRAQ (i...

Microparticles (MP) are small membranous particles (100-1000 nm) released under normal steady-state conditions and are thought to provide a communication network between host cells. Previous studies demonstrated that Mycobacterium tuberculosis (M.tb) infection of macrophages increased the release of MPs, and these MPs induced a proinflammatory resp...

Extensive surface profiles of colorectal cancer (CRC) cells and tumour infiltrating lymphocytes (TIL) have been obtained from 45 surgical resection samples. Live cells were captured on an antibody microarray and stained with fluorescently-labeled antibodies. Minimal panels of 11 CRC antigens (CD13, CD24, CD26, CD49d, CD138, CD166, CA-125, CA19-9, E...

Drugs that impair tumour angiogenesis, i.e. therapeutic antibody anti-vascular endothelial growth factor, bevacizumab (BEV), are becoming standard therapy for recurrent GBM, despite having no impact on overall survival times. Resistance to BEV is fatal, and mechanisms are largely unexplored. With access to exceedingly rare fresh-frozen serial GBM t...

s P17.54. Glioblastoma multiforme (GBM) tumours are diffusely infiltrative making surgical resection virtually impossible. Invasion of brain parenchyma is facil-itated by cell migration and degradation of the extracellular matrix (ECM). Invadopodia are actin-rich organelles that protrude from the ventral side of the plasma membrane in direct contac...

Clinico-pathologic characteristics of 33 AJCC stage III melanoma patients and their tumors. Table S2. Melanoma patient demographics, pathology, lymph node recurrence and survival parameters. Table S3. Differentially abundant proteins between poor and good prognosis AJCC stage IIIc melanoma patients identified using DIGE. Table S4. Summary of iTRAQ...

iTRAQ labeling and 2DLC-MS/MS methods.

SRM methods and survival analysis.

Representative protein map from DIGE analysis of proteins extracted from a lymph node melanoma metastasis. Figure S2. Density plot for the percentage of missing peptides for iTRAQ 2DLC-MS/MS data. Figure S3. Summary of identification of all 2031 proteins and 73 differentially abundant proteins identified bu iTRAQ 2DLC-MS/MS. Figure S4. Cellular loc...

DIGE methods and protein function and network analysis.

List of proteins identified by iTRAQ LC-MS/MS including sequence coverage and the number of peptides matched.

There is an urgent need to identify more accurate prognostic biomarkers in melanoma patients, particularly in those with metastatic disease. This study aimed to identify melanoma and leukocyte surface antigens predictive of survival in a prospective series of AJCC stage IIIb/c melanoma patients (n = 29). Live cell suspensions were prepared from mel...

There is an urgent need to identify more accurate prognostic biomarkers in melanoma patients, particularly in those with metastatic disease. This study aimed to identify melanoma and leukocyte surface antigens predictive of survival in a prospective series of AJCC stage IIIb/c melanoma patients (n=29). Live cell suspensions were prepared from melan...

Citation: Alomari M, Mactier S, Kaufman KL, Best OG, Mulligan SP, et al. (2014) Profiling the Lipid Raft Proteome from Human MEC1 Chronic Lymphocytic Leukemia Cells. J Proteomics Bioinform S7: 005. doi:10.4172/jpb.S7-005 Copyright: © 2014 Alomari M, et al. This is an open-access article distributed under the terms of the Creative Commons Attributio...

The proteomic effects of the Hsp90 inhibitor, SNX-7081, have been determined on the p53-mutated B-cell chronic lymphocytic leukemia (CLL) cell line, MEC1. Following SNX-7081 treatment (500 nM, 24 h), 51 proteins changed abundance by more than 2-fold (p<0.05); 7 proteins increased, while 44 proteins decreased. Proteins identified as differentially a...

The proposed anticancer drug LY294002, inhibits phosphoinositide-3 kinase (PI3K) that initiates a signalling pathway often activated in colorectal cancer (CRC). The effects of LY294002 (10 μM, 48 h) on the cytosolic, mitochondrial and nuclear proteomes of human HT-29 CRC cells have been determined using iTRAQ (isobaric tag for relative and absolute...

Tumour invasion and metastasis are often responsible for a high mortality rate in cancer patients. The spread of cancer involves multiple signalling pathways mediated by cell surface proteins and generally results in the loss of cell-cell adhesion leading to cell migration and invasion of distant tissues. Many transformed cells may be eliminated by...

B-RAF, a serine-threonine protein kinase, is one of the three RAF paralogs in humans. B-RAF participates in the RAS-RAF-MEK-ERK pathway, a conserved protein kinase-signalling cascade that is involved in regulating a number of critical cellular functions. Mutated B-RAF is believed to play a crucial role in the development, maintenance and progressio...

An antibody microarray was developed for profiling the surface proteome of melanoma cells, which may facilitate melanoma sub-classification and provide important prognostic information useful in predicting the clinical behavior of the melanoma (e.g., likely sites of metastatic spread), patient outcome and treatment response. Forty-eight antibodies...

Thiamin, or vitamin B1, is crucial for brain function. In its active form, thiamin pyrophosphate (TPP), it is a co-enzyme for several enzymes, including transketolase. Transketolase is an important enzyme in the non-oxidative branch of the pentose phosphate pathway (PPP), a pathway responsible for generating reducing equivalents, which is essential...

Alcohol abuse and dependence are among the most prevalent and costly health problems in the world from both a social and economic viewpoint. A number of large epidemiological studies have consistently demonstrated the pervasive nature of these disorders. Excessive drinking can lead to impairment of cognitive function and structural changes in the b...

The frontal lobes, particularly the prefrontal region, have been of a great interest to researchers examining human behaviour and the origins of medical conditions involving disturbances in cognitive functions. However, further characterisation of this brain region is necessary to help understand the mechanisms of its disturbance in various disease...

Drugs of abuse, including alcohol, can induce dependency formation and/or brain damage in brain regions important for cognition. 'High-throughput' approaches, such as cDNA microarray and proteomics, allow the analysis of global expression profiles of genes and proteins. These technologies have recently been applied to human brain tissue from patien...

Cerebellar changes are commonly associated with alcoholism and chronic alcohol consumption can produce profound impairments in motor functioning and various aspects of cognition. Although the mechanisms underlying alcohol-induced changes in the cerebellar vermis are poorly understood, observations in the alcoholic vermis are thought to be consequen...

Alcoholic patients commonly experience cognitive decline. It is postulated that cognitive dysfunction is caused by an alcohol-induced region-selective brain damage, particularly to the prefrontal region, and grey and white matter may be affected differently. We used a proteomics-based approach to compare protein expression profiles of the dorsolate...

Neuroimaging and post-mortem studies indicate that chronic alcohol use induces global changes in brain morphology, such as cortical and subcortical atrophy. Recent studies have shown that frontal lobe structures are specifically susceptible to alcohol-related brain damage and shrinkage in this area is largely due to a loss of white matter. This may...