ABSTRACT
BACKGROUND: Fatigue is a prevalent and debilitating symptom in neurological disorders, including spinocerebellar ataxias (SCAs). However, the risk factors of fatigue in the SCAs as well as its impact have not been well investigated. OBJECTIVES: To study the prevalence of fatigue in SCAs, the factors contributing to fatigue, and the influence of fatigue on quality of life. METHODS: Fatigue was assessed in 418 participants with SCA1, SCA2, SCA3, and SCA6 from the Clinical Research Consortium for the Study of Cerebellar Ataxia using the Fatigue Severity Scale. We conducted multi-variable linear regression models to examine the factors contributing to fatigue as well as the association between fatigue and quality of life. RESULTS: Fatigue was most prevalent in SCA3 (52.6%), followed by SCA1 (36.7%), SCA6 (35.7%), and SCA2 (35.6%). SCA cases with fatigue had more severe ataxia and worse depressive symptoms. In SCA3, those with fatigue had a longer disease duration and longer pathological CAG repeat numbers. In multi-variable models, depressive symptoms, but not ataxia severity, were associated with more severe fatigue. Fatigue, independent of ataxia and depression, contributed to worse quality of life in SCA3 and SCA6 at baseline, and fatigue continued affecting quality of life throughout the disease course in all types of SCA. CONCLUSIONS: Fatigue is a common symptom in SCAs and is closely related to depression. Fatigue significantly impacts patients' quality of life. Therefore, screening for fatigue should be considered a part of standard clinical care for SCAs.
Subject(s)
Fatigue , Quality of Life , Spinocerebellar Ataxias , Humans , Quality of Life/psychology , Spinocerebellar Ataxias/psychology , Spinocerebellar Ataxias/complications , Spinocerebellar Ataxias/genetics , Spinocerebellar Ataxias/epidemiology , Male , Fatigue/psychology , Fatigue/epidemiology , Female , Middle Aged , Adult , Aged , Severity of Illness Index , Prevalence , Depression/epidemiology , Depression/psychologyABSTRACT
BACKGROUND: Genetic prion diseases, including Gerstmann-Sträussler-Scheinker disease (GSS), are extremely rare, fatal neurodegenerative disorders, often associated with progressive ataxia and cognitive/neuropsychiatric symptoms. GSS typically presents as a rapidly progressive cerebellar ataxia, associated with cognitive decline. Late-onset cases are rare. OBJECTIVE: To compare a novel GSS phenotype with six other cases and present pathological findings from a single case. METHODS: Case series of seven GSS patients, one proceeding to autopsy. RESULTS: Case 1 developed slowly progressive gait difficulties at age 71, mimicking a spinocerebellar ataxia, with a family history of balance problems in old age. Genome sequencing revealed a heterozygous c.392G > A (p.G131E) pathogenic variant and a c.395A > G resulting in p.129 M/V polymorphism in the PRNP gene. Probability analyses considering family history, phenotype, and a similar previously reported point mutation (p.G131V) suggest p.G131E as a new pathogenic variant. Clinical features and imaging of this case are compared with those six additional cases harboring p.P102L mutations. Autopsy findings of a case are described and were consistent with the prion pathology of GSS. CONCLUSIONS: We describe a patient with GSS with a novel p.G131E mutation in the PRNP gene, presenting with a late-onset, slowly progressive phenotype, mimicking a spinocerebellar ataxia, and six additional cases with the typical P102L mutation.
Subject(s)
Cerebellar Ataxia , Gerstmann-Straussler-Scheinker Disease , Prions , Spinocerebellar Ataxias , Humans , Aged , Gerstmann-Straussler-Scheinker Disease/diagnosis , Prion Proteins/genetics , Prions/genetics , Cerebellar Ataxia/complications , Spinocerebellar Ataxias/diagnosisABSTRACT
Atherosclerosis is a medical condition that causes buildup of plaque in the blood vessels and narrowing of the arteries. Surgeons often treat this condition through angioplasty with catheter placements. Continuum guidewire robots offer significant advantages for catheter placements due to their dexterity. Tracking these guidewire robots and their surrounding workspace under fluoroscopy in real-time can be useful for visualization and accurate control. This paper discusses algorithms and methods to track the shape and orientation of the guidewire and the surrounding workspaces of phantom vasculatures in real-time under C-arm fluoroscopy. The shape of continuum guidewires is found through a semantic segmentation architecture based on MobileNetv2 with a Tversky loss function to deal with class imbalances. This shape is refined through medial axis filtering and parametric curve fitting to quantitatively describe the guidewire's pose. Using a constant curvature assumption for the guidewire's bending segments, the parameters that describe the joint variables are estimated in real-time for a tendon-actuated COaxially Aligned STeerable (COAST) guidewire robot and tracked through its traversal of an aortic bifurcation phantom. The accuracy of the tracking is ~90% and the execution times are within 100ms, and hence this method is deemed suitable for real-time tracking.
ABSTRACT
Cardiovascular diseases are the leading cause of death globally and surgical treatments for these often begin with the manual placement of a long compliant wire, called a guidewire, through different vasculature. To improve procedure outcomes and reduce radiation exposure, we propose steps towards a fully automated approach for steerable guidewire navigation within vessels. In this paper, we utilize fluoroscopic images to fully reconstruct 3-D printed phantom vasculature models by using a shape-from-silhouette algorithm. The reconstruction is subsequently de-noised using a deep learning-based encoder-decoder network and morphological filtering. This volume is used to model the environment for guidewire traversal. Following this, we present a novel method to plan an optimal path for guidewire traversal in three-dimensional vascular models through the use of slice planes and a modified hybrid A-star algorithm. Finally, the developed reconstruction and planning approaches are applied to an ex vivo porcine aorta, and navigation is demonstrated through the use of a tendon-actuated COaxially Aligned STeerable guidewire (COAST).
ABSTRACT
Multiple system atrophy (MSA) is a fatal neurodegenerative disease of unknown etiology characterized by widespread aggregation of the protein alpha-synuclein in neurons and glia. Its orphan status, biological relationship to Parkinson's disease (PD), and rapid progression have sparked interest in drug development. One significant obstacle to therapeutics is disease heterogeneity. Here, we share our process of developing a clinical trial-ready cohort of MSA patients (69 patients in 2 years) within an outpatient clinical setting, and recruiting 20 of these patients into a longitudinal "n-of-few" clinical trial paradigm. First, we deeply phenotype our patients with clinical scales (UMSARS, BARS, MoCA, NMSS, and UPSIT) and tests designed to establish early differential diagnosis (including volumetric MRI, FDG-PET, MIBG scan, polysomnography, genetic testing, autonomic function tests, skin biopsy) or disease activity (PBR06-TSPO). Second, we longitudinally collect biospecimens (blood, CSF, stool) and clinical, biometric, and imaging data to generate antecedent disease-progression scores. Third, in our Mass General Brigham SCiN study (stem cells in neurodegeneration), we generate induced pluripotent stem cell (iPSC) models from our patients, matched to biospecimens, including postmortem brain. We present 38 iPSC lines derived from MSA patients and relevant disease controls (spinocerebellar ataxia and PD, including alpha-synuclein triplication cases), 22 matched to whole-genome sequenced postmortem brain. iPSC models may facilitate matching patients to appropriate therapies, particularly in heterogeneous diseases for which patient-specific biology may elude animal models. We anticipate that deeply phenotyped and genotyped patient cohorts matched to cellular models will increase the likelihood of success in clinical trials for MSA.
ABSTRACT
Impaired cerebellar Purkinje neuron firing resulting from reduced expression of large-conductance calcium-activated potassium (BK) channels is a consistent feature in models of inherited neurodegenerative spinocerebellar ataxia (SCA). Restoring BK channel expression improves motor function and delays cerebellar degeneration, indicating that BK channels are an attractive therapeutic target. Current BK channel activators lack specificity and potency and are therefore poor templates for future drug development. We implemented an automated patch clamp platform for high-throughput drug discovery of BK channel activators using the Nanion SyncroPatch 384PE system. We screened over 15,000 compounds for their ability to increase BK channel current amplitude under conditions of lower intracellular calcium that is present in disease. We identified several novel BK channel activators that were then retested on the SyncroPatch 384PE to generate concentration-response curves (CRCs). Compounds with favorable CRCs were subsequently tested for their ability to improve irregular cerebellar Purkinje neuron spiking, characteristic of BK channel dysfunction in SCA1 mice. We identified a novel BK channel activator, 4-chloro-N-(5-chloro-2-cyanophenyl)-3-(trifluoromethyl)benzene-1-sulfonamide (herein renamed BK-20), that exhibited a more potent half-maximal activation of BK current (pAC50 = 4.64) than NS-1619 (pAC50 = 3.7) at a free internal calcium concentration of 270 nM in a heterologous expression system and improved spiking regularity in SCA1 Purkinje neurons. BK-20 had no activity on small-conductance calcium-activated potassium (SK)1-3 channels but interestingly was a potent blocker of the T-type calcium channel, Cav3.1 (IC50 = 1.05 µM). Our work describes both a novel compound for further drug development in disorders with irregular Purkinje spiking and a unique platform for drug discovery in degenerative ataxias. SIGNIFICANCE STATEMENT: Motor impairment associated with altered Purkinje cell spiking due to dysregulation of large-conductance calcium-activated potassium (BK) channel expression and function is a shared feature of disease in many degenerative ataxias. BK channel activators represent an outstanding therapeutic agent for ataxia. We have developed a high-throughput platform to screen for BK channel activators and identified a novel compound that can serve as a template for future drug development for the treatment of these disabling disorders.
Subject(s)
Cerebellar Ataxia , Potassium Channels, Calcium-Activated , Spinocerebellar Ataxias , Animals , Ataxia , Calcium/metabolism , Cerebellar Ataxia/drug therapy , Large-Conductance Calcium-Activated Potassium Channels , Mice , Potassium/metabolism , Spinocerebellar Ataxias/metabolismABSTRACT
The purpose of this study was to determine whether a regular follow-up schedule with examination by clinicians results in a better detection rate of disease recurrence and eventual better clinical outcomes when compared to patients who present with symptoms to the clinic and are subsequently detected to have a recurrence of oral squamous cell carcinoma. Retrospective data from 642 patients who underwent treatment for oral squamous cell carcinoma at a tertiary level cancer centre were analysed. Of the 642 patients, 197 had recurrences of which 108 were detected on regular follow-up and 87 were detected in patients presenting out of schedule with symptoms; two patients were detected to have recurrence at another centre, but their mode of detection could not be ascertained. There was no difference in the loco-regional recurrence-free survival or disease-free survival between the two groups. A strict follow-up schedule in the first year followed by a more flexible symptom-based schedule in the subsequent years, with supplementation of imaging if clinically indicated, should be an adequate surveillance plan for oral cancer patients.
Subject(s)
Carcinoma, Squamous Cell , Head and Neck Neoplasms , Mouth Neoplasms , Carcinoma, Squamous Cell/diagnostic imaging , Carcinoma, Squamous Cell/therapy , Feasibility Studies , Follow-Up Studies , Humans , Mouth Neoplasms/diagnostic imaging , Mouth Neoplasms/pathology , Mouth Neoplasms/therapy , Neoplasm Recurrence, Local/pathology , Retrospective Studies , Squamous Cell Carcinoma of Head and NeckABSTRACT
BACKGROUND: ANAT is a Cytoscape plugin for the inference of functional protein-protein interaction networks in yeast and human. It is a flexible graphical tool for scientists to explore and elucidate the protein-protein interaction pathways of a process under study. RESULTS: Here we present ANAT3.0, which comes with updated PPI network databases of 544,455 (human) and 155,504 (yeast) interactions, and a new machine-learning layer for refined network elucidation. Together they improve network reconstruction to more than twofold increase in the quality of reconstructing known signaling pathways from KEGG. CONCLUSIONS: ANAT3.0 includes improved network reconstruction algorithms and more comprehensive protein-protein interaction networks than previous versions. ANAT is available for download on the Cytoscape Appstore and at https://www.cs.tau.ac.il/~bnet/ANAT/ .
Subject(s)
Proteins , Software , Algorithms , Humans , Machine Learning , Protein Interaction Mapping , Protein Interaction Maps , Proteins/genetics , Proteins/metabolismABSTRACT
Mitral regurgitation (MR) is a condition caused by a deformity in the mitral valve leading to the backflow of blood into the left atrium. MR can be treated through a minimally invasive procedure and our lab is currently developing a robot that could potentially be used to treat MR. The robot would carry a clip that latches onto the valve's leaflets and closes them to minimize leakage. The robot's accurate localization is needed to navigate the clip to the leaflets successfully. This paper discusses algorithms used to track the clip's position and orientation under real-time using C-arm fluoroscopy. The positions are found through a deep learning semantic segmentation framework and the pose is found by calculating its bending and rotational angles. The robot's bending angle and the clip's rotational angle is found through an equivalent ellipse algorithm and an SVM classifier, respectively, and were validated by comparing orientations obtained from an electromagnetic tracker. The bending angle calculation has an average error of 7.7° and the rotational angle calculation is 76% for classifying them into five classes. Execution times are within 100ms and hence this could be a promising approach in real-time pose estimation.
ABSTRACT
The spinocerebellar ataxias (SCAs) are a group of neurodegenerative disorders inherited in an autosomal dominant fashion. The SCAs result in progressive gait imbalance, incoordination of the limbs, speech changes, and oculomotor dysfunction, among other symptoms. Over the past few decades, significant strides have been made in understanding the pathogenic mechanisms underlying these diseases. Although multiple efforts using a combination of genetics and pharmacology with small molecules have been made towards developing new therapeutics, no FDA approved treatment currently exists. In this review, we focus on SCA1, a common SCA subtype, in which some of the greatest advances have been made in understanding disease biology, and consequently potential therapeutic targets. Understanding of the underlying basic biology and targets of therapy in SCA1 is likely to give insight into treatment strategies in other SCAs. The diversity of the biology in the SCAs, and insight from SCA1 suggests, however, that both shared treatment strategies and specific approaches tailored to treat distinct genetic causes of SCA are likely needed for this group of devastating neurological disorders.
Subject(s)
Ataxin-1/genetics , Clinical Trials as Topic/methods , Drug Delivery Systems/trends , Gene Targeting/trends , Spinocerebellar Ataxias/genetics , Spinocerebellar Ataxias/therapy , Animals , Ataxin-1/antagonists & inhibitors , Ataxin-1/metabolism , Drug Delivery Systems/methods , Excitatory Amino Acid Agents/administration & dosage , Excitatory Amino Acid Agents/metabolism , Gene Targeting/methods , Genetic Therapy/methods , Genetic Therapy/trends , Humans , Oligonucleotides, Antisense/administration & dosage , Oligonucleotides, Antisense/genetics , Oligonucleotides, Antisense/metabolism , RNA Interference/drug effects , RNA Interference/physiology , Spinocerebellar Ataxias/metabolismABSTRACT
Non-small cell lung cancer (NSCLC) is a major form of cancer and is resistant to chemo- and radio-therapy. Vicenin-2 (VCN-2) is a flavonoid obtained from Ocimum sanctum L. and it has been reported to have radioprotective and anti-cancer properties. This study was conducted to check for the radiosensitizing potential of VCN-2 in the NSCLC cell line, NCI-H23. NCI-H23 cells were exposed to VCN-2 singularly, and to X-rays with and without prior VCN-2 treatment. Cytotoxicity assay, cell proliferation assay, caspase-3 activity assay, DNA fragmentation assay and Western blotting for Rad50, MMP-2 and p21 were performed to investigate the radiosensitizing properties of VCN-2. Fibroblast survival assay was performed using HEK293T cells to check for any adverse effects of VCN-2 on normal fibroblast cell line. VCN-2 singularly and in combination with radiation reduced the surviving cancer cells, increased caspase-3 activity, increased DNA fragmentation, increased the levels of Rad50 and lowered levels of MMP-2 and p21 proteins while being non-toxic and radioprotective to the fibroblast cells. VCN-2 showed a potent radiosensitizing property while also showing a chemotherapeutic property against NSCLC cell line NCI-H23.
Subject(s)
Apigenin/pharmacology , Carcinoma, Non-Small-Cell Lung/metabolism , Glucosides/pharmacology , Lung Neoplasms/metabolism , Radiation-Sensitizing Agents/pharmacology , Acid Anhydride Hydrolases , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/therapy , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Proliferation/radiation effects , Cell Survival/drug effects , Cell Survival/radiation effects , Cyclin-Dependent Kinase Inhibitor p21/metabolism , DNA Repair Enzymes/metabolism , DNA-Binding Proteins/metabolism , Gene Expression Regulation, Neoplastic/drug effects , Gene Expression Regulation, Neoplastic/radiation effects , HEK293 Cells , Humans , Lung Neoplasms/genetics , Lung Neoplasms/therapy , Matrix Metalloproteinase 2/metabolismABSTRACT
Heat shock protein 70 (Hsp70) and Hsp90 are molecular chaperones that play essential roles in tumor growth by stabilizing pro-survival client proteins. However, although the development of Hsp90 inhibitors has benefited from the identification of clients, such as Raf-1 proto-oncogene, Ser/Thr kinase (RAF1), that are particularly dependent on this chaperone, no equivalent clients for Hsp70 have been reported. Using chemical probes and MDA-MB-231 breast cancer cells, we found here that the inhibitors of apoptosis proteins, including c-IAP1 and X-linked inhibitor of apoptosis protein (XIAP), are obligate Hsp70 clients that are rapidly (within â¼3-12 h) lost after inhibition of Hsp70 but not of Hsp90. Mutagenesis and pulldown experiments revealed multiple Hsp70-binding sites on XIAP, suggesting that it is a direct, physical Hsp70 client. Interestingly, this interaction was unusually tight (â¼260 nm) for an Hsp70-client interaction and involved non-canonical regions of the chaperone. Finally, we also found that Hsp70 inhibitor treatments caused loss of c-IAP1 and XIAP in multiple cancer cell lines and in tumor xenografts, but not in healthy cells. These results are expected to significantly accelerate Hsp70 drug discovery by providing XIAP as a pharmacodynamic biomarker. More broadly, our findings further suggest that Hsp70 and Hsp90 have partially non-overlapping sets of obligate protein clients in cancer cells.
Subject(s)
HSP70 Heat-Shock Proteins/metabolism , X-Linked Inhibitor of Apoptosis Protein/metabolism , Biomarkers/metabolism , Cell Line, Tumor , HSP70 Heat-Shock Proteins/genetics , HSP90 Heat-Shock Proteins/genetics , HSP90 Heat-Shock Proteins/metabolism , Humans , Protein Binding , Proto-Oncogene Mas , X-Linked Inhibitor of Apoptosis Protein/geneticsABSTRACT
Hsp70 is a molecular chaperone that binds to "client" proteins and protects them from protein degradation. Hsp70 is essential for the survival of many cancer cells, but it is not yet clear which of its clients are involved. Using structurally distinct chemical inhibitors, we found that many of the well-known clients of the related chaperone, Hsp90, are not strikingly responsive to Hsp70 inhibition. Rather, Hsp70 appeared to be important for the stability of the RIP1 (RIPK1) regulators: cIAP1/2 (BIRC1 and BIRC3), XIAP, and cFLIPS/L (CFLAR). These results suggest that Hsp70 limits apoptosis and necroptosis pathways downstream of RIP1. Consistent with this model, MDA-MB-231 breast cancer cells treated with Hsp70 inhibitors underwent apoptosis, while cotreatment with z-VAD.fmk switched the cell death pathway to necroptosis. In addition, cell death in response to Hsp70 inhibitors was strongly suppressed by RIP1 knockdown or inhibitors. Thus, these data indicate that Hsp70 plays a previously unrecognized and important role in suppressing RIP1 activity.Implications: These findings clarify the role of Hsp70 in prosurvival signaling and suggest IAPs as potential new biomarkers for Hsp70 inhibition. Mol Cancer Res; 16(1); 58-68. ©2017 AACR.
Subject(s)
Breast Neoplasms/pathology , HSP70 Heat-Shock Proteins/metabolism , Receptor-Interacting Protein Serine-Threonine Kinases/metabolism , Apoptosis/physiology , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Cell Line, Tumor , Female , HSP70 Heat-Shock Proteins/antagonists & inhibitors , HSP70 Heat-Shock Proteins/genetics , Humans , Jurkat Cells , MCF-7 Cells , Necrosis , Receptor-Interacting Protein Serine-Threonine Kinases/genetics , Signal TransductionABSTRACT
Chronic obstructive pulmonary disease is a major health problem becoming a leading cause of morbidity and mortality worldwide. A large part of these disorders is associated with acute exacerbations resulting from infection by bacteria, such as non-typeable Haemophilus influenzae (NTHi). Our understanding of the pathogenesis of these exacerbations is still elusive. We demonstrate herein that NTHi infection of mice chronically exposed to cigarette smoke (CS), an experimental model of chronic obstructive pulmonary disease (COPD), not only causes acute pulmonary inflammation but also impairs the production of interleukin (IL)-22, a cytokine with potential anti-bacterial activities. We also report that mice lacking IL-22, as well as mice exposed to CS, have a delayed clearance of NTHi bacteria and display enhanced alveolar wall thickening and airway remodeling compared with controls. Supplementation with IL-22 not only boosted bacterial clearance and the production of anti-microbial peptides but also limited lung damages induced by infection both in IL-22-/- and CS-exposed mice. In vitro exposure to CS extract altered the NTHi-induced IL-22 production by spleen cells. This study shows for the first time that a defect in IL-22 is involved in the acute exacerbation induced by NTHi infection during experimental COPD and opens the way to innovative therapeutic strategies.
Subject(s)
Haemophilus Infections/immunology , Haemophilus influenzae/immunology , Interleukins/metabolism , Lung/immunology , Pulmonary Disease, Chronic Obstructive/immunology , Airway Remodeling , Animals , Bacterial Load , Cells, Cultured , Disease Models, Animal , Humans , Interleukins/genetics , Lung/microbiology , Lung/pathology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Pulmonary Disease, Chronic Obstructive/microbiology , Smoking/adverse effects , Interleukin-22ABSTRACT
Heat shock protein 70 (Hsp70) is a chaperone that normally scans the proteome and initiates the turnover of some proteins (termed clients) by linking them to the degradation pathways. This activity is critical to normal protein homeostasis, yet it appears to fail in diseases associated with abnormal protein accumulation. It is not clear why Hsp70 promotes client degradation under some conditions, while sparing that protein under others. Here, we used a combination of chemical biology and genetic strategies to systematically perturb the affinity of Hsp70 for the model client, tau. This approach revealed that tight complexes between Hsp70 and tau were associated with enhanced turnover while transient interactions favored tau retention. These results suggest that client affinity is one important parameter governing Hsp70-mediated quality control.
Subject(s)
Benzothiazoles/pharmacology , HSP70 Heat-Shock Proteins/metabolism , Models, Biological , Tauopathies/drug therapy , Tauopathies/metabolism , Thiazolidines/pharmacology , tau Proteins/metabolism , Benzothiazoles/chemistry , Dose-Response Relationship, Drug , HSP70 Heat-Shock Proteins/chemistry , HeLa Cells , Humans , Molecular Structure , Protein Stability/drug effects , Structure-Activity Relationship , Thiazolidines/chemistry , Tumor Cells, Cultured , tau Proteins/chemistryABSTRACT
Background Selective neck dissection in multimodality treatment protocols is slowly being accepted for the management of N+ neck in many centers. This is because the functional disability is lower than modified radical neck dissection. Objective This study compares the regional recurrence rates between patients who underwent selective neck dissection and patients underwent comprehensive neck dissection for node positive oral squamous cell carcinoma. Method A retrospective study comparing patients with node positive oral squamous cell carcinoma who underwent either selective neck dissection or comprehensive neck dissection between August 2011 and January 2014 was done, with a mean follow up period of 12 months. Regional failures were assessed to whether they were isolated neck failures or associated with a local or distant failure. Result A total of 131 neck dissections were performed which included 93 selective neck dissections and 38 comprehensive neck dissections. A total of 17 patients developed regional recurrence, of which 11 patients had ipsilateral neck recurrence. Of the 11 patients with ipsilateral neck recurrence one patient also had contralateral neck nodes and in two patients there was associated distant metastasis. Conclusion Selective neck dissection for management of node positive neck disease is based on sound scientific principles and a randomised controlled trial comparing it with modified radical neck dissection would probably give the answer regarding the optimal procedure for these patients.
Subject(s)
Carcinoma, Squamous Cell/surgery , Mouth Neoplasms/surgery , Neck Dissection/methods , Adult , Aged , Carcinoma, Squamous Cell/pathology , Female , Follow-Up Studies , Humans , Male , Middle Aged , Mouth Neoplasms/pathology , Neoplasm Metastasis , Neoplasm Recurrence, Local , Retrospective Studies , Treatment OutcomeABSTRACT
BACKGROUND: Gene expression studies are increasingly used to provide valuable information on the diagnosis and prognosis of human cancers. Also, for in vitro and in vivo experimental cancer models gene expression studies are widely used. The complex algorithms of differential gene expression analyses require normalization of data against a reference or normalizer gene, or a set of such genes. For this purpose, mostly invariant housekeeping genes are used. Unfortunately, however, there are no consensus (housekeeping) genes that serve as reference or normalizer for different human cancers. In fact, scientists have employed a wide range of reference genes across different types of cancer for normalization of gene expression data. As a consequence, comparisons of these data and/or data harmonizations are difficult to perform and challenging. In addition, an inadequate choice for a reference gene may obscure genuine changes and/or result in erroneous gene expression data comparisons. METHODS: In our effort to highlight the importance of selecting the most appropriate reference gene(s), we have screened the literature for gene expression studies published since the turn of the century on thirteen of the most prevalent human cancers worldwide. CONCLUSIONS: Based on the analysis of the data at hand, we firstly recommend that in each study the suitability of candidate reference gene(s) should carefully be evaluated in order to yield reliable differential gene expression data. Secondly, we recommend that a combination of PPIA and either GAPDH, ACTB, HPRT and TBP, or appropriate combinations of two or three of these genes, should be employed in future studies, to ensure that results from different studies on different human cancers can be harmonized. This approach will ultimately increase the depth of our understanding of gene expression signatures across human cancers.
Subject(s)
Gene Expression Profiling/standards , Genes, Essential , Neoplasms/genetics , Reference Standards , Algorithms , HumansABSTRACT
Previously, we have shown that shortening of telomeres by telomerase inhibition sensitized cancer cells to cisplatinum, slowed their migration, increased DNA damage and impaired DNA repair. The mechanism behind these effects is not fully characterized. Its clarification could facilitate novel therapeutics development and may obviate the time consuming process of telomere shortening achieved by telomerase inhibition. Here we aimed to decipher the microRNA and proteomic profiling of cancer cells with shortened telomeres and identify the key mediators in telomere shortening-induced damage to those cells. Of 870 identified proteins, 98 were differentially expressed in shortened-telomere cells. 47 microRNAs were differentially expressed in these cells; some are implicated in growth arrest or act as oncogene repressors. The obtained data was used for a network construction, which provided us with nodal candidates that may mediate the shortened-telomere dependent features. These proteins' expression was experimentally validated, supporting their potential central role in this system.