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In JoVE (1)
Other Publications (25)
- American Journal of Physiology. Regulatory, Integrative and Comparative Physiology
- American Journal of Physiology. Regulatory, Integrative and Comparative Physiology
- Neuro-oncology
- Physiological Genomics
- AIDS (London, England)
- Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology
- BMC Cancer
- BMC Complementary and Alternative Medicine
- Neurosurgery
- BMC Medical Genomics
- Journal of Neuro-oncology
- Frontiers in Bioscience (Elite Edition)
- American Journal of Physiology. Regulatory, Integrative and Comparative Physiology
- JAMA : the Journal of the American Medical Association
- JAMA : the Journal of the American Medical Association
- Neurosurgical Focus
- Neurosurgery
- Nutrition & Metabolism
- Journal of Neurosurgery
- Journal of Neuropathology and Experimental Neurology
- The New England Journal of Medicine
- Cancer Immunology, Immunotherapy : CII
- Neuro-oncology
- Journal of Neurosurgery
- Epilepsy Research
Articles by Adrienne C. Scheck in JoVE
JoVE Clinical and Translational Medicine
Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas
1Neuro-Oncology Research, Barrow Neurological Institute of St. Joseph’s Hospital and Medical Center, 2Neurosurgery Research Laboratory, Barrow Neurological Institute of St. Joseph’s Hospital and Medical Center
Intracranial implantation of GL261 cells into C57BL/6 mice produces malignant gliomas that recapitulate many of the hallmarks of human glioblastoma multiforme. We used GL261 cells stably expressing luciferase to allow us to use in vivo imaging to follow tumor progression. The surgery and 3D in vivo imaging are demonstrated.
Other articles by Adrienne C. Scheck on PubMed
Prostaglandin E(2)-synthesizing Enzymes in Fever: Differential Transcriptional Regulation
The febrile response to lipopolysaccharide (LPS) consists of three phases (phases I-III), all requiring de novo synthesis of prostaglandin (PG) E(2). The major mechanism for activation of PGE(2)-synthesizing enzymes is transcriptional upregulation. The triphasic febrile response of Wistar-Kyoto rats to intravenous LPS (50 microg/kg) was studied. Using real-time RT-PCR, the expression of seven PGE(2)-synthesizing enzymes in the LPS-processing organs (liver and lungs) and the brain "febrigenic center" (hypothalamus) was quantified. Phase I involved transcriptional upregulation of the functionally coupled cyclooxygenase (COX)-2 and microsomal (m) PGE synthase (PGES) in the liver and lungs. Phase II entailed robust upregulation of all enzymes of the major inflammatory pathway, i.e., secretory (s) phospholipase (PL) A(2)-IIA --> COX-2 --> mPGES, in both the periphery and brain. Phase III was accompanied by the induction of cytosolic (c) PLA(2)-alpha in the hypothalamus, further upregulation of sPLA(2)-IIA and mPGES in the hypothalamus and liver, and a decrease in the expression of COX-1 and COX-2 in all tissues studied. Neither sPLA(2)-V nor cPGES was induced by LPS. The high magnitude of upregulation of mPGES and sPLA(2)-IIA (1,257-fold and 133-fold, respectively) makes these enzymes attractive targets for anti-inflammatory therapy.
Expression of Genes Controlling Transport and Catabolism of Prostaglandin E2 in Lipopolysaccharide Fever
Prostaglandin (PG) E(2) is a principal downstream mediator of fever and other symptoms of systemic inflammation. Its inactivation occurs in peripheral tissues, primarily the lungs and liver, via carrier-mediated cellular uptake and enzymatic oxidation. We hypothesized that inactivation of PGE(2) is suppressed during LPS fever and that transcriptional downregulation of PGE(2) carriers and catabolizing enzymes contributes to this suppression. Fever was induced in inbred Wistar-Kyoto rats by intravenous LPS (50 microg/kg); the controls received saline. Samples of the liver, lungs, and hypothalamus were harvested 0, 0.5, 1.5, and 5 h postinjection. The expression of the two principal transmembrane PGE(2) carriers (PG transporter and multispecific organic anion transporter) and the two key PGE(2)-inactivating enzymes [15-hydroxy-PG dehydrogenase (15-PGDH) and carbonyl reductase] was quantified by RT-PCR. All four genes of interest were downregulated in peripheral tissues (but not the brain) during fever. Most remarkably, the expression of hepatic 15-PGDH was decreased 26-fold 5 h post-LPS, whereas expression of pulmonary 15-PGDH was downregulated (as much as 18-fold) throughout the entire febrile course. The transcriptional downregulation of several proteins involved in PGE(2) inactivation, first reported here, is an unrecognized mechanism of systemic inflammation. By increasing the blood-brain gradient of PGE(2), this mechanism likely facilitates penetration of PGE(2) into the brain and prevents its elimination from the brain.
Diagnostic and Prognostic Significance of Genetic Regional Heterogeneity in Meningiomas
We analyzed the frequency and regional distribution of cells with genetic abnormalities of chromosomes 1, 14, and 22 in meningiomas. This data was evaluated for correlation to the clinical outcome of the patients. Eight defined areas of each of 77 paraffin-embedded meningioma samples (59 grade I, 13 grade II, and 5 grade III) were analyzed by fluorescent in situ hybridization using bacterial artificial chromosome probes localized to chromosomes 1p36.32, 1q25.3, 14q13.3, 14q32.12, 22q11.2, and 22q12.1-3. Chromosome deletion was considered to be regionally heterogeneous if 7 regions showed cells with chromosome deletions. Deletion of 1p occurred in 35% of the grade I tumors. Distribution of cells with 1p deletion was regionally heterogeneous in 25% and homogeneous in 10% of grade I tumors. Distribution of cells with deletion of 1p was regionally heterogeneous in 23% and homogeneous in 69% of the grade II tumors. All grade III meningiomas had homogeneous distribution of cells with deletion of chromosome 1p. Distribution of cells with deletion of 14q was regionally heterogeneous in 27% and homogeneous in 2% of the grade I meningiomas, heterogeneous in 31% and homogeneous in 62% of the grade II tumors, and heterogeneous in 40% and homogeneous in 60% of the grade III meningiomas. Distribution of cells with deletion of 22q was regionally heterogeneous in 15% and homogeneous in 3% of the grade I tumors, heterogeneous in 15% and homogeneous in 31% of grade II tumors, and homogeneous in 20% of the grade III meningiomas. Distribution of cells with trisomy 22q was regionally heterogeneous in 10% of grade I tumors, heterogeneous in 23% of grade II, and homogeneous in 80% of grade III meningiomas. The proportion of patients with a deletion of 22q (either homogeneous or heterogeneous) who had recurrence was greater than the proportion of those without 22q deletion who had recurrence, and deletion of 22q was significantly associated with radiologically detected recurrence (P < 0.05). We conclude that the appearance of chromosomal aberrations in different areas of the tumor demonstrates the importance of regional heterogeneity in the biological behavior of meningiomas.
Expression of Eph Receptors and Their Ligands, Ephrins, During Lipopolysaccharide Fever in Rats
Erythropoietin-producing hepatocellular (Eph) receptor tyrosine kinases and their ligands, ephrins, are involved in embryogenesis and oncogenesis by mediating cell adhesion and migration. Although ephrins can be induced by bacterial LPS in vitro, whether they are involved in inflammation in vivo is unknown. Using differential mRNA display, we found that a febrigenic dose of LPS (50 microg/kg iv) induces a strong transcriptional upregulation of ephrin-A1 in rat liver. We confirmed this finding by real-time RT-PCR. We then quantified the mRNA expression of different ephrins and Eph receptors at phases 1-3 of LPS fever in different organs. Febrile phases 2 (90 min post-LPS) and 3 (300 min) were characterized by robust upregulation (up to 16-fold) and downregulation (up to 21-fold) of several ephrins and Eph receptors. With the exception of EphA2, which showed upregulation in the brain at phase 2, expressional changes of Eph receptors and ephrins were limited to the LPS-processing organs: liver and lung. Characteristic, counter-directed changes in expressional regulation of Eph receptors and their corresponding ligands were found: upregulation of EphA2, downregulation of ephrin-A1 in the liver and lung at phase 2; downregulation of EphB3, upregulation of ephrin-B2 in the liver at phase 2; downregulation of EphA1 and EphA3, upregulation of ephrins-A1 and -A3 in liver at phase 3. In the liver, transcriptional changes of EphA2 and EphB3 at phase 2 were confirmed at protein level. These coordinated, phase-specific responses suggest that different sets of ephrins and Eph receptors may be involved in cellular events (such as disruption of tissue barriers and leukocyte transmigration) underlying different stages of systemic inflammatory response to LPS.
Emerging Evidence of Hepatitis C Virus Neuroinvasion
It has been reported that hepatitis C virus (HCV) infection is associated with cognitive dysfunction, fatigue and depression, which do not correlate with the severity of liver disease and cannot be accounted for by hepatic encephalopathy or drug abuse. There is also emerging evidence that HCV infection can have negative neurocognitive effects in HIV-infected cohorts. Magnetic resonance spectroscopy has suggested the likely existence of a biological basis for these effects. HCV replicative forms have recently been detected in autopsy brain tissue and the infected cells have been identified as CD68-positive (macrophages/microglia). These findings raise the possibility that HCV infection of the brain could be directly related to the reported neuropsychological and cognitive changes. HCV is not strictly hepatotropic, as it can also replicate in leukocytes, including monocytes/macrophages. The latter cells could provide access of HCV into the central nervous system ('Trojan horse' mechanism) in a process similar to that postulated for HIV-1. In support of this hypothetical mechanism come reports showing a close relationship between HCV sequences present in the brain and cerebrospinal fluid and sequences found in lymph nodes and peripheral blood mononuclear cells. However, despite some similarities there is a fundamental difference between HIV-1 and HCV infection as the latter does not progress into AIDS-type dementia.
Tumor Necrosis Factor-alpha-induced Protein 3 As a Putative Regulator of Nuclear Factor-kappaB-mediated Resistance to O6-alkylating Agents in Human Glioblastomas
Pre-existing and acquired drug resistance are major obstacles to the successful treatment of glioblastomas.
Over-representation of Specific Regions of Chromosome 22 in Cells from Human Glioma Correlate with Resistance to 1,3-bis(2-chloroethyl)-1-nitrosourea
Glioblastoma multiforme is the most malignant form of brain tumor. Despite treatment including surgical resection, adjuvant chemotherapy, and radiation, these tumors typically recur. The recurrent tumor is often resistant to further therapy with the same agent, suggesting that the surviving cells that repopulate the tumor mass have an intrinsic genetic advantage. We previously demonstrated that cells selected for resistance to 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) are near-diploid, with over-representation of part or all of chromosomes 7 and 22. While cells from untreated gliomas often have over-representation of chromosome 7, chromosome 22 is typically under-represented.
Anticancer Activity of Extracts Derived from the Mature Roots of Scutellaria Baicalensis on Human Malignant Brain Tumor Cells
Flavonoid-rich extracts from the mature roots of Scutellaria baicalensis have been shown to exhibit antiproliferative effects on various cancer cell lines. We assessed the ability of an ethanolic extract of S. baicalensis root to inhibit the proliferation of malignant glioma cells.
Fluorescent in Situ Hybridization and Ex Vivo 1H Magnetic Resonance Spectroscopic Examinations of Meningioma Tumor Tissue: is It Possible to Identify a Clinically-aggressive Subset of Benign Meningiomas?
Although histologically benign, Grade I meningiomas can sometimes behave aggressively. The clinically-aggressive subset of Grade I meningiomas is typically indistinguishable from clinically-benign Grade I meningiomas in vivo. We compared molecular genetic and biochemical findings to clinical, pathological, and immunohistochemical information in a series of clinically-aggressive Grade I meningiomas with a series of clinically-benign meningiomas to identify characteristics that may be used to distinguish between these two groups.
Gene Expression Analysis of Glioblastomas Identifies the Major Molecular Basis for the Prognostic Benefit of Younger Age
Glioblastomas are the most common primary brain tumour in adults. While the prognosis for patients is poor, gene expression profiling has detected signatures that can sub-classify GBMs relative to histopathology and clinical variables. One category of GBM defined by a gene expression signature is termed ProNeural (PN), and has substantially longer patient survival relative to other gene expression-based subtypes of GBMs. Age of onset is a major predictor of the length of patient survival where younger patients survive longer than older patients. The reason for this survival advantage has not been clear.
Implicating Chromosomal Aberrations with Meningioma Growth and Recurrence: Results from FISH and MIB-I Analysis of Grades I and II Meningioma Tissue
The fluorescence in situ hybridization (FISH) technique was used in 111 WHO grades I and II meningioma patients. Clinical, radiological, pathological, and immunohistochemical data were compared to aberrations of chromosomes 1p, 14q, and 22q determined by FISH. Significant differences for MIB-1 labeling were found between grades I and II tumors (p < 0.001), and between grade I tumors that recurred and those that did not recur (p < 0.001). Chromosomal aberrations were detected with FISH analysis in nearly 50% of grade I, and in 93% of grade II meningiomas. The numbers of chromosomal aberrations correlated significantly to MIB-1 (p < 0.001), with signs of grossly invasive tumor growth (p < 0.001), and with tumor recurrence (p < 0.01). The findings suggest that adding FISH analysis may allow better prediction of possible meningioma recurrence and may be a useful adjunct for therapy decisions.
Molecular Biological Determinants of Meningioma Progression and Aggressive Behavior
Meningiomas are the most commonly reported brain tumor in the United States. Though these tumors are often surgically curable, even World Health Organization (WHO) grade 1 meningiomas can recur. The variability seen in the clinical behavior of meningiomas suggests that these tumors are genetically heterogeneous. The most common genetic aberrations found in meningiomas are deletions of chromosomes 1p, 14q, and 22q. Fluorescent in situ hybridization (FISH) analyses have demonstrated the presence of intratumor heterogeneity; however, the loss of a single chromosome region was not indicative of aggressive behavior. In fact, tumors of higher grade are less heterogeneous in that all of the cells tend to demonstrate deletion of these chromosome arms. Tumor suppressor genes that map to these chromosomes have been identified but have not been found to play a significant role in the initiation or progression of the disease. The identification of a marker of aggressive behavior would allow the development of improved clinical protocols based on early intervention for those patients likely to experience a recurrence.
Cyclooxygenase-1 or -2--which One Mediates Lipopolysaccharide-induced Hypothermia?
Systemic inflammation is associated with either fever or hypothermia. Fever, a response to mild systemic inflammation, is mediated by cyclooxygenase (COX)-2 and not by COX-1. However, it is still disputed whether COX-2, COX-1, neither, or both mediate(s) responses to severe systemic inflammation, and, in particular, the hypothermic response. We compared the effects of SC-236 (COX-2 inhibitor) and SC-560 (COX-1 inhibitor) on the deep body temperature (T(b)) of rats injected with a lower (10 microg/kg i.v.) or higher (1,000 microg/kg i.v.) dose of LPS at different ambient temperatures (T(a)s). At a neutral T(a) (30 degrees C), the rats responded to LPS with a polyphasic fever (lower dose) or a brief hypothermia followed by fever (higher dose). SC-236 (2.5 mg/kg i.v.) blocked the fever induced by either LPS dose, whereas SC-560 (5 mg/kg i.v.) altered neither the febrile response to the lower LPS dose nor the fever component of the response to the higher dose. However, SC-560 blocked the initial hypothermia caused by the higher LPS dose. At a subneutral T(a) (22 degrees C), the rats responded to LPS with early (70-90 min, nadir) dose-dependent hypothermia. The hypothermic response to either dose was enhanced by SC-236 but blocked by SC-560. The hypothermic response to the higher LPS dose was associated with a fall in arterial blood pressure. This hypotensive response was attenuated by either SC-236 or SC-560. At the onset of LPS-induced hypothermia and hypotension, the functional activity of the COX-1 pathway (COX-1-mediated PGE(2) synthesis ex vivo) increased in the spleen but not liver, lung, kidney, or brain. The expression of splenic COX-1 was unaffected by LPS. We conclude that COX-1, but not COX-2, mediates LPS hypothermia, and that both COX isoforms are required for LPS hypotension.
A Network Model of a Cooperative Genetic Landscape in Brain Tumors
Gliomas, particularly glioblastomas, are among the deadliest of human tumors. Gliomas emerge through the accumulation of recurrent chromosomal alterations, some of which target yet-to-be-discovered cancer genes. A persistent question concerns the biological basis for the coselection of these alterations during gliomagenesis.
Monosomy of Chromosome 10 Associated with Dysregulation of Epidermal Growth Factor Signaling in Glioblastomas
Glioblastomas--uniformly fatal brain tumors--often have both monosomy of chromosome 10 and gains of the epidermal growth factor receptor (EGFR) gene locus on chromosome 7, an association for which the mechanism is poorly understood.
Using Ex Vivo Proton Magnetic Resonance Spectroscopy to Reveal Associations Between Biochemical and Biological Features of Meningiomas
The goal in this study was to determine if proton ((1)H) MR spectroscopy can differentiate meningioma grade and is associated with interpretations of biological behavior; the study was performed using ex vivo high-resolution spectra indicating metabolic characteristics.
Miniaturized Handheld Confocal Microscopy for Neurosurgery: Results in an Experimental Glioblastoma Model
Recent developments in optical science and image processing have miniaturized the components required for confocal microscopy. Clinical confocal imaging applications have emerged, including assessment of colonic mucosal dysplasia during colonoscopy. We present our initial experience with handheld, miniaturized confocal imaging in a murine brain tumor model.
The Ketogenic Diet Reverses Gene Expression Patterns and Reduces Reactive Oxygen Species Levels when Used As an Adjuvant Therapy for Glioma
Malignant brain tumors affect people of all ages and are the second leading cause of cancer deaths in children. While current treatments are effective and improve survival, there remains a substantial need for more efficacious therapeutic modalities. The ketogenic diet (KD) - a high-fat, low-carbohydrate treatment for medically refractory epilepsy - has been suggested as an alternative strategy to inhibit tumor growth by altering intrinsic metabolism, especially by inducing glycopenia.
In Vitro Biological Dosimeter Modeling of the Glioblastoma Response to Radiation Delivered by the Gamma Knife. Laboratory Investigation
The goal of this study was to develop an assay that makes possible the assessment of the glioma cell response to single-fraction high-dose Gamma Knife surgery. In this assay, the isolation of radioresistant cell subpopulations facilitates mechanistic studies of radioresistance.
Identification of Gene Markers Associated with Aggressive Meningioma by Filtering Across Multiple Sets of Gene Expression Arrays
Meningiomas are common intracranial tumors, but relatively little is known about the genetic events responsible for their clinical diversity. Although recent genomic studies have provided clues, the genes identified often differ among publications. We used microarray expression profiling to identify genes that are differentially expressed, with at least a 4-fold change, between grade I and grade III meningiomas. We filtered this initial set of potential biomarkers through a second cohort of meningiomas and then verified the remaining genes by quantitative polymerase chain reaction followed by examination using a third microarray expression cohort. Using this approach, we identified 9 overexpressed (TPX2, RRM2, TOP2A, PI3, BIRC5, CDC2, NUSAP1, DLG7, SOX11) and 2 underexpressed (TIMP3, KCNMA1) genes in grade III versus grade I meningiomas. As a further validation step, we analyzed these genes in a fourth cohort and found that patients with grade II meningiomas with high topoisomerase 2-α protein expressions (>5% labeling index) had shorter times to death than patients with low expressions. We believe that this multistep multi-cohort approach provides a robust method for reducing false-positives while generating a list of reproducible candidate genes that are associated with clinically aggressive meningiomas and are suitable for analysis for their potential prognostic value.
NFKBIA Deletion in Glioblastomas
Amplification and activating mutations of the epidermal growth factor receptor (EGFR) oncogene are molecular hallmarks of glioblastomas. We hypothesized that deletion of NFKBIA (encoding nuclear factor of κ-light polypeptide gene enhancer in B-cells inhibitor-α), an inhibitor of the EGFR-signaling pathway, promotes tumorigenesis in glioblastomas that do not have alterations of EGFR.
CTL Recognition of a Novel HLA-A*0201-binding Peptide Derived from Glioblastoma Multiforme Tumor Cells
Genetic instability of tumor cells can result in translation of proteins that are out of frame, resulting in expression of neopeptides. These neopeptides are not self-proteins and therefore should be immunogenic. By eluting peptides from human glioblastoma multiforme (GBM) tumor cell surfaces and subjecting them to tandem mass spectrometry, we identified a novel peptide (KLWGLTPKVTPS) corresponding to a frameshift in the 3' beta-hydroxysteroid dehydrogenase type 7 (HSD3B7) gene. HLA-binding algorithms predicted that a 9-amino acid sequence embedded in this peptide would bind to HLA-A*0201. We confirmed this prediction using an HLA-A*0201 refolding assay followed by live cell relative affinity assays, but also showed that the 12-mer binds to HLA-A*0201. Based on the 9-mer sequence, optimized peptide ligands (OPL) were designed and tested for their affinities to HLA-A*0201 and their abilities to elicit anti-peptide and CTL capable of killing GBM in vitro. Wild-type peptides as well as OPL induced anti-peptide CTL as measured by IFN-γ ELISPOTS. These CTL also killed GBM tumor cells in chromium-51 release assays. This study reports a new CTL target in GBM and further substantiates the concept that rational design and testing of multiple peptides for the same T-cell epitope elicits a broader response among different individuals than single peptide immunization.
Gene-protein Correlation in Single Cells
We present a novel methodology combining traditional fluorescent in situ hybridization with an in situ protein detection technology called proximity ligation assay. This method has potential to perform a detailed analysis of the relationship between gene status and corresponding protein expression in cells and tissues. We demonstrate that the fluorescent in situ gene protein assay methodology is capable of resolving gene and protein patterns simultaneously on a cell-by-cell basis.
Use of in Vivo Near-infrared Laser Confocal Endomicroscopy with Indocyanine Green to Detect the Boundary of Infiltrative Tumor
Infiltrative tumor resection is based on regional (macroscopic) imaging identification of tumorous tissue and the attempt to delineate invasive tumor margins in macroscopically normal-appearing tissue, while preserving normal brain tissue. The authors tested miniaturized confocal fiberoptic endomicroscopy by using a near-infrared (NIR) imaging system with indocyanine green (ICG) as an in vivo tool to identify infiltrating glioblastoma cells and tumor margins.
The Ketogenic Diet for the Treatment of Glioma: Insights from Genetic Profiling
Seizures, particularly first onset seizures in adults, are a diagnostic hallmark of brain tumors (Giglio and Villano, 2010). Unfortunately, malignant brain tumors are almost uniformly fatal due, in part, to the limitations of available therapies. Improvement in the survival of brain cancer patients requires the design of new therapeutic modalities including those that enhance currently available therapies. One potential strategy is to exploit differences in metabolic regulation between normal cells and tumor cells through dietary approaches. Previous studies have shown that a high-fat, low-carbohydrate ketogenic diet (KD) extends survival in animal models of glioma; however, the mechanism for this effect is not entirely known. We examined the effects of an experimental KD on a mouse model of glioma, and compared patterns of gene expression in tumors versus contralateral non-tumor containing brain from animals fed either a KD or a standard diet. We found that the KD reduced reactive oxygen species (ROS) production in tumor cells. Gene expression profiling demonstrated that the KD induces an overall reversion to expression patterns seen in non-tumor specimens, and a number of genes involved in modulating ROS levels and oxidative stress were altered in tumor cells. In addition, there was reduced expression of genes involved in signal transduction from growth factors known to be involved in glioma growth. These results suggest that the anti-tumor effect of the KD is multifactorial, and elucidation of genes whose expression is altered will help identify mechanisms through which ketones inhibit tumor growth, reduce seizure activity and provide neuroprotection.
