Christopher McPherson

Discrete mechanisms of mTOR and cell cycle regulation by AMPK agonists independent of AMPK

Significance Cancer cells reprogram their metabolism for optimal growth and survival. AMPK-activated protein kinase (AMPK) is a key energy sensor that controls many metabolic pathways including metabolic reprogramming. However, its role in cancer is poorly understood. Some studies claim that it has a tumor suppressor role while others show its protumor role. Two AMPK-activating compounds (including metformin, now in many clinical trials) are widely used to suppress cancer cell proliferation. We found that AMPK is abundantly expressed in high-grade gliomas and, in contrast to popular belief, these two AMPK activators suppressed glioma cell proliferation through unique AMPK-independent mechanisms. The multifunctional AMPK-activated protein kinase (AMPK) is an evolutionarily conserved energy sensor that plays an important role in cell proliferation, growth, and survival. It remains unclear whether AMPK functions as a tumor suppressor or a contextual oncogene. This is because although on one hand active AMPK inhibits mammalian target of rapamycin (mTOR) and lipogenesis—two crucial arms of cancer growth—AMPK also ensures viability by metabolic reprogramming in cancer cells. AMPK activation by two indirect AMPK agonists AICAR and metformin (now in over 50 clinical trials on cancer) has been correlated with reduced cancer cell proliferation and viability. Surprisingly, we found that compared with normal tissue, AMPK is constitutively activated in both human and mouse gliomas. Therefore, we questioned whether the antiproliferative actions of AICAR and metformin are AMPK independent. Both AMPK agonists inhibited proliferation, but through unique AMPK-independent mechanisms and both reduced tumor growth in vivo independent of AMPK. Importantly, A769662, a direct AMPK activator, had no effect on proliferation, uncoupling high AMPK activity from inhibition of proliferation. Metformin directly inhibited mTOR by enhancing PRAS40’s association with RAPTOR, whereas AICAR blocked the cell cycle through proteasomal degradation of the G2M phosphatase cdc25c. Together, our results suggest that although AICAR and metformin are potent AMPK-independent antiproliferative agents, physiological AMPK activation in glioma may be a response mechanism to metabolic stress and anticancer agents.

Results of Contemporary Surgical Management of Radiation Necrosis using Frameless Stereotaxis and Intraoperative Magnetic Resonance Imaging

AbstractObjective: Radiation necrosis is a well-known complication of radiotherapy for malignant brain tumors. Although surgery was once considered the mainstay of treatment, no recent reports have evaluated the use of intraoperative magnetic resonance imaging (IOMRI) and frameless stereotaxis during surgical resection of radiation necrosis. In this retrospective review, we evaluate the effectiveness of surgical resection using frameless stereotaxis and IOMRI for the treatment of radiation necrosis. Methods: From October 1999 through February 2002, 11 patients who had malignant brain tumors underwent surgery for radiation necrosis. The diagnosis of radiation necrosis was based primarily on MRI and clinical suspicion. Frameless stereotaxis was used in all patients and IOMRI was used in nine. All patients underwent at least one radiation treatment before surgery and nine patients had multiple treatments. Patient outcome was based on changes in steroid dose, Karnofsky Performance Score (KPS), and neurologic deficit. Results: Optimal resection as confirmed by IOMRI was achieved in all patients by the use of frameless stereotaxis alone; no additional resection was performed in any patient. For nine patients taking steroids (mean preoperative dose 24 mg/day) before treatment of necrosis, all had a substantial reduction in steroid dosage (mean postoperative dose 8 mg/day) after surgical treatment. Postoperatively, KPS improved in four patients, remained stable in four, and worsened in three. Three complications that resulted from surgery included wound infection, asymptomatic carotid dissection, and pulmonary embolism; thus, overall morbidity including both surgical complications and neurologic deterioration was 54%. Conclusions: In this review, frameless stereotaxis was helpful in guiding the surgeon; however, IOMRI did not provide any additional benefit for the surgical treatment of radiation necrosis. Surgical treatment of radiation necrosis was associated with high risks of complication or neurologic deficit. Given the success of medical therapies, including hyperbaric oxygen, we believe that surgical treatment of radiation necrosis should be reserved for symptomatic patients in whom medical therapy has failed.

Comparing the risks of frameless stereotactic biopsy in eloquent and noneloquent regions of the brain: a retrospective review of 284 cases

OBJECT Frameless stereotactic biopsy has been shown in multiple studies to be a safe and effective tool for the diagnosis of brain lesions. However, no study has directly evaluated its safety in lesions located in eloquent regions in comparison with noneloquent locations. In this study, the authors determine whether an increased risk of neurological decline is associated with biopsy of lesions in eloquent regions of the brain. METHODS Medical records, including imaging studies, were reviewed for 284 cases in which frameless stereotactic biopsy procedures were performed by 19 neurosurgeons at 7 institutions between January 2000 and December 2006. Lesion location was classified as eloquent or noneloquent in each patient. The incidence of neurological decline was calculated for each group. RESULTS During the study period, 160 of the 284 biopsies predominately involved eloquent regions of the brain. In evaluation of the complication rate with respect to biopsy site, neurological decline occurred in 9 (5.6%) of 160 biopsies in eloquent brain areas and 10 (8.1%) of 124 biopsies in noneloquent regions; this difference was not statistically significant (p = 0.416). A higher number of needle passes was associated with the presence of a postoperative hemorrhage at the biopsy site, although not with a change in the result of neurological examination. CONCLUSIONS Frameless stereotactic biopsy of lesions located in eloquent brain regions is as safe and effective as biopsy of lesions in noneloquent regions. Therefore, with careful planning, frameless stereotactic biopsy remains a valuable and safe tool for diagnosis of brain lesions, independent of lesion location.

Flt-3 ligand: a potent dendritic cell stimulator and novel antitumor agent.

Dendritic cell (DC) vaccination has generated intense interest as a potential cancer therapy. However, the rate limiting step has been the generation of DCs. Flt-3 ligand (FL) is a growth factor that was first discovered by its ability to stimulate the proliferation of hematopoietic progenitor cells of both lymphoid and myeloid origin. The remarkable activity of FL to induce large numbers of dendritic cells both in vivo and in vitro soon captured the interest of numerous researchers. In this review, we examine the structure and function of the FL, its antitumor activity in animal models, and its potential as a novel cancer treatment. Key Words: Flt3-L, Dendritic cells, Cell mediated immune response, Cancer therapy

Atypical meningiomas: is postoperative radiotherapy indicated?

OBJECT The role of postoperative radiation therapy after surgery for atypical meningiomas remains controversial. In this retrospective cohort study, the authors examine the recurrence rates for atypical meningiomas after resection (with or without adjuvant radiotherapy) and identify which factors were associated with recurrence. METHODS Of 90 patients with atypical meningiomas who underwent surgery between 1999 and 2009, 71 (79%) underwent gross-total resection (GTR) and 19 (21%) underwent subtotal resection (STR); 31 patients received adjuvant radiotherapy. All tumors were pathology-confirmed WHO Grade II atypical meningiomas. Univariate and multivariate analyses were performed to identify factors associated with recurrence-free survival. RESULTS Among 90 patients, 17 developed tumor recurrence (81% recurrence-free survival at 5 years). In the overall group, adjuvant radiotherapy reduced the recurrence rate to 9% from 19% at 5 years (p = 0.048). After STR, adjuvant radiotherapy significantly reduced recurrence from 91% to 20% (p = 0.0016). However, after GTR, adjuvant radiotherapy did not significantly reduce the recurrence rate (16.7% without radiation therapy vs 11.8% with radiation therapy) (p = 1.00). Five factors independently predictive of tumor recurrence included mitotic index, sheeting, necrosis, nonuse of radiation therapy, and STR. Further recursive partitioning analysis showed significant increases in risk for patients older than 55 years with mitoses and sheeting. CONCLUSIONS Adjuvant radiotherapy was effective at lowering recurrence rates in patients after STR but delivered no significant improvement in patients after GTR. Given that rates after GTR were similar with or without adjuvant radiotherapy, close observation without postoperative radiation therapy may be a viable option for these patients. Patients older than 55 years and those with mitoses noted during pathological examination had a significant risk of recurrence after GTR; for these patients, postoperative radiotherapy is recommended.

Detection of EGFRvIII mutant DNA in the peripheral blood of brain tumor patients.

Glioblastoma multiforme (GBM) is the most aggressive brain tumor in adults and remains incurable despite multimodal intensive treatment regimens including surgical resection, radiation and chemotherapy. EGFRvIII is a truncated extracellular mutant of the EGF receptor (EGFR) found in about a third of GBMs. It confers enhanced tumorigenic behavior and is associated with chemo- and radio-resistance. GBM patients testing positive for EGFRvIII have a bleaker prognosis than those who do not. Targeting EGFRvIII positive tumors via vaccines or antibody–drug-conjugates represents a new challenging therapeutic avenue with potential great clinical benefits. In this study, we developed a strategy to detect EGFRvIII deletion in the circulating tumor DNA. The overall goal is to identify a simple and robust biomarker in the peripheral blood of patients diagnosed with GBM in order to follow their disease status while on treatment. Thirteen patients were included in this study, three of which were found to carry the EGFRvIII deletion. The circulating DNA status for EGFRvIII correlates with the analysis performed on the respective tumor samples, and its level seems to correlate with the extent of the tumor resection. This semi-quantitative blood biomarker may represent a strategy to (1) screen patients for an anti-EGFRvIII therapy and (2) monitor the patients’ response to treatment.

Tumor Resection in a Shared-Resource Magnetic Resonance Operating Room: Experience at the University of Cincinnati

INTRODUCTION At the University of Cincinnati, we have developed a shared-resource magnetic resonance operating suite that facilitates performance of both neurosurgical and diagnostic procedures in a single unit. METHODS The shared-resource magnetic resonance operating suite utilizes a Hitachi AIRIS II, 0.3-T, vertical field, open MRI unit located in the MROR. This magnet can be used for both diagnostic and interventional procedures. The addition of a rotating-operating table permits neurosurgical procedures to be performed outside of the 5-G line using standard neurosurgical equipment and operating microscopes. RESULTS We review our results with the shared-resource magnetic resonance operating room including the tabulated results from 30 transsphenoidal procedures and 63 glioma procedures. In addition, 2832 diagnostic procedures have been performed in the first 4 years of use. CONCLUSION The shared-resource intraoperative MRI facility produces high-quality intraoperative imaging studies, equal to those of high-resolution magnets, and is valuable in enabling the surgeon to achieve the planned degree of resection of glioma and pituitary tumors. The ability to perform diagnostic procedures in a shared unit has been a cost-effective solution for our institution.

Critical Role of Imaging in the Neurosurgical and Radiotherapeutic Management of Brain Tumors

Abbreviations: ADC = apparent diffusion coefficient, AF = arcuate fasciculus, BOLD = blood oxygen level–dependent, CBV = cerebral blood volume, CNS = central nervous system, CST = corticospinal tract, CTV = clinical target volume, DSC = dynamic susceptibility contrast, DTI = diffusion tensor imaging, DW = diffusion-weighted, ESM = electrocortical stimulation mapping, FLAIR = fluid-attenuated inversion recovery, GBM = glioblastoma multiforme, GTV = gross tumor volume, NAA = N-acetylaspartate, TE = echo time

Imaging of brain tumors with paramagnetic vesicles targeted to phosphatidylserine

To investigate paramagnetic saposin C and dioleylphosphatidylserine (SapC‐DOPS) vesicles as a targeted contrast agent for imaging phosphatidylserine (PS) expressed by glioblastoma multiforme (GBM) tumors.

Methylation markers of malignant potential in meningiomas

OBJECT Although most meningiomas are benign, about 20% are atypical (Grade II or III) and have increased mortality and morbidity. Identifying tumors with greater malignant potential can have significant clinical value. This validated genome-wide methylation study comparing Grade I with Grade II and III meningiomas aims to discover genes that are aberrantly methylated in atypical meningiomas. METHODS Patients with newly diagnosed meningioma were identified as part of the Ohio Brain Tumor Study. The Infinium HumanMethylation27 BeadChip (Illumina, Inc.) was used to interrogate 27,578 CpG sites in 14,000 genes per sample for a discovery set of 33 samples (3 atypical). To verify the results, the Infinium HumanMethylation450 BeadChip (Illumina, Inc.) was used to interrogate 450,000 cytosines at CpG loci throughout the genome for a verification set containing 7 replicates (3 atypical), as well as 12 independent samples (6 atypical). A nonparametric Wilcoxon exact test was used to test for difference in methylation between benign and atypical meningiomas in both sets. Heat maps were generated for each set. Methylation results were validated for the 2 probes with the largest difference in methylation intensity by performing Western blot analysis on a set of 20 (10 atypical) samples, including 11 replicates. RESULTS The discovery array identified 95 probes with differential methylation between benign and atypical meningiomas, creating 2 distinguishable groups corresponding to tumor grade when visually examined on a heat map. The validation array evaluated 87 different probes and showed that 9 probes were differentially methylated. On heat map examination the results of this array also suggested the existence of 2 major groups that corresponded to histological grade. IGF2BP1 and PDCD1, 2 proteins that can increase the malignant potential of tumors, were the 2 probes with the largest difference in intensity, and for both of these the atypical meningiomas had a decreased median production of protein, though this was not statistically significant (p = 0.970 for IGF2BP1 and p = 1 for PDCD1). CONCLUSIONS A genome-wide methylation analysis of benign and atypical meningiomas identified 9 genes that were reliably differentially methylated, with the strongest difference in IGF2BP1 and PDCD1. The mechanism why increased methylation of these sites is associated with an aggressive phenotype is not evident. Future research may investigate this mechanism, as well as the utility of IGF2BP1 as a marker for pathogenicity in otherwise benign-appearing meningiomas.