Ali Chahlavi

Glioblastomas Induce T-Lymphocyte Death by Two Distinct Pathways Involving Gangliosides and CD70

Here we report that glioblastoma multiforme (GBM) mediates immunosuppression by promoting T-cell death via tumor-associated CD70 and gangliosides that act through receptor-dependent and receptor-independent pathways, respectively. GBM lines cocultured with T cells induced lymphocyte death. The GBM lines were characterized for their expression of CD70, Fas ligand (FasL), and tumor necrosis factor-alpha (TNF-alpha), and the possible participation of those molecules in T-cell killing was assessed by doing GBM/T cell cocultures in the presence of anti-CD70 antibodies, Fas fusion proteins, or anti-TNF-alpha antibodies. CD70 but not TNF-alpha or FasL is responsible for initiating T-cell death via the receptor-dependent pathway. Of the four GBM cell lines that induced T-cell death, three highly expressed CD70. Two nonapoptogenic GBM lines (CCF3 and U138), on the other hand, had only minimally detectable CD70 expression. Blocking experiments with the anti-CD70 antibody confirmed that elevated CD70 levels were involved in the apoptogenicity of the three GBM lines expressing that molecule. Gangliosides were found to participate in the induction of T-cell apoptosis, because the glucosylceramide synthase inhibitor (PPPP) significantly reduced the abilities of all four apoptogenic lines to kill the lymphocytes. High-performance liquid chromatography (HPLC) and mass spectroscopy revealed that GM2, GM2-like gangliosides, and GD1a were synthesized in abundance by all four apoptogenic GBM lines but not by the two GBMs lacking activity. Furthermore, gangliosides isolated from GBM lines as well as HPLC fractions containing GM2 and GD1a were directly apoptogenic for T cells. Our results indicate that CD70 and gangliosides are both products synthesized by GBMs that may be key mediators of T-cell apoptosis and likely contribute to the T-cell dysfunction observed within the tumor microenvironment.

Impact of Chromosome 1p Status in Response of Oligodendroglioma to Temozolomide: Preliminary Results

In this IRB-approved retrospective study, we analyzed the efficacy of temozolomide on World Health Organization Grade II and III oligodendrogliomas, as well as mixed oligoastrocytomas, to determine if a correlation exists between the tumors 1p status and control of growth by this new oral agent. We assessed six patients with oligodendrogliomas with 1p intact (38%) and 10 patients with 1p loss (62%), who received temozolomide.Chromosome 1p status was significantly associated with response to treatment using temozolomide. While nine of 10 patients (90%) with 1p loss responded to temozolomide, only two of six patients (33%) with 1p intact benefited from this treatment (p = 0.04). Although the number of patients evaluated was small, there was no association between 1p status and gender, age, and tumor grade. Gender, age, and tumor grade were similarly not correlated with response to chemotherapy.This report is the first to find that patients harboring oligodendrogliomas with 1p loss are more likely to be sensitive to treatment with temozolomide than those that retain this chromosomal element. Larger prospective trials are needed to confirm these findings; however, temozolomide should be considered in the management of these tumors.

A feasibility study of optical coherence tomography for guiding deep brain probes

Deep brain simulation (DBS) is effective for the treatment of various diseases including Parkinsons disease and essential tremor. However, anatomical targeting combined with microelectrode mapping of the region requires significant surgical time. Also, the fine-tipped microelectrode imposes a risk of hemorrhage in the event that the trajectory intersects subcortical vessels. To reduce the operation time and the risk of hemorrhage, we propose to use optical coherence tomography (OCT) to guide the insertion of the DBS probe. We conducted in vitro experiments in the rat brain to study the feasibility of this application. The result shows that OCT is able to differentiate structures in the rat brain. White matter tends to have higher peak reflectivity and steeper attenuation rate compared to gray matter. This structural information may help guide DBS probe advance and electrical measurements.

GM2 Expression in Renal Cell Carcinoma: Potential Role in Tumor-Induced T-Cell Dysfunction

Multiple mechanisms have been proposed to account for immune escape by tumors. Although gangliosides have long been known to suppress T-cell immunity, few studies have examined the effect of human tumor-derived gangliosides on immune responses. Here, we show that gangliosides isolated from renal cell carcinoma (RCC) cell lines and clear cell tumor tissue can induce apoptosis in peripheral blood T cells. The RCC tissue-derived gangliosides also suppressed IFN-gamma and, in many cases, interleukin-4 production by CD4+ T cells at concentrations (1 ng/mL-100 pg/mL) well below those that induce any detectable T-cell death (4-20 microg/mL). Additional findings show that GM2 expressed by RCC plays a significant role in promoting T-cell dysfunction. This is supported by the demonstration that all RCC cell lines examined (n = 5) expressed GM2 as did the majority of tumors (15 of 18) derived from patients with clear cell RCC. Furthermore, an antibody specific for GM2 (DMF10.167.4) partially blocked (50-60%) T-cell apoptosis induced by coculturing lymphocytes with RCC cell lines or with RCC tissue-derived gangliosides. DMF10.167.4 also partially blocked the suppression of IFN-gamma production induced by RCC tissue-derived gangliosides, suggesting that GM2 plays a role in down-regulating cytokine production by CD4+ T cells.

World Health Organization Grades II and III meningiomas are rare in the cranial base and spine.

OBJECTIVEThis study was undertaken to assess a possible relationship between the tumor location and the incidence of World Health Organization (WHO) Grades II and III meningiomas. METHODSA retrospective review of 794 consecutive patients who underwent meningioma resection between January 1991 and March 2004 was conducted. Among these, 47 patients (5.9%) with WHO Grade II meningiomas and 16 patients (2%) with Grade III meningiomas were further analyzed. Tumor location was assessed using preoperative magnetic resonance imaging scans and/or operative reports. Histological grading was done according to the WHO 2000 Classification scheme. RESULTSWHO Grade II tumors were found in eight out of 289 (2.8%) cranial base meningiomas and in zero spinal meningiomas, compared with 39 out of 429 (9.1%) non-cranial base meningiomas. Grade III histology was encountered in two (0.7%) cranial base tumors and in one out of 76 (1.3%) spinal tumors, compared with 13 (3%) non-cranial base tumors. The combined incidence of Grades II and III meningiomas was significantly lower in the cranial base (3.5%) and spinal (1.3%) locations compared with non-cranial base locations (12.1%) (P < 0.001). CONCLUSIONWHO Grades II and III meningiomas occur far less frequently in the cranial base and spinal locations. Tumors arising from these locations may have different mechanisms of tumorigenesis and/or progression compared with meningiomas arising from other (non-cranial base) regions.

Intracranial collision tumor mimicking an octreotide-SPECT positive and FDG-PET negative meningioma

UNLABELLEDnOBJECTIVE/IMPORTANCE: Cancer metastasis to a pre-existing intracranial tumor is rare, but several cases have been reported. We report an unusual case of a collision tumor consisting of a renal cell carcinoma metastasis to an intracranial meningioma.nnnCLINICAL PRESENTATIONnA 67-year old male with renal cell carcinoma had an asymptomatic right posterior frontal dural-based lesion identified on a screening CT scan. MRI characteristics of the tumor were consistent with meningioma. On octreotide-SPECT and F-18 fluorodeoxyglucose (FDG)-PET scans, the lesion showed octreotide uptake but did not accumulate FDG, both of which are consistent with a diagnosis of benign meningioma. One week later, he presented with a 1-day history of progressive left-sided weakness. The intracranial tumor was resected, and subacute subdural blood was found overlying a soft, reddish tumor. Microscopic examination was consistent with renal cell carcinoma with a minor portion consisting of meningioma. The meningothelial component was strongly immunoreactive to vimentin and weakly reactive to epithelial membrane antigens. Neither area reacted with glucose transporter-1 (GLUT-1), correlating with low FDG-PET uptake.nnnCONCLUSIONnCollision tumor involving metastatic renal cell carcinoma to an intracranial meningioma is a rare occurrence. Diagnosis by non-invasive means, with use of a combination of octreotide-SPECT and FDG-PET may not accurately reflect the malignant component of such a collision tumor. In this case, the collision tumor also demonstrated a propensity to undergo spontaneous hemorrhage. A high degree of suspicion of intracranial metastasis should be maintained for patients who have known systemic cancer and are found incidentally to have a dural-based mass lesion.

GBM Derived Gangliosides Induce T Cell Apoptosis through Activation of the Caspase Cascade Involving Both the Extrinsic and the Intrinsic Pathway

Previously we demonstrated that human glioblastoma cell lines induce apoptosis in peripheral blood T cells through partial involvement of secreted gangliosides. Here we show that GBM-derived gangliosides induce apoptosis through involvement of the TNF receptor and activation of the caspase cascade. Culturing T lymphocytes with GBM cell line derived gangliosides (10-20μg/ml) demonstrated increased ROS production as early as 18 hrs as indicated by increased uptake of the dye H2DCFDA while western blotting demonstrated mitochondrial damage as evident by cleavage of Bid to t-Bid and by the release of cytochrome-c into the cytosol. Within 48-72 hrs apoptosis was evident by nuclear blebbing, trypan blue positivity and annexinV/7AAD staining. GBM-ganglioside induced activation of the effector caspase-3 along with both initiator caspases (-9 and -8) in T cells while both the caspase-8 and -9 inhibitors were equally effective in blocking apoptosis (60% protection) confirming the role of caspases in the apoptotic process. Ganglioside-induced T cell apoptosis did not involve production of TNF-α since anti-human TNFα antibody was unable to protect T cells from nuclear blebbing and subsequent cell death. However, confocal microscopy demonstrated co-localization of GM2 ganglioside with the TNF receptor and co-immunoprecipitation experiments showed recruitment of death domains FADD and TRADD with the TNF receptor post ganglioside treatment, suggesting direct interaction of gangliosides with the TNF receptor. Further confirmation of the interaction between GM2 and TNFR1 was obtained from confocal microscopy data with wild type and TNFR1 KO (TALEN mediated) Jurkat cells, which clearly demonstrated co-localization of GM2 and TNFR1 in the wild type cells but not in the TNFR1 KO clones. Thus, GBM-ganglioside can mediate T cell apoptosis by interacting with the TNF receptor followed by activation of both the extrinsic and the intrinsic pathway of caspases.

Optical Coherence Tomography and Optical Coherence Domain Reflectometry for Deep Brain Stimulation probe guidance

Deep Brain Stimulation (DBS) is FDA-approved for the treatment of Parkinsons disease and essential tremor. Currently, placement of DBS leads is guided through a combination of anatomical targeting and intraoperative microelectrode recordings. The physiological mapping process requires several hours, and each pass of the microelectrode into the brain increases the risk of hemorrhage. Optical Coherence Domain Reflectometry (OCDR) in combination with current methodologies could reduce surgical time and increase accuracy and safety by providing data on structures some distance ahead of the probe. For this preliminary study, we scanned a rat brain in vitro using polarization-insensitive Optical Coherence Tomography (OCT). For accurate measurement of intensity and attenuation, polarization effects arising from tissue birefringence are removed by polarization diversity detection. A fresh rat brain was sectioned along the coronal plane and immersed in a 5 mm cuvette with saline solution. OCT images from a 1294 nm light source showed depth profiles up to 2 mm. Light intensity and attenuation rate distinguished various tissue structures such as hippocampus, cortex, external capsule, internal capsule, and optic tract. Attenuation coefficient is determined by linear fitting of the single scattering regime in averaged A-scans where Beer’s law is applicable. Histology showed very good correlation with OCT images. From the preliminary study using OCT, we conclude that OCDR is a promising approach for guiding DBS probe placement.