CLR1404 is a cancer-selective alkyl phosphocholine (APC) analog that can be radiolabeled with 124... more CLR1404 is a cancer-selective alkyl phosphocholine (APC) analog that can be radiolabeled with 124I for PET imaging, 131I for targeted radiotherapy and/or SPECT imaging, or 125I for targeted radiotherapy. Studies have demonstrated avid CLR1404 uptake and prolonged retention in a broad spectrum of preclinical tumor models. The purpose of this pilot trial was to demonstrate avidity of 124I-CLR1404 in human brain tumors and develop a framework to evaluate this uptake for use in larger studies. 12 patients (8 men and 4 women; mean age of 43.9 ± 15.1 y; range 23-66 y) with 13 tumors were enrolled. Eleven patients had suspected tumor recurrence and 1 patient had a new diagnosis of high grade tumor. Patients were injected with 185 MBq ± 10% of 124I-CLR1404 followed by PET/CT imaging at 6-, 24-, and 48-hour. 124I-CLR1404 PET uptake was assessed qualitatively and compared with MRI. After PET image segmentation SUV values and tumor to background ratios were calculated. There was no significant uptake of 124I-CLR1404 in normal brain. In tumors, uptake tended to increase to 48 hours. Positive uptake was detected in 9 of 13 lesions: 5/5 high grade tumors, 1/2 low grade tumors, 1/1 meningioma, and 2/4 patients with treatment related changes. 124I-CLR1404 uptake was not detected in 1/2 low grade tumors, 2/4 lesions from treatment related changes, and 1/1 indeterminate lesion. For 6 malignant tumors, the average tumor to background ratios (TBR) were 9.32 ± 4.33 (range 3.46 to 15.42) at 24 hours and 10.04 ± 3.15 (range 5.17 to 13.17) at 48 hours. For 2 lesions from treatment related change, the average TBR were 5.05 ± 0.4 (range 4.76 to 5.33) at 24 hours and 4.88 ± 1.19 (range 4.04 to 5.72) at 48 hours. PET uptake had areas of both concordance and discordance compared with MRI. 124I-CLR1404 PET demonstrated avid tumor uptake in a variety of brain tumors with high tumor-to-background ratios. There were regions of concordance and discordance compared with MRI, which has potential clinical relevance. Expansion of these studies is required to determine the clinical significance of the 124I-CLR1404 PET findings.
Synaptic connectivity between gliomas and adjacent brain was recently implicated in tumorigenesis... more Synaptic connectivity between gliomas and adjacent brain was recently implicated in tumorigenesis. However, the interaction of synapse-related genes (SRGs) with patient survival and with established clinical and molecular glioma subtypes merit further study in a large patient cohort. We characterized differential expression of SRGs in gliomas and investigated SRG expression as putative clinical biomarkers in a large glioma cohort. Expression of 1189 SRGs was interrogated via RNA sequencing analysis in 603 gliomas (LGG n=451, GBM n=152) of The Cancer Genome Atlas. SRG expression patterns partitioned gliomas into two clusters that were more distinctive than priorly identified glioma subtypes. The two glioma clusters showed significantly low (14.9 months) or high median survival (105.1 months; Hazard Ratio = 7.1, 95% CI: [5.32, 9.78], p = 1.29e-46 using a log-rank test). The high survival cluster showed overrepresentation of known pro-neural and neural subtypes and IDH-mutated gliomas ...
BACKGROUND Radiomics and connectome analysis are distinct and non-invasive methods of deriving bi... more BACKGROUND Radiomics and connectome analysis are distinct and non-invasive methods of deriving biologic information from MRI. Radiomics analyzes features intrinsic to the tumor, and connectomics incorporates data regarding the tumor and surrounding neural circuitry. In this study we used both techniques to predict glioma survival. METHODS We retrospectively identified 305 adult patients with histopathologically confirmed WHO grade II–IV gliomas who had presurgical, 3D, T1-weighted brain MRI. Available clinical variables included tumor lobe, hemisphere, multifocal nature grade, histology extent of surgical resection, patient age gender. For connectomics, we calculated nodal efficiencies, network size and degree for all pairs of 33 voxel cubes spanning the entire gray matter volume using similarity-based extraction and graph theory. Radiomic features were extracted using Pyradiomics and subjected to patient-level and population-level clustering (N=172). These clusters were then used t...
BackgroundImmune checkpoint inhibition (ICI) alone is not efficacious for a large number of patie... more BackgroundImmune checkpoint inhibition (ICI) alone is not efficacious for a large number of patients with melanoma brain metastases. We previously established an in situ vaccination (ISV) regimen combining radiation and immunocytokine to enhance response to ICIs. Here, we tested whether ISV inhibits the development of brain metastases in a murine melanoma model.MethodsB78 (GD2+) melanoma ‘primary’ tumors were engrafted on the right flank of C57BL/6 mice. After 3–4 weeks, primary tumors were treated with ISV (radiation (12 Gy, day 1), α-GD2 immunocytokine (hu14.18-IL2, days 6–10)) and ICI (α-CTLA-4, days 3, 6, 9). Complete response (CR) was defined as no residual tumor observed at treatment day 90. Mice with CR were tested for immune memory by re-engraftment with B78 in the left flank and then the brain. To test ISV efficacy against metastases, tumors were also engrafted in the left flank and brain of previously untreated mice. Tumors were analyzed by quantitative reverse transcripti...
INTRODUCTION The median survival of glioblastoma (GBM) patients remains less than two years even ... more INTRODUCTION The median survival of glioblastoma (GBM) patients remains less than two years even with state-of-the-art treatment. Current targeted GBM therapies demonstrate initial therapeutic benefit; however, patients relapse due to therapeutic selection of treatment resistant GBM cellular populations. Therefore, we propose targeting pathologic disruption of the blood brain barrier (BBB) via exposure of neural ECM, rather than disease markers, to overcome therapy-resistant GBM. METHODS We identify Variable Lymphocyte Receptors (VLRs, the antigen recognition system used by lamprey) that demonstrate high specificity for neural ECM. Candidate VLRs underwent further refinement using in vitro binding assays and ex vivo tissue staining. Utilizing pathologic disruption of BBB as an approach for targeting GBM was confirmed in vivo using intracranial murine glioblastoma models. RESULTS The lead neural ECM-binding VLR candidate, named P1C10, demonstrates diffuse binding to parenchymal neura...
n engl j med 358;8 www.nejm.org february 21, 2008 853 consider other imaging options, if they wer... more n engl j med 358;8 www.nejm.org february 21, 2008 853 consider other imaging options, if they were equally effective. No CT scans were canceled or deferred after the parents received the risk information. Critique 4: It will be very difficult to reduce CT usage. We completely agree. Physicians are often subject to significant pressures, from the medical system, the medicolegal system, and the public, to order CT studies, even when they are not really necessary or even when alternatives exist. Our goal was to promote already ongoing dialogues among radiologists, emergency department and other physicians, and indeed the public about practical ways to reduce CT usage and CT doses, without compromising patient care. David J. Brenner. Ph.D., D.Sc. Eric J. Hall, D.Phil., D.Sc.
Many human malignancies such as prostate cancer (CaP) and primary brain tumor (GBM) exhibit high ... more Many human malignancies such as prostate cancer (CaP) and primary brain tumor (GBM) exhibit high oxidative stress. Developing a successful therapeutic drug to treat advanced metastatic CaP and GBM is a major unmet medical need. It has been demonstrated that in certain chemo- and/or radiation resistant cancer cells with high oxidative stress, cells depend on reactive oxygen species (ROS) for survival and proliferation. Therefore, reducing ROS can be a successful strategy for developing chemotherapeutic drugs for use against chemo- and/or radiation-resistant tumors. The mitochondria are a major source of superoxide and other ROS. We focused on targeting spin-trapping nitroxide analogs at the mitochondrial interstitial space with appropriate linker length in order to remove the superoxide in order to reduce the cellular ROS levels without interfering with the mitochondrial electron transport chain. Our lead compound in this new class of drugs is Mito-tempol-C10. The drug successfully b...
A Research Highlight of a recently published paper: "The effects of tumor treating fields (and te... more A Research Highlight of a recently published paper: "The effects of tumor treating fields (and temozolomide in MGMT expressing and non-expressing patient-derived glioblastoma cells" (J Clin Neurosci 2017; 36:120-124) is presented. Introductory background on alternating tumor treating field (TTFields) therapy, an FDA approved therapy for newly diagnosed glioblastoma, is reviewed in the context of standard temozolomide (TMZ) chemotherapy. The highlighted paper evaluated the potential interactions of TMZ and TTFields in vitro, as this could not be readily accomplished clinically. The authors reported a clinical model using 2 different sets of patient-derived GBM stem-like cells (GSCs) including MGMT-expressing (TMZ resistant) GSC and non-MGMT-expressing (TMZ sensitive) GSC. The results demonstrated no interactions, and the inability of TTFields to overcome TMZ resistance. The significant clinical implications of these results, and the rationale for exploring other innovative treatment strategies in combination with TTFields are discussed.
Glioblastoma multiforme (GBM) is a poorly treated human brain cancer with few established clinica... more Glioblastoma multiforme (GBM) is a poorly treated human brain cancer with few established clinically useful molecular prognostic markers. GBM gene expression and microRNA profiling recently revealed developmental subtypes that potentially correlates with prognosis and treatment response. However, the clinically significant subset of glioblastoma stem-like cells (GSC) still lacks detailed molecular characterization and correlation to patient survival. We report that molecular classification of patient-derived GSCs using neural lineage markers show association with differential xenograft invasiveness, and also demonstrate significant correlation to survival in both the mouse model and human patients. Immunoblot array of neural lineage markers classified five independently isolated human GSC lines into three classes exhibiting differential expression of oligodendrocyte progenitor cells (OPC), astrocyte progenitor cells (APC), and neural progenitor cells (NPC) markers. After orthotopic injection into immunodeficient mice, Ki-67 proliferative index independent xenograft infiltration was observed: class I GSCs (OPC and NPC positive) created focal lesions, class II GSCs (NPC positive) formed minimally invasive lesions, and class III GSCs (APC positive) established highly infiltrative lesions. The OPC marker, cyclic nucleotide phosphodiesterase (CNP), exhibited high expression in focal xenografts versus low expression in invasive xenografts. Differential CNP expression significantly correlated with mouse model survival, and CNP assay of a large clinically annotated human GBM tissue microarray also showed differential patient survival. Therefore, GSC molecular characterization with neural lineage markers revealed a novel and potentially useful clinical prognosis marker, and suggests clinical importance for patient-specific GSC subtyping. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5344. doi:1538-7445.AM2012-5344
Purpose: Glioblastoma is the most malignant primary brain tumor, with a median survival of less t... more Purpose: Glioblastoma is the most malignant primary brain tumor, with a median survival of less than 2 years. More effective therapeutic approaches are needed to improve clinical outcomes. Experimental Design: Glioblastoma patient-derived cells (GPDC) were isolated from patient glioblastomas and implanted in mice to form xenografts. IHC was performed for human Ether-ago go Related Gene (hERG) expression and tumor proliferation. Sphere-forming assays with the hERG blocker E-4031 were performed on a high and low hERG-expressing lines. A glioblastoma tissue microarray (TMA; 115 patients) was used to correlate hERG expression with patient survival. Clinical data were analyzed to determine whether patient survival was affected by incidental administration of hERG inhibitory drugs and the correlative effect of patient glioblastoma hERG expression levels. Results: hERG expression was upregulated in glioblastoma xenografts with higher proliferative indices. High hERGexpressing GPDCs showed a reduction in sphere formation when treated with hERG inhibitors compared with low hERG-expressing GPDCs. Glioblastoma TMA analysis showed worse survival for glioblastoma patients with high hERG expression versus low expression-43.5 weeks versus 60.9 weeks, respectively (P ¼ 0.022). Furthermore, patients who received at least one hERG blocker had a better survival rate compared with patients who did not (P ¼ 0.0015). Subgroup analysis showed that glioblastoma patients with high hERG expression who received hERG blockers had improved survival (P ¼ 0.0458). There was no difference in survival for low hERG-expressing glioblastoma patients who received hERG blockers (P ¼ 0.4136). Conclusions: Our findings suggest that hERG is a potential glioblastoma survival marker, and that already approved drugs with non-torsadogenic hERG inhibitory activity may potentially be repurposed as adjuvant glioblastoma therapy in high hERG-expressing glioblastoma patients. Clin Cancer Res; 23(1); 73-80. Ó2016 AACR.
Introduction: Glioblastoma Multiforme (GBM) is the most aggressive type of brain cancer and progr... more Introduction: Glioblastoma Multiforme (GBM) is the most aggressive type of brain cancer and progresses at a rapid rate. The major obstacle in the treatment of GBM is selective survival of a subpopulation of GBM derived stem cells (GSC) which are highly proliferative and resistant to drug induced apoptosis. The proteins involved in the regulation of the cell cycle impact survival potential of GBM and GSCs. Pentraxin 3 (PTX3), an angiogenic and survival factor plays a key role in cell proliferation, angiogenesis and invasion. However, little is known about the association of PTX3 and cell cycle regulation in GBM derived stem cells. Mehods: GBM patient tumor lines, and GSC derived from the same GBM lines were used. These cell lines were exposed to hypoxia and re-oxygenation. Cell cycle profiling and analysis were carried out using flow cytometry and FlowJo Single Cell Data Analysis Software. Xeno-transplanted mouse brain sections were used for PTX3 expression analysis by immunohistochemistry and immunofluorescence staining. Targeted proteomic approach was used to determine PTX3 levels in GBM and GSC. Alterations in the cell cycle regulatory gene expression in GSC and GBM tumor line in presence and absence of PTX3 was identified by transcriptome array analysis. Results: Our targeted proteomic approach using antibody array identified substantially higher levels of PTX3 in GSCs. GBM tumor lines showed higher percent of cells in G1 phase, where as GSCs showed increased number of cells in G2 and S-phase, both under normoxia and hypoxia. GSCs showed increased levels of GADD45A, known to protect GBM cells from apoptosis. Increased PTX3 expression correlated with increased cell cycle progression. Furthermore, exogenous PTX3 increased CyclinD1, a protein required for progression through the G1 phase of the cell cycle. Conclusion: GSC cells show cell cycle progression, presumably with an increased growth advantage. PTX3 expression is associated with GBM malignancy, and may contribute to GBM cell cycle progression and invasion. Overall, these results indicate that PTX3 and the cell cycle pathway are strongly associated with malignant GBM. Citation Format: Umadevi V. Wesley, Paul Clark, Jacob Jaeger, John Kuo, Robert Dempsey. Cell cycle pathway gene regulation in glioblastoma multiforme (GBM) and GBM derived stem cells: Implicating Pentraxin 3 upregulation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4300.
Glioblastoma multiforme (GBM) is highly invasive and resistant to current therapies. The aggressi... more Glioblastoma multiforme (GBM) is highly invasive and resistant to current therapies. The aggressiveness of brain tumor is attributed to GBM stem cells (GSC) that release inflammatory cytokines, and exacerbate inflammation in tumor microenvironment, critically affecting GBM survival and invasiveness. The glycoprotein pentraxin-3 (PTX3), a modulator of tumor-associated inflammation, is known to be correlated with tumor grade and severity of malignancies of some cancers. However, little is known about the role of PTX3 in GBM growth and invasion. In this study, we examined the role of PTX3 in GSCs exhibiting various degree of invasiveness. From patient brain tumors, our team has isolated the small subsets of glioblastoma stem cells (GSCs) and are validated for their self-renewal and multi-lineage potential. Our studies have shown that these GSCs efficiently form orthotopic brain tumors in immune-deficient mice. Our results show that conditioned media from GSCs promote pro-angiogenic tube formation from endothelial cells. GSCs that are grown in the absence of growth factors EGF and bFGF, formed efficient xenografts in immune-deficient mice and showed increased dispersal in the mice brain. Interestingly, polymerase chain reaction (RT-PCR) and immunofluorescence staining showed upregulated expression of PTX3 in these GSCs, and correlated with the degree of invasiveness. Differentiation of GSC into astrocytes in vitro, lead to decreased expression of PTX3. In support of this, recent histochemical study has shown that PTX3 levels correlate with GBM grade and is a candidate biomarker for cancer progression. Thus, PTX3 may represent a new target and marker of inflammation for glioma malignancy. Citation Format: Umadevi V. Wesley, Paul Clark, John S. Kuo, Robert J. Dempsey. Dysregulated expression of pentraxin-3 in glioblastoma (GBM) stem cells: Implications for GBM invasion and progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4773. doi:10.1158/1538-7445.AM2017-4773
Background: Glioblastoma (GBM) is highly invasive and resistant to multimodal treatment due to di... more Background: Glioblastoma (GBM) is highly invasive and resistant to multimodal treatment due to distorted vasculature and aggravated inflammation in a tumor microenvironment rich in dysregulated cytokines and growth factors. This phenomenon is partly attributed to GBM stem cells (GSC) that release angiogenic and inflammatory cytokines. The glycoprotein pentraxin-3 (PTX3), a modulator of inflammation, is correlated with disease severity in some cancers. However, its role in GBM and the molecular mechanisms of PTX3-mediated oncogenesis remain unclear. In this study, we examined the role of PTX3 in GBM growth and invasion, using in vitro and in vivo models. Methods: Patient derived GSC lines were isolated and validated. Tumor tissue micro arrays (TMA) were used to determine the association of PTX3 expression with patient GBM specimens representing various degree of disease. Intracranial tumor xenograft/orthotopic mouse models, cell culture models, proteomic profiling, immunohistochemistry, molecular and biochemical approaches were used. Results: Proteomic profiling identified increased PTX3 expression in GSCs as compared to GBM lines, and correlated with the degree of GSC invasiveness in an orthotopic tumor xenograft model. Increased PTX3 correlated with cell cycle progression and survival under hypoxic micro-environment. Conditioned media obtained from PTX3-over-expressing U87 cell line enhanced pro-angiogenic tube formation from endothelial cells. TMA screening showed that PTX3 expression is substantially increased in more than 70% of GBM specimans. Furthermore, PTX3 over-expression significantly increased survival of U87 cells under hypoxic conditions and resulted in increased tumor growth and invasion in an intracranial tumor xenograft mouse model. Upregulation of PTX3 in these tumors was associated with increased levels of inflammatory and angiogenic markers including interleukin-8 (IL-8) and vascular endothelial growth factor (VEGF), but decreased levels of thrombospondin1, a major anti-angiogenic factor. Downstream activation of PI3K/Akt - NFkB signaling pathways was also observed. Conclusion: PTX3 is dysregulated in GBM, and may augment inflammation and angiogenesis in tumor microenvironment. Thus, PTX3 may serve as an inflammatory marker for GBM microenvironment and also may represent a potential therapeutic target for GBM suppresion. Citation Format: Umadevi V. Wesley, Paul Clark, Ian Sutton, John Kuo, Robert Dempsey. Pentraxin-3 exacerbates glioblastoma invasion and angiogenesis through IL8-VEGF signaling in tumor microenvironment: PTX3 is a potential target for GBM suppression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 101.
CLR1404 is a cancer-selective alkyl phosphocholine (APC) analog that can be radiolabeled with 124... more CLR1404 is a cancer-selective alkyl phosphocholine (APC) analog that can be radiolabeled with 124I for PET imaging, 131I for targeted radiotherapy and/or SPECT imaging, or 125I for targeted radiotherapy. Studies have demonstrated avid CLR1404 uptake and prolonged retention in a broad spectrum of preclinical tumor models. The purpose of this pilot trial was to demonstrate avidity of 124I-CLR1404 in human brain tumors and develop a framework to evaluate this uptake for use in larger studies. 12 patients (8 men and 4 women; mean age of 43.9 ± 15.1 y; range 23-66 y) with 13 tumors were enrolled. Eleven patients had suspected tumor recurrence and 1 patient had a new diagnosis of high grade tumor. Patients were injected with 185 MBq ± 10% of 124I-CLR1404 followed by PET/CT imaging at 6-, 24-, and 48-hour. 124I-CLR1404 PET uptake was assessed qualitatively and compared with MRI. After PET image segmentation SUV values and tumor to background ratios were calculated. There was no significant uptake of 124I-CLR1404 in normal brain. In tumors, uptake tended to increase to 48 hours. Positive uptake was detected in 9 of 13 lesions: 5/5 high grade tumors, 1/2 low grade tumors, 1/1 meningioma, and 2/4 patients with treatment related changes. 124I-CLR1404 uptake was not detected in 1/2 low grade tumors, 2/4 lesions from treatment related changes, and 1/1 indeterminate lesion. For 6 malignant tumors, the average tumor to background ratios (TBR) were 9.32 ± 4.33 (range 3.46 to 15.42) at 24 hours and 10.04 ± 3.15 (range 5.17 to 13.17) at 48 hours. For 2 lesions from treatment related change, the average TBR were 5.05 ± 0.4 (range 4.76 to 5.33) at 24 hours and 4.88 ± 1.19 (range 4.04 to 5.72) at 48 hours. PET uptake had areas of both concordance and discordance compared with MRI. 124I-CLR1404 PET demonstrated avid tumor uptake in a variety of brain tumors with high tumor-to-background ratios. There were regions of concordance and discordance compared with MRI, which has potential clinical relevance. Expansion of these studies is required to determine the clinical significance of the 124I-CLR1404 PET findings.
Synaptic connectivity between gliomas and adjacent brain was recently implicated in tumorigenesis... more Synaptic connectivity between gliomas and adjacent brain was recently implicated in tumorigenesis. However, the interaction of synapse-related genes (SRGs) with patient survival and with established clinical and molecular glioma subtypes merit further study in a large patient cohort. We characterized differential expression of SRGs in gliomas and investigated SRG expression as putative clinical biomarkers in a large glioma cohort. Expression of 1189 SRGs was interrogated via RNA sequencing analysis in 603 gliomas (LGG n=451, GBM n=152) of The Cancer Genome Atlas. SRG expression patterns partitioned gliomas into two clusters that were more distinctive than priorly identified glioma subtypes. The two glioma clusters showed significantly low (14.9 months) or high median survival (105.1 months; Hazard Ratio = 7.1, 95% CI: [5.32, 9.78], p = 1.29e-46 using a log-rank test). The high survival cluster showed overrepresentation of known pro-neural and neural subtypes and IDH-mutated gliomas ...
BACKGROUND Radiomics and connectome analysis are distinct and non-invasive methods of deriving bi... more BACKGROUND Radiomics and connectome analysis are distinct and non-invasive methods of deriving biologic information from MRI. Radiomics analyzes features intrinsic to the tumor, and connectomics incorporates data regarding the tumor and surrounding neural circuitry. In this study we used both techniques to predict glioma survival. METHODS We retrospectively identified 305 adult patients with histopathologically confirmed WHO grade II–IV gliomas who had presurgical, 3D, T1-weighted brain MRI. Available clinical variables included tumor lobe, hemisphere, multifocal nature grade, histology extent of surgical resection, patient age gender. For connectomics, we calculated nodal efficiencies, network size and degree for all pairs of 33 voxel cubes spanning the entire gray matter volume using similarity-based extraction and graph theory. Radiomic features were extracted using Pyradiomics and subjected to patient-level and population-level clustering (N=172). These clusters were then used t...
BackgroundImmune checkpoint inhibition (ICI) alone is not efficacious for a large number of patie... more BackgroundImmune checkpoint inhibition (ICI) alone is not efficacious for a large number of patients with melanoma brain metastases. We previously established an in situ vaccination (ISV) regimen combining radiation and immunocytokine to enhance response to ICIs. Here, we tested whether ISV inhibits the development of brain metastases in a murine melanoma model.MethodsB78 (GD2+) melanoma ‘primary’ tumors were engrafted on the right flank of C57BL/6 mice. After 3–4 weeks, primary tumors were treated with ISV (radiation (12 Gy, day 1), α-GD2 immunocytokine (hu14.18-IL2, days 6–10)) and ICI (α-CTLA-4, days 3, 6, 9). Complete response (CR) was defined as no residual tumor observed at treatment day 90. Mice with CR were tested for immune memory by re-engraftment with B78 in the left flank and then the brain. To test ISV efficacy against metastases, tumors were also engrafted in the left flank and brain of previously untreated mice. Tumors were analyzed by quantitative reverse transcripti...
INTRODUCTION The median survival of glioblastoma (GBM) patients remains less than two years even ... more INTRODUCTION The median survival of glioblastoma (GBM) patients remains less than two years even with state-of-the-art treatment. Current targeted GBM therapies demonstrate initial therapeutic benefit; however, patients relapse due to therapeutic selection of treatment resistant GBM cellular populations. Therefore, we propose targeting pathologic disruption of the blood brain barrier (BBB) via exposure of neural ECM, rather than disease markers, to overcome therapy-resistant GBM. METHODS We identify Variable Lymphocyte Receptors (VLRs, the antigen recognition system used by lamprey) that demonstrate high specificity for neural ECM. Candidate VLRs underwent further refinement using in vitro binding assays and ex vivo tissue staining. Utilizing pathologic disruption of BBB as an approach for targeting GBM was confirmed in vivo using intracranial murine glioblastoma models. RESULTS The lead neural ECM-binding VLR candidate, named P1C10, demonstrates diffuse binding to parenchymal neura...
n engl j med 358;8 www.nejm.org february 21, 2008 853 consider other imaging options, if they wer... more n engl j med 358;8 www.nejm.org february 21, 2008 853 consider other imaging options, if they were equally effective. No CT scans were canceled or deferred after the parents received the risk information. Critique 4: It will be very difficult to reduce CT usage. We completely agree. Physicians are often subject to significant pressures, from the medical system, the medicolegal system, and the public, to order CT studies, even when they are not really necessary or even when alternatives exist. Our goal was to promote already ongoing dialogues among radiologists, emergency department and other physicians, and indeed the public about practical ways to reduce CT usage and CT doses, without compromising patient care. David J. Brenner. Ph.D., D.Sc. Eric J. Hall, D.Phil., D.Sc.
Many human malignancies such as prostate cancer (CaP) and primary brain tumor (GBM) exhibit high ... more Many human malignancies such as prostate cancer (CaP) and primary brain tumor (GBM) exhibit high oxidative stress. Developing a successful therapeutic drug to treat advanced metastatic CaP and GBM is a major unmet medical need. It has been demonstrated that in certain chemo- and/or radiation resistant cancer cells with high oxidative stress, cells depend on reactive oxygen species (ROS) for survival and proliferation. Therefore, reducing ROS can be a successful strategy for developing chemotherapeutic drugs for use against chemo- and/or radiation-resistant tumors. The mitochondria are a major source of superoxide and other ROS. We focused on targeting spin-trapping nitroxide analogs at the mitochondrial interstitial space with appropriate linker length in order to remove the superoxide in order to reduce the cellular ROS levels without interfering with the mitochondrial electron transport chain. Our lead compound in this new class of drugs is Mito-tempol-C10. The drug successfully b...
A Research Highlight of a recently published paper: "The effects of tumor treating fields (and te... more A Research Highlight of a recently published paper: "The effects of tumor treating fields (and temozolomide in MGMT expressing and non-expressing patient-derived glioblastoma cells" (J Clin Neurosci 2017; 36:120-124) is presented. Introductory background on alternating tumor treating field (TTFields) therapy, an FDA approved therapy for newly diagnosed glioblastoma, is reviewed in the context of standard temozolomide (TMZ) chemotherapy. The highlighted paper evaluated the potential interactions of TMZ and TTFields in vitro, as this could not be readily accomplished clinically. The authors reported a clinical model using 2 different sets of patient-derived GBM stem-like cells (GSCs) including MGMT-expressing (TMZ resistant) GSC and non-MGMT-expressing (TMZ sensitive) GSC. The results demonstrated no interactions, and the inability of TTFields to overcome TMZ resistance. The significant clinical implications of these results, and the rationale for exploring other innovative treatment strategies in combination with TTFields are discussed.
Glioblastoma multiforme (GBM) is a poorly treated human brain cancer with few established clinica... more Glioblastoma multiforme (GBM) is a poorly treated human brain cancer with few established clinically useful molecular prognostic markers. GBM gene expression and microRNA profiling recently revealed developmental subtypes that potentially correlates with prognosis and treatment response. However, the clinically significant subset of glioblastoma stem-like cells (GSC) still lacks detailed molecular characterization and correlation to patient survival. We report that molecular classification of patient-derived GSCs using neural lineage markers show association with differential xenograft invasiveness, and also demonstrate significant correlation to survival in both the mouse model and human patients. Immunoblot array of neural lineage markers classified five independently isolated human GSC lines into three classes exhibiting differential expression of oligodendrocyte progenitor cells (OPC), astrocyte progenitor cells (APC), and neural progenitor cells (NPC) markers. After orthotopic injection into immunodeficient mice, Ki-67 proliferative index independent xenograft infiltration was observed: class I GSCs (OPC and NPC positive) created focal lesions, class II GSCs (NPC positive) formed minimally invasive lesions, and class III GSCs (APC positive) established highly infiltrative lesions. The OPC marker, cyclic nucleotide phosphodiesterase (CNP), exhibited high expression in focal xenografts versus low expression in invasive xenografts. Differential CNP expression significantly correlated with mouse model survival, and CNP assay of a large clinically annotated human GBM tissue microarray also showed differential patient survival. Therefore, GSC molecular characterization with neural lineage markers revealed a novel and potentially useful clinical prognosis marker, and suggests clinical importance for patient-specific GSC subtyping. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5344. doi:1538-7445.AM2012-5344
Purpose: Glioblastoma is the most malignant primary brain tumor, with a median survival of less t... more Purpose: Glioblastoma is the most malignant primary brain tumor, with a median survival of less than 2 years. More effective therapeutic approaches are needed to improve clinical outcomes. Experimental Design: Glioblastoma patient-derived cells (GPDC) were isolated from patient glioblastomas and implanted in mice to form xenografts. IHC was performed for human Ether-ago go Related Gene (hERG) expression and tumor proliferation. Sphere-forming assays with the hERG blocker E-4031 were performed on a high and low hERG-expressing lines. A glioblastoma tissue microarray (TMA; 115 patients) was used to correlate hERG expression with patient survival. Clinical data were analyzed to determine whether patient survival was affected by incidental administration of hERG inhibitory drugs and the correlative effect of patient glioblastoma hERG expression levels. Results: hERG expression was upregulated in glioblastoma xenografts with higher proliferative indices. High hERGexpressing GPDCs showed a reduction in sphere formation when treated with hERG inhibitors compared with low hERG-expressing GPDCs. Glioblastoma TMA analysis showed worse survival for glioblastoma patients with high hERG expression versus low expression-43.5 weeks versus 60.9 weeks, respectively (P ¼ 0.022). Furthermore, patients who received at least one hERG blocker had a better survival rate compared with patients who did not (P ¼ 0.0015). Subgroup analysis showed that glioblastoma patients with high hERG expression who received hERG blockers had improved survival (P ¼ 0.0458). There was no difference in survival for low hERG-expressing glioblastoma patients who received hERG blockers (P ¼ 0.4136). Conclusions: Our findings suggest that hERG is a potential glioblastoma survival marker, and that already approved drugs with non-torsadogenic hERG inhibitory activity may potentially be repurposed as adjuvant glioblastoma therapy in high hERG-expressing glioblastoma patients. Clin Cancer Res; 23(1); 73-80. Ó2016 AACR.
Introduction: Glioblastoma Multiforme (GBM) is the most aggressive type of brain cancer and progr... more Introduction: Glioblastoma Multiforme (GBM) is the most aggressive type of brain cancer and progresses at a rapid rate. The major obstacle in the treatment of GBM is selective survival of a subpopulation of GBM derived stem cells (GSC) which are highly proliferative and resistant to drug induced apoptosis. The proteins involved in the regulation of the cell cycle impact survival potential of GBM and GSCs. Pentraxin 3 (PTX3), an angiogenic and survival factor plays a key role in cell proliferation, angiogenesis and invasion. However, little is known about the association of PTX3 and cell cycle regulation in GBM derived stem cells. Mehods: GBM patient tumor lines, and GSC derived from the same GBM lines were used. These cell lines were exposed to hypoxia and re-oxygenation. Cell cycle profiling and analysis were carried out using flow cytometry and FlowJo Single Cell Data Analysis Software. Xeno-transplanted mouse brain sections were used for PTX3 expression analysis by immunohistochemistry and immunofluorescence staining. Targeted proteomic approach was used to determine PTX3 levels in GBM and GSC. Alterations in the cell cycle regulatory gene expression in GSC and GBM tumor line in presence and absence of PTX3 was identified by transcriptome array analysis. Results: Our targeted proteomic approach using antibody array identified substantially higher levels of PTX3 in GSCs. GBM tumor lines showed higher percent of cells in G1 phase, where as GSCs showed increased number of cells in G2 and S-phase, both under normoxia and hypoxia. GSCs showed increased levels of GADD45A, known to protect GBM cells from apoptosis. Increased PTX3 expression correlated with increased cell cycle progression. Furthermore, exogenous PTX3 increased CyclinD1, a protein required for progression through the G1 phase of the cell cycle. Conclusion: GSC cells show cell cycle progression, presumably with an increased growth advantage. PTX3 expression is associated with GBM malignancy, and may contribute to GBM cell cycle progression and invasion. Overall, these results indicate that PTX3 and the cell cycle pathway are strongly associated with malignant GBM. Citation Format: Umadevi V. Wesley, Paul Clark, Jacob Jaeger, John Kuo, Robert Dempsey. Cell cycle pathway gene regulation in glioblastoma multiforme (GBM) and GBM derived stem cells: Implicating Pentraxin 3 upregulation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4300.
Glioblastoma multiforme (GBM) is highly invasive and resistant to current therapies. The aggressi... more Glioblastoma multiforme (GBM) is highly invasive and resistant to current therapies. The aggressiveness of brain tumor is attributed to GBM stem cells (GSC) that release inflammatory cytokines, and exacerbate inflammation in tumor microenvironment, critically affecting GBM survival and invasiveness. The glycoprotein pentraxin-3 (PTX3), a modulator of tumor-associated inflammation, is known to be correlated with tumor grade and severity of malignancies of some cancers. However, little is known about the role of PTX3 in GBM growth and invasion. In this study, we examined the role of PTX3 in GSCs exhibiting various degree of invasiveness. From patient brain tumors, our team has isolated the small subsets of glioblastoma stem cells (GSCs) and are validated for their self-renewal and multi-lineage potential. Our studies have shown that these GSCs efficiently form orthotopic brain tumors in immune-deficient mice. Our results show that conditioned media from GSCs promote pro-angiogenic tube formation from endothelial cells. GSCs that are grown in the absence of growth factors EGF and bFGF, formed efficient xenografts in immune-deficient mice and showed increased dispersal in the mice brain. Interestingly, polymerase chain reaction (RT-PCR) and immunofluorescence staining showed upregulated expression of PTX3 in these GSCs, and correlated with the degree of invasiveness. Differentiation of GSC into astrocytes in vitro, lead to decreased expression of PTX3. In support of this, recent histochemical study has shown that PTX3 levels correlate with GBM grade and is a candidate biomarker for cancer progression. Thus, PTX3 may represent a new target and marker of inflammation for glioma malignancy. Citation Format: Umadevi V. Wesley, Paul Clark, John S. Kuo, Robert J. Dempsey. Dysregulated expression of pentraxin-3 in glioblastoma (GBM) stem cells: Implications for GBM invasion and progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4773. doi:10.1158/1538-7445.AM2017-4773
Background: Glioblastoma (GBM) is highly invasive and resistant to multimodal treatment due to di... more Background: Glioblastoma (GBM) is highly invasive and resistant to multimodal treatment due to distorted vasculature and aggravated inflammation in a tumor microenvironment rich in dysregulated cytokines and growth factors. This phenomenon is partly attributed to GBM stem cells (GSC) that release angiogenic and inflammatory cytokines. The glycoprotein pentraxin-3 (PTX3), a modulator of inflammation, is correlated with disease severity in some cancers. However, its role in GBM and the molecular mechanisms of PTX3-mediated oncogenesis remain unclear. In this study, we examined the role of PTX3 in GBM growth and invasion, using in vitro and in vivo models. Methods: Patient derived GSC lines were isolated and validated. Tumor tissue micro arrays (TMA) were used to determine the association of PTX3 expression with patient GBM specimens representing various degree of disease. Intracranial tumor xenograft/orthotopic mouse models, cell culture models, proteomic profiling, immunohistochemistry, molecular and biochemical approaches were used. Results: Proteomic profiling identified increased PTX3 expression in GSCs as compared to GBM lines, and correlated with the degree of GSC invasiveness in an orthotopic tumor xenograft model. Increased PTX3 correlated with cell cycle progression and survival under hypoxic micro-environment. Conditioned media obtained from PTX3-over-expressing U87 cell line enhanced pro-angiogenic tube formation from endothelial cells. TMA screening showed that PTX3 expression is substantially increased in more than 70% of GBM specimans. Furthermore, PTX3 over-expression significantly increased survival of U87 cells under hypoxic conditions and resulted in increased tumor growth and invasion in an intracranial tumor xenograft mouse model. Upregulation of PTX3 in these tumors was associated with increased levels of inflammatory and angiogenic markers including interleukin-8 (IL-8) and vascular endothelial growth factor (VEGF), but decreased levels of thrombospondin1, a major anti-angiogenic factor. Downstream activation of PI3K/Akt - NFkB signaling pathways was also observed. Conclusion: PTX3 is dysregulated in GBM, and may augment inflammation and angiogenesis in tumor microenvironment. Thus, PTX3 may serve as an inflammatory marker for GBM microenvironment and also may represent a potential therapeutic target for GBM suppresion. Citation Format: Umadevi V. Wesley, Paul Clark, Ian Sutton, John Kuo, Robert Dempsey. Pentraxin-3 exacerbates glioblastoma invasion and angiogenesis through IL8-VEGF signaling in tumor microenvironment: PTX3 is a potential target for GBM suppression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 101.
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