48 results on '"Bakhshinyan, D"'
Search Results
2. Hitting more birds with one stone: CD70 as an actionable immunotherapeutic target in recurrent glioblastoma
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Seyfrid, M, primary, Maich, W, additional, Shaikh, MV, additional, Tatari, N, additional, Upreti, D, additional, Piyasena, D, additional, Subapanditha, M, additional, Savage, N, additional, McKenna, D, additional, Kuhlmann, L, additional, Khoo, A, additional, Salim, SK, additional, Bassey-Archibong, B, additional, Gwynne, W, additional, Chokshi, C, additional, Brown, K, additional, Murtaza, N, additional, Bakhshinyan, D, additional, Vora, P, additional, Venugopal, C, additional, Moffat, J, additional, and Singh, SK, additional
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- 2021
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3. CD133+ brain tumor-initiating cells are dependent on STAT3 signaling to drive medulloblastoma recurrence
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Garg, N, primary, Bakhshinyan, D, additional, Venugopal, C, additional, Mahendram, S, additional, Rosa, D A, additional, Vijayakumar, T, additional, Manoranjan, B, additional, Hallett, R, additional, McFarlane, N, additional, Delaney, K H, additional, Kwiecien, J M, additional, Arpin, C C, additional, Lai, P-S, additional, Gómez-Biagi, R F, additional, Ali, A M, additional, de Araujo, E D, additional, Ajani, O A, additional, Hassell, J A, additional, Gunning, P T, additional, and Singh, S K, additional
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- 2016
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4. OS4 - 161 Activated Wnt Signaling for the Therapeutic Targeting of Treatment-Refractory Medulloblastoma Stem Cells
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Manoranjan, B., primary, Mahendram, S., additional, Bakhshinyan, D., additional, Kameda-Smith, M., additional, Venugopal, C., additional, Doble, B.W., additional, and Singh, S. K., additional
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- 2016
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5. PS1 - 170 Bmi1 is a Therapeutic Target in Recurrent Childhood Medulloblastoma
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Garg, N., primary, Bakhshinyan, D., additional, Manoranjan, B., additional, Venugopal, C., additional, Hallett, R., additional, Mahendram, S., additional, Vijayakumar, T., additional, Subapanditha, M., additional, Qazi, M., additional, Singh, M., additional, McFarlane, N., additional, Mann, A., additional, Vora, P., additional, Davis, T., additional, and Singh, S., additional
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- 2016
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6. OS2 - 166 A Novel Model of Human Lung-to-Brain Metastasis and its Application to the Identification of Essential Metastatic Regulatory Genes
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Singh, M, primary, Venugopal, C, additional, Tokar, T, additional, Brown, K R, additional, McFarlane, N, additional, Bakhshinyan, D, additional, Vijayakumar, T, additional, Manoranjan, B, additional, Mahendram, S, additional, Vora, P, additional, Qazi, M, additional, Dhillon, M, additional, Tong, A, additional, Durrer, K, additional, Murty, N, additional, Hallett, R, additional, Hassell, J A, additional, Kaplan, D, additional, Cutz, JC, additional, Jurisica, I, additional, Moffat, J, additional, and Singh, S K, additional
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- 2016
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7. PS2 - 171 Bmi1 Identifies Treatment-Refractory Stem Cells in Human Glioblastoma
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Vora, P., primary, Qazi, M., additional, Venugopal, C., additional, Subapanditha, M., additional, Mahendram, S., additional, Chokshi, C., additional, Singh, M., additional, Bakhshinyan, D., additional, McFarlane, N., additional, and Singh, S., additional
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- 2016
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8. Modelling Therapy Resistance for the Identification of Treatment-Refractory Cell Population(s) in Human Glioblastoma
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Qazi, M.A., primary, Vora, P., additional, Venugopal, C., additional, Bakhshinyan, D., additional, Nixon, A., additional, McFarlane, N., additional, Subapanditha, M.K., additional, Murty, N.K., additional, Hallett, R.M., additional, Moffat, J., additional, and Singh, S.K., additional
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- 2016
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9. PS1 - 169 Discovering the Treatment Refractory BTIC Population in Group 3 Medulloblastoma through Therapy Adapted Patient-Derived Human Mouse Xenograft (PDX) Model
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Bakhshinyan, D., primary, Vijayakumar, T., additional, Manoranjan, B., additional, McFarlane, N., additional, Venugopal, C., additional, Singh, M., additional, Qazi, M., additional, Vora, P., additional, and Singh, S., additional
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- 2016
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10. MB-14 * DISCOVERING THE TREATMENT REFRACTORY BTIC POPULATION IN GROUP 3 MEDULLOBLASTOMA
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Bakhshinyan, D., primary, Vijayakumar, T., additional, Garg, N., additional, Manoranjan, B., additional, McFarlane, N., additional, Venugopal, C., additional, and Singh, S., additional
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- 2015
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11. MB-15 * ACTIVATED Wnt SIGNALING TARGETS Sox2+ TREATMENT-REFRACTORY Shh-DEPENDENT MEDULLOBLASTOMA STEM CELLS
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Manoranjan, B., primary, Venugopal, C., additional, Mahendram, S., additional, Moreira, S., additional, Hallett, R., additional, Vijayakumar, T., additional, Bakhshinyan, D., additional, McFarlane, N., additional, Hassell, J., additional, Doble, B., additional, and Singh, S., additional
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- 2015
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12. SOX2 IDENTIFIES THE TREATMENT-REFRACTORY STEM CELL POPULATION IN GROUP 2 MEDULLOBLASTOMA
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Singh, S. K., primary, Manoranjan, B., additional, Venugopal, C., additional, Vora, P., additional, McFarlane, N., additional, Garg, N., additional, Singh, M., additional, Mann, A., additional, Bakhshinyan, D., additional, Mahendran, S., additional, and Dunn, S., additional
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- 2014
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13. CD133+brain tumor-initiating cells are dependent on STAT3 signaling to drive medulloblastoma recurrence
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Garg, N, Bakhshinyan, D, Venugopal, C, Mahendram, S, Rosa, D A, Vijayakumar, T, Manoranjan, B, Hallett, R, McFarlane, N, Delaney, K H, Kwiecien, J M, Arpin, C C, Lai, P-S, Gómez-Biagi, R F, Ali, A M, de Araujo, E D, Ajani, O A, Hassell, J A, Gunning, P T, and Singh, S K
- Abstract
Medulloblastoma (MB), the most common malignant paediatric brain tumor, is currently treated using a combination of surgery, craniospinal radiotherapy and chemotherapy. Owing to MB stem cells (MBSCs), a subset of MB patients remains untreatable despite standard therapy. CD133 is used to identify MBSCs although its functional role in tumorigenesis has yet to be determined. In this work, we showed enrichment of CD133 in Group 3 MB is associated with increased rate of metastasis and poor clinical outcome. The signal transducers and activators of transcription-3 (STAT3) pathway are selectively activated in CD133+MBSCs and promote tumorigenesis through regulation of c-MYC, a key genetic driver of Group 3 MB. We screened compound libraries for STAT3 inhibitors and treatment with the selected STAT3 inhibitors resulted in tumor size reduction in vivo. We propose that inhibition of STAT3 signaling in MBSCs may represent a potential therapeutic strategy to treat patients with recurrent MB.
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- 2017
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14. Correction to: Dynamic profiling of medulloblastoma surfaceome.
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Bakhshinyan D, Suk Y, Kuhlmann L, Adile AA, Ignatchenko V, Custers S, Gwynne WD, Macklin A, Venugopal C, Kislinger T, and Singh SK
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- 2023
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15. Dynamic profiling of medulloblastoma surfaceome.
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Bakhshinyan D, Suk Y, Kuhlmann L, Adile AA, Ignatchenko V, Custers S, Gwynne WD, Macklin A, Venugopal C, Kislinger T, and Singh SK
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- Humans, Child, Brain, Medulloblastoma therapy, Brain Neoplasms, Cerebellar Neoplasms therapy
- Abstract
Medulloblastoma (MB) is the most common type of malignant pediatric brain cancer. The current standard of care (SOC) involves maximal safe resection and chemoradiotherapy in individuals older than 3 years, often leading to devastating neurocognitive and developmental deficits. Out of the four distinct molecular subgroups, Group 3 and 4 have the poorest patient outcomes due to the aggressive nature of the tumor and propensity to metastasize and recur post therapy. The toxicity of the SOC and lack of response in specific subtypes to the SOC underscores the urgent need for developing and translating novel treatment options including immunotherapies. To identify differentially enriched surface proteins that could be evaluated for potential future immunotherapeutic interventions, we leveraged N-glycocapture surfaceome profiling on Group 3 MB cells from primary tumor, through therapy, to recurrence using our established therapy-adapted patient derived xenograft model. Integrin 𝛼5 (ITGA5) was one of the most differentially enriched targets found at recurrence when compared to engraftment and untreated timepoints. In addition to being enriched at recurrence, shRNA-mediated knockdown and small molecule inhibition of ITGA5 have resulted in marked decrease in proliferation and self-renewal in vitro and demonstrated a survival advantage in vivo. Together, our data highlights the value of dynamic profiling of cells as they evolve through therapy and the identification of ITGA5 as a promising therapeutic target for recurrent Group 3 MB., (© 2023. The Author(s).)
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- 2023
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16. An effective kinase inhibition strategy for metastatic recurrent childhood medulloblastoma.
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Adile AA, Bakhshinyan D, Suk Y, Uehling D, Saini M, Aman A, Magolan J, Subapanditha MK, McKenna D, Chokshi C, Savage N, Kameda-Smith MM, Venugopal C, and Singh SK
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- Humans, Child, Mice, Animals, Adolescent, Xenograft Model Antitumor Assays, Neoplasm Recurrence, Local drug therapy, Disease Models, Animal, Protein Kinase Inhibitors pharmacology, Protein Kinase Inhibitors therapeutic use, Cell Line, Tumor, Medulloblastoma pathology, Brain Neoplasms drug therapy, Cerebellar Neoplasms pathology
- Abstract
Purpose: Medulloblastomas (MBs) constitute the most common malignant brain tumor in children and adolescents. MYC-amplified Group 3 MBs are characterized by disease recurrence, specifically in the leptomeninges, whereby patients with these metastatic tumors have a mortality rate nearing 100%. Despite limited research on such tumors, studies on MB metastases at diagnosis suggest targeting kinases to be beneficial., Methods: To identify kinase inhibitors that eradicate cells driving therapy evasion and tumor dissemination, we utilized our established patient-derived xenograft (PDX) mouse-adapted therapy platform that models human MB metastatic recurrences following standard chemoradiotherapy. High-throughput screens of 640 kinase inhibitors were conducted against cells isolated from mouse spines in the PDX model and human fetal neural stem cells to reveal compounds that targeted these treatment-refractory, metastatic cells, whilst sparing healthy cells. Blood-brain barrier permeability assays and additional in vitro experimentation helped select top candidates for in vivo studies., Results: Recurrent Group 3 MB PDX spine cells were therapeutically vulnerable to a selective checkpoint kinase 1 (CHK1) inhibitor and small molecular inhibitor of platelet-derived growth factor receptor beta (PDGFRβ). Inhibitor-treated cells showed a significant reduction in MB stem cell properties associated with treatment failure. Mice also demonstrated survival advantage when treated with a CHK1 inhibitor ex vivo., Conclusion: We identified CHK1 and PDGFRβ inhibitors that effectively target MB cells fueling treatment-refractory metastases. With limited research on effective therapies for Group 3 MB metastatic recurrences, this work highlights promising therapeutic options to treat these aggressive tumors. Additional studies are warranted to investigate these inhibitors' mechanisms and recommended in vivo administration., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
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- 2023
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17. Author Correction: Characterization of an RNA binding protein interactome reveals a context-specific post-transcriptional landscape of MYC-amplified medulloblastoma.
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Kameda-Smith MM, Zhu H, Luo EC, Suk Y, Xella A, Yee B, Chokshi C, Xing S, Tan F, Fox RG, Adile AA, Bakhshinyan D, Brown K, Gwynne WD, Subapanditha M, Miletic P, Picard D, Burns I, Moffat J, Paruch K, Fleming A, Hope K, Provias JP, Remke M, Lu Y, Reya T, Venugopal C, Reimand J, Wechsler-Reya RJ, Yeo GW, and Singh SK
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- 2023
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18. Cancer-selective metabolic vulnerabilities in MYC-amplified medulloblastoma.
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Gwynne WD, Suk Y, Custers S, Mikolajewicz N, Chan JK, Zador Z, Chafe SC, Zhai K, Escudero L, Zhang C, Zaslaver O, Chokshi C, Shaikh MV, Bakhshinyan D, Burns I, Chaudhry I, Nachmani O, Mobilio D, Maich WT, Mero P, Brown KR, Quaile AT, Venugopal C, Moffat J, Montenegro-Burke JR, and Singh SK
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- Child, Humans, Dihydroorotate Dehydrogenase, Cell Line, Tumor, Neoplasm Recurrence, Local, Pyrimidines therapeutic use, Medulloblastoma drug therapy, Medulloblastoma genetics, Medulloblastoma metabolism, Cerebellar Neoplasms drug therapy, Cerebellar Neoplasms genetics, Cerebellar Neoplasms metabolism
- Abstract
MYC-driven medulloblastoma (MB) is an aggressive pediatric brain tumor characterized by therapy resistance and disease recurrence. Here, we integrated data from unbiased genetic screening and metabolomic profiling to identify multiple cancer-selective metabolic vulnerabilities in MYC-driven MB tumor cells, which are amenable to therapeutic targeting. Among these targets, dihydroorotate dehydrogenase (DHODH), an enzyme that catalyzes de novo pyrimidine biosynthesis, emerged as a favorable candidate for therapeutic targeting. Mechanistically, DHODH inhibition acts on target, leading to uridine metabolite scarcity and hyperlipidemia, accompanied by reduced protein O-GlcNAcylation and c-Myc degradation. Pyrimidine starvation evokes a metabolic stress response that leads to cell-cycle arrest and apoptosis. We further show that an orally available small-molecule DHODH inhibitor demonstrates potent mono-therapeutic efficacy against patient-derived MB xenografts in vivo. The reprogramming of pyrimidine metabolism in MYC-driven medulloblastoma represents an unappreciated therapeutic strategy and a potential new class of treatments with stronger cancer selectivity and fewer neurotoxic sequelae., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 Elsevier Inc. All rights reserved.)
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- 2022
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19. Characterization of an RNA binding protein interactome reveals a context-specific post-transcriptional landscape of MYC-amplified medulloblastoma.
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Kameda-Smith MM, Zhu H, Luo EC, Suk Y, Xella A, Yee B, Chokshi C, Xing S, Tan F, Fox RG, Adile AA, Bakhshinyan D, Brown K, Gwynne WD, Subapanditha M, Miletic P, Picard D, Burns I, Moffat J, Paruch K, Fleming A, Hope K, Provias JP, Remke M, Lu Y, Reya T, Venugopal C, Reimand J, Wechsler-Reya RJ, Yeo GW, and Singh SK
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- Animals, Mice, Humans, Proteomics, RNA-Binding Proteins genetics, Nerve Tissue Proteins, Medulloblastoma genetics, Brain Neoplasms, Cerebellar Neoplasms genetics
- Abstract
Pediatric medulloblastoma (MB) is the most common solid malignant brain neoplasm, with Group 3 (G3) MB representing the most aggressive subgroup. MYC amplification is an independent poor prognostic factor in G3 MB, however, therapeutic targeting of the MYC pathway remains limited and alternative therapies for G3 MB are urgently needed. Here we show that the RNA-binding protein, Musashi-1 (MSI1) is an essential mediator of G3 MB in both MYC-overexpressing mouse models and patient-derived xenografts. MSI1 inhibition abrogates tumor initiation and significantly prolongs survival in both models. We identify binding targets of MSI1 in normal neural and G3 MB stem cells and then cross referenced these data with unbiased large-scale screens at the transcriptomic, translatomic and proteomic levels to systematically dissect its functional role. Comparative integrative multi-omic analyses of these large datasets reveal cancer-selective MSI1-bound targets sharing multiple MYC associated pathways, providing a valuable resource for context-specific therapeutic targeting of G3 MB., (© 2022. The Author(s).)
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- 2022
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20. Characterization of the minimal residual disease state reveals distinct evolutionary trajectories of human glioblastoma.
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Qazi MA, Salim SK, Brown KR, Mikolajewicz N, Savage N, Han H, Subapanditha MK, Bakhshinyan D, Nixon A, Vora P, Desmond K, Chokshi C, Singh M, Khoo A, Macklin A, Khan S, Tatari N, Winegarden N, Richards L, Pugh T, Bock N, Mansouri A, Venugopal C, Kislinger T, Goyal S, Moffat J, and Singh SK
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- Humans, Neoplasm Recurrence, Local genetics, Neoplasm Recurrence, Local pathology, Neoplasm, Residual genetics, Neoplastic Stem Cells pathology, Proteomics, Brain Neoplasms pathology, Glioblastoma genetics, Glioblastoma pathology
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Recurrence of solid tumors renders patients vulnerable to advanced, treatment-refractory disease state with mutational and oncogenic landscape distinctive from initial diagnosis. Improving outcomes for recurrent cancers requires a better understanding of cell populations that expand from the post-therapy, minimal residual disease (MRD) state. We profile barcoded tumor stem cell populations through therapy at tumor initiation, MRD, and recurrence in our therapy-adapted, patient-derived xenograft models of glioblastoma (GBM). Tumors show distinct patterns of recurrence in which clonal populations exhibit either a pre-existing fitness advantage or an equipotency fitness acquired through therapy. Characterization of the MRD state by single-cell and bulk RNA sequencing reveals a tumor-intrinsic immunomodulatory signature with prognostic significance at the transcriptomic level and in proteomic analysis of cerebrospinal fluid (CSF) collected from patients with GBM. Our results provide insight into the innate and therapy-driven dynamics of human GBM and the prognostic value of interrogating the MRD state in solid cancers., Competing Interests: Declaration of interests S.K. Singh and J.M. are consultants for and own shares in Century Therapeutics, Inc., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
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- 2022
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21. CD70 as an actionable immunotherapeutic target in recurrent glioblastoma and its microenvironment.
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Seyfrid M, Maich WT, Shaikh VM, Tatari N, Upreti D, Piyasena D, Subapanditha M, Savage N, McKenna D, Mikolajewicz N, Han H, Chokshi C, Kuhlmann L, Khoo A, Salim SK, Archibong-Bassey B, Gwynne W, Brown K, Murtaza N, Bakhshinyan D, Vora P, Venugopal C, Moffat J, Kislinger T, and Singh S
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- Animals, Brain Neoplasms immunology, Cell Proliferation, Glioblastoma immunology, Humans, Male, Mice, Inbred NOD, Mice, SCID, Neoplasm Recurrence, Local, Prognosis, Mice, Brain Neoplasms therapy, CD27 Ligand metabolism, Glioblastoma therapy, Immunotherapy methods, Proteomics methods, Transcriptome genetics, Tumor Microenvironment immunology
- Abstract
Purpose: Glioblastoma (GBM) patients suffer from a dismal prognosis, with standard of care therapy inevitably leading to therapy-resistant recurrent tumors. The presence of cancer stem cells (CSCs) drives the extensive heterogeneity seen in GBM, prompting the need for novel therapies specifically targeting this subset of tumor-driving cells. Here, we identify CD70 as a potential therapeutic target for recurrent GBM CSCs., Experimental Design: In the current study, we identified the relevance and functional influence of CD70 on primary and recurrent GBM cells, and further define its function using established stem cell assays. We use CD70 knockdown studies, subsequent RNAseq pathway analysis, and in vivo xenotransplantation to validate CD70's role in GBM. Next, we developed and tested an anti-CD70 chimeric antigen receptor (CAR)-T therapy, which we validated in vitro and in vivo using our established preclinical model of human GBM. Lastly, we explored the importance of CD70 in the tumor immune microenvironment (TIME) by assessing the presence of its receptor, CD27, in immune infiltrates derived from freshly resected GBM tumor samples., Results: CD70 expression is elevated in recurrent GBM and CD70 knockdown reduces tumorigenicity in vitro and in vivo . CD70 CAR-T therapy significantly improves prognosis in vivo . We also found CD27 to be present on the cell surface of multiple relevant GBM TIME cell populations, notably putative M1 macrophages and CD4 T cells., Conclusion: CD70 plays a key role in recurrent GBM cell aggressiveness and maintenance. Immunotherapeutic targeting of CD70 significantly improves survival in animal models and the CD70/CD27 axis may be a viable polytherapeutic avenue to co-target both GBM and its TIME., Competing Interests: Competing interests: DU, DB and PV are employees of Century Therapeutics Canada. The other authors declare no competing interests., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)
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- 2022
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22. Temporal profiling of therapy resistance in human medulloblastoma identifies novel targetable drivers of recurrence.
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Bakhshinyan D, Adile AA, Liu J, Gwynne WD, Suk Y, Custers S, Burns I, Singh M, McFarlane N, Subapanditha MK, Qazi MA, Vora P, Kameda-Smith MM, Savage N, Desmond KL, Tatari N, Tran D, Seyfrid M, Hope K, Bock NA, Venugopal C, Bader GD, and Singh SK
- Abstract
Medulloblastoma (MB) remains a leading cause of cancer-related mortality among children. The paucity of MB samples collected at relapse has hindered the functional understanding of molecular mechanisms driving therapy failure. New models capable of accurately recapitulating tumor progression in response to conventional therapeutic interventions are urgently needed. In this study, we developed a therapy-adapted PDX MB model that has a distinct advantage of generating human MB recurrence. The comparative gene expression analysis of MB cells collected throughout therapy led to identification of genes specifically up-regulated after therapy, including one previously undescribed in the setting of brain tumors, bactericidal/permeability-increasing fold-containing family B member 4 ( BPIFB4 ). Subsequent functional validation resulted in a markedly diminished in vitro proliferation, self-renewal, and longevity of MB cells, translating into extended survival and reduced tumor burden in vivo. Targeting endothelial nitric oxide synthase, a downstream substrate of BPIFB4, impeded growth of several patient-derived MB lines at low nanomolar concentrations.
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- 2021
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23. The Strange Case of Jekyll and Hyde: Parallels Between Neural Stem Cells and Glioblastoma-Initiating Cells.
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Bakhshinyan D, Savage N, Salim SK, Venugopal C, and Singh SK
- Abstract
During embryonic development, radial glial precursor cells give rise to neural lineages, and a small proportion persist in the adult mammalian brain to contribute to long-term neuroplasticity. Neural stem cells (NSCs) reside in two neurogenic niches of the adult brain, the hippocampus and the subventricular zone (SVZ). NSCs in the SVZ are endowed with the defining stem cell properties of self-renewal and multipotent differentiation, which are maintained by intrinsic cellular programs, and extrinsic cellular and niche-specific interactions. In glioblastoma, the most aggressive primary malignant brain cancer, a subpopulation of cells termed glioblastoma stem cells (GSCs) exhibit similar stem-like properties. While there is an extensive overlap between NSCs and GSCs in function, distinct genetic profiles, transcriptional programs, and external environmental cues influence their divergent behavior. This review highlights the similarities and differences between GSCs and SVZ NSCs in terms of their gene expression, regulatory molecular pathways, niche organization, metabolic programs, and current therapies designed to exploit these differences., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Bakhshinyan, Savage, Salim, Venugopal and Singh.)
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- 2021
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24. Strategies to Enhance the Efficacy of T-Cell Therapy for Central Nervous System Tumors.
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Upreti D, Bakhshinyan D, Bloemberg D, Vora P, Venugopal C, and Singh SK
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- Animals, Humans, Brain Neoplasms immunology, Brain Neoplasms pathology, Brain Neoplasms therapy, Glioblastoma immunology, Glioblastoma pathology, Glioblastoma therapy, Immunotherapy, Adoptive, Neoplasms, Experimental immunology, Neoplasms, Experimental pathology, Neoplasms, Experimental therapy
- Abstract
Mortality rates in patients diagnosed with central nervous system (CNS) tumors, originating in the brain or spinal cord, continue to remain high despite the advances in multimodal treatment regimens, including surgery, radiation, and chemotherapy. Recent success of adoptive cell transfer immunotherapy treatments using chimeric antigen receptor (CAR) engineered T cells against in chemotherapy resistant CD19 expressing B-cell lymphomas, has provided the foundation for investigating efficacy of CAR T immunotherapies in the context of brain tumor. Although significant efforts have been made in developing and translating the novel CAR T therapies for CNS tumors, including glioblastoma (GBM), researchers are yet to achieve a similar level of success as with liquid malignancies. In this review, we discuss strategies and considerations essential for developing robust preclinical models for the translation of T cell-based therapies for CNS tumors. Some of the key considerations include route of delivery, increasing persistence of T cells in tumor environment, remodeling of myeloid environment, establishing the window of treatment opportunity, harnessing endogenous immune system, designing multiple antigen targeting T cells, and rational combination of immunotherapy with the current standard of care. Although this review focuses primarily on CAR T therapies for GBM, similar strategies, and considerations are applicable to all CNS tumors in general., (Copyright © 2020 Upreti, Bakhshinyan, Bloemberg, Vora, Venugopal and Singh.)
- Published
- 2020
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25. De novo necroptosis creates an inflammatory environment mediating tumor susceptibility to immune checkpoint inhibitors.
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Workenhe ST, Nguyen A, Bakhshinyan D, Wei J, Hare DN, MacNeill KL, Wan Y, Oberst A, Bramson JL, Nasir JA, Vito A, El-Sayes N, Singh SK, McArthur AG, and Mossman KL
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- Animals, Antineoplastic Agents, Cell Line, Tumor, Female, Gene Deletion, Humans, Mammary Neoplasms, Animal, Mice, Mice, Transgenic, Necroptosis, Osteosarcoma metabolism, Immune Checkpoint Inhibitors pharmacology, Inflammation metabolism, Oncolytic Virotherapy, Oncolytic Viruses, Tumor Microenvironment
- Abstract
Cancer immunotherapies using monoclonal antibodies to block inhibitory checkpoints are showing durable remissions in many types of cancer patients, although the majority of breast cancer patients acquire little benefit. Human melanoma and lung cancer patient studies suggest that immune checkpoint inhibitors are often potent in patients that already have intratumoral T cell infiltrate; although it remains unknown what types of interventions can result in an intratumoral T cell infiltrate in breast cancer. Using non-T cell-inflamed mammary tumors, we assessed what biological processes and downstream inflammation can overcome the barriers to spontaneous T cell priming. Here we show a specific type of combination therapy, consisting of oncolytic virus and chemotherapy, activates necroptosis and limits tumor growth in autochthonous tumors. Combination therapy activates proinflammatory cytokines; intratumoral influx of myeloid cells and cytotoxic T cell infiltrate in locally treated and distant autochthonous tumors to render them susceptible to immune checkpoint inhibitors.
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- 2020
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26. Wnt activation as a therapeutic strategy in medulloblastoma.
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Manoranjan B, Venugopal C, Bakhshinyan D, Adile AA, Richards L, Kameda-Smith MM, Whitley O, Dvorkin-Gheva A, Subapanditha M, Savage N, Tatari N, McKenna D, Bassey-Archibong B, Winegarden N, Hallett R, Provias JP, Yarascavitch B, Ajani O, Fleming A, Bader GD, Pugh TJ, Doble BW, and Singh SK
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- Animals, Carcinogenesis, Cell Line, Tumor, Cell Proliferation, Cerebellar Neoplasms pathology, Disease Models, Animal, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, Heterografts, Humans, Medulloblastoma genetics, Medulloblastoma pathology, Mice, Stem Cells, Wnt Proteins genetics, Wnt Signaling Pathway, beta Catenin therapeutic use, Cerebellar Neoplasms therapy, Medulloblastoma therapy, Wnt Proteins pharmacology, Wnt Proteins therapeutic use
- Abstract
Medulloblastoma (MB) is defined by four molecular subgroups (Wnt, Shh, Group 3, Group 4) with Wnt MB having the most favorable prognosis. Since prior reports have illustrated the antitumorigenic role of Wnt activation in Shh MB, we aimed to assess the effects of activated canonical Wnt signaling in Group 3 and 4 MBs. By using primary patient-derived MB brain tumor-initiating cell (BTIC) lines, we characterize differences in the tumor-initiating capacity of Wnt, Group 3, and Group 4 MB. With single cell RNA-seq technology, we demonstrate the presence of rare Wnt-active cells in non-Wnt MBs, which functionally retain the impaired tumorigenic potential of Wnt MB. In treating MB xenografts with a Wnt agonist, we provide a rational therapeutic option in which the protective effects of Wnt-driven MBs may be augmented in Group 3 and 4 MB and thereby support emerging data for a context-dependent tumor suppressive role for Wnt/β-catenin signaling.
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- 2020
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27. The Rational Development of CD133-Targeting Immunotherapies for Glioblastoma.
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Vora P, Venugopal C, Salim SK, Tatari N, Bakhshinyan D, Singh M, Seyfrid M, Upreti D, Rentas S, Wong N, Williams R, Qazi MA, Chokshi C, Ding A, Subapanditha M, Savage N, Mahendram S, Ford E, Adile AA, McKenna D, McFarlane N, Huynh V, Wylie RG, Pan J, Bramson J, Hope K, Moffat J, and Singh S
- Subjects
- AC133 Antigen, Animals, Humans, Immunotherapy, Mice, Neoplastic Stem Cells, Brain Neoplasms therapy, Glioblastoma therapy
- Abstract
CD133 marks self-renewing cancer stem cells (CSCs) in a variety of solid tumors, and CD133+ tumor-initiating cells are known markers of chemo- and radio-resistance in multiple aggressive cancers, including glioblastoma (GBM), that may drive intra-tumoral heterogeneity. Here, we report three immunotherapeutic modalities based on a human anti-CD133 antibody fragment that targets a unique epitope present in glycosylated and non-glycosylated CD133 and studied their effects on targeting CD133+ cells in patient-derived models of GBM. We generated an immunoglobulin G (IgG) (RW03-IgG), a dual-antigen T cell engager (DATE), and a CD133-specific chimeric antigen receptor T cell (CAR-T): CART133. All three showed activity against patient-derived CD133+ GBM cells, and CART133 cells demonstrated superior efficacy in patient-derived GBM xenograft models without causing adverse effects on normal CD133+ hematopoietic stem cells in humanized CD34+ mice. Thus, CART133 cells may be a therapeutically tractable strategy to target CD133+ CSCs in human GBM or other treatment-resistant primary cancers., Competing Interests: Declaration of Interests P.V., C.V., R.W., J.M., and S.S. have patents around CD133 binding agents and uses thereof. These patents include Canadian Patent Application no. 2,962,157; Chinese Patent Application no. 2017800782373; European Patent Application no. EP17863201; Japanese Patent Application no. 2019-521046; and U.S. Patent Application no. 16/342,807. P.V., C.V., J.M., and S.S. are also shareholders of Empirica Therapeutics, which has an exclusive license to the IP as mentioned above. P.V. and D.B. are employees of Empirica Therapeutics., (Copyright © 2020 Elsevier Inc. All rights reserved.)
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- 2020
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28. Salvage therapy for progressive, treatment-refractory or recurrent pediatric medulloblastoma: a systematic review protocol.
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Adile AA, Kameda-Smith MM, Bakhshinyan D, Banfield L, Salim SK, Farrokhyar F, and Fleming AJ
- Subjects
- Child, Humans, Quality of Life, Salvage Therapy, Systematic Reviews as Topic, Brain Neoplasms, Cerebellar Neoplasms therapy, Medulloblastoma therapy
- Abstract
Background: Central nervous system tumors remain the leading cause of cancer-related mortality amongst children with solid tumors, with medulloblastoma (MB) representing the most common pediatric brain malignancy. Despite best current therapies, patients with recurrent MB experience have an alarmingly high mortality rate and often have limited therapeutic options beyond inadequate chemotherapy or experimental clinical trials. Therefore, a systematic review of the literature regarding treatment strategies employed in recurrent pediatric MB will evaluate previous salvage therapies in order to guide future clinical trials. The aim of this systematic review will be to investigate the efficacy and safety of salvage therapies for the management of children with progressive, treatment-refractory, or recurrent MB., Methods: We will conduct literature searches (from 1995 onwards) in MEDLINE, EMBASE, ClinicalTrials.gov, WHO International Clinical Trials Registry Platform, and Cochrane Central Register of Controlled Trials. Studies examining the survival and toxicity of therapies administered to treatment-refractory pediatric MB patients will be included. Two reviewers will independently assess the search results based on predefined selection criteria, complete data abstraction, and quality assessment. The primary outcomes of this review will be overall and progression-free survival. Secondary outcomes will include safety and toxicity of each therapy administered. The study methodological quality (or bias) will be appraised using an appropriate tool. Due to the nature of the research question and published literature, we expect large inter-study heterogeneity and therefore will use random effects regression analysis to extract the combined effect. In additional analyses, we will investigate the role of re-irradiation and mono- vs. poly-therapy in recurrent disease, and whether molecular subgrouping of MB influences salvage therapy., Discussion: This systematic review will provide an overview of the current literature regarding salvage therapies for relapsed MB patients. Investigation of clinically tested therapies for children with recurrent MB has significant implications for clinical practice. By reviewing the efficacy and toxicity of MB salvage therapies, this study will identify effective therapeutic strategies administered to recurrent MB patients and can inform future clinical trials aimed to improve patient survivorship and quality of life., Systematic Review Registration: PROSPERO CRD42020167421.
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- 2020
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29. Deciphering brain tumor heterogeneity, one cell at a time.
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Qazi MA, Bakhshinyan D, and Singh SK
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- Brain Neoplasms pathology, Glioblastoma pathology, Humans, Medulloblastoma pathology, RNA Interference, Single-Cell Analysis, Brain Neoplasms genetics, Genetic Heterogeneity, Glioblastoma genetics, Medulloblastoma genetics
- Published
- 2019
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30. Bmi1 regulates human glioblastoma stem cells through activation of differential gene networks in CD133+ brain tumor initiating cells.
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Vora P, Seyfrid M, Venugopal C, Qazi MA, Salim S, Isserlin R, Subapanditha M, O'Farrell E, Mahendram S, Singh M, Bakhshinyan D, Chokshi C, McFarlane N, Dvorkin-Gheva A, Brown KR, Murty N, Moffat J, Bader GD, and Singh SK
- Subjects
- AC133 Antigen genetics, Animals, Apoptosis, Brain Neoplasms genetics, Brain Neoplasms metabolism, Cell Proliferation, Glioblastoma genetics, Glioblastoma metabolism, Humans, Mice, Mice, Inbred NOD, Mice, SCID, Neoplastic Stem Cells metabolism, Polycomb Repressive Complex 1 genetics, Tumor Cells, Cultured, Xenograft Model Antitumor Assays, AC133 Antigen metabolism, Brain Neoplasms pathology, Gene Expression Regulation, Neoplastic, Gene Regulatory Networks, Glioblastoma pathology, Neoplastic Stem Cells pathology, Polycomb Repressive Complex 1 metabolism
- Abstract
Purpose: Glioblastoma (GBM) is the most aggressive adult brain cancer, with a 15 month median survivorship attributed to the existence of treatment-refractory brain tumor initiating cells (BTICs). In order to better understand the mechanisms regulating the tumorigenic properties of this population, we studied the role of the polycomb group member BMI1 in our patient-derived GBM BTICs and its relationship with CD133, a well-established marker of BTICs., Methods: Using gain and loss-of-function studies for Bmi1 in neural stem cells (NSCs) and patient-derived GBM BTICs respectively, we assessed in vitro self-renewal and in vivo tumor formation in these two cell populations. We further explored the BMI1 transcriptional regulatory network through RNA sequencing of different GBM BTIC populations that were knocked down for Bmi1., Results: There is a differential role of BMI1 in CD133-positive cells, notably involving cell metabolism. In addition, we identified pivotal targets downstream of BMI1 in CD133+ cells such as integrin alpha 2 (ITGA2), that may contribute to regulating GBM stem cell properties., Conclusions: Our work sheds light on the association of three genes with CD133-BMI1 circuitry, their importance as downstream effectors of the BMI1 signalling pathway, and their potential as future targets for tackling GBM treatment-resistant cell populations.
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- 2019
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31. Salvage Therapy for Childhood Medulloblastoma: A Single Center Experience.
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Kameda-Smith MM, Wang A, Abdulhadi N, Voth R, Sergeant A, Maharaj A, Bakhshinyan D, Adile AA, Pai AM, Ajani O, Yarascavitch B, Alyman MC, Duckworth J, Samaan MC, Farrokhyar F, Singh SK, and Fleming A
- Subjects
- Adolescent, Cerebellar Neoplasms mortality, Child, Child, Preschool, Female, Humans, Infant, Male, Medulloblastoma mortality, Neoplasm Recurrence, Local mortality, Retrospective Studies, Salvage Therapy mortality, Treatment Outcome, Cerebellar Neoplasms therapy, Medulloblastoma therapy, Neoplasm Recurrence, Local therapy, Salvage Therapy methods
- Abstract
Introduction: Children diagnosed with medulloblastoma (MB) who are refractory to upfront therapy or experience recurrence have very poor prognoses. Although phase I and phase II trials exist, these treatments bear significant treatment-related morbidity and mortality., Methods: A retrospective review of children diagnosed with a recurrence of MB from 2002 to 2015 at McMaster University was undertaken., Results: Recurrent disease in 10 patients involved leptomeningeal dissemination, with 3 experiencing local recurrence. In three recurrent patients the disease significantly progressed, and the children were palliated. The remaining 10 children underwent some form of salvage therapy, including surgical re-resection, radiation, and chemotherapy, either in isolation or in varying combinations. Of the 13 children experiencing treatment-refractory or recurrent disease, 4 are currently alive with a median follow-up of 38.5 months (75.5 months). Of the eight patients with molecular subgrouping data, none of the Wnt MB experienced recurrence., Conclusion: Recurrent MB carried a poor prognosis with a 5-year overall survival (OS) of 18.2% despite the administration of salvage therapy. The upfront therapy received, available treatment, and tolerability of the proposed salvage therapy resulted in significant heterogeneity in the treatment of our recurrent cohort.
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- 2019
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32. Identification and Characterization of AES-135, a Hydroxamic Acid-Based HDAC Inhibitor That Prolongs Survival in an Orthotopic Mouse Model of Pancreatic Cancer.
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Shouksmith AE, Shah F, Grimard ML, Gawel JM, Raouf YS, Geletu M, Berger-Becvar A, de Araujo ED, Luchman HA, Heaton WL, Bakhshinyan D, Adile AA, Venugopal C, O'Hare T, Deininger MW, Singh SK, Konieczny SF, Weiss S, Fishel ML, and Gunning PT
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- Animals, Apoptosis drug effects, Benzamides chemistry, Benzamides pharmacokinetics, Benzamides therapeutic use, Cell Line, Tumor, Cell Proliferation drug effects, Coculture Techniques, Disease Models, Animal, Histone Deacetylase Inhibitors chemistry, Histone Deacetylase Inhibitors pharmacokinetics, Histone Deacetylase Inhibitors therapeutic use, Humans, Hydrocarbons, Fluorinated chemistry, Hydrocarbons, Fluorinated pharmacokinetics, Hydrocarbons, Fluorinated therapeutic use, Mice, Pancreatic Neoplasms pathology, Sulfonamides chemistry, Sulfonamides pharmacokinetics, Sulfonamides therapeutic use, Benzamides pharmacology, Histone Deacetylase Inhibitors pharmacology, Hydrocarbons, Fluorinated pharmacology, Hydroxamic Acids chemistry, Pancreatic Neoplasms drug therapy, Sulfonamides pharmacology
- Abstract
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive, incurable cancer with a 20% 1 year survival rate. While standard-of-care therapy can prolong life in a small fraction of cases, PDAC is inherently resistant to current treatments, and novel therapies are urgently required. Histone deacetylase (HDAC) inhibitors are effective in killing pancreatic cancer cells in in vitro PDAC studies, and although there are a few clinical studies investigating combination therapy including HDAC inhibitors, no HDAC drug or combination therapy with an HDAC drug has been approved for the treatment of PDAC. We developed an inhibitor of HDACs, AES-135, that exhibits nanomolar inhibitory activity against HDAC3, HDAC6, and HDAC11 in biochemical assays. In a three-dimensional coculture model, AES-135 kills low-passage patient-derived tumor spheroids selectively over surrounding cancer-associated fibroblasts and has excellent pharmacokinetic properties in vivo. In an orthotopic murine model of pancreatic cancer, AES-135 prolongs survival significantly, therefore representing a candidate for further preclinical testing.
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- 2019
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33. BMI1 is a therapeutic target in recurrent medulloblastoma.
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Bakhshinyan D, Venugopal C, Adile AA, Garg N, Manoranjan B, Hallett R, Wang X, Mahendram S, Vora P, Vijayakumar T, Subapanditha M, Singh M, Kameda-Smith MM, Qazi M, McFarlane N, Mann A, Ajani OA, Yarascavitch B, Ramaswamy V, Farooq H, Morrissy S, Cao L, Sydorenko N, Baiazitov R, Du W, Sheedy J, Weetall M, Moon YC, Lee CS, Kwiecien JM, Delaney KH, Doble B, Cho YJ, Mitra S, Kaplan D, Taylor MD, Davis TW, and Singh SK
- Subjects
- Animals, Cell Line, Tumor, Cell Proliferation drug effects, Cell Survival drug effects, Cerebellar Neoplasms genetics, Cerebellar Neoplasms metabolism, Child, Epigenesis, Genetic, Gene Expression Regulation, Neoplastic drug effects, Humans, Medulloblastoma genetics, Medulloblastoma metabolism, Mice, Neoplastic Stem Cells cytology, Neoplastic Stem Cells metabolism, Polycomb Repressive Complex 1 genetics, Small Molecule Libraries pharmacology, Treatment Outcome, Up-Regulation drug effects, Xenograft Model Antitumor Assays, Cerebellar Neoplasms drug therapy, Medulloblastoma drug therapy, Neoplastic Stem Cells drug effects, Polycomb Repressive Complex 1 antagonists & inhibitors, Small Molecule Libraries administration & dosage
- Abstract
Medulloblastoma (MB) is the most frequent malignant pediatric brain tumor, representing 20% of newly diagnosed childhood central nervous system malignancies. Although advances in multimodal therapy yielded a 5-year survivorship of 80%, MB still accounts for the leading cause of childhood cancer mortality. In this work, we describe the epigenetic regulator BMI1 as a novel therapeutic target for the treatment of recurrent human Group 3 MB, a childhood brain tumor for which there is virtually no treatment option beyond palliation. Current clinical trials for recurrent MB patients based on genomic profiles of primary, treatment-naive tumors will provide limited clinical benefit since recurrent metastatic MBs are highly genetically divergent from their primary tumor. Using a small molecule inhibitor against BMI1, PTC-028, we were able to demonstrate complete ablation of self-renewal of MB stem cells in vitro. When administered to mice xenografted with patient tumors, we observed significant reduction in tumor burden in both local and metastatic compartments and subsequent increased survival, without neurotoxicity. Strikingly, serial in vivo re-transplantation assays demonstrated a marked reduction in tumor initiation ability of recurrent MB cells upon re-transplantation of PTC-028-treated cells into secondary recipient mouse brains. As Group 3 MB is often metastatic and uniformly fatal at recurrence, with no current or planned trials of targeted therapy, an efficacious targeted agent would be rapidly transitioned to clinical trials.
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- 2019
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34. In Vitro Self-Renewal Assays for Brain Tumor Stem Cells.
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Seyfrid M, Bobrowski D, Bakhshinyan D, Tatari N, Venugopal C, and Singh SK
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- Cell Line, Tumor, Humans, Regression Analysis, Biological Assay methods, Brain Neoplasms pathology, Cell Self Renewal, Neoplastic Stem Cells pathology
- Abstract
Early development of human organisms relies on stem cells, a population of non-specialized cells that can divide symmetrically to give rise to two identical daughter cells, or divide asymmetrically to produce one identical daughter cell and another more specialized cell. The capacity to undergo cellular divisions while maintaining an undifferentiated state is termed self-renewal and is responsible for the maintenance of stem cell populations during development. In addition, self-renewal plays a crucial role in the homeostasis of developed organism through replacement of defective cells.Similar to their non-malignant counterparts, it has been postulated that tumor cells follow a differentiation hierarchy, with the least differentiated cells termed cancer stem cells (CSCs) at the apex. These tumor stem cells possess the ability to self-renew, have a higher capacity to initiate tumor growth when xenografted into an animal model, and can recapitulate the cell heterogeneity of the tumor they originate from. Hence, further investigation of mechanisms governing the self-renewal in cancer can lead to development of novel therapies targeting CSCs.In this chapter, we described the soft agar assay and the limiting dilution assay (LDA) as two easy-to-implement and inexpensive assays to measure the stemness properties of brain tumor stem cells (BTSCs). These techniques constitute useful tools for the preclinical evaluation of therapeutic strategies targeting BTSCs clonogenicity.
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- 2019
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35. Introduction to Brain Tumor Stem Cells.
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Yelle N, Bakhshinyan D, Venugopal C, and Singh SK
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- Animals, Humans, Neural Stem Cells pathology, Brain Neoplasms pathology, Neoplastic Stem Cells pathology
- Abstract
From stem cells, to the cancer stem cell hypothesis and intratumoral heterogeneity, the following introductory chapter on brain tumor stem cells explores the history of normal and cancerous stem cells, and their implication in the current model of brain tumor development. The origins of stem cells date back to the 1960s, when they were first described as cells capable of self-renewal, extensive proliferation, and differentiation. Since then, many advances have been made and adult stem cells are now known to be present in a very wide variety of tissues. Neural stem cells were subsequently discovered 30 years later, which was shortly followed by the discovery of cancer stem cells in leukemia and in brain tumors over the next decade, effectively enabling a new understanding of cancer. Since then, many markers including CD133, brain cancer stem cells have been implicated in a variety of phenomena including intratumoral heterogeneity on the genomic, cellular, and functional levels, tumor initiation, chemotherapy-resistance, radiation-resistance, and are believed to be ultimately responsible for tumor relapse. Understanding this small and rare population of cells could be the key to solving the great enigma that is cancer.
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- 2019
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36. In Vitro Assays for Screening Small Molecules.
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Adile AA, Bakhshinyan D, Venugopal C, and Singh SK
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- Cell Line, Cell Proliferation, Cell Self Renewal, Humans, Inhibitory Concentration 50, Suspensions, Biological Assay methods, Small Molecule Libraries analysis
- Abstract
Traditionally anti-cancer therapeutics have been designed to target rapidly proliferating cells causing DNA damage and inducing apoptosis. However, with the development of the cancer stem cell (CSC) hypothesis, it has been postulated that a rare, slow dividing tumor cell population is able to escape therapy and contribute to tumor relapse and metastasis. The advances in characterization of CSCs across multiple cancer subtypes have allowed for development of targeted therapies using small molecule inhibitors. In this chapter, we describe two in vitro assays measuring proliferation and secondary sphere formation, which have become gold-standard assays to evaluate the effects of targeted therapies against CSCs. Together these assays constitute a rapid, inexpensive, and highly reproducible pipeline for testing small molecule inhibitors prior to more resource demanding in vivo studies.
- Published
- 2019
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37. Therapeutic Targeting of the Premetastatic Stage in Human Lung-to-Brain Metastasis.
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Singh M, Venugopal C, Tokar T, McFarlane N, Subapanditha MK, Qazi M, Bakhshinyan D, Vora P, Murty NK, Jurisica I, and Singh SK
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- Apomorphine pharmacology, Apoptosis drug effects, Blood-Brain Barrier drug effects, Brain drug effects, Brain pathology, Brain Neoplasms genetics, Brain Neoplasms pathology, Brain Neoplasms secondary, Cell Line, Tumor, Cell Proliferation drug effects, Dopamine metabolism, Dopamine Agonists pharmacology, Gene Expression Regulation, Neoplastic drug effects, Humans, Intracellular Signaling Peptides and Proteins genetics, Kinesins genetics, Lung pathology, Lung Neoplasms genetics, Lung Neoplasms pathology, Neoplasm Metastasis genetics, Neoplasm Metastasis pathology, Protein Serine-Threonine Kinases genetics, Selenoprotein W genetics, Brain Neoplasms drug therapy, Lung Neoplasms drug therapy, Molecular Targeted Therapy, Neoplasm Metastasis drug therapy
- Abstract
Brain metastases (BM) result from the spread of primary tumors to the brain and are a leading cause of cancer mortality in adults. Secondary tissue colonization remains the main bottleneck in metastatic development, yet this "premetastatic" stage of the metastatic cascade, when primary tumor cells cross the blood-brain barrier and seed the brain before initiating a secondary tumor, remains poorly characterized. Current studies rely on specimens from fully developed macrometastases to identify therapeutic options in cancer treatment, overlooking the potentially more treatable "premetastatic" phase when colonizing cancer cells could be targeted before they initiate the secondary brain tumor. Here we use our established brain metastasis initiating cell (BMIC) models and gene expression analyses to characterize premetastasis in human lung-to-BM. Premetastatic BMIC engaged invasive and epithelial developmental mechanisms while simultaneously impeding proliferation and apoptosis. We identified the dopamine agonist apomorphine to be a potential premetastasis-targeting drug. In vivo treatment with apomorphine prevented BM formation, potentially by targeting premetastasis-associated genes KIF16B, SEPW1 , and TESK2 Low expression of these genes was associated with poor survival of patients with lung adenocarcinoma. These results illuminate the cellular and molecular dynamics of premetastasis, which is subclinical and currently impossible to identify or interrogate in human patients with BM. These data present several novel therapeutic targets and associated pathways to prevent BM initiation. Significance: These findings unveil molecular features of the premetastatic stage of lung-to-brain metastases and offer a potential therapeutic strategy to prevent brain metastases. Cancer Res; 78(17); 5124-34. ©2018 AACR ., (©2018 American Association for Cancer Research.)
- Published
- 2018
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38. Transforming the prostatic tumor microenvironment with oncolytic virotherapy.
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Atherton MJ, Stephenson KB, Tzelepis F, Bakhshinyan D, Nikota JK, Son HH, Jirovec A, Lefebvre C, Dvorkin-Gheva A, Ashkar AA, Wan Y, Stojdl DF, Belanger EC, Breau RH, Bell JC, Saad F, Singh SK, Diallo JS, and Lichty BD
- Abstract
Prostate cancer (PCa) was estimated to have the second highest global incidence rate for male non-skin tumors and is the fifth most deadly in men thus mandating the need for novel treatment options. MG1-Maraba is a potent and versatile oncolytic virus capable of lethally infecting a variety of prostatic tumor cell lines alongside primary PCa biopsies and exerts direct oncolytic effects against large TRAMP-C2 tumors in vivo . An oncolytic immunotherapeutic strategy utilizing a priming vaccine and intravenously administered MG1-Maraba both expressing the human six-transmembrane antigen of the prostate (STEAP) protein generated specific CD8+ T-cell responses against multiple STEAP epitopes and resulted in functional breach of tolerance. Treatment of mice with bulky TRAMP-C2 tumors using oncolytic STEAP immunotherapy induced an overt delay in tumor progression, marked intratumoral lymphocytic infiltration with an active transcriptional profile and up-regulation of MHC class I. The preclinical data generated here offers clear rationale for clinically evaluating this approach for men with advanced PCa.
- Published
- 2018
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39. Erratum to: Introduction to Cancer Stem Cells: Past, Present, and Future.
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Bakhshinyan D, Adile AA, Qazi MA, Singh M, Kameda-Smith MM, Yelle N, Chokshi C, Venugopal C, and Singh SK
- Published
- 2018
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40. Introduction to Cancer Stem Cells: Past, Present, and Future.
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Bakhshinyan D, Adile AA, Qazi MA, Singh M, Kameda-Smith MM, Yelle N, Chokshi C, Venugopal C, and Singh SK
- Subjects
- CRISPR-Cas Systems genetics, CRISPR-Cas Systems physiology, Cell Differentiation genetics, Cell Differentiation physiology, Drug Resistance, Neoplasm genetics, Drug Resistance, Neoplasm physiology, Humans, Immunotherapy methods, Leukemia therapy, Neoplastic Stem Cells metabolism
- Abstract
The Cancer Stem Cell (CSC) hypothesis postulates the existence of a small population of cancer cells with intrinsic properties allowing for resistance to conventional radiochemotherapy regiments and increased metastatic potential. Clinically, the aggressive nature of CSCs has been shown to correlate with increased tumor recurrence, metastatic spread, and overall poor patient outcome across multiple cancer subtypes. Traditionally, isolation of CSCs has been achieved through utilization of cell surface markers, while the functional differences between CSCs and remaining tumor cells have been described through proliferation, differentiation, and limiting dilution assays. The generated insights into CSC biology have further highlighted the importance of studying intratumoral heterogeneity through advanced functional assays, including CRISPR-Cas9 screens in the search of novel targeted therapies. In this chapter, we review the discovery and characterization of cancer stem cells populations within several major cancer subtypes, recent developments of novel assays used in studying therapy resistant tumor cells, as well as recent developments in therapies targeted at cancer stem cells.
- Published
- 2018
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41. RNAi screen identifies essential regulators of human brain metastasis-initiating cells.
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Singh M, Venugopal C, Tokar T, Brown KR, McFarlane N, Bakhshinyan D, Vijayakumar T, Manoranjan B, Mahendram S, Vora P, Qazi M, Dhillon M, Tong A, Durrer K, Murty N, Hallet R, Hassell JA, Kaplan DR, Cutz JC, Jurisica I, Moffat J, and Singh SK
- Subjects
- Adenocarcinoma genetics, Adenocarcinoma pathology, Adenocarcinoma physiopathology, Adult, Aged, Aged, 80 and over, Animals, Brain Neoplasms physiopathology, Carcinoma, Non-Small-Cell Lung genetics, Carcinoma, Non-Small-Cell Lung pathology, Carcinoma, Non-Small-Cell Lung physiopathology, Cell Line, Tumor, Female, Humans, Male, Mice, Inbred NOD, Mice, SCID, Middle Aged, Neoplasm Transplantation, Prospective Studies, Proteoglycans genetics, Proteoglycans metabolism, RNA Interference, Repressor Proteins genetics, Repressor Proteins metabolism, Twist-Related Protein 1 genetics, Twist-Related Protein 1 metabolism, Brain Neoplasms genetics, Brain Neoplasms secondary, Gene Expression Regulation, Neoplastic, Neoplasm Metastasis genetics, Neoplasm Metastasis physiopathology
- Abstract
Brain metastases (BM) are the most common brain tumor in adults and are a leading cause of cancer mortality. Metastatic lesions contain subclones derived from their primary lesion, yet their functional characterization is limited by a paucity of preclinical models accurately recapitulating the metastatic cascade, emphasizing the need for a novel approach to BM and their treatment. We identified a unique subset of stem-like cells from primary human patient brain metastases, termed brain metastasis-initiating cells (BMICs). We now establish a BMIC patient-derived xenotransplantation (PDXT) model as an investigative tool to comprehensively interrogate human BM. Using both in vitro and in vivo RNA interference screens of these BMIC models, we identified SPOCK1 and TWIST2 as essential BMIC regulators. SPOCK1 in particular is a novel regulator of BMIC self-renewal, modulating tumor initiation and metastasis from the lung to the brain. A prospective cohort of primary lung cancer specimens showed that SPOCK1 was overexpressed only in patients who ultimately developed BM. Protein-protein interaction network mapping between SPOCK1 and TWIST2 identified novel pathway interactors with significant prognostic value in lung cancer patients. Of these genes, INHBA, a TGF-β ligand found mutated in lung adenocarcinoma, showed reduced expression in BMICs with knockdown of SPOCK1. In conclusion, we have developed a useful preclinical model of BM, which has served to identify novel putative BMIC regulators, presenting potential therapeutic targets that block the metastatic process, and transform a uniformly fatal systemic disease into a locally controlled and eminently more treatable one.
- Published
- 2017
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42. Preclinical Modeling and Therapeutic Avenues for Cancer Metastasis to the Central Nervous System.
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Singh M, Bakhshinyan D, Venugopal C, and Singh SK
- Abstract
Metastasis is the dissemination of cells from the primary tumor to other locations within the body, and continues to be the predominant cause of death among cancer patients. Metastatic progression within the adult central nervous system is 10 times more frequent than primary brain tumors. Metastases affecting the brain parenchyma and leptomeninges are associated with grave prognosis, and even after successful control of the primary tumor the median survival is a dismal 2-3 months with treatment options typically limited to palliative care. Current treatment options for brain metastases (BM) and disseminated brain tumors are scarce, and the improvement of novel targeted therapies requires a broader understanding of the biological complexity that characterizes metastatic progression. In this review, we provide insight into patterns of BM progression and leptomeningeal spread, outlining the development of clinically relevant in vivo models and their contribution to the discovery of innovative cancer therapies. In vivo models paired with manipulation of in vitro methods have expanded the tools available for investigators to develop agents that can be used to prevent or treat metastatic disease. The knowledge gained from the use of such models can ultimately lead to the prevention of metastatic dissemination and can extend patient survival by transforming a uniformly fatal systemic disease into a locally controlled and eminently more treatable one.
- Published
- 2017
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43. CD133 + brain tumor-initiating cells are dependent on STAT3 signaling to drive medulloblastoma recurrence.
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Garg N, Bakhshinyan D, Venugopal C, Mahendram S, Rosa DA, Vijayakumar T, Manoranjan B, Hallett R, McFarlane N, Delaney KH, Kwiecien JM, Arpin CC, Lai PS, Gómez-Biagi RF, Ali AM, de Araujo ED, Ajani OA, Hassell JA, Gunning PT, and Singh SK
- Subjects
- AC133 Antigen immunology, Animals, Brain Neoplasms immunology, Cell Line, Tumor, Cell Proliferation physiology, Female, Heterografts, Humans, Male, Medulloblastoma immunology, Mice, Neoplasm Recurrence, Local immunology, Neoplastic Stem Cells immunology, Neoplastic Stem Cells metabolism, STAT3 Transcription Factor metabolism, Signal Transduction, Small Molecule Libraries pharmacology, Up-Regulation, AC133 Antigen biosynthesis, Brain Neoplasms drug therapy, Brain Neoplasms pathology, Medulloblastoma drug therapy, Medulloblastoma pathology, Neoplasm Recurrence, Local pathology, Neoplastic Stem Cells pathology, STAT3 Transcription Factor antagonists & inhibitors
- Abstract
Medulloblastoma (MB), the most common malignant paediatric brain tumor, is currently treated using a combination of surgery, craniospinal radiotherapy and chemotherapy. Owing to MB stem cells (MBSCs), a subset of MB patients remains untreatable despite standard therapy. CD133 is used to identify MBSCs although its functional role in tumorigenesis has yet to be determined. In this work, we showed enrichment of CD133 in Group 3 MB is associated with increased rate of metastasis and poor clinical outcome. The signal transducers and activators of transcription-3 (STAT3) pathway are selectively activated in CD133
+ MBSCs and promote tumorigenesis through regulation of c-MYC, a key genetic driver of Group 3 MB. We screened compound libraries for STAT3 inhibitors and treatment with the selected STAT3 inhibitors resulted in tumor size reduction in vivo. We propose that inhibition of STAT3 signaling in MBSCs may represent a potential therapeutic strategy to treat patients with recurrent MB.- Published
- 2017
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44. Pyrvinium Targets CD133 in Human Glioblastoma Brain Tumor-Initiating Cells.
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Venugopal C, Hallett R, Vora P, Manoranjan B, Mahendram S, Qazi MA, McFarlane N, Subapanditha M, Nolte SM, Singh M, Bakhshinyan D, Garg N, Vijayakumar T, Lach B, Provias JP, Reddy K, Murty NK, Doble BW, Bhatia M, Hassell JA, and Singh SK
- Subjects
- AC133 Antigen, Animals, Antigens, CD genetics, Antigens, CD metabolism, Brain Neoplasms drug therapy, Brain Neoplasms genetics, Brain Neoplasms mortality, Cell Proliferation, Cell Self Renewal drug effects, Cell Self Renewal genetics, Disease Models, Animal, Gene Expression, Gene Expression Profiling, Gene Expression Regulation, Neoplastic drug effects, Gene Knockdown Techniques, Gene Regulatory Networks, Glioblastoma drug therapy, Glioblastoma genetics, Glioblastoma mortality, Glycoproteins genetics, Glycoproteins metabolism, Humans, Peptides genetics, Peptides metabolism, Prognosis, Signal Transduction drug effects, Spheroids, Cellular, Tumor Cells, Cultured, Xenograft Model Antitumor Assays, Antineoplastic Agents pharmacology, Brain Neoplasms metabolism, Glioblastoma metabolism, Glycoproteins antagonists & inhibitors, Neoplastic Stem Cells drug effects, Neoplastic Stem Cells metabolism, Peptides antagonists & inhibitors, Pyrvinium Compounds pharmacology
- Abstract
Purpose: Clonal evolution of cancer may be regulated by determinants of stemness, specifically self-renewal, and current therapies have not considered how genetic perturbations or properties of stemness affect such functional processes. Glioblastoma-initiating cells (GICs), identified by expression of the cell surface marker CD133, are shown to be chemoradioresistant. In the current study, we sought to elucidate the functional role of CD133 in self-renewal and identify compounds that can specifically target this CD133(+) treatment-refractory population., Experimental Design: Using gain/loss-of-function studies for CD133 we assessed the in vitro self-renewal and in vivo tumor formation capabilities of patient-derived glioblastoma cells. We generated a CD133 signature combined with an in silico screen to find compounds that target GICs. Self-renewal and proliferation assays on CD133-sorted samples were performed to identify the preferential action of hit compounds. In vivo efficacy of the lead compound pyrvinium was assessed in intracranial GIC xenografts and survival studies. Lastly, microarray analysis was performed on pyrvinium-treated GICs to discover core signaling events involved., Results: We discovered pyrvinium, a small-molecule inhibitor of GIC self-renewal in vitro and in vivo, in part through inhibition of Wnt/β-catenin signaling and other essential stem cell regulatory pathways. We provide a therapeutically tractable strategy to target self-renewing, chemoradioresistant, and functionally important CD133(+) stem cells that drive glioblastoma relapse and mortality., Conclusions: Our study provides an integrated approach for the eradication of clonal populations responsible for cancer progression, and may apply to other aggressive and heterogeneous cancers., (©2015 American Association for Cancer Research.)
- Published
- 2015
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45. Protein expression strategies in Tobacco necrosis virus-D.
- Author
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Chkuaseli T, Newburn LR, Bakhshinyan D, and White KA
- Subjects
- 5' Untranslated Regions, Base Sequence, Cucumis sativus virology, Genome, Viral, Molecular Sequence Data, Nucleic Acid Conformation, Plant Diseases virology, Protein Biosynthesis, RNA, Viral chemistry, RNA, Viral genetics, RNA, Viral metabolism, Tombusviridae chemistry, Tombusviridae metabolism, Viral Proteins chemistry, Viral Proteins metabolism, Gene Expression Regulation, Viral, Tombusviridae genetics, Viral Proteins genetics
- Abstract
Tobacco necrosis virus (TNV-D) has a plus-strand RNA genome that is neither 5' capped nor 3' poly-adenylated. Instead, it utilizes a 3' cap-independent translational enhancer (3'CITE) located in its 3' untranslated region (UTR) for translation of its proteins. We have examined the protein expression strategies used by TNV-D and our results indicate that: (i) a base pairing interaction between conserved ACCA and UGGU motifs in the genomic 5'UTR and 3'CITE, respectively, is not required for efficient plant cell infection, (ii) similar potential 5'UTR-3'CITE interactions in the two viral subgenomic mRNAs are not needed for efficient translation of viral proteins in vitro, (iii) a small amount of capsid protein is translated from the viral genome by a largely 3'CITE-independent mechanism, (iv) the larger of two possible forms of capsid protein is efficiently translated, and (v) p7b is translated from subgenomic mRNA1 by a leaky scanning mechanism., (Copyright © 2015 Elsevier Inc. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
46. STAT3 pathway regulates lung-derived brain metastasis initiating cell capacity through miR-21 activation.
- Author
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Singh M, Garg N, Venugopal C, Hallett R, Tokar T, McFarlane N, Mahendram S, Bakhshinyan D, Manoranjan B, Vora P, Qazi M, Arpin CC, Page B, Haftchenary S, Rosa DA, Lai PS, Gómez-Biagi RF, Ali AM, Lewis A, Geletu M, Murty NK, Hassell JA, Jurisica I, Gunning PT, and Singh SK
- Subjects
- Animals, Cell Line, Tumor, Cell Movement, Genes, Regulator, Humans, Inhibitory Concentration 50, Mice, Mice, Inbred NOD, Mice, SCID, Neoplasm Metastasis, Neoplasm Transplantation, Protein Interaction Mapping, Proteomics, RNA, Small Interfering metabolism, Stem Cells cytology, Brain Neoplasms secondary, Gene Expression Regulation, Neoplastic, Lung Neoplasms pathology, MicroRNAs metabolism, STAT3 Transcription Factor metabolism
- Abstract
Brain metastases (BM) represent the most common tumor to affect the adult central nervous system. Despite the increasing incidence of BM, likely due to consistently improving treatment of primary cancers, BM remain severely understudied. In this study, we utilized patient-derived stem cell lines from lung-to-brain metastases to examine the regulatory role of STAT3 in brain metastasis initiating cells (BMICs). Annotation of our previously described BMIC regulatory genes with protein-protein interaction network mapping identified STAT3 as a novel protein interactor. STAT3 knockdown showed a reduction in BMIC self-renewal and migration, and decreased tumor size in vivo. Screening of BMIC lines with a library of STAT3 inhibitors identified one inhibitor to significantly reduce tumor formation. Meta-analysis identified the oncomir microRNA-21 (miR-21) as a target of STAT3 activity. Inhibition of miR-21 displayed similar reductions in BMIC self-renewal and migration as STAT3 knockdown. Knockdown of STAT3 also reduced expression of known downstream targets of miR-21. Our studies have thus identified STAT3 and miR-21 as cooperative regulators of stemness, migration and tumor initiation in lung-derived BM. Therefore, STAT3 represents a potential therapeutic target in the treatment of lung-to-brain metastases.
- Published
- 2015
- Full Text
- View/download PDF
47. MicroRNA Regulation of Brain Tumour Initiating Cells in Central Nervous System Tumours.
- Author
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Garg N, Vijayakumar T, Bakhshinyan D, Venugopal C, and Singh SK
- Abstract
CNS tumours occur in both pediatric and adult patients and many of these tumours are associated with poor clinical outcome. Due to a paradigm shift in thinking for the last several years, these tumours are now considered to originate from a small population of stem-like cells within the bulk tumour tissue. These cells, termed as brain tumour initiating cells (BTICs), are perceived to be regulated by microRNAs at the posttranscriptional/translational levels. Proliferation, stemness, differentiation, invasion, angiogenesis, metastasis, apoptosis, and cell cycle constitute some of the significant processes modulated by microRNAs in cancer initiation and progression. Characterization and functional studies on oncogenic or tumour suppressive microRNAs are made possible because of developments in sequencing and microarray techniques. In the current review, we bring recent knowledge of the role of microRNAs in BTIC formation and therapy. Special attention is paid to two highly aggressive and well-characterized brain tumours: gliomas and medulloblastoma. As microRNA seems to be altered in the pathogenesis of many human diseases, "microRNA therapy" may now have potential to improve outcomes for brain tumour patients. In this rapidly evolving field, further understanding of miRNA biology and its contribution towards cancer can be mined for new therapeutic tools.
- Published
- 2015
- Full Text
- View/download PDF
48. The role of stem cells in pediatric central nervous system malignancies.
- Author
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Manoranjan B, Garg N, Bakhshinyan D, and Singh SK
- Subjects
- Central Nervous System Neoplasms classification, Child, Ependymoma pathology, Glioblastoma pathology, Glioma pathology, Humans, Medulloblastoma pathology, Neuroectodermal Tumors pathology, Rhabdoid Tumor pathology, Teratoma pathology, Central Nervous System Neoplasms pathology, Neoplastic Stem Cells physiology, Neural Stem Cells physiology
- Abstract
Representing the leading cause of childhood cancer mortality, pediatric brain tumors are comprised of diverse histological features, genetic perturbations, cellular populations, treatment protocols, and clinical outcomes. In this chapter we discuss recent and emerging data that implicate cancer stem cells (also known as brain tumor-initiating cells) in initiating and maintaining the growth of a number of pediatric brain tumors including: medulloblastoma, supratentorial primitive neuroectodermal tumor, atypical teratoid/rhabdoid tumor, ependymoma, low-grade glioma, glioblastoma, diffuse intrinsic pontine glioma, germ cell tumor, and craniopharyngioma. The development of a stem cell framework for the study and treatment of these tumors will enable future clinical approaches to harness the heterogeneous cellular and genomic landscape of these solid tumors as an avenue for developing targeted patient-oriented therapies, thereby improving the overall survivorship for the most lethal childhood cancer.
- Published
- 2015
- Full Text
- View/download PDF
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