40 results on '"Hingtgen SD"'
Search Results
2. Combining the constitutive TRAIL-secreting induced neural stem cell therapy with the novel anti-cancer drug TR-107 in glioblastoma.
- Author
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Thang M, Mellows C, Kass LE, Daglish S, Fennell EMJ, Mann BE, Mercer-Smith AR, Valdivia A, Graves LM, and Hingtgen SD
- Abstract
Tumor-homing neural stem cell (NSC) therapy is emerging as a promising treatment for aggressive cancers of the brain. Despite their success, developing tumor-homing NSC therapy therapies that maintain durable tumor suppression remains a challenge. Herein, we report a synergistic combination regimen where the novel small molecule TR-107 augments NSC-tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) therapy (hiNeuroS-TRAIL) in models of the incurable brain cancer glioblastoma (GBM) in vitro . We report that the combination of hiNeuroS-TRAIL and TR-107 synergistically upregulated caspase markers and restored sensitivity to the intrinsic apoptotic pathway by significantly downregulating inhibitory pathways associated with chemoresistance and radioresistance in the TRAIL-resistant LN229 cell line. This combination also showed robust tumor suppression and enhanced survival of mice bearing human xenografts of both solid and invasive GBMs. These findings elucidate a novel combination regimen and suggest that the combination of these clinically relevant agents may represent a new therapeutic option with increased efficacy for patients with GBM., Competing Interests: S.D.H. has ownership interest as the CSO of Falcon Therapeutics., (© 2024 The Authors. Published by Elsevier Inc. on behalf of The American Society of Gene and Cell Therapy.)
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- 2024
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3. Longitudinal 3-D Visualization of Microvascular Disruption and Perfusion Changes in Mice During the Evolution of Glioblastoma Using Super-Resolution Ultrasound.
- Author
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McCall JR, DeRuiter R, Ross M, Santibanez F, Hingtgen SD, Pinton GF, and Dayton PA
- Subjects
- Mice, Humans, Animals, Microvessels diagnostic imaging, Ultrasonography methods, Brain blood supply, Perfusion, Microbubbles, Glioblastoma diagnostic imaging
- Abstract
Glioblastoma is an aggressive brain cancer with a very poor prognosis in which less than 6% of patients survive more than five-year post-diagnosis. The outcome of this disease for many patients may be improved by early detection. This could provide clinicians with the information needed to take early action for treatment. In this work, we present the utilization of a non-invasive, fully volumetric ultrasonic imaging method to assess microvascular change during the evolution of glioblastoma in mice. Volumetric ultrasound localization microscopy (ULM) was used to observe statistically significant ( ) reduction in the appearance of functional vasculature over the course of three weeks. We also demonstrate evidence suggesting the reduction of vascular flow for vessels peripheral to the tumor. With an 82.5% consistency rate in acquiring high-quality vascular images, we demonstrate the possibility of volumetric ULM as a longitudinal method for microvascular characterization of neurological disease.
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- 2023
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4. Cell-based therapies for glioblastoma: Promising tools against tumor heterogeneity.
- Author
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Nehama D, Woodell AS, Maingi SM, Hingtgen SD, and Dotti G
- Subjects
- Humans, Treatment Outcome, Immunotherapy, Tumor Microenvironment, Glioblastoma pathology, Brain Neoplasms therapy
- Abstract
Glioblastoma (GBM) is a highly aggressive tumor with a devastating impact on quality-of-life and abysmal survivorship. Patients have very limited effective treatment options. The successes of targeted small molecule drugs and immune checkpoint inhibitors seen in various solid tumors have not translated to GBM, despite significant advances in our understanding of its molecular, immune, and microenvironment landscapes. These discoveries, however, have unveiled GBM's incredible heterogeneity and its role in treatment failure and survival. Novel cellular therapy technologies are finding successes in oncology and harbor characteristics that make them uniquely suited to overcome challenges posed by GBM, such as increased resistance to tumor heterogeneity, modularity, localized delivery, and safety. Considering these advantages, we compiled this review article on cellular therapies for GBM, focusing on cellular immunotherapies and stem cell-based therapies, to evaluate their utility. We categorize them based on their specificity, review their preclinical and clinical data, and extract valuable insights to help guide future cellular therapy development., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2023
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5. Utilizing induced neural stem cell-based delivery of a cytokine cocktail to enhance chimeric antigen receptor-modified T-cell therapy for brain cancer.
- Author
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Woodell AS, Landoni E, Valdivia A, Buckley A, Ogunnaike EA, Dotti G, and Hingtgen SD
- Abstract
Chimeric antigen receptor (CAR)-modified T-cell therapy has shown enormous clinical promise against blood cancers, yet efficacy against solid tumors remains a challenge. Here, we investigated the potential of a new combination cell therapy, where tumor-homing induced neural stem cells (iNSCs) are used to enhance CAR-T-cell therapy and achieve efficacious suppression of brain tumors. Using in vitro and in vivo migration assays, we found iNSC-secreted RANTES/IL-15 increased CAR-T-cell migration sixfold and expansion threefold, resulting in greater antitumor activity in a glioblastoma (GBM) tumor model. Furthermore, multimodal imaging showed iNSC delivery of RANTES/IL-15 in combination with intravenous administration of CAR-T cells reduced established orthotopic GBM xenografts 2538-fold within the first week, followed by durable tumor remission through 60 days post-treatment. By contrast, CAR-T-cell therapy alone only partially controlled tumor growth, with a median survival of only 19 days. Together, these studies demonstrate the potential of combined cell therapy platforms to improve the efficacy of CAR-T-cell therapy for brain tumors., Competing Interests: Shawn D. Hingtgen has an ownership interest in Falcon Therapeutics, Inc., which has licensed aspects of iNSC technology from UNC‐CH. All remaining 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., (© 2023 The Authors. Bioengineering & Translational Medicine published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers.)
- Published
- 2023
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6. Injectable pH Thermo-Responsive Hydrogel Scaffold for Tumoricidal Neural Stem Cell Therapy for Glioblastoma Multiforme.
- Author
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King JL, Maturavongsadit P, Hingtgen SD, and Benhabbour SR
- Abstract
Glioblastoma multiforme (GBM) is the most common malignant brain tumor in adults and despite recent advances in treatment modalities, GBM remains incurable. Injectable hydrogel scaffolds are a versatile delivery system that can improve delivery of drug and cell therapeutics for GBM. In this report, we investigated an injectable nanocellulose/chitosan-based hydrogel scaffold for neural stem cell encapsulation and delivery. Hydrogels were prepared using thermogelling beta-glycerophosphate (BGP) and hydroxyethyl cellulose (HEC), chitosan (CS), and cellulose nanocrystals (CNCs). We evaluated the impact of neural stem cells on hydrogel gelation kinetics, microstructures, and degradation. Furthermore, we investigated the biomaterial effects on cell viability and functionality. We demonstrated that the incorporation of cells at densities of 1, 5 and 10 million does not significantly impact rheological and physical properties CS scaffolds. However, addition of CNCs significantly prolonged hydrogel degradation when cells were seeded at 5 and 10 million per 1 mL hydrogel. In vitro cell studies demonstrated high cell viability, release of TRAIL at therapeutic concentrations, and effective tumor cell killing within 72 h. The ability of these hydrogel scaffolds to support stem cell encapsulation and viability and maintain stem cell functionality makes them an attractive cell delivery system for local treatment of post-surgical cancers.
- Published
- 2022
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7. Next-generation Tumor-homing Induced Neural Stem Cells as an Adjuvant to Radiation for the Treatment of Metastatic Lung Cancer.
- Author
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Mercer-Smith AR, Buckley A, Valdivia A, Jiang W, Thang M, Bell N, Kumar RJ, Bomba HN, Woodell AS, Luo J, Floyd SR, and Hingtgen SD
- Subjects
- Animals, Apoptosis, Cell Line, Tumor, Humans, Mice, RNA, Messenger, TNF-Related Apoptosis-Inducing Ligand genetics, TNF-Related Apoptosis-Inducing Ligand metabolism, TNF-Related Apoptosis-Inducing Ligand pharmacology, Antineoplastic Agents pharmacology, Carcinoma, Non-Small-Cell Lung genetics, Carcinoma, Non-Small-Cell Lung radiotherapy, Lung Neoplasms genetics, Lung Neoplasms radiotherapy, Neural Stem Cells metabolism
- Abstract
The spread of non-small cell lung cancer (NSCLC) to the leptomeninges is devastating with a median survival of only a few months. Radiation offers symptomatic relief, but new adjuvant therapies are desperately needed. Spheroidal, human induced neural stem cells (hiNeuroS) secreting the cytotoxic protein, TRAIL, have innate tumoritropic properties. Herein, we provide evidence that hiNeuroS-TRAIL cells can migrate to and suppress growth of NSCLC metastases in combination with radiation. In vitro cell tracking and post-mortem tissue analysis showed that hiNeuroS-TRAIL cells migrate to NSCLC tumors. Importantly, isobolographic analysis suggests that TRAIL with radiation has a synergistic cytotoxic effect on NSCLC tumors. In vivo, mice treated with radiation and hiNeuroS-TRAIL showed significant (36.6%) improvements in median survival compared to controls. Finally, bulk mRNA sequencing analysis showed both NSCLC and hiNeuroS-TRAIL cells showed changes in genes involved in migration following radiation. Overall, hiNeuroS-TRAIL cells +/- radiation have the capacity to treat NSCLC metastases., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
- Published
- 2022
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8. Use of FLOSEAL® as a scaffold and its impact on induced neural stem cell phenotype, persistence, and efficacy.
- Author
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Bomba HN, Carey-Ewend A, Sheets KT, Valdivia A, Goetz M, Findlay IA, Mercer-Smith A, Kass LE, Khagi S, and Hingtgen SD
- Abstract
Induced neural stem cells (iNSCs) have emerged as a promising therapeutic platform for glioblastoma (GBM). iNSCs have the innate ability to home to tumor foci, making them ideal carriers for antitumor payloads. However, the in vivo persistence of iNSCs limits their therapeutic potential. We hypothesized that by encapsulating iNSCs in the FDA-approved, hemostatic matrix FLOSEAL®, we could increase their persistence and, as a result, therapeutic durability. Encapsulated iNSCs persisted for 95 days, whereas iNSCs injected into the brain parenchyma persisted only 2 weeks in mice. Two orthotopic GBM tumor models were used to test the efficacy of encapsulated iNSCs. In the GBM8 tumor model, mice that received therapeutic iNSCs encapsulated in FLOSEAL® survived 30 to 60 days longer than mice that received nonencapsulated cells. However, the U87 tumor model showed no significant differences in survival between these two groups, likely due to the more solid and dense nature of the tumor. Interestingly, the interaction of iNSCs with FLOSEAL® appears to downregulate some markers of proliferation, anti-apoptosis, migration, and therapy which could also play a role in treatment efficacy and durability. Our results demonstrate that while FLOSEAL® significantly improves iNSC persistence, this alone is insufficient to enhance therapeutic durability., Competing Interests: Shawn D. Hingtgen has an ownership interest in Falcon Therapeutics, Inc., which has licensed aspects of iNSC technology from the University of North Carolina at Chapel Hill. Hunter N. Bomba, Abigail Carey‐Ewend, Kevin T. Sheets, Alain Valdivia, Morgan Goetz, Ingrid A. Findlay, Lauren E. Kass, and Simon Khagi have no competing interests to disclose., (© 2021 The Authors. Bioengineering & Translational Medicine published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers.)
- Published
- 2022
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9. Intravenously Infused Stem Cells for Cancer Treatment.
- Author
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Mercer-Smith AR, Findlay IA, Bomba HN, and Hingtgen SD
- Subjects
- Humans, Stem Cells, Mesenchymal Stem Cells, Neoplasms therapy, Oncolytic Virotherapy methods, Oncolytic Viruses
- Abstract
Despite the recent influx of immunotherapies and small molecule drugs to treat tumors, cancer remains a leading cause of death in the United States, in large part due to the difficulties of treating metastatic cancer. Stem cells, which are inherently tumoritropic, provide a useful drug delivery vehicle to target both primary and metastatic tumors. Intravenous infusions of stem cells carrying or secreting therapeutic payloads show significant promise in the treatment of cancer. Stem cells may be engineered to secrete cytotoxic products, loaded with oncolytic viruses or nanoparticles containing small molecule drugs, or conjugated with immunotherapies. Herein we describe these preclinical and clinical studies, discuss the distribution and migration of stem cells following intravenous infusion, and examine both the limitations of and the methods to improve the migration and therapeutic efficacy of tumoritropic, therapeutic stem cells., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
- Published
- 2021
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10. Cytotoxic Engineered Induced Neural Stem Cells as an Intravenous Therapy for Primary Non-Small Cell Lung Cancer and Triple-Negative Breast Cancer.
- Author
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Mercer-Smith AR, Jiang W, Bago JR, Valdivia A, Thang M, Woodell AS, Montgomery SA, Sheets KT, Anders CK, and Hingtgen SD
- Subjects
- Animals, Humans, Mice, Carcinoma, Non-Small-Cell Lung drug therapy, Lung Neoplasms drug therapy, Neural Stem Cells transplantation, Triple Negative Breast Neoplasms drug therapy
- Abstract
Converting human fibroblasts into personalized induced neural stem cells (hiNSC) that actively seek out tumors and deliver cytotoxic agents is a promising approach for treating cancer. Herein, we provide the first evidence that intravenously-infused hiNSCs secreting cytotoxic agent home to and suppress the growth of non-small cell lung cancer (NSCLC) and triple-negative breast cancer (TNBC). Migration of hiNSCs to NSCLC and TNBC in vitro was investigated using time-lapse motion analysis, which showed directional movement of hiNSCs to both tumor cell lines . In vivo , migration of intravenous hiNSCs to orthotopic NSCLC or TNBC tumors was determined using bioluminescent imaging (BLI) and immunofluorescent post-mortem tissue analysis, which indicated that hiNSCs colocalized with tumors within 3 days of intravenous administration and persisted through 14 days. In vitro , efficacy of hiNSCs releasing cytotoxic TRAIL (hiNSC-TRAIL) was monitored using kinetic imaging of co-cultures, in which hiNSC-TRAIL therapy induced rapid killing of both NSCLC and TNBC. Efficacy was determined in vivo by infusing hiNSC-TRAIL or control cells intravenously into mice bearing orthotopic NSCLC or TNBC and tracking changes in tumor volume using BLI. Mice treated with intravenous hiNSC-TRAIL showed a 70% or 72% reduction in NSCLC or TNBC tumor volume compared with controls within 14 or 21 days, respectively. Safety was assessed by hematology, blood chemistry, and histology, and no significant changes in these safety parameters was observed through 28 days. These results indicate that intravenous hiNSCs-TRAIL seek out and kill NSCLC and TNBC tumors, suggesting a potential new strategy for treating aggressive peripheral cancers., (©2021 American Association for Cancer Research.)
- Published
- 2021
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11. Developing Bioinspired Three-Dimensional Models of Brain Cancer to Evaluate Tumor-Homing Neural Stem Cell Therapy.
- Author
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Carey-Ewend AG, Hagler SB, Bomba HN, Goetz MJ, Bago JR, and Hingtgen SD
- Subjects
- Animals, Apoptosis, Brain diagnostic imaging, Mice, Brain Neoplasms therapy, Glioblastoma therapy, Neural Stem Cells
- Abstract
Engineered neural stem cells (NSCs) have recently emerged as a promising therapy. Acting as a tumor-homing drug-delivery system, NSCs migrate through brain tissue to seek out primary and invasive tumor foci. NSCs can deliver therapeutic agents, such as TNFα-related apoptosis-inducing ligand, directly to the tumor and suppress glioblastoma (GBM) in murine models. While the mainstays for evaluating NSC migration and efficacy have been two-dimensional chemotaxis assays and mouse models, these low-throughput and small-scale systems limit our ability to implant and track these cells for human translation. To circumvent these challenges, we developed a three-dimensional culture system using a matrix of poly-l-lactic acid 6100 microfibers suspended in agar. These bioinspired brain matrices were used to model tumor growth, NSC migration, and efficacy of NSC therapy at small and human scale. Kinetic fluorescent imaging confirmed growth of tumors in both small and human-sized bioinspired brain matrix. Tumors proliferated 50-fold and 3-fold for GBM and human metastatic breast cancer, respectively, over 7 days. We next explored the impact of tumor location on NSC migration. When NSCs were implanted 2 mm lateral from the tumor foci, NSCs colocalized with the GBM within 7 days. In models of multifocal disease, NSCs were found to colocalize with multiple tumors, preferentially migrating to tumor foci closest to the site of NSC implantation. Lastly, therapeutic NSCs were implanted at increasing distances (0, 2, 5, or 10 mm) laterally from GBM foci to investigate the effects of distance on NSC efficacy. Serial imaging showed reduced fluorescence at tumor sites, implicating GBM apoptosis across all distances. NSCs coinjected with tumor induced a near-complete response in <10 days, while NSCs implanted 10 mm laterally from the tumor induced a near-complete response by day 30. Lastly, GBM foci were established in each hemisphere of the model and control or therapeutic NSCs were implanted adjacent to tumor cells in the right hemisphere. Kinetic imaging showed that NSC therapy attenuated progression of GBM foci, while GBM cells treated with control NSC expanded rapidly over 21 days. In conclusion, we developed a new bioinspired model that supports growth of human brain cancer cells and enables rapid tracking of NSC therapy. Impact statement Tumor-homing and tumor-killing-engineered neural stem cell (NSC) therapies have shown immense promise in both preclinical and clinical trials. However, as cell therapies continue to evolve, cost-effective and high-throughput screening assays are needed to assess the proliferation, migration, and efficacy of these cells. In this study, we developed a bioinspired brain matrix for the evaluation of engineered NSCs. Importantly, this matrix is easy to fabricate, scalable, and allows for sterile real-time, noninvasive imaging using our custom bioreactor. We then utilized the bioinspired brain matrix system to answer key questions around the tumor-homing migration and efficacy of engineered NSC therapies that are challenging to address with traditional models.
- Published
- 2021
- Full Text
- View/download PDF
12. Development of next-generation tumor-homing induced neural stem cells to enhance treatment of metastatic cancers.
- Author
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Jiang W, Yang Y, Mercer-Smith AR, Valdivia A, Bago JR, Woodell AS, Buckley AA, Marand MH, Qian L, Anders CK, and Hingtgen SD
- Subjects
- Cell Line, Tumor, Fibroblasts, Humans, Neoplastic Stem Cells pathology, Neural Stem Cells pathology, Triple Negative Breast Neoplasms pathology
- Abstract
Engineered tumor-homing neural stem cells (NSCs) have shown promise in treating cancer. Recently, we transdifferentiated skin fibroblasts into human-induced NSCs (hiNSC) as personalized NSC drug carriers. Here, using a SOX2 and spheroidal culture-based reprogramming strategy, we generated a new hiNSC variant, hiNeuroS, that was genetically distinct from fibroblasts and first-generation hiNSCs and had significantly enhanced tumor-homing and antitumor properties. In vitro, hiNeuroSs demonstrated superior migration to human triple-negative breast cancer (TNBC) cells and in vivo rapidly homed to TNBC tumor foci following intracerebroventricular (ICV) infusion. In TNBC parenchymal metastasis models, ICV infusion of hiNeuroSs secreting the proapoptotic agent TRAIL (hiNeuroS-TRAIL) significantly reduced tumor burden and extended median survival. In models of TNBC leptomeningeal carcinomatosis, ICV dosing of hiNeuroS-TRAIL therapy significantly delayed the onset of tumor formation and extended survival when administered as a prophylactic treatment, as well as reduced tumor volume while prolonging survival when delivered as established tumor therapy., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)
- Published
- 2021
- Full Text
- View/download PDF
13. Developing Bioinspired Three-Dimensional Models of Brain Cancer to Evaluate Tumor-Homing Neural Stem Cell Therapy.
- Author
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Carey-Ewend AG, Hagler SB, Bomba HN, Goetz MJ, Bago JR, and Hingtgen SD
- Abstract
Engineered neural stem cells (NSCs) have recently emerged as a promising therapy. Acting as a tumor-homing drug-delivery system, NSCs migrate through brain tissue to seek out primary and invasive tumor foci. NSCs can deliver therapeutic agents, such as TNFα-related apoptosis-inducing ligand, directly to the tumor and suppress glioblastoma (GBM) in murine models. While the mainstays for evaluating NSC migration and efficacy have been two-dimensional chemotaxis assays and mouse models, these low-throughput and small-scale systems limit our ability to implant and track these cells for human translation. To circumvent these challenges, we developed a three-dimensional culture system using a matrix of poly-l-lactic acid 6100 microfibers suspended in agar. These bioinspired brain matrices were used to model tumor growth, NSC migration, and efficacy of NSC therapy at small and human scale. Kinetic fluorescent imaging confirmed growth of tumors in both small and human-sized bioinspired brain matrix. Tumors proliferated 50-fold and 3-fold for GBM and human metastatic breast cancer, respectively, over 7 days. We next explored the impact of tumor location on NSC migration. When NSCs were implanted 2 mm lateral from the tumor foci, NSCs colocalized with the GBM within 7 days. In models of multifocal disease, NSCs were found to colocalize with multiple tumors, preferentially migrating to tumor foci closest to the site of NSC implantation. Lastly, therapeutic NSCs were implanted at increasing distances (0, 2, 5, or 10 mm) laterally from GBM foci to investigate the effects of distance on NSC efficacy. Serial imaging showed reduced fluorescence at tumor sites, implicating GBM apoptosis across all distances. NSCs coinjected with tumor induced a near-complete response in <10 days, while NSCs implanted 10 mm laterally from the tumor induced a near-complete response by day 30. Lastly, GBM foci were established in each hemisphere of the model and control or therapeutic NSCs were implanted adjacent to tumor cells in the right hemisphere. Kinetic imaging showed that NSC therapy attenuated progression of GBM foci, while GBM cells treated with control NSC expanded rapidly over 21 days. In conclusion, we developed a new bioinspired model that supports growth of human brain cancer cells and enables rapid tracking of NSC therapy. Impact statement Tumor-homing and tumor-killing-engineered neural stem cell (NSC) therapies have shown immense promise in both preclinical and clinical trials. However, as cell therapies continue to evolve, cost-effective and high-throughput screening assays are needed to assess the proliferation, migration, and efficacy of these cells. In this study, we developed a bioinspired brain matrix for the evaluation of engineered NSCs. Importantly, this matrix is easy to fabricate, scalable, and allows for sterile real-time, noninvasive imaging using our custom bioreactor. We then utilized the bioinspired brain matrix system to answer key questions around the tumor-homing migration and efficacy of engineered NSC therapies that are challenging to address with traditional models.
- Published
- 2020
- Full Text
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14. Personalized-induced neural stem cell therapy: Generation, transplant, and safety in a large animal model.
- Author
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Bomba HN, Sheets KT, Valdivia A, Khagi S, Ruterbories L, Mariani CL, Borst LB, Tokarz DA, and Hingtgen SD
- Abstract
In this study, we take an important step toward clinical translation by generating the first canine-induced neural stem cells (iNSCs). We explore key aspects of scale-up, persistence, and safety of personalized iNSC therapy in autologous canine surgery models. iNSCs are a promising new approach to treat aggressive cancers of the brain, including the deadly glioblastoma. Created by direct transdifferentiation of fibroblasts, iNSCs are known to migrate through the brain, track down invasive cancer foci, and deliver anticancer payloads that significantly reduce tumor burden and extend survival of tumor-bearing mice. Here, skin biopsies were collected from canines and converted into the first personalized canine iNSCs engineered to carry TNFα-related apoptosis-inducing ligand (TRAIL) and thymidine kinase (TK), as well as magnetic resonance imaging (MRI) contrast agents for in vivo tracking. Time-lapse analysis showed canine iNSCs efficiently migrate to human tumor cells, and cell viability assays showed both TRAIL and TK monotherapy markedly reduced tumor growth. Using intraoperative navigation and two delivery methods to closely mimic human therapy, canines received autologous iNSCs either within postsurgical cavities in a biocompatible matrix or via a catheter placed in the lateral ventricle. Both strategies were well tolerated, and serial MRI showed hypointense regions at the implant sites that remained stable through 86 days postimplant. Serial fluid sample testing following iNSC delivery showed the bimodal personalized therapy was well tolerated, with no iNSC-induced abnormal tissue pathology. Overall, this study lays an important foundation as this promising personalized cell therapy advances toward human patient testing., Competing Interests: K. T. S. and S. D. H have an ownership interest in Falcon Therapeutics, Inc., which has licensed aspects of iNSC technology from the University of North Carolina at Chapel Hill. H. N. B., A. V., S. K., C. L. M., L. R., L. B. B., and D. A. T. have no conflicts to disclose., (© 2020 The Authors. Bioengineering & Translational Medicine published by Wiley Periodicals LLC on behalf of The American Institute of Chemical Engineers.)
- Published
- 2020
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15. Polymeric Biomaterial Scaffolds for Tumoricidal Stem Cell Glioblastoma Therapy.
- Author
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Moore KM, Murthy AB, Graham-Gurysh EG, Hingtgen SD, Bachelder EM, and Ainslie KM
- Subjects
- Biocompatible Materials, Humans, Neoplasm Recurrence, Local, Stem Cells, Brain Neoplasms therapy, Glioblastoma therapy
- Abstract
Glioblastoma (GBM) is the most common primary brain tumor and has a poor prognosis; as such, there is an urgent need to develop innovative new therapies. Tumoricidal stem cells are an emerging therapy that has the potential to combat limitations of traditional local and systemic chemotherapeutic strategies for GBM by providing a source for high, sustained concentrations of tumoricidal agents locally to the tumor. One major roadblock for tumoricidal stem cell therapy is that the persistence of tumoricidal stem cells injected as a cell suspension into the GBM surgical resection cavity is limited. Polymeric biomaterial scaffolds have been utilized to enhance the delivery of tumoricidal stem cells in the surgical resection cavity and extend their persistence in the brain, ultimately increasing their therapeutic efficacy against GBM. In this review, we examine three main scaffold categories explored for tumoricidal stem cell therapy: microcapsules, hydrogels, and electrospun scaffolds. Furthermore, considering the significant impact of surgery on the brain and recurrent GBM, we survey a brief history of orthotopic models of GBM surgical resection.
- Published
- 2020
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16. Synergistic drug combinations for a precision medicine approach to interstitial glioblastoma therapy.
- Author
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Graham-Gurysh EG, Murthy AB, Moore KM, Hingtgen SD, Bachelder EM, and Ainslie KM
- Subjects
- Animals, Cell Line, Tumor, Drug Combinations, Drug Synergism, Humans, Mice, Mice, Nude, Precision Medicine, Xenograft Model Antitumor Assays, Brain Neoplasms drug therapy, Glioblastoma drug therapy
- Abstract
Glioblastoma (GBM) is a highly aggressive and heterogeneous form of brain cancer. Genotypic and phenotypic heterogeneity drives drug resistance and tumor recurrence. Combination chemotherapy could overcome drug resistance; however, GBM's location behind the blood-brain barrier severely limits chemotherapeutic options. Interstitial therapy, delivery of chemotherapy locally to the tumor site, via a biodegradable polymer implant can overcome the blood-brain barrier and increase the range of drugs available for therapy. Ideal drug candidates for interstitial therapy are those that are potent against GBM and work in combination with both standard-of-care therapy and new precision medicine targets. Herein we evaluated paclitaxel for interstitial therapy, investigating the effect of combination with both temozolomide, a clinical standard-of-care chemotherapy for GBM, and everolimus, a mammalian target of rapamycin (mTOR) inhibitor that modulates aberrant signaling present in >80% of GBM patients. Tested against a panel of GBM cell lines in vitro, paclitaxel was found to be effective at nanomolar concentrations, complement therapy with temozolomide, and synergize strongly with everolimus. The strong synergism seen with paclitaxel and everolimus was then explored in vivo. Paclitaxel and everolimus were separately formulated into fibrous scaffolds composed of acetalated dextran, a biodegradable polymer with tunable degradation rates, for implantation in the brain. Acetalated dextran degradation rates were tailored to attain matching release kinetics (~3% per day) of both paclitaxel and everolimus to maintain a fixed combination ratio of the two drugs. Combination interstitial therapy of both paclitaxel and everolimus significantly reduced GBM growth and improved progression free survival in two clinically relevant orthotopic models of GBM resection and recurrence. This work illustrates the advantages of synchronized interstitial therapy of paclitaxel and everolimus for post-surgical tumor control of GBM., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
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17. Generation and Profiling of Tumor-Homing Induced Neural Stem Cells from the Skin of Cancer Patients.
- Author
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Buckley A, Hagler SB, Lettry V, Bagó JR, Maingi SM, Khagi S, Ewend MG, Miller CR, and Hingtgen SD
- Subjects
- Animals, Cell Line, Tumor, Cell Movement, Cell Proliferation, Cell Survival, Cell Transdifferentiation, Cells, Cultured, Female, Fibroblasts cytology, Glioblastoma genetics, Humans, Induced Pluripotent Stem Cells cytology, Male, Mice, Neural Stem Cells cytology, Primary Cell Culture, Rats, Survival Analysis, Xenograft Model Antitumor Assays, Glioblastoma therapy, Induced Pluripotent Stem Cells transplantation, Membrane Glycoproteins genetics, Neural Stem Cells transplantation, Semaphorins genetics, Skin cytology
- Abstract
The conversion of human fibroblasts into personalized induced neural stem cells (iNSCs) that actively seek out tumors and deliver cytotoxic agents is a highly promising approach for treating various types of cancer. However, the ability to generate iNSCs from the skin of cancer patients has not been explored. Here, we take an important step toward clinical application by generating iNSCs from skin biopsies of human patients undergoing treatment for the aggressive brain cancer, glioblastoma (GBM). We then utilized a panel of functional and genomic studies to investigate the efficacy and tumor-homing capacity of these patient-derived cells, as well as genomic analysis, to characterize the impact of interpatient variability on this personalized cell therapy. From the skin-tissue biopsies, we established fibroblasts and transdifferentiated the cells into iNSCs. Genomic and functional testing revealed marked variability in growth rates, therapeutic agent production, and gene expression during fibroblast-to-iNSC conversion among patient lines. In vivo testing showed patient-derived iNSCs home to tumors, yet rates and expression of homing-related pathways varied among patients. With the use of surgical-resection mouse models of invasive human cluster of differentiation 133
+ (CD133+ ) GBM cells and serial kinetic imaging, we found that "high-performing" patient-derived iNSC lines reduced the volume of GBM cells 60-fold and extended survival from 28 to 45 days. Treatment with "low-performing" patient lines had minimal effect on tumor growth, but the anti-tumor effect could be rescued by increasing the intracavity dose. Together, these data show, for the first time, that tumor-homing iNSCs can be generated from the skin of cancer patients and efficaciously suppress tumor growth. We also begin to define genetic markers that could be used to identify cells that will contain the most effective attributes for tumor homing and kill in human patients, including high gene expression of the semaphorin-3B (SEMA3B), which is known to be involved in neuronal cell migration. These studies should serve as an important guide toward clinical GBM therapy, where the personalized nature of optimized iNSC therapy has the potential to avoid transplant rejection and maximize treatment durability., (Copyright © 2020 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)- Published
- 2020
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18. Impact of composite scaffold degradation rate on neural stem cell persistence in the glioblastoma surgical resection cavity.
- Author
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Moore KM, Graham-Gurysh EG, Bomba HN, Murthy AB, Bachelder EM, Hingtgen SD, and Ainslie KM
- Subjects
- Acetylation, Animals, Cell Line, Cell Survival, Cross-Linking Reagents chemistry, Dextrans chemistry, Female, Gelatin chemistry, Mice, Nude, Temperature, Brain Neoplasms pathology, Glioblastoma pathology, Neural Stem Cells pathology, Tissue Scaffolds chemistry
- Abstract
Tumoricidal neural stem cells (NSCs) are an emerging therapy to combat glioblastoma (GBM). This therapy employs genetically engineered NSCs that secrete tumoricidal agents to seek out and kill tumor foci remaining after GBM surgical resection. Biomaterial scaffolds have previously been utilized to deliver NSCs to the resection cavity. Here, we investigated the impact of scaffold degradation rate on NSC persistence in the brain resection cavity. Composite acetalated dextran (Ace-DEX) gelatin electrospun scaffolds were fabricated with two distinct degradation profiles created by changing the ratio of cyclic to acyclic acetal coverage of Ace-DEX. In vitro, fast degrading scaffolds were fully degraded by one week, whereas slow degrading scaffolds had a half-life of >56 days. The scaffolds also retained distinct degradation profiles in vivo. Two different NSC lines readily adhered to and remained viable on Ace-DEX gelatin scaffolds, in vitro. Therapeutic NSCs secreting tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) had the same TRAIL output as tissue culture treated polystyrene (TCPS) when seeded on both scaffolds. Furthermore, secreted TRAIL was found to be highly potent against the human derived GBM cell line, GBM8, in vitro. Firefly luciferase expressing NSCs were seeded on scaffolds, implanted in a surgical resection cavity and their persistence in the brain was monitored by bioluminescent imaging (BLI). NSC loaded scaffolds were compared to a direct injection (DI) of NSCs in suspension, which is the current clinical approach to NSC therapy for GBM. Fast and slow degrading scaffolds enhanced NSC implantation efficiency 2.87 and 3.08-fold over DI, respectively. Interestingly, scaffold degradation profile did not significantly impact NSC persistence. However, persistence and long-term survival of NSCs was significantly greater for both scaffolds compared to DI, with scaffold implanted NSCs still detected by BLI at day 120 in most mice. Overall, these results highlight the benefit of utilizing a scaffold for application of tumoricidal NSC therapy for GBM., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Drs. Ainslie and Bachelder serve on the advisory board for IMMvention Therapeutix, Inc. Although a financial conflict of interest was identified for management based on the overall scope of the project and its potential benefit to IMMvention Therapeutix, Inc., the research findings included in this publication may not necessarily related to the interests of IMMvention Therapeutix, Inc. Dr. Shawn Hingtgen is the Founder and CSO of Falcon Therapeutics which has exclusively licensed aspects of scaffold-based technology for GBM therapy. The terms of this arrangement have been reviewed and approved by the University of North Carolina at Chapel Hill in accordance with its policy on objectivity in research., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
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19. Tumor Responsive and Tunable Polymeric Platform for Optimized Delivery of Paclitaxel to Treat Glioblastoma.
- Author
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Graham-Gurysh EG, Moore KM, Schorzman AN, Lee T, Zamboni WC, Hingtgen SD, Bachelder EM, and Ainslie KM
- Subjects
- Animals, Antineoplastic Agents pharmacokinetics, Cell Line, Tumor, Drug Liberation, Female, Humans, Hydrogen-Ion Concentration, Mice, Nude, Neoplasm Metastasis drug therapy, Paclitaxel pharmacokinetics, Secondary Prevention methods, Tumor Microenvironment physiology, Xenograft Model Antitumor Assays, Antineoplastic Agents therapeutic use, Dextrans chemistry, Drug Carriers chemistry, Glioblastoma drug therapy, Paclitaxel therapeutic use, Polyesters chemistry
- Abstract
Current interstitial therapies for glioblastoma can overcome the blood-brain barrier but fail to optimally release therapy at a rate that stalls cancer reoccurrence. To address this lapse, acetalated dextran (Ace-DEX) nanofibrous scaffolds were used for their unique degradation rates that translate to a broad range of drug release kinetics. A distinctive range of drug release rates was illustrated via electrospun Ace-DEX or poly(lactic acid) (PLA) scaffolds. Scaffolds composed of fast, medium, and slow degrading Ace-DEX resulted in 14.1%, 2.9%, and 1.3% paclitaxel released per day. To better understand the impact of paclitaxel release rate on interstitial therapy, two clinically relevant orthotopic glioblastoma mouse models were explored: (1) a surgical model of resection and recurrence (resection model) and (2) a distant metastasis model. The effect of unique drug release was illustrated in the resection model when a 78% long-term survival was observed with combined fast and slow release scaffolds, in comparison to a survival of 20% when the same dose is delivered at a medium release rate. In contrast, only the fast release rate scaffold displayed treatment efficacy in the distant metastasis model. Additionally, the acid-sensitive Ace-DEX scaffolds were shown to respond to the lower pH conditions associated with GBM tumors, releasing more paclitaxel in vivo when a tumor was present in contrast to nonacid sensitive PLA scaffolds. The unique range of tunable degradation and stimuli-responsive nature makes Ace-DEX a promising drug delivery platform to improve interstitial therapy for glioblastoma.
- Published
- 2020
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20. Developing Implantable Scaffolds to Enhance Neural Stem Cell Therapy for Post-Operative Glioblastoma.
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Sheets KT, Ewend MG, Mohiti-Asli M, Tuin SA, Loboa EG, Aboody KS, and Hingtgen SD
- Subjects
- Animals, Brain Neoplasms pathology, Brain Neoplasms surgery, Cell Line, Tumor, Combined Modality Therapy, Ganciclovir pharmacology, Glioblastoma pathology, Glioblastoma surgery, Humans, Luminescent Measurements, Mice, Neural Stem Cells metabolism, Polyesters chemistry, Prodrugs administration & dosage, Prodrugs pharmacology, Tissue Scaffolds chemistry, Treatment Outcome, Xenograft Model Antitumor Assays, Brain Neoplasms therapy, Ganciclovir administration & dosage, Glioblastoma therapy, Neural Stem Cells transplantation, Thymidine Kinase metabolism
- Abstract
Pre-clinical and clinical studies have shown that engineered tumoricidal neural stem cells (tNSCs) are a promising treatment strategy for the aggressive brain cancer glioblastoma (GBM). Yet, stabilizing human tNSCs within the surgical cavity following GBM resection is a significant challenge. As a critical step toward advancing engineered human NSC therapy for GBM, we used a preclinical variant of the clinically utilized NSC line HB1.F3.CD and mouse models of human GBM resection/recurrence to identify a polymeric scaffold capable of maximizing the transplant, persistence, and tumor kill of NSC therapy for post-surgical GBM. Using kinetic bioluminescence imaging, we found that tNSCs delivered into the mouse surgical cavity wall by direct injection persisted only 3 days. We found that delivery of tNSCs into the cavity on nanofibrous electrospun poly-l-lactic acid scaffolds extended tNSC persistence to 8 days. Modifications to fiber surface coating, diameter, and morphology of the scaffold failed to significantly extend tNSC persistence in the cavity. In contrast, tNSCs delivered into the post-operative cavity on gelatin matrices (GEMs) persisted 8-fold longer as compared to direct injection. GEMs remained permissive to tumor-tropic homing, as tNSCs migrated off the scaffolds and into invasive tumor foci both in vitro and in vivo. To mirror envisioned human brain tumor trials, we engineered tNSCs to express the prodrug/enzyme thymidine kinase (tNSCs
tk ) and transplanted the therapeutic cells in the post-operative cavity of mice bearing resected orthotopic patient-derived GBM xenografts. Following administration of the prodrug ganciclovir, residual tumor volumes in mice receiving GEM/tNSCs were reduced by 10-fold at day 35, and median survival was extended from 31 to 46 days. Taken together, these data begin to define design parameters for effective scaffold/tNSC composites and suggest a new approach to maximizing the efficacy of tNSC therapy in human patient trials., (Published by Elsevier Inc.)- Published
- 2020
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21. Engineered Mesenchymal Stem Cell/Nanomedicine Spheroid as an Active Drug Delivery Platform for Combinational Glioblastoma Therapy.
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Suryaprakash S, Lao YH, Cho HY, Li M, Ji HY, Shao D, Hu H, Quek CH, Huang D, Mintz RL, Bagó JR, Hingtgen SD, Lee KB, and Leong KW
- Subjects
- Cell Engineering trends, Cell Movement drug effects, Combined Modality Therapy, Glioblastoma genetics, Glioblastoma pathology, Humans, Mesenchymal Stem Cells cytology, Nanomedicine trends, Spheroids, Cellular chemistry, Viral Tropism drug effects, Drug Delivery Systems, Glioblastoma drug therapy, Mesenchymal Stem Cells chemistry, Spheroids, Cellular transplantation
- Abstract
Mesenchymal stem cell (MSC) has been increasingly applied to cancer therapy because of its tumor-tropic capability. However, short retention at target tissue and limited payload option hinder the progress of MSC-based cancer therapy. Herein, we proposed a hybrid spheroid/nanomedicine system, comprising MSC spheroid entrapping drug-loaded nanocomposite, to address these limitations. Spheroid formulation enhanced MSC's tumor tropism and facilitated loading of different types of therapeutic payloads. This system acted as an active drug delivery platform seeking and specifically targeting glioblastoma cells. It enabled effective delivery of combinational protein and chemotherapeutic drugs by engineered MSC and nanocomposite, respectively. In an in vivo migration model, the hybrid spheroid showed higher nanocomposite retention in the tumor tissue compared with the single MSC approach, leading to enhanced tumor inhibition in a heterotopic glioblastoma murine model. Taken together, this system integrates the merits of cell- and nanoparticle- mediated drug delivery with the tumor-homing characteristics of MSC to advance targeted combinational cancer therapy.
- Published
- 2019
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22. Image-Guided Resection of Glioblastoma and Intracranial Implantation of Therapeutic Stem Cell-seeded Scaffolds.
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Sheets KT, Bagó JR, Paulk IL, and Hingtgen SD
- Subjects
- Animals, Brain Neoplasms pathology, Glioblastoma pathology, Humans, Mesenchymal Stem Cells metabolism, Mice, Stem Cells pathology, Brain Neoplasms diagnostic imaging, Brain Neoplasms surgery, Glioblastoma diagnostic imaging, Glioblastoma surgery, Stem Cells metabolism, Tissue Scaffolds chemistry
- Abstract
Glioblastoma (GBM), the most common and aggressive primary brain cancer, carries a life expectancy of 12-15 months. The short life expectancy is due in part to the inability of the current treatment, consisting of surgical resection followed by radiation and chemotherapy, to eliminate invasive tumor foci. Treatment of these foci may be improved with tumoricidal human mesenchymal stem cells (MSCs). MSCs exhibit potent tumor tropism and can be engineered to express therapeutic proteins that kill tumor cells. Advancements in preclinical models indicate that surgical resection induces premature MSC loss and reduces therapeutic efficacy. Efficacy of MSC treatment can be improved by seeding MSCs on a biodegradable poly(lactic acid) (PLA) scaffold. MSC delivery into the surgical resection cavity on a PLA scaffold restores cell retention, persistence, and tumor killing. To study the effects of MSC-seeded PLA implantation on GBM, an accurate preclinical model is needed. Here we provide a preclinical surgical protocol for image-guided tumor resection of GBM in immune-deficient mice followed by MSC-seeded scaffold implantation. MSCs are engineered with lentiviral constructs to constitutively express and secrete therapeutic TNFα-related apoptosis-inducing ligand (TRAIL) as well as green fluorescent protein (GFP) to allow fluorescent tracking. Similarly, the U87 tumor cells are engineered to express mCherry and firefly luciferase, providing dual fluorescent/luminescent tracking. While currently used for investigating stem cell mediated delivery of therapeutics, this protocol could be modified to investigate the impact of surgical resection on other GBM interventions.
- Published
- 2018
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23. Intra-cavity stem cell therapy inhibits tumor progression in a novel murine model of medulloblastoma surgical resection.
- Author
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Okolie O, Irvin DM, Bago JR, Sheets K, Satterlee A, Carey-Ewend AG, Lettry V, Dumitru R, Elton S, Ewend MG, Miller CR, and Hingtgen SD
- Subjects
- Animals, Brain pathology, Brain surgery, Brain Neoplasms mortality, Brain Neoplasms pathology, Brain Neoplasms surgery, Cell Differentiation, Cell Movement, Disease Models, Animal, Enzyme Therapy methods, Epithelial Cells cytology, Ganciclovir pharmacology, Humans, Injections, Intralesional, Medulloblastoma mortality, Medulloblastoma pathology, Medulloblastoma surgery, Mice, Neoplasm Recurrence, Local mortality, Neoplasm Recurrence, Local pathology, Neoplasm Recurrence, Local surgery, Neural Stem Cells cytology, Prodrugs pharmacology, Skin cytology, Survival Analysis, Thymidine Kinase genetics, Thymidine Kinase metabolism, Brain Neoplasms therapy, Cell- and Tissue-Based Therapy methods, Medulloblastoma therapy, Neoplasm Recurrence, Local prevention & control, Neural Stem Cells transplantation, Surgery, Computer-Assisted methods
- Abstract
Background: Cytotoxic neural stem cells (NSCs) have emerged as a promising treatment for Medulloblastoma (MB), the most common malignant primary pediatric brain tumor. The lack of accurate pre-clinical models incorporating surgical resection and tumor recurrence limits advancement in post-surgical MB treatments. Using cell lines from two of the 5 distinct MB molecular sub-groups, in this study, we developed an image-guided mouse model of MB surgical resection and investigate intra-cavity NSC therapy for post-operative MB., Methods: Using D283 and Daoy human MB cells engineered to express multi-modality optical reporters, we created the first image-guided resection model of orthotopic MB. Brain-derived NSCs and novel induced NSCs (iNSCs) generated from pediatric skin were engineered to express the pro-drug/enzyme therapy thymidine kinase/ganciclovir, seeded into the post-operative cavity, and used to investigate intra-cavity therapy for post-surgical MB., Results: We found that surgery reduced MB volumes by 92%, and the rate of post-operative MB regrowth increased 3-fold compared to pre-resection growth. Real-time imaging showed NSCs rapidly homed to MB, migrating 1.6-fold faster and 2-fold farther in the presence of tumors, and co-localized with MB present in the contra-lateral hemisphere. Seeding of cytotoxic NSCs into the post-operative surgical cavity decreased MB volumes 15-fold and extended median survival 133%. As an initial step towards novel autologous therapy in human MB patients, we found skin-derived iNSCs homed to MB cells, while intra-cavity iNSC therapy suppressed post-surgical tumor growth and prolonged survival of MB-bearing mice by 123%., Conclusions: We report a novel image-guided model of MB resection/recurrence and provide new evidence of cytotoxic NSCs/iNSCs delivered into the surgical cavity effectively target residual MB foci., Competing Interests: SH and MG are co-founders of Falcon Therapeutics. This does not alter our adherence to PLoS ONE policies on sharing data and materials.
- Published
- 2018
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24. Sustained Delivery of Doxorubicin via Acetalated Dextran Scaffold Prevents Glioblastoma Recurrence after Surgical Resection.
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Graham-Gurysh E, Moore KM, Satterlee AB, Sheets KT, Lin FC, Bachelder EM, Miller CR, Hingtgen SD, and Ainslie KM
- Subjects
- Acetals chemistry, Animals, Antibiotics, Antineoplastic pharmacokinetics, Brain pathology, Brain surgery, Brain Neoplasms pathology, Brain Neoplasms surgery, Cell Line, Tumor, Cell Survival drug effects, Delayed-Action Preparations administration & dosage, Delayed-Action Preparations pharmacokinetics, Dextrans chemistry, Disease Progression, Doxorubicin pharmacokinetics, Drug Liberation, Glioblastoma pathology, Glioblastoma surgery, Humans, Hydrogen-Ion Concentration, Mice, Mice, Inbred BALB C, Mice, Nude, Treatment Outcome, Xenograft Model Antitumor Assays, Antibiotics, Antineoplastic administration & dosage, Brain Neoplasms drug therapy, Doxorubicin administration & dosage, Drug Delivery Systems methods, Glioblastoma drug therapy, Neoplasm Recurrence, Local prevention & control
- Abstract
The primary cause of mortality for glioblastoma (GBM) is local tumor recurrence following standard-of-care therapies, including surgical resection. With most tumors recurring near the site of surgical resection, local delivery of chemotherapy at the time of surgery is a promising strategy. Herein drug-loaded polymer scaffolds with two distinct degradation profiles were fabricated to investigate the effect of local drug delivery rate on GBM recurrence following surgical resection. The novel biopolymer, acetalated dextran (Ace-DEX), was compared with commercially available polyester, poly(l-lactide) (PLA). Steady-state doxorubicin (DXR) release from Ace-DEX scaffolds was found to be faster when compared with scaffolds composed of PLA, in vitro. This increased drug release rate translated to improved therapeutic outcomes in a novel surgical model of orthotopic glioblastoma resection and recurrence. Mice treated with DXR-loaded Ace-DEX scaffolds (Ace-DEX/10DXR) resulted in 57% long-term survival out to study completion at 120 days compared with 20% survival following treatment with DXR-loaded PLA scaffolds (PLA/10DXR). Additionally, all mice treated with PLA/10DXR scaffolds exhibited disease progression by day 38, as defined by a 5-fold growth in tumor bioluminescent signal. In contrast, 57% of mice treated with Ace-DEX/10DXR scaffolds displayed a reduction in tumor burden, with 43% exhibiting complete remission. These results underscore the importance of polymer choice and drug release rate when evaluating local drug delivery strategies to improve prognosis for GBM patients undergoing tumor resection.
- Published
- 2018
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25. Delivery of Cytotoxic Mesenchymal Stem Cells with Biodegradable Scaffolds for Treatment of Postoperative Brain Cancer.
- Author
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Sheets KT, Bagó JR, and Hingtgen SD
- Subjects
- Animals, Brain Neoplasms surgery, Cell Death, Cell Line, Tumor, Humans, Mice, Nude, Biocompatible Materials pharmacology, Brain Neoplasms therapy, Mesenchymal Stem Cell Transplantation methods, Mesenchymal Stem Cells cytology, Tissue Scaffolds chemistry
- Abstract
Engineered stem cells have recently entered clinical trials as therapeutic agents for treating glioblastoma foci that remain after primary brain tumor resection. However, efficient delivery of anti-cancer mesenchymal stem cells (MSCs) into the resection cavity remains a potential obstacle to therapeutic efficacy in humans. Direct injection quickly leads to significant stem cell loss and poor tumor killing. Recent reports have shown that biodegradable scaffolds improve MSC persistence and restore therapeutic potential. Here, we describe a method for the delivery of therapeutic MSCs on biodegradable fibrin scaffolds into the resection cavity to treat postoperative brain cancer.
- Published
- 2018
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26. Tumor-homing cytotoxic human induced neural stem cells for cancer therapy.
- Author
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Bagó JR, Okolie O, Dumitru R, Ewend MG, Parker JS, Werff RV, Underhill TM, Schmid RS, Miller CR, and Hingtgen SD
- Subjects
- Animals, Cell Movement, Cell Transdifferentiation, Drug Delivery Systems, Fibroblasts cytology, Humans, Mice, Neural Stem Cells transplantation, Skin cytology, Spheroids, Cellular, Xenograft Model Antitumor Assays, Brain Neoplasms therapy, Glioblastoma therapy, Neural Stem Cells cytology, TNF-Related Apoptosis-Inducing Ligand administration & dosage
- Abstract
Engineered neural stem cells (NSCs) are a promising approach to treating glioblastoma (GBM). The ideal NSC drug carrier for clinical use should be easily isolated and autologous to avoid immune rejection. We transdifferentiated (TD) human fibroblasts into tumor-homing early-stage induced NSCs (h-iNSC
TE ), engineered them to express optical reporters and different therapeutic gene products, and assessed the tumor-homing migration and therapeutic efficacy of cytotoxic h-iNSCTE in patient-derived GBM models of surgical and nonsurgical disease. Molecular and functional analysis revealed that our single-factor SOX2 TD strategy converted human skin fibroblasts into h-iNSCTE that were nestin+ and expressed pathways associated with tumor-homing migration in 4 days. Time-lapse motion analysis showed that h-iNSCTE rapidly migrated to human GBM cells and penetrated human GBM spheroids, a process inhibited by blockade of CXCR4. Serial imaging showed that h-iNSCTE delivery of the proapoptotic agent tumor necrosis factor-α-related apoptosis-inducing ligand (TRAIL) reduced the size of solid human GBM xenografts 250-fold in 3 weeks and prolonged median survival from 22 to 49 days. Additionally, h-iNSCTE thymidine kinase/ganciclovir enzyme/prodrug therapy (h-iNSCTE -TK) reduced the size of patient-derived GBM xenografts 20-fold and extended survival from 32 to 62 days. Mimicking clinical NSC therapy, h-iNSCTE -TK therapy delivered into the postoperative surgical resection cavity delayed the regrowth of residual GBMs threefold and prolonged survival from 46 to 60 days. These results suggest that TD of human skin into h-iNSCTE is a platform for creating tumor-homing cytotoxic cell therapies for cancer, where the potential to avoid carrier rejection could maximize treatment durability in human trials., (Copyright © 2017, American Association for the Advancement of Science.)- Published
- 2017
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27. Reactive astrocytes potentiate tumor aggressiveness in a murine glioma resection and recurrence model.
- Author
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Okolie O, Bago JR, Schmid RS, Irvin DM, Bash RE, Miller CR, and Hingtgen SD
- Subjects
- Allografts, Animals, Astrocytes metabolism, Brain Neoplasms metabolism, Brain Neoplasms pathology, Cell Line, Tumor, Cell Movement, Cell Proliferation, Coculture Techniques, Glioblastoma metabolism, Glioblastoma pathology, Mice, Neoplasm Recurrence, Local metabolism, Transcriptome, Astrocytes physiology, Brain Neoplasms physiopathology, Disease Models, Animal, Glioblastoma physiopathology, Neoplasm Recurrence, Local physiopathology, Tumor Microenvironment
- Abstract
Background: Surgical resection is a universal component of glioma therapy. Little is known about the postoperative microenvironment due to limited preclinical models. Thus, we sought to develop a glioma resection and recurrence model in syngeneic immune-competent mice to understand how surgical resection influences tumor biology and the local microenvironment., Methods: We genetically engineered cells from a murine glioma mouse model to express fluorescent and bioluminescent reporters. Established allografts were resected using image-guided microsurgery. Postoperative tumor recurrence was monitored by serial imaging, and the peritumoral microenvironment was characterized by histopathology and immunohistochemistry. Coculture techniques were used to explore how astrocyte injury influences tumor aggressiveness in vitro. Transcriptome and secretome alterations in injured astrocytes was examined by RNA-seq and Luminex., Results: We found that image-guided resection achieved >90% reduction in tumor volume but failed to prevent both local and distant tumor recurrence. Immunostaining for glial fibrillary acidic protein and nestin showed that resection-induced injury led to temporal and spatial alterations in reactive astrocytes within the peritumoral microenvironment. In vitro, we found that astrocyte injury induced transcriptome and secretome alterations and promoted tumor proliferation, as well as migration., Conclusions: This study demonstrates a unique syngeneic model of glioma resection and recurrence in immune-competent mice. Furthermore, this model provided insights into the pattern of postsurgical tumor recurrence and changes in the peritumoral microenvironment, as well as the impact of injured astrocytes on glioma growth and invasion. A better understanding of the postsurgical tumor microenvironment will allow development of targeted anticancer agents that improve surgery-mediated effects on tumor biology., (© The Author(s) 2016. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2016
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28. Electrospun nanofibrous scaffolds increase the efficacy of stem cell-mediated therapy of surgically resected glioblastoma.
- Author
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Bagó JR, Pegna GJ, Okolie O, Mohiti-Asli M, Loboa EG, and Hingtgen SD
- Subjects
- Animals, Antineoplastic Agents therapeutic use, Brain pathology, Brain surgery, Brain Neoplasms pathology, Brain Neoplasms surgery, Cell Line, Cell Line, Tumor, Glioblastoma pathology, Glioblastoma surgery, Humans, Mice, Nude, Nanofibers ultrastructure, Polyesters chemistry, Stem Cells cytology, Antineoplastic Agents administration & dosage, Brain Neoplasms therapy, Drug Delivery Systems methods, Glioblastoma therapy, Nanofibers chemistry, Stem Cell Transplantation methods, Tissue Scaffolds chemistry
- Abstract
Engineered stem cell (SC)-based therapy holds enormous promise for treating the incurable brain cancer glioblastoma (GBM). Retaining the cytotoxic SCs in the surgical cavity after GBM resection is one of the greatest challenges to this approach. Here, we describe a biocompatible electrospun nanofibrous scaffold (bENS) implant capable of delivering and retaining tumor-homing cytotoxic stem cells that suppress recurrence of post-surgical GBM. As a new approach to GBM therapy, we created poly(l-lactic acid) (PLA) bENS bearing drug-releasing human mesenchymal stem cells (hMSCs). We discovered that bENS-based implant increased hMSC retention in the surgical cavity 5-fold and prolonged persistence 3-fold compared to standard direct injection using our mouse model of GBM surgical resection/recurrence. Time-lapse imaging showed cytotoxic hMSC/bENS treatment killed co-cultured human GBM cells, and allowed hMSCs to rapidly migrate off the scaffolds as they homed to GBMs. In vivo, bENS loaded with hMSCs releasing the anti-tumor protein TRAIL (bENS(sTR)) reduced the volume of established GBM xenografts 3-fold. Mimicking clinical GBM patient therapy, lining the post-operative GBM surgical cavity with bENS(sTR) implants inhibited the re-growth of residual GBM foci 2.3-fold and prolonged post-surgical median survival from 13.5 to 31 days in mice. These results suggest that nanofibrous-based SC therapies could be an innovative new approach to improve the outcomes of patients suffering from terminal brain cancer., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2016
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29. Neural stem cell therapy for cancer.
- Author
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Bagó JR, Sheets KT, and Hingtgen SD
- Subjects
- Animals, Cell Culture Techniques, Cell Engineering, Cell Movement, Cellular Reprogramming, Humans, Neural Stem Cells physiology, Neoplasms therapy, Neural Stem Cells transplantation
- Abstract
Cancers of the brain remain one of the greatest medical challenges. Traditional surgery and chemo-radiation therapy are unable to eradicate diffuse cancer cells and tumor recurrence is nearly inevitable. In contrast to traditional regenerative medicine applications, engineered neural stem cells (NSCs) are emerging as a promising new therapeutic strategy for cancer therapy. The tumor-homing properties allow NSCs to access both primary and invasive tumor foci, creating a novel delivery platform. NSCs engineered with a wide array of cytotoxic agents have been found to significantly reduce tumor volumes and markedly extend survival in preclinical models. With the recent launch of new clinical trials, the potential to successfully manage cancer in human patients with cytotoxic NSC therapy is moving closer to becoming a reality., (Copyright © 2015. Published by Elsevier Inc.)
- Published
- 2016
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30. Development of exosome-encapsulated paclitaxel to overcome MDR in cancer cells.
- Author
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Kim MS, Haney MJ, Zhao Y, Mahajan V, Deygen I, Klyachko NL, Inskoe E, Piroyan A, Sokolsky M, Okolie O, Hingtgen SD, Kabanov AV, and Batrakova EV
- Subjects
- Animals, Antineoplastic Agents, Phytogenic pharmacokinetics, Antineoplastic Agents, Phytogenic therapeutic use, Cell Line, Cell Line, Tumor, Dogs, Drug Delivery Systems, Drug Resistance, Neoplasm, Female, Lung pathology, Lung Neoplasms pathology, Macrophages chemistry, Mice, Mice, Inbred C57BL, Paclitaxel pharmacokinetics, Paclitaxel therapeutic use, Sonication, Antineoplastic Agents, Phytogenic administration & dosage, Drug Carriers chemistry, Exosomes chemistry, Lung drug effects, Lung Neoplasms drug therapy, Lung Neoplasms secondary, Paclitaxel administration & dosage
- Abstract
Exosomes have recently come into focus as "natural nanoparticles" for use as drug delivery vehicles. Our objective was to assess the feasibility of an exosome-based drug delivery platform for a potent chemotherapeutic agent, paclitaxel (PTX), to treat MDR cancer. Herein, we developed different methods of loading exosomes released by macrophages with PTX (exoPTX), and characterized their size, stability, drug release, and in vitro antitumor efficacy. Reformation of the exosomal membrane upon sonication resulted in high loading efficiency and sustained drug release. Importantly, incorporation of PTX into exosomes increased cytotoxicity more than 50 times in drug resistant MDCKMDR1 (Pgp+) cells. Next, our studies demonstrated a nearly complete co-localization of airway-delivered exosomes with cancer cells in a model of murine Lewis lung carcinoma pulmonary metastases, and a potent anticancer effect in this mouse model. We conclude that exoPTX holds significant potential for the delivery of various chemotherapeutics to treat drug resistant cancers., From the Clinical Editor: Exosomes are membrane-derived natural vesicles of ~40 - 200 nm size. They have been under extensive research as novel drug delivery vehicles. In this article, the authors developed exosome-based system to carry formulation of PTX and showed efficacy in the treatment of multi-drug resistant cancer cells. This novel system may be further developed to carry other chemotherapeutic agents in the future., (Published by Elsevier Inc.)
- Published
- 2016
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31. Fibrin matrices enhance the transplant and efficacy of cytotoxic stem cell therapy for post-surgical cancer.
- Author
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Bagó JR, Pegna GJ, Okolie O, and Hingtgen SD
- Subjects
- Animals, Brain Neoplasms surgery, Cell Death drug effects, Cell Line, Tumor, Cell Movement drug effects, Disease Progression, Glioblastoma surgery, Humans, Mice, Nude, Spheroids, Cellular drug effects, Spheroids, Cellular pathology, Tissue Scaffolds chemistry, Treatment Outcome, Brain Neoplasms therapy, Fibrin pharmacology, Glioblastoma therapy, Stem Cell Transplantation
- Abstract
Tumor-homing cytotoxic stem cell (SC) therapy is a promising new approach for treating the incurable brain cancer glioblastoma (GBM). However, problems of retaining cytotoxic SCs within the post-surgical GBM resection cavity are likely to significantly limit the clinical utility of this strategy. Here, we describe a new fibrin-based transplant approach capable of increasing cytotoxic SC retention and persistence within the resection cavity, yet remaining permissive to tumoritropic migration. This fibrin-based transplant can effectively treat both solid and post-surgical human GBM in mice. Using our murine model of image-guided model of GBM resection, we discovered that suspending human mesenchymal stem cells (hMSCS) in a fibrin matrix increased initial retention in the surgical resection cavity 2-fold and prolonged persistence in the cavity 3-fold compared to conventional delivery strategies. Time-lapse motion analysis revealed that cytotoxic hMSCs in the fibrin matrix remain tumoritropic, rapidly migrating from the fibrin matrix to co-localize with cultured human GBM cells. We encapsulated hMSCs releasing the cytotoxic agent TRAIL (hMSC-sTR) in fibrin, and found hMSC-sTR/fibrin therapy reduced the viability of multiple 3-D human GBM spheroids and regressed established human GBM xenografts 3-fold in 11 days. Mimicking clinical therapy of surgically resected GBM, intra-cavity seeding of therapeutic hMSC-sTR encapsulated in fibrin reduced post-surgical GBM volumes 6-fold, increased time to recurrence 4-fold, and prolonged median survival from 15 to 36 days compared to control-treated animals. Fibrin-based SC therapy could represent a clinically compatible, viable treatment to suppress recurrence of post-surgical GBM and other lethal cancer types., (Published by Elsevier Ltd.)
- Published
- 2016
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32. Therapeutically engineered induced neural stem cells are tumour-homing and inhibit progression of glioblastoma.
- Author
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Bagó JR, Alfonso-Pecchio A, Okolie O, Dumitru R, Rinkenbaugh A, Baldwin AS, Miller CR, Magness ST, and Hingtgen SD
- Subjects
- Animals, Cell Differentiation, Cell Movement, Cell Proliferation, Cell Survival, Cells, Cultured, Fibroblasts, Humans, Induced Pluripotent Stem Cells, Mice, Neoplasms, Experimental, Astrocytes, Glioblastoma, Neural Stem Cells cytology, Neural Stem Cells physiology
- Abstract
Transdifferentiation (TD) is a recent advancement in somatic cell reprogramming. The direct conversion of TD eliminates the pluripotent intermediate state to create cells that are ideal for personalized cell therapy. Here we provide evidence that TD-derived induced neural stem cells (iNSCs) are an efficacious therapeutic strategy for brain cancer. We find that iNSCs genetically engineered with optical reporters and tumouricidal gene products retain the capacity to differentiate and induced apoptosis in co-cultured human glioblastoma cells. Time-lapse imaging shows that iNSCs are tumouritropic, homing rapidly to co-cultured glioblastoma cells and migrating extensively to distant tumour foci in the murine brain. Multimodality imaging reveals that iNSC delivery of the anticancer molecule TRAIL decreases the growth of established solid and diffuse patient-derived orthotopic glioblastoma xenografts 230- and 20-fold, respectively, while significantly prolonging the median mouse survival. These findings establish a strategy for creating autologous cell-based therapies to treat patients with aggressive forms of brain cancer.
- Published
- 2016
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33. Engineering toxin-resistant therapeutic stem cells to treat brain tumors.
- Author
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Stuckey DW, Hingtgen SD, Karakas N, Rich BE, and Shah K
- Subjects
- Animals, Brain Neoplasms metabolism, Brain Neoplasms pathology, Cell Line, Genetic Engineering, Heterografts, Humans, Mice, Pseudomonas genetics, Stem Cells pathology, Bacterial Proteins genetics, Bacterial Proteins metabolism, Brain Neoplasms therapy, Drug Resistance genetics, Exotoxins genetics, Exotoxins metabolism, Interleukin-13 genetics, Interleukin-13 metabolism, Peptide Elongation Factor 2 genetics, Peptide Elongation Factor 2 metabolism, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Stem Cell Transplantation, Stem Cells metabolism
- Abstract
Pseudomonas exotoxin (PE) potently blocks protein synthesis by catalyzing the inactivation of elongation factor-2 (EF-2). Targeted PE-cytotoxins have been used as antitumor agents, although their effective clinical translation in solid tumors has been confounded by off-target delivery, systemic toxicity, and short chemotherapeutic half-life. To overcome these limitations, we have created toxin-resistant stem cells by modifying endogenous EF-2, and engineered them to secrete PE-cytotoxins that target specifically expressed (interleukin-13 receptor subunit alpha-2) or overexpressed (epidermal growth factor receptor) in glioblastomas (GBM). Molecular analysis correlated efficacy of PE-targeted cytotoxins with levels of cognate receptor expression, and optical imaging was applied to simultaneously track the kinetics of protein synthesis inhibition and GBM cell viability in vivo. The release of IL13-PE from biodegradable synthetic extracellular matrix (sECM) encapsulated stem cells in a clinically relevant GBM resection model led to increased long-term survival of mice compared to IL13-PE protein infusion. Moreover, multiple patient-derived GBM lines responded to treatment, underscoring its clinical relevance. In sum, integrating stem cell-based engineering, multimodal imaging, and delivery of PE-cytotoxins in a clinically relevant GBM model represents a novel strategy and a potential advancement in GBM therapy., (© 2014 AlphaMed Press.)
- Published
- 2015
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34. Macrophages offer a paradigm switch for CNS delivery of therapeutic proteins.
- Author
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Klyachko NL, Haney MJ, Zhao Y, Manickam DS, Mahajan V, Suresh P, Hingtgen SD, Mosley RL, Gendelman HE, Kabanov AV, and Batrakova EV
- Subjects
- Animals, Catalase metabolism, Cattle, Cell Line, Chemistry, Pharmaceutical, Cross-Linking Reagents, Drug Carriers chemistry, Encephalitis drug therapy, Encephalitis pathology, Encephalitis physiopathology, Enzyme Stability, Macrophage Activation, Macrophages cytology, Macrophages enzymology, Male, Mice, Mice, Inbred C57BL, Nanocapsules chemistry, Nanocapsules ultrastructure, Nanomedicine, Particle Size, Blood-Brain Barrier metabolism, Catalase administration & dosage, Drug Delivery Systems, Macrophages physiology, Nanocapsules administration & dosage
- Abstract
Aims: Active targeted transport of the nanoformulated redox enzyme, catalase, in macrophages attenuates oxidative stress and as such increases survival of dopaminergic neurons in animal models of Parkinson's disease. Optimization of the drug formulation is crucial for the successful delivery in living cells. We demonstrated earlier that packaging of catalase into a polyion complex micelle ('nanozyme') with a synthetic polyelectrolyte block copolymer protected the enzyme against degradation in macrophages and improved therapeutic outcomes. We now report the manufacture of nanozymes with superior structure and therapeutic indices., Methods: Synthesis, characterization and therapeutic efficacy of optimal cell-based nanoformulations are evaluated., Results: A formulation design for drug carriers typically works to avoid entrapment in monocytes and macrophages focusing on small-sized nanoparticles with a polyethylene glycol corona (to provide a stealth effect). By contrast, the best nanozymes for delivery in macrophages reported in this study have a relatively large size (≈ 200 nm), which resulted in improved loading capacity and release from macrophages. Furthermore, the cross-linking of nanozymes with the excess of a nonbiodegradable linker ensured their low cytotoxicity, and efficient catalase protection in cell carriers. Finally, the 'alternatively activated' macrophage phenotype (M2) utilized in these studies did not promote further inflammation in the brain, resulting in a subtle but statistically significant effect on neuronal regeneration and repair in vivo., Conclusion: The optimized cross-linked nanozyme loaded into macrophages reduced neuroinflammatory responses and increased neuronal survival in mice. Importantly, the approach for nanoformulation design for cell-mediated delivery is different from the common requirements for injectable formulations.
- Published
- 2014
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35. Specific transfection of inflamed brain by macrophages: a new therapeutic strategy for neurodegenerative diseases.
- Author
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Haney MJ, Zhao Y, Harrison EB, Mahajan V, Ahmed S, He Z, Suresh P, Hingtgen SD, Klyachko NL, Mosley RL, Gendelman HE, Kabanov AV, and Batrakova EV
- Subjects
- Animals, Anti-Inflammatory Agents pharmacology, Brain drug effects, Catalase genetics, Catalase therapeutic use, Cell Line, Disease Models, Animal, Exosomes drug effects, Exosomes metabolism, Genes, Reporter, Green Fluorescent Proteins metabolism, Humans, Kinetics, Macrophages drug effects, Male, Mice, Mice, Inbred BALB C, Models, Biological, Neurons drug effects, Neurons metabolism, Neurons pathology, Neuroprotective Agents pharmacology, Parkinson Disease pathology, Parkinson Disease therapy, Tissue Distribution drug effects, Brain pathology, Genetic Therapy, Inflammation pathology, Macrophages metabolism, Neurodegenerative Diseases therapy, Transfection methods
- Abstract
The ability to precisely upregulate genes in inflamed brain holds great therapeutic promise. Here we report a novel class of vectors, genetically modified macrophages that carry reporter and therapeutic genes to neural cells. Systemic administration of macrophages transfected ex vivo with a plasmid DNA (pDNA) encoding a potent antioxidant enzyme, catalase, produced month-long expression levels of catalase in the brain resulting in three-fold reductions in inflammation and complete neuroprotection in mouse models of Parkinson's disease (PD). This resulted in significant improvements in motor functions in PD mice. Mechanistic studies revealed that transfected macrophages secreted extracellular vesicles, exosomes, packed with catalase genetic material, pDNA and mRNA, active catalase, and NF-κb, a transcription factor involved in the encoded gene expression. Exosomes efficiently transfer their contents to contiguous neurons resulting in de novo protein synthesis in target cells. Thus, genetically modified macrophages serve as a highly efficient system for reproduction, packaging, and targeted gene and drug delivery to treat inflammatory and neurodegenerative disorders.
- Published
- 2013
- Full Text
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36. Superoxide scavenging and Akt inhibition in myocardium ameliorate pressure overload-induced NF-κB activation and cardiac hypertrophy.
- Author
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Hingtgen SD, Li Z, Kutschke W, Tian X, Sharma RV, and Davisson RL
- Subjects
- Adenoviridae genetics, Animals, Disease Models, Animal, Enzyme Activation, Gene Transfer Techniques, Genes, Reporter, Genetic Vectors, Hypertrophy, Left Ventricular enzymology, Hypertrophy, Left Ventricular genetics, Hypertrophy, Left Ventricular pathology, Luminescent Measurements, Male, Mice, Mice, Inbred C57BL, Myocardium pathology, Proto-Oncogene Proteins c-akt antagonists & inhibitors, Proto-Oncogene Proteins c-akt genetics, Signal Transduction, Superoxide Dismutase genetics, Superoxides metabolism, Time Factors, Genetic Therapy, Hypertrophy, Left Ventricular prevention & control, Myocardium enzymology, NF-kappa B metabolism, Oxidative Stress, Proto-Oncogene Proteins c-akt metabolism, Superoxide Dismutase metabolism
- Abstract
Recent studies from our laboratory and others have shown that increases in cytoplasmic superoxide (O(2)(·-)) levels and Akt activation play a key role in agonist-stimulated NF-κB activation and cardiomyocyte hypertrophy in vitro. In this study, we tested the hypothesis that adenovirus (Ad)-mediated intramyocardial gene transfer of cytoplasmic superoxide dismutase (AdCu/ZnSOD) or a dominant-negative form of Akt (AdDNAkt) in mice would attenuate pressure overload-induced increases in activation of the redox-sensitive transcription factor NF-κB and cardiac hypertrophy. Adult C57BL/6 mice were subjected to thoracic aortic banding (TAB) or sham surgery, and intramyocardial injections of viral vectors (AdCu/ZnSOD, AdDNAkt, or control) were performed. There was robust transgene expression in the heart, which peaked 6-7 days after injection and then declined to undetectable levels by 12-14 days. In mice injected with AdBgL II, TAB caused a significant increase in O(2)(·-) generation and cardiac mass at 1 wk, and these responses were markedly attenuated by AdCu/ZnSOD. In addition, TAB induced time-dependent activation of NF-κB in the myocardium as measured longitudinally by in vivo bioluminescent imaging of NF-κB-dependent luciferase expression. This was also abolished by intracardiac AdCu/ZnSOD or AdDNAkt, but not the control vector. The inhibition of Akt and O(2)(·-)-mediated NF-κB activation in TAB hearts was associated with an attenuation of cardiac hypertrophy. Since a direct cause-and-effect relationship between NF-κB activation and cardiomyocyte hypertrophy has been established previously, our data support the hypothesis that increased O(2)(·-) generation and Akt activation are key signaling intermediates in pressure overload-induced activation of NF-κB and cardiac hypertrophy.
- Published
- 2010
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37. A novel molecule integrating therapeutic and diagnostic activities reveals multiple aspects of stem cell-based therapy.
- Author
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Hingtgen SD, Kasmieh R, van de Water J, Weissleder R, and Shah K
- Subjects
- Animals, Apoptosis, Cell Line, Coculture Techniques, Genetic Therapy, Glioblastoma genetics, Glioblastoma surgery, Glioblastoma therapy, Humans, Mice, Xenograft Model Antitumor Assays, Genes, Reporter, Stem Cell Transplantation
- Abstract
Stem cells are promising therapeutic delivery vehicles; however pre-clinical and clinical applications of stem cell-based therapy would benefit significantly from the ability to simultaneously determine therapeutic efficacy and pharmacokinetics of therapies delivered by engineered stem cells. In this study, we engineered and screened numerous fusion variants that contained therapeutic (TRAIL) and diagnostic (luciferase) domains designed to allow simultaneous investigation of multiple events in stem cell-based therapy in vivo. When various stem cell lines were engineered with the optimized molecule, SRL(O)L(2)TR, diagnostic imaging showed marked differences in the levels and duration of secretion between stem cell lines, while the therapeutic activity of the molecule showed the different secretion levels translated to significant variability in tumor cell killing. In vivo, simultaneous diagnostic and therapeutic monitoring revealed that stem cell-based delivery significantly improved pharmacokinetics and anti-tumor effectiveness of the therapy compared to intravenous or intratumoral delivery. As treatment for highly malignant brain tumor xenografts, tracking SRL(O)L(2)TR showed stable stem cell-mediated delivery significantly regressed peripheral and intracranial tumors. Together, the integrated diagnostic and therapeutic properties of SRL(O)L(2)TR answer critical questions necessary for successful utilization of stem cells as novel therapeutic vehicles.
- Published
- 2010
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38. Nox2-containing NADPH oxidase and Akt activation play a key role in angiotensin II-induced cardiomyocyte hypertrophy.
- Author
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Hingtgen SD, Tian X, Yang J, Dunlay SM, Peek AS, Wu Y, Sharma RV, Engelhardt JF, and Davisson RL
- Subjects
- Adenoviridae genetics, Animals, Animals, Newborn, Blotting, Western, Cell Line, Cells, Cultured, Enzyme Activation drug effects, Female, Genetic Vectors genetics, Humans, Hypertrophy, Myocytes, Cardiac drug effects, Myocytes, Cardiac pathology, NADPH Oxidases genetics, Proto-Oncogene Proteins c-akt genetics, RNA, Small Interfering genetics, Rats, Rats, Sprague-Dawley, Reactive Oxygen Species metabolism, Superoxide Dismutase genetics, Superoxide Dismutase metabolism, Transfection, rac1 GTP-Binding Protein genetics, rac1 GTP-Binding Protein metabolism, Angiotensin II pharmacology, Myocytes, Cardiac metabolism, NADPH Oxidases metabolism, Proto-Oncogene Proteins c-akt metabolism
- Abstract
Angiotensin II (ANG II) has profound effects on the development and progression of pathological cardiac hypertrophy; however, the intracellular signaling mechanisms are not fully understood. In this study, we used genetic tools to test the hypothesis that increased formation of superoxide (O2-*) radicals from a Rac1-regulated Nox2-containing NADPH oxidase is a key upstream mediator of ANG II-induced activation of serine-threonine kinase Akt, and that this signaling cascade plays a crucial role in ANG II-dependent cardiomyocyte hypertrophy. ANG II caused a significant time-dependent increase in Rac1 activation and O2-* production in primary neonatal rat cardiomyocytes, and these responses were abolished by adenoviral (Ad)-mediated expression of a dominant-negative Rac1 (AdN17Rac1) or cytoplasmic Cu/ZnSOD (AdCu/ZnSOD). Moreover, both AdN17Rac1 and AdCu/ZnSOD significantly attenuated ANG II-stimulated increases in cardiomyocyte size. Quantitative real-time PCR analysis demonstrated that Nox2 is the homolog expressed at highest levels in primary neonatal cardiomyocytes, and small interference RNA (siRNA) directed against it selectively decreased Nox2 expression by >95% and abolished both ANG II-induced O2-* generation and cardiomyocyte hypertrophy. Finally, ANG II caused a time-dependent increase in Akt activity via activation of AT(1) receptors, and this response was abolished by Ad-mediated expression of cytosolic human O2-* dismutase (AdCu/ZnSOD). Furthermore, pretreatment of cardiomyocytes with dominant-negative Akt (AdDNAkt) abolished ANG II-induced cellular hypertrophy. These findings suggest that O2-* generated by a Nox2-containing NADPH oxidase is a central mediator of ANG II-induced Akt activation and cardiomyocyte hypertrophy, and that dysregulation of this signaling cascade may play an important role in cardiac hypertrophy.
- Published
- 2006
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39. Ganglionic action of angiotensin contributes to sympathetic activity in renin-angiotensinogen transgenic mice.
- Author
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Ma X, Sigmund CD, Hingtgen SD, Tian X, Davisson RL, Abboud FM, and Chapleau MW
- Subjects
- Angiotensin II Type 1 Receptor Blockers, Angiotensinogen genetics, Animals, Blood Pressure drug effects, Chlorisondamine pharmacology, Ganglia, Sympathetic drug effects, Ganglionic Blockers pharmacology, Humans, Hypertension genetics, Kidney innervation, Losartan pharmacology, Mice, Mice, Transgenic, Renin genetics, Renin-Angiotensin System drug effects, Sympathetic Nervous System physiopathology, Angiotensin II pharmacology, Ganglia, Sympathetic physiopathology, Hypertension physiopathology, Renin-Angiotensin System physiology
- Abstract
In addition to central nervous system actions, angiotensin (Ang) II may increase sympathetic nerve activity (SNA) via a direct action on sympathetic ganglia. We hypothesized that sympathetic ganglionic actions of endogenous Ang II contribute to SNA in transgenic mice that overexpress renin and angiotensinogen (R+A+ mice). Renal SNA and arterial pressure were recorded in anesthetized R+A+ and littermate control mice before and after ganglionic blockade, and after additional blockade of angiotensin type 1 (AT1) receptors with losartan. Ganglionic blockade essentially abolished SNA in control mice, but only reduced SNA to 47+/-18% of baseline in R+A+ mice. The residual SNA remaining after ganglionic blockade in R+A+ mice was reduced from 47+/-18% to 8+/-6% of baseline by losartan (P<0.05). The sympathoinhibitory response to losartan was accompanied by an enhanced decrease in arterial pressure in R+A+ mice compared with that observed in control mice. AT1 receptor expression in sympathetic ganglia, as measured by real-time reverse transcription-polymerase chain reaction, was increased approximately 3-fold in R+A+ versus control mice. The results demonstrate that, as anticipated, essentially all of the renal postganglionic SNA in control mice is driven by preganglionic input. The major new finding is that Ang II-evoked ganglionic activity accounts for approximately 40% of total SNA in R+A+ mice. The significant contribution of the direct ganglionic action of Ang II in R+A+ mice likely reflects both increased levels of Ang II and upregulation of AT1 receptors in sympathetic ganglia.
- Published
- 2004
- Full Text
- View/download PDF
40. Gene therapeutic approaches to oxidative stress-induced cardiac disease: principles, progress, and prospects.
- Author
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Hingtgen SD and Davisson RL
- Subjects
- Animals, Cardiomegaly etiology, Cardiomegaly therapy, Forecasting, Genetic Vectors, Heart Diseases etiology, Heart Failure etiology, Humans, Mice, Models, Biological, Myocardial Reperfusion Injury etiology, Myocardial Reperfusion Injury therapy, Oxidation-Reduction, Viruses genetics, Genetic Therapy methods, Heart Diseases therapy, Oxidative Stress
- Abstract
Heart and vascular diseases continue to rank among the most frequent and devastating disorders to affect adults in many parts of the world. Increasing evidence from a variety of experimental models indicates that reactive oxygen species can play a key role in the development of myocardial damage from ischemia/reperfusion, the development of cardiac hypertrophy, and the transition of hypertrophy to cardiac failure. The recent dramatic increase in availability of genomic data has included information on the genetic modulation of reactive oxygen species and the antioxidant systems that normally prevent damage from these radicals. Nearly simultaneously, progressively more sophisticated and powerful methods for altering the genetic complement of selected tissues and cells have permitted application of gene therapeutic methods to understand better the pathophysiology of reactive oxygen species-mediated myocardial damage and to attenuate or treat that damage. Although exciting and promising, gene therapy approaches to these common disorders are still in the experimental and developmental stages. Improved understanding of pathophysiology, better gene delivery systems, and specific gene therapeutic strategies will be needed before gene therapy of oxyradical-mediated myocardial damage becomes a clinical reality.
- Published
- 2001
- Full Text
- View/download PDF
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