Consequences of ionizing radiation-induced damage in human neural stem cells

Acharya, Munjal M.; Lan, Mary L.; Kan, Vickie H.; Patel, Neal H.; Giedzinski, Erich; Tseng, Bertrand P.; Limoli, Charles L.

doi:10.1016/j.freeradbiomed.2010.08.021

Cited by 114 publications

(105 citation statements)

References 27 publications

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“…NSCs are routinely validated by the manufacturer for high levels of expression of the multipotent markers nestin and Sox2, and low-level expression of the pluripotent marker Oct-4, along with the ability to differentiate into multiple neuronal phenotypes and to maintain a normal karyotype after multiple passages. Under our growth conditions, these NSCs displayed abundant expression of Sox2 and nestin and retained their ability to differentiate, as described previously 1 . NSCs were grown on poly-L-ornithine (20 μg/ml) and laminin (5 μg/ml) coated tissue culture treated flasks.…”

Section: Protocolsupporting

confidence: 79%

“…Growth and preparation of human neural stem cells (NSCs) for transplantation 1. The EnStem-A cell line (EMD Millipore) was used in this study.…”

Section: Protocolmentioning

confidence: 99%

“…Cells were grown in neural expansion medium (Millipore) supplemented with L-glutamine (2 mM) and basic fibroblast growth factor (bFGF; 20 ng/ml). Adherent monolayers of NSCs were passaged every other day (1:2) with accutase as the dissociation agent 1 . For transplantation studies, NSCs were used at passages below 10.…”

Section: Protocolmentioning

confidence: 99%

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Stem Cell Transplantation Strategies for the Restoration of Cognitive Dysfunction Caused by Cranial Radiotherapy

Acharya

Roa

Bosch

et al. 2011

JoVE

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Radiotherapy often provides the only clinical recourse for those afflicted with primary or metastatic brain tumors. While beneficial, cranial irradiation can induce a progressive and debilitating decline in cognition that may, in part, be caused by the depletion of neural stem cells. Given the increased survival of patients diagnosed with brain cancer, quality of life in terms of cognitive health has become an increasing concern, especially in the absence of any satisfactory long-term treatments.To address this serious health concern we have used stem cell replacement as a strategy to combat radiation-induced cognitive decline. Our model utilizes athymic nude rats subjected to cranial irradiation. The ionizing radiation is delivered as either whole brain or as a highly focused beam to the hippocampus via linear accelerator (LINAC) based stereotaxic radiosurgery. Two days following irradiation, human neural stem cells (hNSCs) were stereotaxically transplanted into the hippocampus. Rats were then assessed for changes in cognition, grafted cell survival and for the expression of differentiation-specific markers 1 and 4-months after irradiation. Our cognitive testing paradigms have demonstrated that animals engrafted with hNSCs exhibit significant improvements in cognitive function. Unbiased stereology reveals significant survival (10-40%) of the engrafted cells at 1 and 4-months after transplantation, dependent on the amount and type of cells grafted. Engrafted cells migrate extensively, differentiate along glial and neuronal lineages, and express a range of immature and mature phenotypic markers.Our data demonstrate direct cognitive benefits derived from engrafted human stem cells, suggesting that this procedure may one day afford a promising strategy for the long-term functional restoration of cognition in individuals subjected to cranial radiotherapy. To promote the dissemination of the critical procedures necessary to replicate and extend our studies, we have provided written and visual documentation of several key steps in our experimental plan, with an emphasis on stereotaxic radiosurgey and transplantation. Video LinkThe video component of this article can be found at https://www.jove.com/video/3107/ ProtocolOur experimental plan is schematically diagrammed in Figure 1. Growth and preparation of human neural stem cells (NSCs) for transplantation1. The EnStem-A cell line (EMD Millipore) was used in this study. NSCs are routinely validated by the manufacturer for high levels of expression of the multipotent markers nestin and Sox2, and low-level expression of the pluripotent marker Oct-4, along with the ability to differentiate into multiple neuronal phenotypes and to maintain a normal karyotype after multiple passages. Under our growth conditions, these NSCs displayed abundant expression of Sox2 and nestin and retained their ability to differentiate, as described previously 1 . NSCs were grown on poly-L-ornithine (20 μg/ml) and laminin (5 μg/ml) coated tissue culture treated flasks. Cells were grown ...

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Section: Protocolsupporting

confidence: 79%

“…Growth and preparation of human neural stem cells (NSCs) for transplantation 1. The EnStem-A cell line (EMD Millipore) was used in this study.…”

Section: Protocolmentioning

confidence: 99%

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Stem Cell Transplantation Strategies for the Restoration of Cognitive Dysfunction Caused by Cranial Radiotherapy

Acharya

Roa

Bosch

et al. 2011

JoVE

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“…The use of hNSCs was approved by the Institutional Human Stem Cell Research Oversight Committee. The validation, expansion, and characterization of hNSCs (ENStem-A; EMD Millipore) followed previously published procedures (4,28). MV were isolated and purified from conditioned hNSC culture medium by ultracentrifugation (29) and characterized using a NanoSight 3000 (Malvern Instruments).…”

Section: Methodsmentioning

confidence: 99%

Cranial grafting of stem cell-derived microvesicles improves cognition and reduces neuropathology in the irradiated brain

Baulch

Acharya

Allen

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Cancer survivors face a variety of challenges as they cope with disease recurrence and a myriad of normal tissue complications brought on by radio-and chemotherapeutic treatment regimens. For patients subjected to cranial irradiation for the control of CNS malignancy, progressive and debilitating cognitive dysfunction remains a pressing unmet medical need. Although this problem has been recognized for decades, few if any satisfactory long-term solutions exist to resolve this serious unintended side effect of radiotherapy. Past work from our laboratory has demonstrated the neurocognitive benefits of human neural stem cell (hNSC) grafting in the irradiated brain, where intrahippocampal transplantation of hNSC ameliorated radiation-induced cognitive deficits. Using a similar strategy, we now provide, to our knowledge, the first evidence that cranial grafting of microvesicles secreted from hNSC affords similar neuroprotective phenotypes after headonly irradiation. Cortical-and hippocampal-based deficits found 1 mo after irradiation were completely resolved in animals cranially grafted with microvesicles. Microvesicle treatment was found to attenuate neuroinflammation and preserve host neuronal morphology in distinct regions of the brain. These data suggest that the neuroprotective properties of microvesicles act through a trophic support mechanism that reduces inflammation and preserves the structural integrity of the irradiated microenvironment.radiation-induced cognitive dysfunction | microvesicles | dendritic complexity | human neural stem cells | neuroinflammation W ith improved diagnosis and treatment, cancer survivorship continues to rise but often at the cost of quality of life. The unintended neurocognitive sequelae resulting from cranial irradiation used to treat primary and secondary malignancies of the brain are both progressive and debilitating (1, 2). Despite the recognition and prevalence of these adverse side effects, relatively few, if any, long-term satisfactory solutions exist for this unmet medical need. Past work from our laboratory has optimized transplantation parameters and established many of the long-term benefits of human stem cell-based therapies for the treatment of radiationinduced cognitive dysfunction (3-5). Cranially grafted stem cells have been shown to impart persistent improvements in behavioral performance in irradiated rats over extended postirradiation intervals (1-8 mo) using short-and long-term cognitive testing paradigms (4, 6, 7). These studies have shown that our stem cell-based approaches improve the functional plasticity of the host brain through a variety of mechanisms including (i) the suppression of neuroinflammation (5), (ii) the addition of new cells to active hippocampal circuits (4), and (iii) a long-term trophic support mechanism that facilitates the expression of activity-regulated cytoskeletonassociated protein that functions in multiple ways as a molecular determinant of memory (7). Moreover, using a distinctly different injury paradigm, stem cell grafting pres...

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“…Especially, proteins involved in mitochondrial respiratory chain have been identified in our study; they produce energy and generate reactive oxygen species. Acharya et al have shown that human NSCs significantly increased their oxidative and nitrosative stresses after radiation (Acharya, et al 2010). The importance of endogenous oxygen reactive species to control NSC proliferation has been reported as well (Le Belle, et al 2010).…”

Section: Resultsmentioning

confidence: 98%

Identification of Factors Involved in Neurogenesis Recovery After Irradiation of the Adult Mouse Subventricular Zone: A Preliminary Study

Chevalier¹,

Chicheportiche²,

Daynac³

et al. 2012

Proteomics - Human Diseases and Protein Functions

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Proteomics -Human Diseases and Protein Functions 328(A) In adult mouse brain, neurons are continuously produced in two restricted regions: the subgranular zone (SGZ) of the dentate gyrus in the hippocampus and the subventricular zone (SVZ) lining the lateral ventricles (LV). The former produces neurons that functionally integrate into the granular cell layer of the hippocampus, whereas the SVZ produces neuroblasts, which migrate along the rostral migratory stream (RMS) to integrate the olfactory bulbs (OB) and differentiate into interneurons. (B) The SVZ is an highly organized neurogenic area which contains three cell types of neural cells: neural stem cells (NSCs), transit-amplifying progenitors (TAPs) and neuroblasts. Ependymal cells that contact the NSCs are organized as a single layer of epithelial cells separating the SVZ from the ventricles. (C) NSCs are defined as slow dividing cells having capacities to self-renew and to generate multiple cell types (neurons, astrocytes and oligodendrocytes). NSCs give rise to highly proliferating TAPs, which differentiate mostly in neuroblasts. NSCs are also able to produce oligodendrocytes and astrocytes according to development stages, in response to brain injury or in vitro conditions. The length of the cell cycle for NSCs and TAPs lasts 22 days and 12 h, respectively (Doetsch, et al.,1999, Morshead, et al.,1994. BV (blood vessel).

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Consequences of ionizing radiation-induced damage in human neural stem cells

Cited by 114 publications

References 27 publications

Stem Cell Transplantation Strategies for the Restoration of Cognitive Dysfunction Caused by Cranial Radiotherapy

Stem Cell Transplantation Strategies for the Restoration of Cognitive Dysfunction Caused by Cranial Radiotherapy

Cranial grafting of stem cell-derived microvesicles improves cognition and reduces neuropathology in the irradiated brain

Identification of Factors Involved in Neurogenesis Recovery After Irradiation of the Adult Mouse Subventricular Zone: A Preliminary Study

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