Volatolomics to Decrypt the Monophagous Nature of a Rice Pest, Scirpophaga Incertulas (Walker).

Scirpophaga incertulas Walker (Lepidoptera: Crambidae, yellow stem borer, YSB) is a monophagous insect pest that causes significant yield loss in rice (Oryza staiva L.). Semiochemical based pest management is being sought as an alternate to chemical pesticides to reduce pesticide footprints. We hypothesized differential release of volatiles from host rice and two companion non-host weeds, Echinochloa colona and Echinochloa crus-galli could be responsible for oviposition and biology of YSB and these chemicals could be used for YSB management. Number of eggs laid, and number of larvae hatched were significantly higher in rice plant as compared to weeds. YSB could only form dead hearts in rice plants. YSB significantly preferred host-plant volatiles compared to the non-host plants both in choice and no-choice tests in an Y-tube olfactometer. 2-Hexenal, hexanal, 2,4-hexadienal, benzaldehyde, nonanal, methyl salicylate and decanal were found in the leaf volatolomes of both the host and non-host plants in HS-SPME-GC-MS (Headspace-Solid phase micro extraction-Gas chromatography-Mass spectrometer). Pentene-3-one, 2-pentyl furan, 2,4-heptadienal, 2-octenal, 2-octenol and menthol were present only in the non-host plants. Fourteen rice unique compounds were also detected. The built-in PCA (Principal Component Analysis) and PLS-DA (Partial least squares-discriminant analysis) analysis in the MS-DIAL tool showed that the volatiles emitted from TN1 formed a cluster distinct from Echinochloa spp. and 2-octenal was identified as a unique compound. Olfactometer bioassays using synthetic compounds showed that rice unique compounds, like xylene, hexanal served as attractants whereas non-host unique compounds, like 2-pentylfuran, 2-octenal acted as repellent. The results indicate that the rice unique compounds xylene, hexanal along with other volatile compounds could be responsible for higher preference of YSB towards rice plants. Similarly, the non-host unique compounds 2-pentylfuran, 2-octenal could possibly be responsible for lower preference and defence against YSB. These compounds could be utilised for devising traps for YSB monitoring and management.

Functional response of an egg parasitoid, Trichogramma chilonis Ishii to sublethal imidacloprid exposure.

BACKGROUND: The effectiveness of a biological control agent depends on how well it can control pests and how compatible it is with pesticides. Therefore, we reported the multigenerational effect of a commonly used insecticide, imidacloprid, on the functional response of a widely acclaimed egg parasitoid, Trichogramma chilonis Ishii, to different densities of the host Corcyra cephalonica Stainton eggs. The study investigated the outcomes of the median lethal concentration (LC50 ) and sublethal concentrations (LC5 , LC30 ), along with control treatments for five continuous generations (F1 to F5 ). RESULTS: The results showed that the F5 generation of LC30 , both of the F1 and F5 generations of LC50 , and the control all had a Type II functional response. A Type I functional response was exhibited for the F1 generation of LC30 and both generations of LC5 . The attack rate on host eggs treated with LC5 and LC30 did not change (decrease) with the shift in the type of functional response as compared to the control. A significant increase in the searching efficiency (a) was observed in the later generation (F5 ) under the exposure of LC5 and LC30 imidacloprid concentrations. A lower handling time (Th ) in both generations of the LC5 followed by LC30 treated individuals was observed when compared with the control and LC50 treatments. The per capita parasitization efficiency (1/Th ) and the rate of parasitization per handling time (a/Th ) were also considerably higher in both the generations of LC5 and LC30 than in the control and LC50 , thereby implying positive effects of imidacloprid on the parasitization potential of T. chilonis. CONCLUSION: Altogether, these multigenerational outcomes on the functional response of T. chilonis could be leveraged to control the intractable lepidopteran pests under the mild exposure of imidacloprid in integrated pest management (IPM) programs as well as in the mass rearing of the parasitoid, T. chilonis. © 2023 Society of Chemical Industry.

Sublethal phosphine fumigation induces transgenerational hormesis in a factitious host, Corcyra cephalonica.

BACKGROUND: The rice moth, Corcyra cephalonica (Stainton) (Lepidoptera: Pyralidae) is a pest of stored grains and widely used as a factitious host during the mass rearing of several natural enemies of crop pests. Hormesis is well-documented in pest insects, to some extent in natural enemies of pests. RESULTS: We report transgenerational stimulatory effects of the widely used fumigant, phosphine. The study reports the consequences of sublethal, low lethal and median lethal concentrations (LC5 , LC25 and LC50 ) and untreated control for two sequential generations of the species (G1 to G2 ). In this study, we investigated the life-history traits, nutrient reserves (protein, lipid and carbohydrate) and larval gut microbiome (using 16 s rRNA V3-V4 metagenomics sequencing) of C. cephalonica. Stimulatory effects were observed for various biological traits of C. cephalonica, notably adult longevity, emergence and increased egg hatchability when exposed to LC5 of phosphine. The total protein, lipid and carbohydrate contents of C. cephalonica also were found to be significantly increased by LC5 in both generations. The microbial diversity of LC5 treated larval gut was higher and found to be different from the rest of the treatments. This is the first report showing hormesis to a fumigant insecticide. CONCLUSION: Our findings increase knowledge on the interaction between hormesis, nutrient reserves and gut bacteria in C. cephalonica exposed to insecticides. Overall, the present study establishes phosphine-induced hormesis at LC5 in the host C. cephalonica, which might help improve the quality of mass rearing of various natural enemies. © 2023 Society of Chemical Industry.

Genome Organization and Comparative Evolutionary Mitochondriomics of Brown Planthopper, Nilaparvata lugens Biotype 4 Using Next Generation Sequencing (NGS).

Nilaparvata lugens is the main rice pest in India. Until now, the Indian N. lugens mitochondrial genome has not been sequenced, which is a very important basis for population genetics and phylogenetic evolution studies. An attempt was made to sequence two examples of the whole mitochondrial genome of N. lugens biotype 4 from the Indian population for the first time. The mitogenomes of N. lugens are 16,072 and 16,081 bp long with 77.50% and 77.45% A + T contents, respectively, for both of the samples. The mitochondrial genome of N. lugens contains 37 genes, including 13 protein-coding genes (PCGs) (cox1-3, atp6, atp8, nad1-6, nad4l, and cob), 22 transfer RNA genes, and two ribosomal RNA (rrnS and rrnL) subunits genes, which are typical of metazoan mitogenomes. However, both samples of N. lugens mitogenome in the present study retained one extra copy of the trnC gene. Additionally, we also found 93 bp lengths for the atp8 gene in both of the samples, which were 60-70 bp less than that of the other sequenced mitogenomes of hemipteran insects. The phylogenetic analysis of the 19 delphacids mitogenome dataset yielded two identical topologies when rooted with Ugyops sp. in one clade, and the remaining species formed another clade with P. maidis and M. muiri being sisters to the remaining species. Further, the genus Nilaparvata formed a separate subclade with the other genera (Sogatella, Laodelphax, Changeondelphax, and Unkanodes) of Delphacidae. Additionally, the relationship among the biotypes of N. lugens was recovered as the present study samples (biotype-4) were separated from the three biotypes reported earlier. The present study provides the reference mitogenome for N. lugens biotype 4 that may be utilized for biotype differentiation and molecular-aspect-based future studies of N. lugens.

Comparative Proteomic Analysis of Rhizoctonia solani Isolates Identifies the Differentially Expressed Proteins with Roles in Virulence.

Sheath blight of rice is a destructive disease that could be calamitous to rice cultivation. The significant objective of this study is to contemplate the proteomic analysis of the high virulent and less virulent isolate of Rhizoctonia solani using a quantitative LC-MS/MS-based proteomic approach to identify the differentially expressed proteins promoting higher virulence. Across several rice-growing regions in Odisha, Eastern India, 58 Rhizoctonia isolates were obtained. All the isolates varied in their pathogenicity. The isolate RS15 was found to be the most virulent and RS22 was identified as the least virulent. The PCR amplification confirmed that the RS15 and RS22 belonged to the Rhizoctonia subgroup of AG1-IA with a specific primer. The proteomic information generated has been deposited in the PRIDE database with PXD023430. The virulent isolate consisted of 48 differentially abundant proteins, out of which 27 proteins had higher abundance, while 21 proteins had lower abundance. The analyzed proteins acquired functionality in fungal development, sporulation, morphology, pathogenicity, detoxification, antifungal activity, essential metabolism and transcriptional activities, protein biosynthesis, glycolysis, phosphorylation and catalytic activities in fungi. A Quantitative Real-Time PCR (qRT-PCR) was used to validate changes in differentially expressed proteins at the mRNA level for selected genes. The abundances of proteins and transcripts were positively correlated. This study provides the role of the proteome in the pathogenicity of R. solani AG1-IA in rice and underpins the mechanism behind the pathogen's virulence in causing sheath blight disease.

Residue dynamics and bio-efficacy of triflumezopyrim against Nilaparvata lugens and non-targeted effect on natural enemies in a rice ecosystem.

Triflumezopyrim (TMP), a mesoionic insecticide, is commonly used for controlling planthoppers in rice. However, the relationship between the TMP residue and toxicity against brown planthoppers (BPHs) has not been studied in detail. We are reporting the dissipation of TMP from rice plant and soil under field conditions. The median lethal dose and median lethal concentration were 0.036 ng per insect and 0.525 mg L-1, respectively. TMP at recommended dose (25 g a.i. ha-1) recorded 1.25 live BPH per hill as against 25.5 per hill in control at 14 days after treatment. TMP was considered to be harmless to the natural enemies, namely, Cyrtorhinus lividipennis and Lycosa pseudoannulata in the rice ecosystem. The residue of TMP from rice plant and soil was estimated using the QuEChERS method using three different doses (12.5, 25, and 50 g a.i. ha-1). The limit of quantitation (LOQ) of TMP in plant and soil was 5 µg kg-1 and 1 µg kg-1, respectively. The maximum content of TMP in soil was less than 1% that of plant content on day 1. The dissipation pattern of TMP both from plant and soil was better explained by the first-order double-exponential decay model (FODED) as compared to the first-order kinetic model. Overall, the half-lives of TMP were ranged from 2.21 to 3.02 days in plant tissues and 3.78 to 4.79 days in soil as per the FODED model. Based on the persistence and toxicity of TMP, we could conclude that TMP will be effective against BPH up to 7-10 days after application. Triflumezopyrim with reasonable persistence and high efficacy could be recommended as an alternate pesticide in BPH management in rice.

Performance of Trichogramma japonicum under field conditions as a function of the factitious host species used for mass rearing.

Different factitious hosts were used to mass rear Trichogramma japonicum Ashmead in different parts of the globe because thorough details were lacking in both the laboratory and the field. The objective of this study was to compare, parasitoid, T. japonicum reared in different factitious hosts. Three commonly used factitious host eggs, Corcyra cephalonica (Stainton), Ephestia kuehniella Zeller and Sitotroga cerealella Olivier were tested under laboratory conditions and then in the field over a yellow stem borer, Scirpophaga incertulus (Walker) of rice. The highest parasitism by T. japonicum was observed on E. kuehniella eggs. The parasitoid's highest emergence (88.99%) was observed on S. cerealella eggs at 24 h exposure, whereas at 48 h it was on E. kuehniella eggs (94.66%). Trichogramma japonicum females that emerged from E. kuehniella eggs were significantly long-lived. The days of oviposition by hosts and the host species were significant individually, but not their interaction. Higher proportions of flying T. japonicum were observed when reared on E. kuehniella and C. cephalonica eggs. Field results showed that T. japonicum mass-reared on E. kuehniella showed higher parasitism of its natural host, S. incertulus eggs. Hence, by considering these biological characteristics and field results, E. kuehniella could be leveraged for the mass rearing of quality parasitoids of T. japonicum in India, the Asian continent and beyond.

Green silver nano-particles: synthesis using rice leaf extract, characterization, efficacy, and non-target effects.

Green synthesis of silver nano-particles (AgNPs) from silver nitrate was carried out using purple-colored rice leaves' extracts containing higher phenols, anthocyanins, and flavonoids. The efficacy of synthesized AgNPs was tested against rice diseases and investigation was carried out to check negative effect of AgNPs on soil microbes. Substantial reduction of total anthocyanins, total phenols, and total flavonoids was observed in reaction mixture during AgNP formation indicating the role of secondary metabolites on AgNP formation and stabilization. Scanning electron microscopy coupled with energy-dispersive spectroscopic images and FTIR spectral analysis of AgNPs confirmed the presence of elemental silver encapped by biomolecules. The optimized reaction parameters for synthesis of AgNPs from silver nitrate were (a) 48 h of incubation, (b) 9:1 (v/v) 1 mM AgNO3:plant extract, and (c) room temperature at 20-30 °C. Zeta potential and hydrodynamic particle sizes of synthesized AgNPs were ranged between - 16.61 to - 29.45 mV and 36-107 nm, respectively, at different time of incubation. AgNPs could control effectively Rhizoctonia solani and Xanthomonas oryzae pv. Oryzae and Helminthosporium oryzae. AgNPs at higher concentration could cause negative effect on microbial biomass carbon and soil enzymes for distant future. But the negative effects of AgNP solution (10% of 1 mM AgNPs) were comparable to commercial fungicide, carbendazim. The synthesized AgNPs with desirable characters were effective against a number of disease-causing pathogens in rice, and it can be recommended as broad-spectrum pesticide.

Crizotinib and erlotinib inhibits growth of c-Met+/EGFRvIII+ primary human glioblastoma xenografts.

OBJECTIVES: Receptor tyrosine kinases (RTK), such as c-Met and epidermal growth factor receptor (EGFR), are implicated in the malignant progression of glioblastoma. Studies show that RTK systems can co-modulate distinct and overlapping oncogenic downstream signaling pathways. EGFRvIII, a constitutively activated EGFR deletion mutant variant, leads to increased tumor growth and diminishes the tumor growth response to HGF: c-Met pathway inhibitor therapy. Conversely, activation of the c-Met pathway diminishes the tumor growth response to EGFR pathway inhibitors. Previously we reported that EGFRvIII and c-Met pathway inhibitors synergize to inhibit tumor growth in isogenic GBM cell lines engineered to express EGFRvIII. More recently, studies suggest that despite targeting RTK signaling in glioblastoma multiforme, a subpopulation of stem-like tumor-propagating cells can persist to replenish the tumor cell population leading to tumor recurrence. PATIENTS AND METHODS: Mayo 39 and Mayo 59 xenograft lines were cultured and xenografts were maintained. Subcutaneous xenograft lines were serially passaged in nude mice to generate subcutaneous xenografts. Xenografts were implanted in 6-8 week old nude mice. Once tumors reached a substantial size (150 mm3), mice were randomly divided into 4 groups: 1) control vehicle, 2) Crizotinib (crizo), 3) Erlotinib (erlot), or 4) Crizotinib + Erlotinib, (n = 5 per group). RESULTS: Crizotinib (c-Met pathway inhibitor) and Erlotinib (EGFR pathway inhibitor) in combination significantly inhibited tumor growth, phospho-EGFRvIII, phospho-Met, phospho-AKT, phospho-MAPK, and neurosphere growth in Mayo 39 and Mayo 59 primary GBM subcutaneous xenografts. The expression of the stem cell markers Nestin, Musashi, Olig 2 and Sox2 were also significantly down-regulated by c-Met inhibition, but no additive down-regulation was seen by co-treatment with Erlotinib. CONCLUSIONS: These results are consistent with and corroborate our previous findings demonstrating that targeting these two parallel pathways with c-Met and EGFR inhibitor therapy provides substantial anti-tumor activity in glioblastoma models.

Non-target effect of continuous application of chlorpyrifos on soil microbes, nematodes and its persistence under sub-humid tropical rice-rice cropping system.

Application of pesticide in agricultural fields is "unnecessary evil" for non-target microflora and fauna. Hence, to identify the safer pesticide molecules against non-target microbes, a long-term pesticide experiment was initiated at National Rice Research Institute, Cuttack, India. In the present study, the effect of continuous application of chlorpyrifos (0.5kgha-1) in rice fields on non-target groups of soil microbes and nematodes was studied for seven seasons (four wet and three dry seasons) during 2009-2013. Treatments were arranged in a randomized complete block design with four replications of chlorpyrifos-treated (0.5kg a.i. ha-1) (CT) and untreated control (UT) plots. During seven seasons of experimentation, regular application of chlorpyrifos had no significant effect on population of heterotrophic aerobic, anaerobic, oligotrophic and copiotrophic bacteria in CT compared to UT, whereas, population of asymbiotic aerobic nitrogen fixer, nitrifiers, denitrifiers, gram positive and spore-forming bacteria were significantly reduced by nearly 0.25-2 fold in CT than UT. However, comparatively less deviation in population of actinomycetes, fungi, phosphate solubilizing and sulfur oxidizing bacteria were observed in CT than UT. Significant interactions were found between effects of chlorpyrifos with time in population dynamics of microbes. In plant parasitic nematode species, Meloidogyne graminicola (RRKN) and Hirschmanniella spp. (RRN), were significantly lower (p<0.01) in CT compared to UT after first year onwards. The overall observation of five years data indicated that the RRKN population showed a decreasing trend (R2=0.644) whereas RRN showed increasing trend (R2=0.932) in CT. The drastic chlorpyrifos dissipation was noticed after 15 days of application from the initial residue of 0.25mgkg-1 soil, which indicated that chlorpyrifos residue in rice field soil was not persistent and its half-life was found to be 4.02 days. Overall, the present findings revealed that non-target effect of repetitive application of chloropyrifos (0.5kgha-1) on soil microbes and nematodes was found less under rice-rice cropping system.

Identification of Global DNA Methylation Signatures in Glioblastoma-Derived Cancer Stem Cells.

Glioblastoma (GBM) is the most common and most aggressive primary brain tumor in adults. The existence of a small population of stem-like tumor cells that efficiently propagate tumors and resist cytotoxic therapy is one proposed mechanism leading to the resilient behavior of tumor cells and poor prognosis. In this study, we performed an in-depth analysis of the DNA methylation landscape in GBM-derived cancer stem cells (GSCs). Parallel comparisons of primary tumors and GSC lines derived from these tumors with normal controls (a neural stem cell (NSC) line and normal brain tissue) identified groups of hyper- and hypomethylated genes that display a trend of either increasing or decreasing methylation levels in the order of controls, primary GBMs, and their counterpart GSC lines, respectively. Interestingly, concurrent promoter hypermethylation and gene body hypomethylation were observed in a subset of genes including MGMT, AJAP1 and PTPRN2. These unique DNA methylation signatures were also found in primary GBM-derived xenograft tumors indicating that they are not tissue culture-related epigenetic changes. Integration of GSC-specific epigenetic signatures with gene expression analysis further identified candidate tumor suppressor genes that are frequently down-regulated in GBMs such as SPINT2, NEFM and PENK. Forced re-expression of SPINT2 reduced glioma cell proliferative capacity, anchorage independent growth, cell motility, and tumor sphere formation in vitro. The results from this study demonstrate that GSCs possess unique epigenetic signatures that may play important roles in the pathogenesis of GBM.

In Vivo c-Met Pathway Inhibition Depletes Human Glioma Xenografts of Tumor-Propagating Stem-Like Cells.

Solid malignancies contain sphere-forming stem-like cells that are particularly efficient in propagating tumors. Identifying agents that target these cells will advance the development of more effective therapies. Recent converging evidence shows that c-Met expression marks tumor-initiating stem-like cells and that c-Met signaling drives human glioblastoma multiforme (GBM) cell stemness in vitro. However, the degree to which tumor-propagating stem-like cells depend on c-Met signaling in histologically complex cancers remains unknown. We examined the effects of in vivo c-Met pathway inhibitor therapy on tumor-propagating stem-like cells in human GBM xenografts. Animals bearing pre-established tumor xenografts expressing activated c-Met were treated with either neutralizing anti- hepatocyte growth factor (HGF) monoclonal antibody L2G7 or with the c-Met kinase inhibitor PF2341066 (Crizotinib). c-Met pathway inhibition inhibited tumor growth, depleted tumors of sphere-forming cells, and inhibited tumor expression of stem cell markers CD133, Sox2, Nanog, and Musashi. Withdrawing c-Met pathway inhibitor therapy resulted in a substantial rebound in stem cell marker expression concurrent with tumor recurrence. Cells derived from xenografts treated with anti-HGF in vivo were depleted of tumor-propagating potential as determined by in vivo serial dilution tumor-propagating assay. Furthermore, daughter xenografts that did form were 12-fold smaller than controls. These findings show that stem-like tumor-initiating cells are dynamically regulated by c-Met signaling in vivo and that c-Met pathway inhibitors can deplete tumors of their tumor-propagating stem-like cells.

Regulation of glioblastoma multiforme stem-like cells by inhibitor of DNA binding proteins and oligodendroglial lineage-associated transcription factors.

Tumor-initiating stem cells (also referred to as cancer stem cells, CSCs) are a subpopulation of cancer cells that play unique roles in tumor propagation, therapeutic resistance and tumor recurrence. It is increasingly important to understand how molecular signaling regulates the self-renewal and differentiation of CSCs. Basic helix-loop-helix (bHLH) transcription factors are critical for the differentiation of normal stem cells, yet their roles in neoplastic stem cells are not well understood. In glioblastoma neurosphere cultures that contain cancer stem cells (GBM-CSCs), the bHLH family member inhibitors of DNA binding protein 2 and 4 (Id2 and Id4) were found to be upregulated during the differentiation of GBM-CSCs in response to histone deacetylase inhibitors. In this study, we examined the functions of Id2 and Id4 in GBM neurosphere cells and identified Id proteins as efficient differentiation regulators of GBM-CSCs. Overexpression of Id2 and Id4 promoted the lineage-specific differentiation of GBM neurosphere cells as evidenced by the induction of neuronal/astroglial differentiation markers Tuj1 and GFAP and the inhibition of the oligodendroglial marker GalC. Id protein overexpression also reduced both stem cell marker expression and neurosphere formation potential, a biological marker of cancer cell "stemness." We further showed that Id2 and Id4 regulated GBM neurosphere differentiation through downregulating of another bHLH family member, the oligodendroglial lineage-associated transcription factors (Olig) 1 and 2. Our results provide evidence for distinct functions of Id proteins in neoplastic stem cells, which supports Id proteins and their downstream targets as potential candidates for differentiation therapy in CSCs.

Isolation and characterization of a population of stem-like progenitor cells from an atypical meningioma.

The majority of meningiomas are benign tumors associated with favorable outcomes; however, the less common aggressive variants with unfavorable outcomes often recur and may be due to subpopulations of less-differentiated cells residing within the tumor. These subpopulations of tumor cells have tumor-initiating properties and may be isolated from heterogeneous tumors when sorted or cultured in defined medium. We report the isolation and characterization of a population of tumor-initiating cells derived from an atypical meningioma. We identify a tumor-initiating population from an atypical meningioma, termed meningioma-initiating cells (MICs). These MICs self-renew, differentiate, and can recapitulate the histological characteristics of the parental tumor when transplanted at 1000 cells into the flank regions of athymic nude mice. Immunohistochemistry reveals stem-like protein expression patterns similar to neural stem and progenitor cells (NSPCs) while genomic profiling verified the isolation of cancer cells (with defined meningioma chromosomal aberrations) from the bulk tumor. Microarray and pathway analysis identifies biochemical processes and gene networks related to aberrant cell cycle progression, particularly the loss of heterozygosity of tumor suppressor genes CDKN2A (p16(INK4A)), p14(ARF), and CDKN2B (p15(INK4B)). Flow cytometric analysis revealed the expression of CD44 and activated leukocyte adhesion molecule (ALCAM/CD166); these may prove to be markers able to identify this cell type. The isolation and identification of a tumor-initiating cell population capable of forming meningiomas demonstrates a useful model for understanding meningioma development. This meningioma model may be used to study the cell hierarchy of meningioma tumorogenesis and provide increased understanding of malignant progression.

Developmental cues and persistent neurogenic potential within an in vitro neural niche.

BACKGROUND: Neurogenesis, the production of neural cell-types from neural stem cells (NSCs), occurs during development as well as within select regions of the adult brain. NSCs in the adult subependymal zone (SEZ) exist in a well-categorized niche microenvironment established by surrounding cells and their molecular products. The components of this niche maintain the NSCs and their definitive properties, including the ability to self-renew and multipotency (neuronal and glial differentiation). RESULTS: We describe a model in vitro NSC niche, derived from embryonic stem cells, that produces many of the cells and products of the developing subventricular zone (SVZ) and adult SEZ NSC niche. We demonstrate a possible role for apoptosis and for components of the extracellular matrix in the maintenance of the NSC population within our niche cultures. We characterize expression of genes relevant to NSC self-renewal and the process of neurogenesis and compare these findings to gene expression produced by an established neural-induction protocol employing retinoic acid. CONCLUSIONS: The in vitro NSC niche shows an identity that is distinct from the neurally induced embryonic cells that were used to derive it. Molecular and cellular components found in our in vitro NSC niche include NSCs, neural progeny, and ECM components and their receptors. Establishment of the in vitro NSC niche occurs in conjunction with apoptosis. Applications of this culture system range from studies of signaling events fundamental to niche formation and maintenance as well as development of unique NSC transplant platforms to treat disease or injury.

Stem cells as vectors to deliver HSV/tk gene therapy for malignant gliomas.

The prognosis of patients diagnosed with malignant gliomas including glioblastoma multiforme (GBM) is poor and there is an urgent need to develop and translate novel therapies into the clinic. Neural stem cells display remarkable tropism toward GBMs and thus may provide a platform to deliver oncolytic agents to improve survival. First we provide a brief review of clinical trials that have used intra-tumoral herpes simplex virus thymidine kinase (HSV/tk) gene therapy to treat brain tumors. Then, we review recent evidence that neural stem cells can be used to deliver HSV/tk to GBMs in animal models. While previous clinical trials used viruses or non-migratory vector-producing cells to deliver HSV/tk, the latter approaches were not effective in humans, primarily because of satellite tumor cells that escaped surgical resection and survived due to low efficiency delivery of HSV/tk. To enhance delivery of HSV/tk to kill gliomas cells, recent animal studies have focused on the ability of neural stem cells, transduced with HSV/tk, to migrate efficiently and selectively to regions occupied by GBM cells. This approach holds the promise of targeting GBM cells that have infiltrated the brain well beyond the original site of the tumor epicenter.