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BackgroundThere is limited information about the mortality and neurodevelopmental outcomes of very preterm infants (ObjectiveTo explore the association between SIP and neurodevelopmental outcomes and mortality in very preterm infants.Data sourcesMedline, EMBASE, Cochrane Library, EMCARE and MedNar.Study selectionDatabases were searched until September 2021. Studies comparing outcomes of ‘SIP’ versus ‘no SIP or necrotising enterocolitis (NEC)’ were included.Data extractionNeurodevelopmental outcomes at ≥1 year corrected age were extracted as the main outcome measure. Data were pooled separately for adjusted and unadjusted ORs using the random-effects model. The evidence level was assessed using the GRADE (Grading of Recommendations, Assessments, Development and Evaluations) framework.ResultsEighteen cohort studies (13 606 infants) were included. Meta-analysis of unadjusted ORs showed that SIP was significantly associated with increased odds of mortality, cerebral palsy, composite outcome of death or disability, visual impairment and hearing impairment. However, pooling of adjusted ORs (aOR) found significant associations only for mortality (aOR (95% CI) 2.27 (2.07 to 2.49); I2: 0%; four studies (n=10 695)), severe disability (aOR (95% CI) 2.06 (1.38 to 3.08); I2: 0%; two studies (n=321)) and composite outcome of ‘death or disability’ (aOR (95% CI) 2.18 (1.55 to 3.06); I2: 0%; two studies (n=321)). The level of evidence was ‘low’ or ‘very low’.LimitationsLack of information on aORs from many studies.ConclusionsSIP in very preterm infants is associated with higher odds of mortality, severe disability, anddeath or disability.

ObjectiveTo evaluate whether diffuse excessive high signal intensity (DEHSI) on term equivalent age MRI (TEA-MRI) predicts disability in preterm infants.DesignThis is a systematic review and meta-analysis. Medline, EMBASE, Cochrane Library, EMCARE, Google Scholar and MedNar databases were searched in July 2019. Studies comparing developmental outcomes of isolated DEHSI on TEA-MRI versus normal TEA-MRI were included. Two reviewers independently extracted data and assessed the risk of bias. Meta-analysis was undertaken where data were available in a format suitable for pooling.Main outcome measuresNeurodevelopmental outcomes ≥1 year of corrected age based on validated tools.ResultsA total of 15 studies (n=1832) were included, of which data from 9 studies were available for meta-analysis. The pooled estimate (n=7) for sensitivity of DEHSI in predicting cognitive/mental disability was 0.58 (95% CI 0.34 to 0.79) and for specificity was 0.46 (95% CI 0.20 to 0.74). The summary area under the receiver operating characteristics (ROC) curve was low at 0.54 (CI 0.50 to 0.58). A pooled diagnostic OR (DOR) of 1 indicated that DEHSI does not discriminate preterm infants with and without mental disability. The pooled estimate (n=8) for sensitivity of DEHSI in predicting cerebral palsy (CP) was 0.57 (95% CI 0.37 to 0.75) and for specificity was 0.41 (95% CI 0.24 to 0.62). The summary area under the ROC curve was low at 0.51 (CI 0.46 to 0.55). A pooled DOR of 1 indicated that DEHSI does not discriminate between preterm infants with and without CP.ConclusionsDEHSI on TEA-MRI did not predict future development of cognitive/mental disabilities or CP.PROSPERO registration numberCRD42019130576.

ObjectiveTo explore the association between hyperglycaemia and adverse outcomes in very preterm infants.DesignSystematic review and meta-analysis. Data were pooled separately for adjusted and unadjusted odds ratios (ORs) using random-effects model. Subgroup analysis was conducted based on study design (cohort and case control).Main outcome measuresAssociation between hyperglycaemia in preterm neonates (FindingsForty-six studies (30 cohort and 16 case control) with data from 34 527 infants were included. Meta-analysis of unadjusted ORs from cohort studies found hyperglycaemia to be significantly associated with mortality, any-grade intraventricular haemorrhage (IVH), severe IVH, any-stage retinopathy of prematurity (ROP), severe ROP, sepsis, chronic lung disease and disability. However, pooling of adjusted ORs found significant associations only for mortality (adjusted OR (CI): 2.37 (1.40 to 4.01); I2: 36%; 6 studies), ‘Any grade IVH’ (adjusted OR (CI): 2.60 (1.09 to 6.20); I2: 0%; 2 studies) and ‘Any stage ROP’ (adjusted OR (CI): 3.70 (1.55 to 8.84); I2: 0%; 2 studies). Meta-regression analysis found glucose levels >10 mmol/L to be associated with increased odds of mortality compared with ConclusionHyperglycaemia in very preterm infants is associated with higher odds of mortality, any-grade IVH and any-stage ROP. A limitation was lack of availability of adjusted ORs from many of the included studies.PROSPERO registration numberCRD42020193016.

Paratuberculosis is caused by Mycobacterium avium subsp. Paratuberculosis (MAP) and is a chronic, intestinal tract infection in the ruminant sector globally. A total of 122 EDTA mixed blood samples, 121 serum and 16 pooled faecal samples were collected from farms of 4 different districts i.e. Nayagarh, Cuttack, Khordha and Angul and a blind review was conducted at the Animal Health Division, CIRG, Mathura. Microscopic examination of 16 pooled faecal samples revealed +2 reactivity to Acid-Fast Bacilli. All the serum samples were subjected to indirect ELISA. Out of them, 23 (19.01%), 85 (70.25%), shows strongly positive, positive, antibody titre respectively. EDTA blood samples of 23 ELISA-strongly positive were subjected to 413 bp IS900 PCR and 11 (9%) of them were found positive for Mycobacterium avium subsp. Paratuberculosis (MAP). MAP isolates were further subjected to genotyping using 608 bpIS1311 PCR and restriction endonuclease analysis (IS1311 PCR-REA) and 2 (1.64%) of them matched with “Indian Bison Type”. Genotyping of the isolates using IS1311 PCR-REA revealed that goat population of Odisha are primarily infected with “Indian Bison Type” strains.

Aim: An experiment was designed to evaluate the role of Vitamin E and glutathione in improving the seminal parameters during hypothermic storage of liquid semen at 4°C for 72 h. Materials and Methods: Thirty-six semen ejaculates were collected by artificial vagina from 6 bucks (Beetal) during the normal reproduction season (September to November) at weekly interval. The samples were centrifuged, and the seminal plasma was removed. The sperm pellet was diluted with Tris-based extender and divided into three groups. Group T1: Control samples without antioxidants, Group T2: Samples supplemented with tocopherol at 3 mM, and Group T3: Samples supplemented with glutathione at 1 mM. The samples were evaluated for progressive motility, percent liveability, percent abnormal spermatozoa, and acrosome integrity after liquid preservation for 0, 24, 48, and 72 h. The level of lipid peroxidation and antioxidant enzymes, namely, glutathione peroxidase (GPx) and superoxide dismutase (SOD) were estimated after liquid preservation for 0 and 72 h. Results: It was observed that, after storage of semen at 4°C up to 72 h, the progressive sperm motility, percent liveability, percent abnormal spermatozoa, and percent intact acrosomes were significantly (p

Health Insurance in India emerges in the year 1999 with the introduction of IRDA bill in the floor of Parliament. In the post-privatization era, health insurance segment developed slowly and steadily. The penetration of insurance sector in Non-Life insurance is kept on increasing since 2001. In order to portray the journey of health insurance sector in India, it is required to show the growth and development of this sector in the country. The present study is intended to evaluate the growth and development of the health insurance sector in India in the post-privatization era. Data are collected mainly from secondary sources. Such data are analyzed and represented suitable through the help of tables, diagrams and charts. From the study, it is concluded that there is a significant upward trend in the growth of health insurance industry in India both at public and private sector after privatization. If this trend continues by keeping other factors constant, then the health insurance business would touch to Rs 20000 crores in the financial year 2016-17 contributed at least 60% by public sector and rest 40% by both private insurers and standalone health insurers.

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.

Mahatma Gandhi National Rural Employment Guarantee Scheme (MGNREGS) played a very vital role by providing wage employment for one hundred days in a year to each rural household whose adult member volunteer to do unskilled manual work. It acts as a fall back measures during these lean seasons by becoming an alternative source of income for these poor household, acting as a social safety net for them. This study analyses the impact of the program on the rural development . Rural development is the overall development of rural areas with respect to food security, economic security, habitat security, health security, nutritional security etc. So, respective index are being constructed to find the impact on rural development. Randomized selection of beneficiary and non-beneficiary communities has been done. A randomisation impact evaluation design was used with households eligible and the rural development index (RDI) is constructed. It has been found that there is significant impact of the program on the rural development.

The present study focused on the development of an effective oral vaccine delivery system of poly(vinyl alcohol)-coated chitosan microparticles-based recombinant hepatitis B vaccine. Chitosan microparticles were prepared by ionotropic gelation technique; they were loaded with recombinant hepatitis B vaccine and coated with poly(vinyl alcohol). The average sizes of the microparticles were measured in the range of 100-410 nm. The optimal loading capacity and loading efficiency were recorded around 3.4 and 74%, respectively. In vitro release study shows that the prepared microparticles release the antigen in a sustained manner. Moreover, the microparticles were resistant to simulated gastric environment and release the antigen in the targeted intestinal milieu. Furthermore, oral immunisation of rats with poly(vinyl alcohol)-coated chitosan hepatitis-B microparticles vaccine shows comparable seroprotective immune response to presently practiced intramuscular vaccination. The results demonstrated that poly(vinyl alcohol)-coated chitosan microparticles have the potential for being used as an oral vaccine delivery system for hepatitis B vaccine and may be a suitable alternative for needle-based vaccination.

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.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 mmCrizotinib (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.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.

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.

Objective: RmlC (dTDP-6-deoxy-D-xylo-4-hexulose 3, 5-epimerase) is a crucial enzyme for cell wall biosynthesis in Mycobacterium tuberculosis. It’s absence in human host attest it as a valid target for drug designing. In the presented study an in-silico method is employed to find out the potential phytochemical inhibitors of RmlC . Methods: AutoDock 4.2 is used to study the binding affinity of ligands in the active site of the protein. The drug-likeness and oral toxicity evaluation is done using the online tools Molsoft and ProTox respectively. Results: Chrysophanol has binding affinity of -9.24 kcal/mole to RmlC active site. PASS tool predicts chrysophanol as antitubercular compound. Its druglikeness is 0.1, and toxicity class is 5 measured by ProTox. Hence, chrysophanol emerges as a lead molecule among the phytochemicals in the database. Conclusions: Crysophanol is the lead inhibitor against RmlC target. The lead molecule may work as a successful drug in future for tuberculosis treatment.

Lantana camara Linn. (Family: Verbenaceae), an ornamental shrub, a noxious weed grows in many tropical and subtropical parts of world. The common name of this ornamental shrub is lantana, wild/red sage, bunch berry, locally known as “Barophulnoo”. The genus Lantana (Verbenaceae) as described by Linnaeus in 1753 contained seven species, six from South America and one from Ethiopia.

Meningiomas are a common form of adult brain tumors. Although most meningiomas are benign tumors that are associated with favorable outcomes, a small group of patients develop more aggressive manifestations which currently are more difficult to treat. It has increasingly been recognized that tumor stem-like cells play critical roles in tumor recurrence, angiogenesis, and invasion in malignant brain tumors. Several recent studies identified the stem-like features of sphere-forming cells in human meningiomas. These meningioma sphere cells expressed various progenitor cell biomarkers and can undergo differentiation if appropriate stimuli are applied. Moreover, these meningioma stem-like cells are more resistant to irradiation treatment, and tumorigenic in in vivo xenograft models. These new findings could lead to a better understanding of the development and etiology of meningioma formation and suggest that meningioma stem-like cells may serve as a novel target in therapeutically resistant meningiomas.

Nippostrongylus brasiliensis is a natural parasite of rat, closely related to human hookworm. Experimental host in which the parasite will attain sexual maturity include mouse, hamster, rabbit, chinchilla and to lesser extend in a cotton rat. The parasite does not occur naturally in laboratory rodents and its primary importance is as a model of immunology, host parasite interaction and anthelmintic testing.

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. (Cancer Sci 2012; 103: 1028–1037)

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.

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.

Glioblastoma (GBM) is the most common and most aggressive brain tumor in adults. Its frequent recurrence after resection and dismal prognosis are thought to be due to a small population of stem-like tumor cells that efficiently propagate tumors and resist cytotoxic therapy. 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 GBM tumors and GSC lines derived from these tumors with normal controls (a neural stem cell (NSC) line and normal brain tissue) identified two groups hyper- and hypomethylated of 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 be found in primary GBM-derived xenograft tumors suggesting that they were 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. The comparison between the NSC line and the normal brain tissue sample leads to the identification of a group of NSC-specific differentially methylated regions (nsDMRs). Cluster analysis using nsDMRs revealed the presence of stem cell-specific DNA methylation signatures in both primary GBM and GSC lines. Higher expression of genes associated with stem cell-specific DNA methylation signatures such as DNMT3A, STAT5A, CSTB, PMEPA1 and G6PD in GBM patients was found to be associated with poor patient survival. The results from this study demonstrate the utility of using cancer stem cell models for advancing understanding of the pathobiology of gliomas. Citation Format: Eun Joon Lee, Prakash Rath, Jimei Liu, Dungsheng Ryu, Alan Free, Lirong Pei, Douglas C Anthony, Suash Sharma, Mark D Kirk, John J. Laterra, Duck Hwan Ryu, Jeong-Hyeon Choi, Huidong Shi, Douglas C. Miller, N. Scott Litofsky, Qi Feng. Identification of global DNA methylation signatures in glioblastoma-derived cancer stem cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1379. doi:10.1158/1538-7445.AM2014-1379

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.

Glioblastoma multiforme (GBM) are the most frequent and aggressive primary brain tumors in adults. Uncontrolled proliferation and abnormal cell migration are two prominent spatially and temporally disassociated characteristics of GBMs. In this study, we investigated the role of the receptor tyrosine kinase EphB2 in controlling the proliferation/migration dichotomy of GBM. We studied EphB2 gain-of-function and loss-of function in glioblastoma-derived stem-like neurospheres (GBM-SCs), whose in vivo growth pattern closely replicates human GBM. EphB2 expression stimulated GBM neurosphere cell migration and invasion, and inhibited neurosphere cell proliferation in vitro. In parallel, EphB2 silencing increased tumor cell proliferation and decreased tumor cell migration. EphB2 was found to increase tumor cell invasion in vivo using an internally controlled dual-fluorescent xenograft model. Xenografts derived from EphB2 overexpressing GBM neurospheres also showed decreased cellular proliferation. The non-receptor tyrosine kinase focal adhesion kinase (FAK) was found to be co-associated with and highly activated by EphB2 expression and FAK activation facilitated focal adhesion formation, cytoskeleton structure change and cell migration in EphB2-expression GBM neurosphere cells. Taken together, our findings indicate that EphB2 has pro-invasive and anti-proliferative actions in GBM stem-like neurospheres mediated, in part, by interactions between EphB2 receptors and FAK. These novel findings suggest that tumor cell invasion can be therapeutically targeted by inhibiting EphB2 signaling and that optimal anti-tumor responses to EphB2 targeting may require the concurrent use of anti-proliferative agents. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4311. doi:1538-7445.AM2012-4311

Hepatocyte Growth Factor (HGF) and its tyrosine kinase receptor c-Met are overexpressed in glioblastomas (GBM) and expression levels correlate with poor prognosis. Previously, we showed that anti-HGF therapy using the monoclonal antibody L2G7 inhibits the progression of orthotopic xenografts derived from HGF+/c-Met+ GBM cell lines by inducing apoptosis, and inhibiting tumor cell proliferation and angiogenesis. Increasingly, there is an interest in cancer therapies targeting the stem-like tumor-initiating pool of cells within GBMs (GBM-TICs), as this subset of cells may be particularly resistant to conventional therapies and associated with tumor recurrence. This study examined the effects of c-Met pathway inhibition on the pool of stem-like cells in GBM xenografts. We used the HGF/c-Met dependant U87 cell line as a GBM model system. Wild-type U87 cells were passaged subcutaneously (s.c.) in nude mice for 2-3 weeks. Tumors (>200mm3) were removed and primary tumor-derived cells were cultured in vitro in serum-free stem cell medium containing EGF and FGF to study neurosphere formation, a marker of stem-like cells. We confirmed the expression of stem/progenitor cell markers Sox2, CD133, Nestin, and Musashi in neurosphere-forming cells (U87-NS). Under conditions of forced differentiation, U87-NS cells revealed multipotent capacities and expressed lineage specific differentiation markers such as Neurofilament, BIII-tubulin, O1, and GFAP. Animals bearing intracranial or s.c. xenografts were treated with +/- L2G7 prior to tumor removal and isolation of tumor cells. Overall, L2G7 treated tumors were substantially smaller than control in both intracranial (p Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3309. doi:10.1158/1538-7445.AM2011-3309