274 research outputs found
Increased Efficacy of Histone Methyltransferase G9a Inhibitors Against <i>MYCN</i>-Amplified Neuroblastoma.
Targeted inhibition of proteins modulating epigenetic changes is an increasingly important priority in cancer therapeutics, and many small molecule inhibitors are currently being developed. In the case of neuroblastoma (NB), a pediatric solid tumor with a paucity of intragenic mutations, epigenetic deregulation may be especially important. In this study we validate the histone methyltransferase G9a/EHMT2 as being associated with indicators of poor prognosis in NB. Immunological analysis of G9a protein shows it to be more highly expressed in NB cell-lines with MYCN amplification, which is a primary determinant of dismal outcome in NB patients. Furthermore, G9a protein in primary tumors is expressed at higher levels in poorly differentiated/undifferentiated NB, and correlates with high EZH2 expression, a known co-operative oncoprotein in NB. Our functional analyses demonstrate that siRNA-mediated G9a depletion inhibits cell growth in all NB cell lines, but, strikingly, only triggers apoptosis in NB cells with MYCN amplification, suggesting a synthetic lethal relationship between G9a and MYCN. This pattern of sensitivity is also evident when using small molecule inhibitors of G9a, UNC0638, and UNC0642. The increased efficacy of G9a inhibition in the presence of MYCN-overexpression is also demonstrated in the SHEP-21N isogenic model with tet-regulatable MYCN. Finally, using RNA sequencing, we identify several potential tumor suppressor genes that are reactivated by G9a inhibition in NB, including the CLU, FLCN, AMHR2, and AKR1C1-3. Together, our study underlines the under-appreciated role of G9a in NB, especially in MYCN-amplified tumors
Computer Aided Classification of Neuroblastoma Histological Images Using Scale Invariant Feature Transform with Feature Encoding
Neuroblastoma is the most common extracranial solid malignancy in early childhood. Optimal management of neuroblastoma depends on many factors, including histopathological classification. Although histopathology study is considered the gold standard for classification of neuroblastoma histological images, computers can help to extract many more features some of which may not be recognizable by human eyes. This paper, proposes a combination of Scale Invariant Feature Transform with feature encoding algorithm to extract highly discriminative features. Then, distinctive image features are classified by Support Vector Machine classifier into five clinically relevant classes. The advantage of our model is extracting features which are more robust to scale variation compared to the Patched Completed Local Binary Pattern and Completed Local Binary Pattern methods. We gathered a database of 1043 histologic images of neuroblastic tumours classified into five subtypes. Our approach identified features that outperformed the state-of-the-art on both our neuroblastoma dataset and a benchmark breast cancer dataset. Our method shows promise for classification of neuroblastoma histological images
LGR5 regulates pro-survival MEK/ERK and proliferative Wnt/ÎČ-catenin signalling in neuroblastoma.
LGR5 is a marker of normal and cancer stem cells in various tissues where it functions as a receptor for R-spondins and increases canonical Wnt signalling amplitude. Here we report that LGR5 is also highly expressed in a subset of high grade neuroblastomas. Neuroblastoma is a clinically heterogenous paediatric cancer comprising a high proportion of poor prognosis cases (~40%) which are frequently lethal. Unlike many cancers, Wnt pathway mutations are not apparent in neuroblastoma, although previous microarray analyses have implicated deregulated Wnt signalling in high-risk neuroblastoma. We demonstrate that LGR5 facilitates high Wnt signalling in neuroblastoma cell lines treated with Wnt3a and R-spondins, with SK-N-BE(2)-C, SK-N-NAS and SH-SY5Y cell-lines all displaying strong Wnt induction. These lines represent MYCN-amplified, NRAS and ALK mutant neuroblastoma subtypes respectively. Wnt3a/R-Spondin treatment also promoted nuclear translocation of ÎČ-catenin, increased proliferation and activation of Wnt target genes. Strikingly, short-interfering RNA mediated knockdown of LGR5 induces dramatic Wnt-independent apoptosis in all three cell-lines, accompanied by greatly diminished phosphorylation of mitogen/extracellular signal-regulated kinases (MEK1/2) and extracellular signal-regulated kinases (ERK1/2), and an increase of BimEL, an apoptosis facilitator downstream of ERK. Akt signalling is also decreased by a Rictor dependent, PDK1-independent mechanism. LGR5 expression is cell cycle regulated and LGR5 depletion triggers G1 cell-cycle arrest, increased p27 and decreased phosphorylated retinoblastoma protein. Our study therefore characterises new cancer-associated pathways regulated by LGR5, and suggest that targeting of LGR5 may be of therapeutic benefit for neuroblastomas with diverse etiologies, as well as other cancers expressing high LGR5
Correction: LGR5 regulates pro-survival MEK/ERK and proliferative Wnt/ÎČ-catenin signalling in neuroblastoma.
Present: The originally supplied Figure 5 contains duplicate total-ERK panels. Correct: The proper Figure 5 appears below. The authors sincerely apologize for this error
Spinocerebellar ataxia types 1, 2, 3, and 6: disease severity and nonataxia symptoms.
OBJECTIVE: To identify factors that determine disease severity and clinical
phenotype of the most common spinocerebellar ataxias (SCAs), we studied 526
patients with SCA1, SCA2, SCA3. or SCA6.
METHODS: To measure the severity of ataxia we used the Scale for the Assessment
and Rating of Ataxia (SARA). In addition, nonataxia symptoms were assessed with
the Inventory of Non-Ataxia Symptoms (INAS). The INAS count denotes the number of
nonataxia symptoms in each patient.
RESULTS: An analysis of covariance with SARA score as dependent variable and
repeat lengths of the expanded and normal allele, age at onset, and disease
duration as independent variables led to multivariate models that explained 60.4%
of the SARA score variance in SCA1, 45.4% in SCA2, 46.8% in SCA3, and 33.7% in
SCA6. In SCA1, SCA2, and SCA3, SARA was mainly determined by repeat length of the
expanded allele, age at onset, and disease duration. The only factors determining
the SARA score in SCA6 were age at onset and disease duration. The INAS count was
5.0 +/- 2.3 in SCA1, 4.6 +/- 2.2 in SCA2, 5.2 +/- 2.5 in SCA3, and 2.0 +/- 1.7 in
SCA6. In SCA1, SCA2, and SCA3, SARA score and disease duration were the strongest
predictors of the INAS count. In SCA6, only age at onset and disease duration had
an effect on the INAS count.
CONCLUSIONS: Our study suggests that spinocerebellar ataxia (SCA) 1, SCA2, and
SCA3 share a number of common biologic properties, whereas SCA6 is distinct in
that its phenotype is more determined by age than by disease-related factors
Protein arginine methyltransferase 5 is a key regulator of the MYCN oncoprotein in neuroblastoma cells.
Approximately half of poor prognosis neuroblastomas (NBs) are characterized by pathognomonic MYCN gene amplification and MYCN over-expression. Here we present data showing that short-interfering RNA mediated depletion of the protein arginine methyltransferase 5 (PRMT5) in cell-lines representative of NBs with MYCN gene amplification leads to greatly impaired growth and apoptosis. Growth suppression is not apparent in the MYCN-negative SH-SY5Y NB cell-line, or in two immortalized human fibroblast cell-lines. Immunoblotting of NB cell-lines shows that high PRMT5 expression is strongly associated with MYCN-amplification (P < 0.004, Mann-Whitney U-test) and immunohistochemical analysis of primary NBs reveals that whilst PRMT5 protein is ubiquitously expressed in the cytoplasm of most cells, MYCN-amplified tumours exhibit pronounced nuclear PRMT5 staining. PRMT5 knockdown in MYCN-overexpressing cells, including the SHEP-21N cell-line with inducible MYCN expression leads to a dramatic decrease in MYCN protein and MYCN-associated cell-death in SHEP-21N cells. Quantitative gene expression analysis and cycloheximide chase experiments suggest that PRMT5 regulates MYCN at a post-transcriptional level. Reciprocal co-immunoprecipitation experiments demonstrated that endogenous PRMT5 and MYCN interact in both SK-N-BE(2)C and NGP cell lines. By using liquid chromatography - tandem mass spectrometry (LC-MS/MS) analysis of immunoprecipitated MYCN protein, we identified several potential sites of arginine dimethylation on the MYCN protein. Together our studies implicate PRMT5 in a novel mode of MYCN post-translational regulation and suggest PRMT5 plays a major role in NB tumorigenesis. Small-molecule inhibitors of PRMT5 may therefore represent a novel therapeutic strategy for neuroblastoma and other cancers driven by the MYCN oncogene
Reconstructing Roma History from Genome-Wide Data
The Roma people, living throughout Europe and West Asia, are a diverse population linked by the Romani language and culture. Previous linguistic and genetic studies have suggested that the Roma migrated into Europe from South Asia about 1,000â1,500 years ago. Genetic inferences about Roma history have mostly focused on the Y chromosome and mitochondrial DNA. To explore what additional information can be learned from genome-wide data, we analyzed data from six Roma groups that we genotyped at hundreds of thousands of single nucleotide polymorphisms (SNPs). We estimate that the Roma harbor about 80% West Eurasian ancestryâderived from a combination of European and South Asian sourcesâand that the date of admixture of South Asian and European ancestry was about 850 years before present. We provide evidence for Eastern Europe being a major source of European ancestry, and North-west India being a major source of the South Asian ancestry in the Roma. By computing allele sharing as a measure of linkage disequilibrium, we estimate that the migration of Roma out of the Indian subcontinent was accompanied by a severe founder event, which appears to have been followed by a major demographic expansion after the arrival in Europe.OrszaÌgos TudomaÌnyos KutataÌsi Alapprogramok (OTKA K 103983)OrszaÌgos TudomaÌnyos KutataÌsi Alapprogramok (OTKA 73430)National Science Foundation (U.S.) (HOMINID grant 1032255)National Institutes of Health (U.S.) (grant GM100233
Early symptoms in spinocerebellar ataxia type 1, 2, 3, and 6.
Abstract: Onset of genetically determined neurodegenerative
diseases is difficult to specify because of their insidious and
slowly progressive nature. This is especially true for spinocerebellar
ataxia (SCA) because of varying affection of many
parts of the nervous system and huge variability of symptoms.
We investigated early symptoms in 287 patients with
SCA1, SCA2, SCA3, or SCA6 and calculated the influence
of CAG repeat length on age of onset depending on (1) the
definition of disease onset, (2) people defining onset, and (3)
duration of symptoms. Gait difficulty was the initial symptom
in two-thirds of patients. Double vision, dysarthria, impaired
hand writing, and episodic vertigo preceded ataxia in 4% of
patients, respectively. Frequency of other early symptoms did
not differ from controls and was regarded unspecific. Data
about disease onset varied between patients and relatives for
1 year or more in 44% of cases. Influence of repeat length
on age of onset was maximum when onset was defined as
beginning of permanent gait disturbance and cases with
symptoms for more than 10 years were excluded. Under
these conditions, CAG repeat length determined 64% of
onset variability in SCA1, 67% in SCA2, 46% in SCA3, and
41% in SCA6 demonstrating substantial influence of nonrepeat
factors on disease onset in all SCA subtypes. Identification
of these factors is of interest as potential targets for
disease modifying compounds. In this respect, recognition of
early symptoms that develop before onset of ataxia is mandatory
to determine the shift from presymptomatic to affected
status in SCA
- âŠ