Establishment and Comprehensive Characterization of a Novel Preclinical Platform of Metastatic Retinoblastoma for Therapeutic Developments.

Purpose: Although there have been improvements in the management of metastatic retinoblastoma, most patients do not survive, and all patients suffer from multiple short- and long-term treatment toxicities. Reliable and informative models to assist clinicians are needed. Thus we developed and comprehensively characterized a novel preclinical platform of primary cell cultures and xenograft models of metastatic retinoblastoma to provide insights into the molecular biology underlying metastases and to perform drug screening for the identification of hit candidates with the highest potential for clinical translation. Methods: Orbital tumor, bone marrow, cerebrospinal fluid, and lymph node tumor infiltration specimens were obtained from seven patients with metastatic retinoblastoma at diagnosis, disease progression, or relapse. Tumor specimens were engrafted in immunodeficient animals, and primary cell lines were established. Genomic, immunohistochemical/immunocytochemical, and pharmacological analysis were performed. Results: We successfully established five primary cell lines: two derived from leptomeningeal, two from orbital, and one from lymph node tumor dissemination. After the intravitreal or intraventricular inoculation of these cells, we established cell-derived xenograft models. Both primary cell lines and xenografts accurately retained the histological and genomic features of the tumors from which they were derived and faithfully recapitulated the dissemination patterns and pharmacological sensitivity observed in the matched patients. Conclusions: Ours is an innovative and thoroughly characterized preclinical platform of metastatic retinoblastoma developed for the understanding of tumor biology of this highly aggressive tumor and has the potential to identify drug candidates to treat patients who currently lack effective treatment options.

The spectrum of morphological findings in pediatric central nervous system MN1-fusion-positive neuroepithelial tumors.

PURPOSE: Central nervous system high-grade neuroepithelial tumor with MN1 alteration (CNS-HGNET-MN1) is a rare entity defined by its DNA methylation pattern and pathologically considered to be high-grade with mixed patterns, stromal hyalinization, and with astrocytic differentiation. Our aim was to present six pediatric cases to contribute to the characterization of this group of tumors. MATERIAL AND METHODS: Six female patients aged 4 to 12 years with CNS tumors with MN1 alteration identified using genome-wide methylation arrays and/or RT-PCR were included. Clinicopathological, morphological, immunohistochemical, and molecular findings were analyzed. RESULTS: Tumor location was the parietal lobe in four and the intramedullary spinal cord in two. Two were morphologically diagnosed as ependymomas, one as gliofibroma, one as a HGNET-MN1 altered and the other two were difficult to classify. All were well-defined tumors, with a cystic component in three. Only two tumors had extensive stromal hyalinization, three had pseudopapillary formations, and four had other patterns. Multinucleated, clear, and rhabdoid cells were present. Necrosis and histiocyte clusters were also observed. Proliferative index was >10 in four. GFAP, EMA, CK, and SYN were variable, while Olig2 staining was mostly positive. Four of six patients with supratentorial tumors and complete resections were alive and tumor free after 2 to 10 years of follow-up. The two cases with medullary involvement and incomplete resections were alive and undergoing treatment 2 years after surgery. CONCLUSION: Neuroepithelial-MN1 tumors are challenging and suspicion requires molecular confirmation. Our pediatric data contribute to the knowledge for accurate diagnosis. Although further studies with a larger number of cases should be conducted in order to draw more robust conclusions regarding clinico-pathological features, here we present valuable pediatric data to increase the knowledge that may lead to the accurate management of this group of tumors.

Tractional retinal detachment and juxtapapillary retinal capillary hemangioma in a 6-year-old girl: A case report.

A 6-year-old girl with visual impairment in the right eye (OD) was referred for an eye evaluation. The fundus of the OD showed a fibrotic orange endophytic lesion located adjacent to the optic disc. In retinal optical coherence tomography, a local tractional retinal detachment and choroidal neovascular membrane were observed together also with the presence of subretinal fluid. Due to the vision of the OD evolved to nonlight perception in the following exam, enucleation was performed. The pathology report was correlated with hemangioblastoma. Herein, we describe a case of a young girl with a retinal hemangioblastoma with quick evolution and without prior systemic diagnosis.

A high-risk retinoblastoma subtype with stemness features, dedifferentiated cone states and neuronal/ganglion cell gene expression.

Retinoblastoma is the most frequent intraocular malignancy in children, originating from a maturing cone precursor in the developing retina. Little is known on the molecular basis underlying the biological and clinical behavior of this cancer. Here, using multi-omics data, we demonstrate the existence of two retinoblastoma subtypes. Subtype 1, of earlier onset, includes most of the heritable forms. It harbors few genetic alterations other than the initiating RB1 inactivation and corresponds to differentiated tumors expressing mature cone markers. By contrast, subtype 2 tumors harbor frequent recurrent genetic alterations including MYCN-amplification. They express markers of less differentiated cone together with neuronal/ganglion cell markers with marked inter- and intra-tumor heterogeneity. The cone dedifferentiation in subtype 2 is associated with stemness features including low immune and interferon response, E2F and MYC/MYCN activation and a higher propensity for metastasis. The recognition of these two subtypes, one maintaining a cone-differentiated state, and the other, more aggressive, associated with cone dedifferentiation and expression of neuronal markers, opens up important biological and clinical perspectives for retinoblastomas.

Recurrent Somatic Chromosomal Abnormalities in Relapsed Extraocular Retinoblastoma.

Most reports about copy number alterations (CNA) in retinoblastoma relate to patients with intraocular disease and features of children with extraocular relapse remain unknown, so we aimed to describe the CNA in this population. We evaluated 23 patients and 27 specimens from 4 centers. Seventeen cases had extraocular relapse after initial enucleation and six cases after an initial preservation attempt. We performed an analysis of CNA and BCOR gene alteration by SNP array (Single Nucleotide Polymorfism array), whole-exome sequencing, IMPACT panel and CGH array (Array-based comparative genomic hybridization). All cases presented CNA at a higher prevalence than those reported in previously published studies for intraocular cases. CNA previously reported for intraocular retinoblastoma were found at a high frequency in our cohort: gains in 1q (69.5%), 2p (60.9%) and 6p (86.9%), and 16q loss (78.2%). Other, previously less-recognized, CNA were found including loss of 11q (34.8%), gain of 17q (56.5%), loss of 19q (30.4%) and BCOR alterations were present in 72.7% of our cases. A high number of CNA including 11q deletions, 17q gains, 19q loss, and BCOR alterations, are more common in extraocular retinoblastoma. Identification of these features may be correlated with a more aggressive tumor warranting consideration for patient management.

Clinical, Genomic, and Pharmacological Study of MYCN-Amplified RB1 Wild-Type Metastatic Retinoblastoma.

An uncommon subgroup of unilateral retinoblastomas with highly aggressive histological features, lacking aberrations in RB1 gene with high-level amplification of MYCN (MCYNamplRB1+/+) has only been described as intra-ocular cases treated with initial enucleation. Here, we present a comprehensive clinical, genomic, and pharmacological analysis of two cases of MCYNamplRB1+/+ with orbital and cervical lymph node involvement, but no central nervous system spread, rapidly progressing to fatal disease due to chemoresistance. Both patients showed in common MYCN high amplification and chromosome 16q and 17p loss. A somatic mutation in TP53, in homozygosis by LOH, and high chromosomal instability leading to aneuploidy was identified in the primary ocular tumor and sites of dissemination of one patient. High-throughput pharmacological screening was performed in a primary cell line derived from the lymph node dissemination of one case. This cell line showed resistance to broad spectrum chemotherapy consistent with the patient's poor response but sensitivity to the synergistic effects of panobinostat-bortezomib and carboplatin-panobinostat associations. From these cells we established a cell line derived xenograft model that closely recapitulated the tumor dissemination pattern of the patient and served to evaluate whether triple chemotherapy significantly prolonged survival of the animals. We report novel genomic alterations in two cases of metastatic MCYNamplRB1+/+ that may be associated with chemotherapy resistance and in vitro/in vivo models that serve as basis for tailoring therapy in these cases.

Bridging the treatment gap in infant medulloblastoma: molecularly informed outcomes of a globally feasible regimen.

BACKGROUND: Infant medulloblastoma represents an enormous challenge in neuro-oncology, due to their simultaneous high-risk of recurrence and high risk of severe neurodevelopmental sequelae with craniospinal irradiation. Currently infant medulloblastoma are treated with intensified protocols, either comprising intraventricular methotrexate or autologous transplant, both of which carry significant morbidity and are not feasible in the majority of the world. We sought to evaluate the molecular predictors of outcome in a cohort of infants homogeneously treated with induction chemotherapy, focal radiation and maintenance chemotherapy. METHODS: In a retrospective analysis, 29 young children treated with a craniospinal irradiation sparing strategy from Hospital Garrahan in Buenos Aires were profiled using Illumina HumanMethylationEPIC arrays, and correlated with survival. RESULTS: Twenty-nine children (range, 0.3-4.6 y) were identified, comprising 17 sonic hedgehog (SHH), 10 Group 3/4, and 2 non-medulloblastomas. Progression-free survival (PFS) across the entire cohort was 0.704 (95% CI: 0.551-0.899). Analysis by t-distributed stochastic neighbor embedding revealed 3 predominant groups, SHHß, SHHγ, and Group 3. Survival by subtype was highly prognostic with SHHγ having an excellent 5-year PFS of 100% (95% CI: 0.633-1) and SHHß having a PFS of 0.56 (95% CI: 0.42-1). Group 3 had a PFS of 0.50 (95% CI: 0.25-1). Assessment of neurocognitive outcome was performed in 11 patients; the majority of survivors fell within the low average to mild intellectual disability, with a median IQ of 73.5. CONCLUSIONS: We report a globally feasible and effective strategy avoiding craniospinal radiation in the treatment of infant medulloblastoma, including a robust molecular correlation along with neurocognitive outcomes.

Melanotic neuroectodermal tumor of infancy arising from the skull: report of an unusual case, review of the literature, and a diagnostic approach.

INTRODUCTION: Melanotic neuroectodermal tumor of infancy is a rare neoplasm mainly seen in children under 1 year of life. The most common location of the tumor is the maxilla followed by the cranial vault. Surgery is the treatment of choice and outcome mainly depends on extent of resection. OBJECTIVES: To report an atypical case of an 8-year-old patient with a melanotic neuroectodermal tumor of infancy, to review the cases with melanotic neuroectodermal tumor of infancy arising from the skull published over the last 13 years, and to provide a diagnostic approach that may allow recognition of a pattern in these rare neoplastic lesions. METHODS: A case is reported with a description of the clinical, radiological, surgical, and histopathological features. Additionally, the literature was reviewed to identify reports of patients with melanotic neuroectodermal tumor of infancy arising from the cranial vault and all cases published in PubMed over the last 13 years were included. Only studies that evaluated clinical, radiological, surgical, and histopathological findings were included. CONCLUSION: Melanotic neuroectodermal tumor of infancy is a rare entity that may present with unusual features, but nevertheless has an identifiable pattern that allows the tumor to be considered in the differential diagnosis of intracranial space-occupying lesions in children.

Impact of temporomandibular disorders on oral health-related quality of life of institutionalized elderly people / Impacto da disfunção temporomandibular na qualidade de vida relacionada à saúde bucal em idosos institucionalizados

Objective: to assess the characteristics associated withtemporomandibular disorders (TMD) and their impacton oral health-related quality of life (OHRQoL) of institutionalizedelderly people. Subjects and method: itwas a cross-sectional study performed in 30 private institutionsfor the elderly. The data collected included theTMD questionnaire and clinical evaluation assessed bythe Research Diagnostic Criteria/TemporomandibularDisorders (RDC/TMD), as well as the OHIP-49 questionnairefor OHRQoL. Results: twenty-four institutionsaccepted to participate in the study (n = 375 with 133fit for answering both questionnaires). Considering thecharacteristics, tinnitus was the most frequent symptomreported, while 98.5% of the research participants didnot present myofascial pain. Disc displacement wasobserved in 26.3%, while 93% of individuals did notpresent chronic pain. Severe depression was found in23.3% of the elderly with 24.1% presenting moderatelevels of non-specific physical symptoms, includingpain. Poisson regression analysis showed that the characteristicsreported were associated with a negativeimpact on OHRQoL. Conclusion: the domain-specificanalysis showed that all domains were affected negativelyby TMD characteristics and higher levels of depressionare associated with a negative impact on OHRQoL. (AU)

Objetivo: avaliar as características associadas às disfunções temporomandibulares (DTM) e seu impacto na qualidade de vida relacionada a saúde bucal (OHRQoL) em idosos institucionalizados. Sujeitos e método: este foi um estudo transversal, realizado em trinta instituições privadas de idosos. Os dados coletados incluíram um questionário e uma avaliação clínica de DTM por meio do instrumento Research Diagnostic Criteria/ Temporomandibular Disorders (RDC/TMD) e do questionário OHIP 49 para OHRQoL. Resultados: vinte e quatro instituições autorizaram a realização do estudo, (n = 375 com 133 idosos aptos a responder ambos os questionários). Dentre as características, o agravo mais frequentemente relatado foi a ocorrência de zumbido no ouvido, enquanto 98,5% dos participantes da pesquisa não apresentaram dor miofascial. Deslocamento de disco foi observado em 26,3%, enquanto 93% dos indivíduos não apresentaram dor crônica. Depressão severa foi encontrada em 23,3% dos idosos avaliados, com 24,1% apresentando grau moderado de sintomas físicos não específicos, incluindo dor. Análise da regressão de Poisson apresentou que as características relatadas estiveram associadas a impacto negativo na qualidade de vida relacionada à saúde bucal. Conclusão: realizando a análise por domínios, todos os domínios foram negativamente afetados pelas características da DTM, e os níveis maiores de depressão estão associados a um impacto negativo na OHRQoL. (AU)

Uso de sal de iodônio em agentes resinosos para redução do tempo de fotoativação na colagem de braquetes ortodônticos: um estudo piloto / Use of iodomium salt in resin agents to reduce the photoactivation time for bonding of orthodontic brackets: a pilot study

Objetivo: Avaliar o efeito da incorporação de um sal de iodônio em cimentos resinosos experimentais em sua eficiência como agentes de fixação de braquetes ao esmalte. Materiais e Métodos: Uma mistura comonomérica de dimetacrilatos foi carregada com 35% de partículas de vidro. Três agentes foram obtidos pelo acréscimo do hexafluorfosfato de ditoliliodônio nas concentrações 0 (controle), 1 (R1) ou 2mol% (R2). Braquetes de aço inoxidável foram colados na face vestibular de incisivos bovinos, sendo fotoativados com fonte de luz de lâmpada halógena com irradiância de 400mW/cm2. Os cimentos foram fotoativados com duas exposições de luz (nas faces cervical e incisal do braquete), sendo testados dois tempos de fotoativação: 5s ou 20s. O teste de cisalhamento foi realizado 10min após a colagem. Os valores de resistência de união foram calculados em MPa e os dados submetidos a ANOVA de 2 critérios e teste de Student-Newman-Keuls (5%). O Índice de Adesivo Remanescente (IAR) foi avaliado sob aumento e submetido ao teste de Kruskal- Wallis (5%). Resultados: Para o tempo 20s houve diferenças entre todos os materiais: R1 > R2 > controle, enquanto para o tempo 5s R1 e R2 foram similares entre si, e superiores ao controle. A resistência de união para o tempo 5s foi inferior a o tempo 20s para os grupos controle e R1, e similar para R2. A redução dos valores na comparação entre os tempos 20s e 5s foi de 55% (controle), 45% (R1) e 15% (R2). Não houve diferenças significativas entre os grupos nos escores IAR. Conclusão: O desenvolvimento de agentes de fixação contendo sais de iodônio pode possibilitar a colagem de braquetes utilizando tempos reduzidos de fotoativação.(AU)

Aim: To evaluate the impact of adding an iodonium salt to experimental resin cements on their efficiency as agents for bonding brackets to enamel. Materials and Methods: A dimethacrylate co-monomer blend was loaded with 35% glass particles. Three agents were obtained by adding concentrations of 0 (control), 1 (R1) or 2mol% (R2) of ditolyliodonium hexafluorophosphate. Stainless steel brackets were bonded to the buccal faces of bovine incisors; photoactivation was carried out using a quartz-tungsten-halogen unit with 400mW/ cm2-irradiance. The cements were photoactivated using two exposures (on the cervical and incisal faces of the bracket), testing two photoactivation times: 5s or 20s. The shear test was carried out 10min after bonding. Bond strength values were calculated in MPa, and the data were submitted to a two-way ANOVA and Student-Newman-Keuls' test (5%). The Adhesive Remnant Index (ARI) was evaluated under magnification and submitted to the Kruskal-Wallis test (5%). Results: For the time 20s, significant differences among all materials were observed: R1 > R2 > control, whereas for the time 5s, R1 and R2 were similar, showing higher bond strength than the control. The bond strength for the time 5s was lower than for the time 20s for control and R1, but similar for R2. The reduction in bond strength values comparing the times 20s and 5s was 55% (control), 45% (R1) and 15% (R2). Conclusion: The development of bonding agents containing iodonium salts may produce a favorable atmosphere for bonding brackets using reduced photoactivation times.(AU)

HTLV type 1 genetic types among native descendants in Argentina.

The province of San Salvador de Jujuy, located in the northwest of Argentina, is a highly endemic area for HTLV-1 infection and a foci of tropical spastic paraparesis/HTLV-1-associated myelopathy (HAM/TSP). Therefore, to better understand this, we carried out a genetic characterization of a large set of HTLV-1 strains (n = 65) of descendants of Amerindians from this region. The LTR and env regions were analyzed. The genetic analysis showed that all of these new HTLV-1 isolates from Argentina belong to the Transcontinental subgroup A of the HTLV-1a Cosmopolitan subtype, with the exception of three isolates that cluster within the Japanese subgroup B. Interestingly, the majority of the sequences from Jujuy province belonged to a distinct cluster within the Latin America Transcontinental subgroup, referred to here as the Jujuy subcluster, and were characterized by specific signatures in the LTR. Given that the samples analyzed in this study belong to the Amerindian population and the high prevalence of HTLV-1 in Jujuy in contrast to the low prevalence of this virus in the country, it could be that HTLV-1aA was spread in Argentina from the Amerindians to the cosmopolitan population. Moreover, this is the first report of an HTLV-1aB or Japanese subgroup in descendants of non-Japanese people in South America.