Harnessing gene expression profiles for the identification of ex vivo drug response genes in pediatric acute myeloid leukemia

Novel treatment strategies are of paramount importance to improve clinical outcomes in pediatric AML. Since chemotherapy is likely to remain the cornerstone of curative treatment of AML, insights in the molecular mechanisms that determine its cytotoxic effects could aid further treatment optimization. To assess which genes and pathways are implicated in tumor drug resistance, we correlated ex vivo drug response data to genome-wide gene expression profiles of 73 primary pediatric AML samples obtained at initial diagnosis. Ex vivo response of primary AML blasts towards cytarabine (Ara C), daunorubicin (DNR), etoposide (VP16), and cladribine (2-CdA) was associated with the expression of 101, 345, 206, and 599 genes, respectively (p < 0.001, FDR 0.004–0.416). Microarray based expression of multiple genes was technically validated using qRT-PCR for a selection of genes. Moreover, expression levels of BRE, HIF1A, and CLEC7A were confirmed to be significantly (p < 0.05) associated with ex vivo drug response in an independent set of 48 primary pediatric AML patients. We present unique data that addresses transcriptomic analyses of the mechanisms underlying ex vivo drug response of primary tumor samples. Our data suggest that distinct gene expression profiles are associated with ex vivo drug response, and may confer a priori drug resistance in leukemic cells. The described associations represent a fundament for the development of interventions to overcome drug resistance in AML, and maximize the benefits of current chemotherapy for sensitive patients

Transcriptomics-Based Phenotypic Screening Supports Drug Discovery in Human Glioblastoma Cells.

We have used three established human glioblastoma (GBM) cell lines-U87MG, A172, and T98G-as cellular systems to examine the plasticity of the drug-induced GBM cell phenotype, focusing on two clinical drugs, the phosphodiesterase PDE10A inhibitor Mardepodect and the multi-kinase inhibitor Regorafenib, using genome-wide drug-induced gene expression (DIGEX) to examine the drug response. Both drugs upregulate genes encoding specific growth factors, transcription factors, cellular signaling molecules, and cell surface proteins, while downregulating a broad range of targetable cell cycle and apoptosis-associated genes. A few upregulated genes encode therapeutic targets already addressed by FDA approved drugs, but the majority encode targets for which there are no approved drugs. Amongst the latter, we identify many novel druggable targets that could qualify for chemistry-led drug discovery campaigns. We also observe several highly upregulated transmembrane proteins suitable for combined drug, immunotherapy, and RNA vaccine approaches. DIGEX is a powerful way of visualizing the complex drug response networks emerging during GBM drug treatment, defining a phenotypic landscape which offers many new diagnostic and therapeutic opportunities. Nevertheless, the extreme heterogeneity we observe within drug-treated cells using this technique suggests that effective pan-GBM drug treatment will remain a significant challenge for many years to come

Síntese, caracterização e avaliação citotóxica de análogos tiossubstituídos do geraniol em células tumorais da linhagem U87MG

The cancer is a serious public health problem defined by a set of more than 100 diseases associated with disordered cell growth, which affects various organs and tissues of the body. This disease is responsible for about nine million deaths annually worldw ide and, according to estimates, this number will rise to 11.5 million in 2030. Of this mortality rate, 70% is concentrated in developing countries, such as the Brazil. Therefore, in view of the high number of deaths caused annually by this disease, it bec omes extremely important to develop new alternatives for the treatment of cancer. In this sense, the healing potential of medicinal plants, more specifically the substances found in the essential oils that are extracted from them, have shown to be good pos sibilities in the development of drugs for diseases in general. Among these essential oils, a compound called geraniol showed promising results in recent studies, in vitro and in vivo, evaluating the antineoplastic potential. Among the various types of can cer, glioblastoma multiforme (GBM) is considered one of the rarest and most lethal types, which belongs to the category of central nervous system (CNS) tumors. In general, patients diagnosed with this disease have a very low life expectancy. A significant challenge related to GBM lies in the scarcity of therapeutic options available, since only one drug has been approved by the Food and Drug Administration (FDA) and is used for the treatment of this disease. Considering the demand for new therapeutic altern atives in the treatment of cancer, especially in the case of glioblastoma, and motivated by the promising results obtained with geraniol, an attempt was made to develop derivatives with better properties, aiming at their improved effectiveness in inhibitin g neoplastic cells. Therefore, this work describes the synthesis, characterization and evaluation of the cytotoxic activity of 11 thiosubstituted analogues of this monoterpene against glioblastoma tumor cells of the U87MG cell line. Of the 11 derivatives s ynthesized so far, only 9 were evaluated for cytotoxicity. The compounds were obtained in low to moderate yields ranging from 10 to 69%. The derivatives were characterized by Infrared (FT IR), 13 C and 1 H NMR, DEPT 135 and high resolution mass spectrometry ESI HRMS (ElectroSpray Ionization High Resolution Mass Spectrometry). The evaluation of the cytotoxic activity of the 9 analogues was carried out in relation to cells of the U87MG lineage. IC 50 values were compared to geraniol and Temozolomide, the standard drug used in the treatment of GBM. Notably, compound 3e stood out as the most active among all evaluated compounds, demonstrating an IC 50 of 25 μ M, being at least four times more effective th an the monoterpenoid geraniol and approximately twenty times more potent than the reference drug used in the treatment of Glioblastoma U87MG. Conversely, compounds 3c, 3d, 3dsub, 3esub, and 5c yielded IC 50 values ranging from 57,8 to 86,6 μ M, whereas the r emaining compounds exhibited values exceeding 100 μ M.O câncer é um grave problema de saúde pública definido por um conjunto de mais de 100 doenças associadas ao crescimento desordenado das células, que afeta diversos órgãos e tecidos do corpo. Essa doença é responsável por cerca de nove milhões de óbitos anualmente em todo o mundo e, segundo estimativas, esse número se elevará para 11,5 milhões em 2030. Dessa taxa de mortalidade apresentada, 70% está concentrada nos países em desenvolvimento, como o Brasil. Portanto, em vista do elevado número de óbitos causados anualmente por este mal, torna-se de suma importância o desenvolvimento de novas alternativas para o tratamento do câncer. Nesse sentido, o potencial curativo de plantas medicinais, mais especificamente das substâncias encontradas nos óleos essenciais que são extraídos delas, demonstraram-se boas possibilidades no desenvolvimento de fármacos para enfermidades em geral. Dentre esses óleos essenciais, um composto denominado geraniol apresentou resultados promissores em estudos recentes, in vitro e in vivo, de avaliação do potencial antineoplásico. Dentre os diversos tipos de câncer, o glioblastoma multiforme (GBM) é considerado um dos tipos mais raros e letais, sendo este pertencente à categoria dos tumores do sistema nervoso central (SNC). Em geral, os pacientes diagnosticados com essa doença possuem uma expectativa de vida muito baixa. Um desafio significativo relacionado ao GBM reside na escassez de opções terapêuticas disponíveis, uma vez que apenas um fármaco foi aprovado pelo Food and Drug Administration (FDA) e são utilizados para o tratamento dessa enfermidade. Considerando a demanda por novas alternativas terapêuticas no tratamento do câncer, especialmente no caso do glioblastoma, e motivados pelos resultados promissores obtidos com o geraniol, buscou-se desenvolver derivados com melhores propriedades, visando sua eficácia aprimorada na inibição de células neoplásicas. Sendo assim, esse trabalho descreve a síntese, caracterização e avaliação da atividade citotóxica de 11 análogos tiossubstituídos desse monoterpeno contra células tumorais de glioblastoma da linhagem celular U87MG. Dentre os 12 derivados sintetizados até o momento, realizou-se a avaliação citotóxica de apenas 9. Os compostos foram obtidos em rendimentos baixos a moderados que variaram de 10 a 69%. Os derivados foram caracterizados por Infravermelho (FT-IR), RMN de 13C e de 1H, DEPT-135 e espectrometria de massas de alta resolução ESI-HRMS (ElectroSpray Ionization – High Resolution Mass Spectrometry). A avaliação da atividade citotóxica dos 9 análogos foi realizada em células da linhagem U87MG. Os valores de IC50 foram comparados ao geraniol e a Temozolomida, fármaco padrão utilizado no tratamento do GBM. Notavelmente, o composto 3e se destacou como o mais ativo entre todos os compostos avaliados, demonstrando um IC50 de 25 μM, sendo, pelo menos, quatro vezes mais eficaz do que o monoterpeno geraniol e aproximadamente vinte vezes mais potente do que o fármaco de referência utilizado no tratamento do Glioblastoma U87MG. Por outro lado, os compostos 3c, 3d, 3dsub, 3esub e 5c obtiveram valores de IC50 variando de 57,8 a 86,6 μM, enquanto os demais compostos exibiram valores superiores a 100 μM

Novel Treatment Strategies for Glioblastoma

This book is a compilation of articles that brings together current knowledge from an international team of contributors who are dedicated investigators exploring novel strategies for the treatment of glioblastoma. These articles describe some of the latest concepts that will provide students, researchers and clinicians with an overview of the therapeutic approaches being developed in the field of neuro-oncology to combat this deadly disease

Sterol biosynthesis pathway is part of the interferon host defence response

Recently, cholesterol metabolism has been shown to modulate the infection of several viruses and there is growing evidence that inflammatory response to infection also modulates lipid metabolism. However little is known about the role of inflammatory processes in modulating lipid metabolism and their consequences for the viral infection. This study investigates host-lipid viral interaction pathways using mouse cytomegalovirus, a large double-stranded DNA genome, which represents one of the few models for a natural infection of its natural host. In this study, transcriptomic and lipidomic profiling of macrophages shows that there is a specific coordinated regulation of the sterol pathways upon viral infection or treatment with IFNγ or β (but not TNFα, IL1β or IL6) resulting in the decrease of free cellular cholesterol. Furthermore, we show that pharmacological and RNAi inhibition of the sterol pathway augments protection against infection in vitro and in vivo and we identified that the prenylation branch of the sterol metabolic network was involved in the protective response. Finally, we show that genetic knock out of IFNβ results in a partial reduction while genetic knock out of Ifnar1 completely abolishes the reduction of the sterol biosynthetic activity upon infection. Overall these results support a role for part of the sterol metabolic network in protective immunity and show that type 1 IFN signalling is both necessary and sufficient for reducing the sterol metabolic network upon infection; thereby linking the sterol pathway with IFN defence responses