Tropomyosin Rezeptor Kinasen (Trks) – ein neues Target in der molekular zielgerichteten Tumortherapie

In der hier vorgelegten Arbeit wurde die Expression von Tropomyosin Rezeptor Kinasen (Trks) in humanen neuroendokrinen Tumoren des GastroEnteroPankreatischen (GEP) Systems und die mögliche Rolle der Trks als neues Target für eine molekular zielgerichtete Therapie von neuroendokrinen Tumoren untersucht. Aus Tumorgewebeproben von Patienten aus dem NeoExNET-Register wurde ein Tissue Microarray (TMA) generiert. In der immunhistochemischen Untersuchung des TMA zeigten 33% (11/33) der NET des Pankreas, aber nur 1-2% (1/65) der NET des Dünndarms eine Expression von Trks. Die humane neuroendokrine Tumorzelllinie GOT1 zeigte die Expression eines funktionellen TrkA Rezeptors. Am Modell der GOT1-Zelllinie konnte gezeigt werden, dass die Inkubation mit dem panTrk-Inhibitor GNF-5837 antiproliferative und Apoptose-induzierende Effekte bewirkte. Erste panTrk-Inhibitoren wie Larotrectinib [73, 74] oder Entrectinib [151, 152] sind bereits für verschiedene Karzinome mit seltenen NTRK-Genfusionen zugelassen [70-72]. Inwieweit aufgrund der in der hier vorgelegten Untersuchung berichteten immunhistochemischen Expressionsdaten von TrkA in 33% der untersuchten neuroendokrinen Tumoren des Pankreas möglicherweise auch ein größeres vorselektioniertes Patientenkollektiv mit neuroendokrinen Tumoren mit Expression von Trk von einer molekular zielgerichteten Therapie mit panTrk-Inhibitoren profitieren könnte, muss erst in weiteren präklinischen und klinischen Studien untersucht werden

Cabozantinib and Tivantinib, but Not INC280, Induce Antiproliferative and Antimigratory Effects in Human Neuroendocrine Tumor Cells in vitro: Evidence for 'Off-Target' Effects Not Mediated by c-Met Inhibition

Background/Aims: The hepatocyte growth factor/transmembrane tyrosine kinase receptor c-Met has been defined as a potential target in antitumoral treatment of various carcinomas. We aimed to investigate the direct effect of c-Met inhibition on neuroendocrine tumor cells in vitro. Methods: The effects of the multi-tyrosine kinase inhibitors cabozantinib and tivantinib and of the highly specific c-Met inhibitor INC280 were investigated in human pancreatic neuroendocrine BON1, bronchopulmonary NCI-H727 and midgut GOT1 cells in vitro. Results: INC280, cabozantinib and tivantinib inhibited c-Met phosphorylation, respectively. However, while equimolar concentrations (10 mu M) of cabozantinib and tivantinib inhibited cell viability and cell migration, INC280 had no inhibitory effect. Knockdown experiments with c-Met siRNA also did not demonstrate effects on cell viability. Cabozantinib and tivantinib caused a G2 arrest in neuroendocrine tumor cells. Conclusions: Our in vitro data suggest that c-Met inhibition alone is not sufficient to exert direct antitumoral or antimigratory effects in neuroendocrine tumor cells. The multi-tyrosine kinase inhibitors cabozantinib and tivantinib show promising antitumoral and antimigratory effects in neuroendocrine tumor cells, which are most probably 'off-target' effects, not mediated by c-Met. (C) 2015 S. Karger AG, Base

Atenção à saúde no primeiro ano de vida de uma coorte prospectiva de lactentes prematuros tardios e a termo de Botucatu, São Paulo, 2015-2017

Objective: To compare late preterm and full-term infants and investigate the association between late preterm birth and the use of reference services. Methods: Prospective cohort study, with data collected from the first to twelfth months of life. Maternal and birth characteristics were compared between term and premature live births. The effect of late preterm birth on the usage of outpatient/emergency services, hospitalizations and intensive care unit admissions were evaluated by calculating odds ratios adjusted for potential confounding factors. Results: The 41 late preterm and 540 full-term infants differed in the frequencies of low weight at birth and in non-permanence in rooming-in wards, which were both higher in late-preterm infants. This group had a higher chance of being hospitalized in a neonatal intensive care unit (OR=6.85 – CI95% 2.56;18.34). Late preterm birth was not associated with the use of other health services. Conclusion: Negative effects of late preterm delivery were only found in the neonatal stage.Objetivo: Avaliar associação entre prematuridade tardia e utilização de serviços de referência no primeiro ano de vida. Estudo de coorte prospectiva, com dados coletados no 1o, 3o, 6o, 9o e 12o meses dos lactentes. Características maternas e de nascimento foram comparadas entre nascidos a termo e prematuros tardios. Avaliou-se o efeito da prematuridade tardia sobre a utilização de ambulatório especializado e unidade de pronto-socorro/pronto atendimento, internação em unidade de terapia intensiva (UTI) e hospitalização, calculando-se razões de chances ajustadas. Resultados: Os 41 prematuros tardios e 540 nascidos a termo diferiram nas frequências de baixo peso ao nascer e não permanência em alojamento conjunto, maiores nos prematuros tardios, estes também com mais chance de internação em UTI neonatal (OR=6,85 – IC95% 2,56;18,34), condição que não se associou à utilização dos demais serviços de referência. Conclusão: Prematuridade tardia não se associou à maior utilização de serviços de referência após alta da maternidade

The HDM2 (MDM2) Inhibitor NVP-CGM097 inhibits tumor cell proliferation and shows additive effects with 5-fluorouracil on the p53 - p21 - Rb - E2F1 cascade in the p53wildtype neuroendocrine tumor cell line GOT1

Background/aims: The tumor suppressor p53 is depleted in many tumor cells by the E3 ubiquitin ligase mouse double minute 2 homolog (MDM2) through MDM2/p53 interaction. A novel target for inhibiting p53 degradation and for causing reexpression of p53wild type is inhibition of MDM2. The small molecule NVP-CGM097 is a novel MDM2 inhibitor. We investigated MDM2 inhibition as a target in neuroendocrine tumor cells in vitro. Methods: Human neuroendocrine tumor cell lines from the pancreas (BON1), lung (NCI-H727), and midgut (GOT1) were incubated with the MDM2 inhibitor NVP-CGM097 (Novartis) at concentrations from 4 to 2,500 nM. Results: While p53wild type GOT1 cells were sensitive to NVP-CGM097, p53mutated BON1 and p53mutated NCI-H727 cells were resistant to NVP-CGM097. Incubation of GOT1 cells with NVP-CGM097 at 100, 500, and 2,500 nM for 96 h caused a significant decline in cell viability to 84.9 ± 9.2% (p < 0.05), 77.4 ± 6.6% (p < 0.01), and 47.7 ± 9.2% (p < 0.01). In a Western blot analysis of GOT1 cells, NVP-CGM097 caused a dose-dependent increase in the expression of p53 and p21 tumor suppressor proteins and a decrease in phospho-Rb and E2F1. Experiments of co-incubation of NVP-CGM097 with 5-fluorouracil, temozolomide, or everolimus each showed additive antiproliferative effects in GOT1 cells. NVP-CGM097 and 5-fluorouracil increased p53 and p21 expression in an additive manner. Conclusions: MDM2 inhibition seems a promising novel therapeutic target in neuroendocrine tumors harboring p53wild type. Further investigations should examine the potential role of MDM2 inhibitors in neuroendocrine tumor treatment. Keywords: 5-Fluorouracil; Everolimus; GOT1; MDM2 inhibitor; NVP-CGM097; Neuroendocrine tumor; Temozolomide; p21; p53

Avaliação de intervenção educativa sobre aleitamento materno dirigida a agentes comunitários de saúde

OBJETIVOS: avaliar ação educativa visando à capacitação de agentes comunitários de saúde em promoção e apoio ao aleitamento materno. MÉTODOS: estudo do processo e adequação de intervenção, com comparação de conhecimentos e práticas de 54 agentes, antes e dois meses após a ação. A intervenção, quatro oficinas teórico-práticas com oito horas cada, fundamentou-se em princípios da Educação Crítico-Reflexiva e em dois cursos de capacitação de equipes de maternidade. Os resultados foram expressos pelas diferenças dos escores médios de conhecimentos, práticas e total, adotando-se p<0,05 como nível crítico. RESULTADOS: houve aumento do escore médio de conhecimentos (21,0 para 26,6; p<0,001) e de escores de conhecimentos classificados como bons (37,0% para 88,8%, p<0,001). Não houve mudanças no escore de participação dos agentes em específicas ações de saúde, como grupos. O efeito da intervenção mais apontado pelos agentes foi sentir-se mais aceito e em melhores condições de participar da equipe multiprofissional em situações envolvendo cuidados com crianças. Mais da metade relatou melhora na qualidade e quantidade das orientações prestadas nas visitas domiciliarias. CONCLUSÕES: a intervenção tem efeitos positivos sobre conhecimentos e a prática dos agentes junto às famílias, mas não promove aumento da sua participação em específicas ações de saúde.OBJECTIVES: to evaluate the educational intervention aiming to train community health visitors to promote and support maternal breastfeeding. METHODS: a study of the process of improving the intervention, comparing the knowledge and practices of 54 agents, before and two months after the training. Four theoretical and practical workshops were carried out based on the principles of Critical-Reflective Education and two training courses were run for the maternity teams. The results were expressed in terms of the differences in mean scores for knowledge, practices and the total, with statistical significance set at p<0.05. RESULTS: there was an increase in the mean score for knowledge (from 21.0 to 26.6; p<0.001) and in the scores for particular items of knowledge classified as good (from 37.0% to 88.8%, p<0.001). There were no changes in the score for participation of health visitors as groups in specific health actions. The effect of the intervention most frequently cited by the health visitors themselves was feeling more accepted and better prepared to participate in the multiprofessional team in situations involving childcare. More than half reported an improvement in the quality and quantity of the advice they were able to dispense during home visits. CONCLUSIONS: the intervention has a positive effect on the knowledge and practices of health visitors dealing with families but does not give rise to an increase in their participation in specific health actions

Tropomyosin receptor kinase: a novel target in screened neuroendocrine tumors

Tropomyosin receptor kinase (Trk) inhibitors are investigated as a novel targeted therapy in various cancers. We investigated the effects of the pan-Trk inhibitor GNF-5837 in human neuroendocrine tumor (NET) cells. The human neuroendocrine pancreatic BON1, bronchopulmonary NCI-H727 and ileal GOT1 cell lines were treated with GNF-5837 alone and in combination with everolimus. Cell viability decreased in a time- and dose-dependent manner in GOT1 cells in response to GNF-5837 treatment, while treatment in BON1 and NCI-H727 cells showed no effect on cellular viability. Trk receptor expression determined GNF-5837 sensitivity. GNF-5837 caused downregulation of PI3K-Akt-mTOR signaling, Ras-Raf-MEK-ERK signaling, the cell cycle and increased apoptotic cell death. The combinational treatment of GNF-5837 with everolimus showed a significant enhancement in inhibition of cell viability vs single substance treatments, due to a cooperative PI3K-Akt-mTOR and Ras-Raf-MEK-ERK pathway downregulation, as well as an enhanced cell cycle component downregulation. Immunohistochemical staining for Trk receptors were performed using a tissue microarray containing 107 tumor samples of gastroenteropancreatic NETs. Immunohistochemical staining with TrkA receptor and pan-Trk receptor antibodies revealed a positive staining in pancreatic NETs in 24.2% (8/33) and 33.3% (11/33), respectively. We demonstrated that the pan-Trk inhibitor GNF-5837 has promising anti-tumoral properties in human NET cell lines expressing the TrkA receptor. Immunohistochemical or molecular screening for Trk expression particularly in pancreatic NETs might serve as predictive marker for molecular targeted therapy with Trk inhibitors