Polymorphism AGT2 (rs4762) is involved in the development of dermatologic events: Proof-of-concept in hepatocellular carcinoma patients treated with sorafenib.

BACKGROUND: Dermatologic adverse events (DAEs) are associated with a better outcome in patients with hepatocellular carcinoma (HCC) irrespective of the therapeutic agent received. The exact mechanisms associated with the development of DAEs are unknown although several studies point to direct toxicity of tyrosine kinase inhibitors (TKIs) to the skin or an immune-mediated reaction triggered by the oncologic treatment. As is the case in other conditions, individual genetic variants may partially explain a higher risk of DAEs. AIM: To evaluate the contribution of several gene variants to the risk of developing DAEs in HCC patients treated with TKIs. METHODS: We first analyzed 27 single-nucleotide polymorphisms (SNPs) from 12 genes selected as potential predictors of adverse event (AE) development in HCC patients treated with sorafenib [Barcelona Clinic Liver Cancer 1 (BCLC1) cohort]. Three additional cohorts were analyzed for AGT1 (rs699) and AGT2 (rs4762) polymorphisms-initially identified as predictors of DAEs: BCLC2 (n = 79), Northern Italy (n = 221) and Naples (n = 69) cohorts, respectively. The relation between SNPs and DAEs and death were assessed by univariate and multivariate Cox regression models, and presented with hazard ratios and their 95% confidence intervals (95%CI). RESULTS: The BCLC1 cohort showed that patients with arterial hypertension (AHT) (HR = 1.61; P value = 0.007) and/or AGT SNPs had an increased risk of DAEs. Thereafter, AGT2 (rs4762) AA genotype was found to be linked to a statistically significant increased probability of DAEs (HR = 5.97; P value = 0.0201, AA vs GG) in the Northern Italy cohort by multivariate analysis adjusted for BCLC stage, ECOG-PS, diabetes and AHT. The value of this genetic marker was externally validated in the cohort combining the BCLC1, BCLC2 and Naples cohorts [HR = 3.12 (95%CI: 1.2-8.14), P value = 0.0199, AGT2 (rs4762) AA vs AG genotype and HR = 2.73 (95%CI: 1.18-6.32) P value = 0.0188, AGT2 (rs4762) AA vs GG genotype]. None of the other gene variants tested were found to be associated with the risk of DAE development. CONCLUSION: DAE development in HCC patients receiving TKIs could be explained by the AGT2 (rs4762) gene variant. If validated in other anti-oncogenic treatments, it might be considered a good prognosis marker.

Mutational profile of skin lesions in hepatocellular carcinoma patients under tyrosine kinase inhibition: a repercussion of a wide-spectrum activity.

BACKGROUND/AIM: Dermatological adverse events (DAE) in hepatocellular carcinoma (HCC) patients treated with sorafenib predicts better outcome. Some turn into skin lesions (SL) requiring pathology examination. We describe incidence, characteristics and molecular profile of SL in HCC patients treated with sorafenib. MATERIALS AND METHODS: SL were prospectively collected in 311 HCC patients who started sorafenib. SL from sorafenib cohort were compared to those from a control patient group selected to match SL type and demographics. HRAS, KRAS and BRAF mutations were analyzed by CAST-PCR, mutated p53 and MAPK pathway activation by immunohistochemistry and immune infiltration by hematoxylin-eosin staining. RESULTS: Eighty-eight out of 311 patients developed DAE and 7.4% SL required histological assessment. Most frequent lesions were keratoacanthomas (n = 4), squamous-cell carcinomas (SCC)(n = 5), basal-cell carcinomas (BCC)(n = 3) and seborrheic keratosis (n = 5). HRAS and KRAS mutations were detected in 4 SL, while no mutations showed in control SL. Nuclear pERK immunostaining was identified in 33.3% of cases versus 5.3% of controls. Most SL (90%) from patients with DAE were proliferative with intense immune infiltration (73%). CONCLUSIONS: The onset of SL and their molecular profile did not impact negatively on patient's prognosis, but intense proliferation of SL may reflect compensatory activation of MAPK pathway and warrants their close monitoring.

Activated Lymphocytes and Increased Risk of Dermatologic Adverse Events during Sorafenib Therapy for Hepatocellular Carcinoma.

Advanced hepatocellular carcinoma patients treated with sorafenib who develop early dermatologic adverse events (eDAEs) have a better prognosis. This may be linked to immune mechanisms, and thus, it is relevant to assess the association between peripheral immunity and the probability of developing eDAEs. Peripheral blood mononuclear cells of 52 HCC patients treated with sorafenib were analyzed at baseline and throughout the first eight weeks of therapy. T, B, Natural Killer cells, and their immune checkpoints expression data were characterized by flow cytometry. Cytokine release and immune-suppression assays were carried out ex vivo. Cox baseline and time-dependent regression models were applied to evaluate the probability of increased risk of eDAEs. DNAM-1, PD-1, CD69, and LAG-3 in T cells, plus CD16 and LAG-3 in NK cells, are significantly associated with the probability of developing eDAEs. While NK DNAM-1+ cells express activation markers, T DNAM-1+ cells induce immune suppression and show immune exhaustion. This is the first study to report an association between immune checkpoints expression in circulating immune cells and the increased incidence of eDAEs. Our results support the hypothesis for an off-target role of sorafenib in immune modulation. We also describe a novel association between DNAM-1 and immune exhaustion in T cells.

Pharmacokinetics and pharmacogenetics of sorafenib in patients with hepatocellular carcinoma: Implications for combination trials.

BACKGROUND & AIMS: Sorafenib and lenvatinib are the first-line treatments approved in hepatocellular carcinoma (HCC), but information is lacking about the relationships between their pharmacokinetics, patients pharmacogenetic profiles, adverse events (AE) and overall survival. We aimed to elucidate these relationships of tyrosine Kinase Inhibitors, such as sorafenib, in order to improve the design of trials testing it in combination with checkpoint inhibitors. METHODS: We assessed the pharmacokinetics of sorafenib and its N-oxide metabolite at day-0, day-7, day-30, day-60, day-90, day-120, day-150 and day-180 and nine single-nucleotide polymorphisms (SNP) in five genes related to sorafenib metabolism/transport to identify the best point for starting the combination between tyrosine kinases and checkpoint inhibitors. RESULTS: We prospectively included 49 patients (96% cirrhotic, 37% hepatitis-C, 82% Child-Pugh-A and 59% BCLC-C). Pharmacokinetic values peaked at day-7 and progressively declined until day-60. In the 16 patients without further dose modifications after day-60, pharmacokinetic values remained stable through day-180 (sorafenib P = .90; N-oxide P = .93). Pharmacokinetic values were higher in patients with early dermatological adverse events and lower in patients with early diarrhoea. Sorafenib and N-oxide pharmacokinetic values varied linearly with different alleles of MRP2*3972. CONCLUSIONS: Sorafenib's pharmacokinetics is heterogeneous across HCC patients. This heterogeneity affects adverse events development and must be taken into account in setting the dose and timing of its combination with checkpoint inhibitors.

Sexualidade durante todas as fases da gravidez: experiências de gestantes / Sexualidad durante todas las fases del embarazo: experiencia de gestantes / Sexuality throughout all the stages of pregnancy: Experiences of expectant mothers

Resumo Objetivo Explorar e compreender as experiências sexuais de gestantes durante a gravidez. Métodos O estudo foi realizado em dois centros de saúde no Distrito Sanitário de Almería, sul da Espanha. Os participantes incluíram gestantes que receberam atendimento pré-natal e/ou educação para maternidade. Os critérios de inclusão foram estar grávida, manter atividade sexual e concordar em participar do estudo. Os critérios de exclusão foram ter limitações na atividade sexual por prescrição médica. A amostra foi composta por 15 gestantes selecionadas por meio de amostra de conveniência, das quais cinco participaram de grupo focal (GF) e 10 de entrevistas em profundidade (EP). Os dados foram coletados entre os meses de junho e dezembro de 2016. Os participantes foram contatados pelo pesquisador principal e foi realizada uma consulta para conduzir o GF ou EP. Resultados Três categorias principais emergiram: Falsas crenças e uma abordagem holística da sexualidade durante a gravidez, que está relacionada ao conceito de sexualidade, falsas crenças e aconselhamento sexual limitado durante a gravidez. Limitações: Do medo no início à dificuldade física no final, referindo-se às flutuações no desejo sexual, bem como às mudanças físicas que limitam a atividade sexual. Adaptação às mudanças: práticas seguras e satisfação com a imagem corporal, que engloba preocupações com os riscos e a relação entre imagem corporal e autoestima. Conclusão A falta de aconselhamento sexual durante a gravidez leva à criação de falsas crenças, que, juntamente com mudanças físicas, preocupações com o risco e flutuações no desejo e interesse sexual, provocam uma diminuição na atividade sexual. Mas a sexualidade permanece um aspecto importante da gravidez, em relação ao qual os participantes devem adotar uma abordagem mais ampla e não limitada ao ato sexual, além de adotar práticas adaptadas às mudanças físicas e emocionais que ocorrem durante esse período.

Resumen Objetivo Explorar y comprender las experiencias sexuales de gestantes durante el embarazo. Métodos Estudio realizado en dos centros de salud del Distrito Sanitario de Almería, Sur de España. Dentro de los participantes se incluyó a gestantes que recibieron atención prenatal y/o educación para la maternidad. Los criterios de inclusión fueron: estar embarazada, mantener actividad sexual y aceptar participar del estudio. Los criterios de exclusión fueron tener limitaciones de actividad sexual por prescripción médica. Muestra compuesta por 15 embarazadas seleccionadas mediante muestra de conveniencia, cinco de las cuales participaron del grupo focal (GF) y 10 de entrevistas en profundidad (EP). Datos recolectados entre junio y diciembre de 2016. Las participantes fueron contactadas por el investigador principal, realizando una consulta para incluirlas en el GF o en el EP. Resultados Surgieron tres categorías principales: Falsas creencias y un abordaje holístico de la sexualidad durante el embarazo, relacionada al concepto de sexualidad; falsas creencias y asesoramiento sexual limitado durante el embarazo. Limitaciones: Del miedo inicial a la dificultad física al final, refiriéndose a las fluctuaciones en el deseo sexual, así como a los cambios físicos limitantes de la actividad sexual. Adaptación a los cambios: prácticas seguras y satisfacción con la imagen corporal, que incluye preocupaciones con los riesgos y la relación entre imagen corporal y autoestima. Conclusión La falta de asesoramiento sexual durante el embarazo lleva a crear falsas creencias, que, conjuntamente con los cambios físicos, preocupaciones por riesgos y fluctuaciones del deseo e interés sexual, provocan una disminución de la actividad sexual. Pero la sexualidad continúa siendo un aspecto importante del embarazo, respecto del cual los participantes deben adoptar un abordaje más amplio y no limitado al acto sexual, además de adoptar prácticas adecuadas a los cambios físicos y emocionales típicos del período.

Abstract Objective To explore and understand the sexual experiences of expectant mothers during their pregnancy. Methods The study was carried out in two healthcare centers in the Almería Health District, in southern Spain. The participants included pregnant women who received prenatal care and/or maternity education. The inclusion criteria were being pregnant, maintaining sexual activity and agreeing to participate in the study. The exclusion criteria were having limitations on sexual activity by medical prescription. The sample consisted of 15 expectant women selected using a convenience sample, of which 5 took part in a focus group (FG) and 10 in in-depth interviews (IDI). Data was collected between the months of June and December 2016. Participants were contacted by the main researcher and an appointment was made to carry out the FGs or the IDIs. Results Three main categories emerged: False beliefs and a holistic approach to sexuality during pregnancy, which is related to the concept of sexuality, false beliefs, and limited sexual counseling during pregnancy. Limitations: From fear at the beginning to physical difficulty at the end, referring to the fluctuations in sexual desire as well as the physical changes that limit sexual activity. Adapting to changes: safe practices and satisfaction with one's body image, which encompasses concerns about the risks and the relationship between body image and self-esteem. Conclusion A lack of sexual counseling during pregnancy leads to the creation of false beliefs, which, together with physical changes, concerns about the risk, and fluctuations in sexual desire and interest, bring about a decrease in sexual activity. But sexuality remains an important aspect of pregnancy, toward which the participants must adopt a broader approach, not limited to intercourse, and adopt sexual practices that are adapted to the physical and emotional changes that happen during this time.

Zeb1 in Stromal Myofibroblasts Promotes Kras-Driven Development of Pancreatic Cancer.

The transcription factor Zeb1 has been identified as a crucial player in Kras-dependent oncogenesis. In pancreatic ductal adenocarcinoma (PDAC), Zeb1 is highly expressed in myofibroblasts and correlates with poor prognosis. As Kras mutations are key drivers in PDAC, we aimed here to assess the necessity of Zeb1 for Kras-driven PDAC and to define the role of Zeb1-expressing myofibroblasts in PDAC development. Genetically engineered mice with conditional pancreatic KrasG12D and Trp53 mutations (KPC) were crossed with Zeb1 haploinsufficient mice (Z+/-). Extensive PDAC was prominent in all 20-week-old KPC;Z+/+ mice, whereas only low-grade precursor lesions were detected in age-matched KPC;Z+/- littermates, with PDAC developing eventually in KPC;Z+/- aged animals. Zeb1 expression in myofibroblasts occurred early in tumorigenesis and Zeb1 haploinsufficiency retarded native expansion of stromal myofibroblasts during precursor-to-cancer progression. Zeb1 downregulation in mPSC repressed their activated gene profile, impaired their migratory and proliferative activity, and attenuated their tumor-supporting features. Conditioned media from Z+/+ mouse-activated (myofibroblast-like) pancreatic stellate cells (mPSC) boosted Ras activity in pancreatic cancer cells carrying mutant Kras; this effect was not observed when using conditioned media from Z+/- mPSC, revealing a paracrinal cooperative axis between Zeb1-expressing PSC and oncogenic Kras-bearing tumor cells. We conclude that Zeb1-expressing stromal myofibroblasts enable a heterotypic collaboration with the Kras-fated epithelial compartment, thus supporting pancreatic malignancy.Significance: Zeb1 expression in stromal myofibroblasts supports PDAC development via collaboration with the epithelial compartment bearing oncogenic Kras mutations. Cancer Res; 78(10); 2624-37. ©2018 AACR.