Ligand-dependent Hedgehog pathway activation in Rhabdomyosarcoma : the oncogenic role of the ligands

Altres ajuts: This work was supported by grants from Institut Català d'Oncologia (ICO), Instituto de Salud Carlos III (RTICC-RD12/0036/0016, /0020, /0035, /0057; and PI14/00647), Fundació A BOSCH, Fundació Amics Joan Petit, ajuts predoctorals del VHIR and RIS3CAT grants COMRDI15-1-0014 (ACCIÓ and FEDER).Rhabdomyosarcoma (RMS) is the most common type of soft tissue sarcoma in children. The Hedgehog (HH) pathway is known to develop an oncogenic role in RMS. However, the molecular mechanism that drives activation of the pathway in RMS is not well understood. The expression of HH ligands was studied by qPCR, western blot and immunohistochemistry. Functional and animal model studies were carried out with cells transduced with shRNAs against HH ligands or treated with HH-specific inhibitors (Vismodegib and MEDI-5304). Finally, the molecular characterisation of an off-target effect of Vismodegib was also made. The results showed a prominent expression of HH ligands supporting an autocrine ligand-dependent activation of the pathway. A comparison of pharmacologic Smoothened inhibition (Vismodegib) and HH ligand blocking (MEDI-5304) is also provided. Interestingly, a first description of pernicious off-target effect of Vismodegib is also reported. The clarification of the HH pathway activation mechanism in RMS opens a door for targeted therapies against HH ligands as a possible alternative in the future development of better treatment protocols. Moreover, the description of a pernicious off-target effect of Vismodegib, via unfolded protein response activation, may mechanistically explain its previously reported inefficiency in several ligand-dependent cancers

INCREASED LEVELS OF INTERLEUKIN-10 EXPRESSION COMPARED TO INTERLEUKIN-6 IN LEUKOCYTES OF HEALTHY SUBJECTS. COULD IT BE USEFUL IN THE FUTURE OF THE DEPRESSION DIAGNOSIS?

It is a well known, that the physical activity promotes mental health. Physically active people relatively rarely suffer from psychosomatic and depressive disorders. It is possible that the differences in gene activity in peripheral blood leukocytes may be associated with depression, especially genes participating in inflammatory response. Thus, the aim of our study was to investigate the levels of IL6 and IL10 mRNA and IL10/IL6 ratio in peripheral blood leukocytes in healthy, physically active individuals. One hundred healthy young men (20-23 years old) participated in this study. All of them declared regular physical activity. Participants were non-smokers, and consumed alcohol occasionally. To access genes expression, 2 ml of venous blood was collected. RNA isolation was performed and then the relative expression of IL6 and IL10 was calculated using quantitative real-time polymerase chain reaction (Q-RT-PCR). Pro and anti-inflammatory balance was calculated as 2^relative of IL10/ 2^relative expression of IL6. Low expression of tested genes was found in healthy young men. Mean expression for IL6 was 2^0.051 (n=90) and for IL10 2^0.08 (n=98). Mean ratio IL10/IL6 was 2^1.58. Higher expression of IL10 compared to IL6 may be essential not only for the physical performance but also for the mental health. Diverse reports in the literature may be associated with choosing various control groups, i.e. sedentary or older individuals. It is possible, that measuring the expression of IL6 and IL10 (especially the ratio IL10/IL6) in peripheral blood leukocytes may be useful in the assessment of depressive disorders. Thus, molecular study of active young men can confirm the need for physical activity among people suffering from depression, but further studies are needed, particularly among people with psychosomatic disorders