Search

PDF - 60K, Linear correlation observed between tumor growth inhibition (% T/C) or tumor regression and the fraction of time over the in vivo S473P-Akt IC80 in different cancer cell line-derived tumor xenografts implanted in nude mice (represented as dots) and nude rats (represented as triangles) following NVP-BYL719 treatment (R2=0.77, p

Somatic PIK3CA mutations are frequently found in solid tumors, raising the hypothesis that selective inhibition of PI3Kα may have robust efficacy in PIK3CA-mutant cancers while sparing patients the side-effects associated with broader inhibition of the class I phosphoinositide 3-kinase (PI3K) family. Here, we report the biologic properties of the 2-aminothiazole derivative NVP-BYL719, a selective inhibitor of PI3Kα and its most common oncogenic mutant forms. The compound selectivity combined with excellent drug-like properties translates to dose- and time-dependent inhibition of PI3Kα signaling in vivo, resulting in robust therapeutic efficacy and tolerability in PIK3CA-dependent tumors. Novel targeted therapeutics such as NVP-BYL719, designed to modulate aberrant functions elicited by cancer-specific genetic alterations upon which the disease depends, require well-defined patient stratification strategies in order to maximize their therapeutic impact and benefit for the patients. Here, we also describe the application of the Cancer Cell Line Encyclopedia as a preclinical platform to refine the patient stratification strategy for NVP-BYL719 and found that PIK3CA mutation was the foremost positive predictor of sensitivity while revealing additional positive and negative associations such as PIK3CA amplification and PTEN mutation, respectively. These patient selection determinants are being assayed in the ongoing NVP-BYL719 clinical trials. Mol Cancer Ther; 13(5); 1117–29. ©2014 AACR.

PDF - 129K, Legends for Supplementary Figures 1 through 4 and Supplementary Tables 1 through 7. Supplementary Table 1. Determination of NVP-BYL719 in vitro effects on human protein kinases. Supplementary Table 2. Activity profile of NVP-BYL719 in the Invitrogen Kinase panel. Supplementary Table 3. Activity profile of NVP-BYL719 in the Ambit Kinase panel. Supplementary Table 4. NVP-BYL719 Kd (nmol/L) determination in the Ambit Kinase panel for the hits found to be inhibited >90%. Supplementary Table 5. NVP-BYL719 anti-tumor effects in diverse cancer cell line-derived xenograft models. Supplementary Table 6. CCLE cell lines responsive to NVP-BYL719. Supplementary Table 7 Anti-tumor effect of NVP-BYL719 in patient-derived xenograft models carrying PIK3CA genetic alterations.

PDF - 71K, Rat1-myr-p110alpha (blue), beta (green) or delta (red) cells were treated with increasing concentrations of NVP-BYL719 for 30 minutes. Levels of S473P-Akt in cell extracts were quantified by Reverse Phase Protein Array as described in (18) and plotted as percentage of untreated control cells. The graph illustrates n=3 independent experiments. IC50s (plus/minus) SD and IC80s (plus/minus) SD of n=3 independent experiments are reported..

PDF - 92K, NVP-BYL719 does not inhibit mTOR and PIKKs involved in DNA damage-repair processes. A. TSC1 -/- MEFs cells were grown in a 96-well format and treated for 1 h with increased concentrations of RAD001 or NVP-BYL719 (from 0.5 nmol/L to 10 ?mol/L in 1 third dilution steps) and immediately fixed. S235/236P-RPS6 levels were measured and IC50 determined with the Excel module XLfit. Background (no primary Ab incubated); BL, Baseline. B: TSC1 -/- MEFs cells were treated with increasing concentrations of NVP-BYL719 as indicated or RAD001 at 500 nmol/L or an equivalent DMSO concentration for 30 minutes. Levels of S235/236P-RPS6 and total RPS6 in protein- normalized lysates were detected by Western blot analyzis using an activation-state specific antibody, followed by incubation with species- specific HRP-labeled secondary antibody and signal development by ECL. C: 24 h post seeding, A549 cells were treated at the same time with Actinomycin D (Act D) at a concentration of 5 ?mol/L (an agent used to induce DNA damage), and with increasing concentrations of NVP-BYL719 as indicated or with the vehicle control (DMSO) for 1 h. Levels of S15P-p53 and tubulin in protein-normalized lysates were detected by Western blot analysis using an activation-state specific antibody, followed by incubation with species- specific HRP-labeled secondary antibody and signal development by ECL. D: 24 h post seeding, U2OS cells were pre-treated for 1 h with increased concentrations of NVP-BYL719 or KU55933 a specific small molecular mass inhibitor of ATM (Supplementary reference 1) at a concentration of 10 ?mol/L or with the vehicle control (DMSO). The cells were then irradiated with 15 Gy and re-incubated at 37 degrees C for 1 h and then lysed. Levels of S1981P-ATM in protein- normalized lysates were detected by Western blot analysis using an activation-state specific antibody, followed by incubation with species- specific HRP-labeled secondary antibody and signal development by ECL.

PDF - 37K, NVP-BYL719 was tested at 10 different concentrations (from 0.5 nmol/L to 10 μmol/L in 1 third dilution steps) against CLK2 (A) and LRRK2 (B). SelectScreenTM Kinase Profiling Service uses XLfit from IDBS to determine the IC50 values (IC50 CLK2=621 nmol/L; IC50 LRRK2=3'220 nmol/L). The dose response curve is curve fit to model number 205 (sigmoidal dose-response model).

Purpose: The myeloproliferative neoplasm myelofibrosis is characterized by frequent deregulation of Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling, and JAK inhibitors were shown to reduce splenomegaly and ameliorate disease-related symptoms. However, the mutant clone and bone marrow fibrosis persist in the majority of patients. Using preclinical models, we explored whether JAK and pan-deacetylase inhibitor combination yielded additional benefits.Experimental Design: The combination of the JAK1/2 inhibitor ruxolitinib and panobinostat was investigated using two different mouse models of JAK2V617F-driven disease. A Ba/F3 JAK2V617F cell–driven leukemic disease model was used to identify tolerated and efficacious doses. The drugs were then evaluated alone and in combination in a mouse model of myeloproliferative neoplasm–like disease based on transplantation of bone marrow transduced with a retrovirus expressing JAK2V617F. Exposures were determined in blood and tissues, and phosphorylated STAT5 and acetylated histone H3 pharmacodynamic readouts were assessed in spleen and bone marrow. Histologic analysis was conducted on spleen and bone marrow, including staining of reticulin fibers in the latter organ.Results: The combination of ruxolitinib and panobinostat was found to have a more profound effect on splenomegaly, as well as on bone marrow and spleen histology, compared with either agent alone, and the analysis of pharmacodynamic readouts showed that ruxolitinib and panobinostat have nonoverlapping and complementary effects.Conclusion: Combining JAK1/2 and pan-deacetylase inhibitors was fairly well tolerated and resulted in improved efficacy in mouse models of JAK2V617F-driven disease compared with the single agents. Thus, the combination of ruxolitinib and panobinostat may represent a promising novel therapeutic modality for myeloproliferative neoplasms. Clin Cancer Res; 19(22); 6230–41. ©2013 AACR.

PDF file 1093K, Supplementary Table S1. Activity and tolerability of ruxolitinib (RUX) and panobinostat (PAN), alone and in combination, in a mouse model of Ba/F3 JAK2V617F cells-driven leukemic disease. Supplementary Table S2. Levels of panobinostat (PAN) and ruxolitinib (RUX) in blood and tissues post-final dose in a mouse model of Ba/F3 JAK2V617F cells-driven leukemic disease. Supplementary Table S3. Exposure to panobinostat (PAN) and ruxolitinib (RUX) in blood and tissues in a mouse model of Ba/F3 JAK2V617F cells-driven leukemic disease. Supplementary Table S4. Levels of panobinostat (PAN) and ruxolitinib (RUX) in blood and tissues post-final dose in a mouse model of JAK2V617F-driven MPN-like disease. Supplementary Table S5. Exposure to panobinostat (PAN) and ruxolitinib (RUX) in blood and tissues in a mouse model of JAK2V617F-driven MPN-like disease. Supplementary Figure S1. Tolerability of ruxolitinib (RUX) and panobinostat (PAN), alone and in combination, in a mouse model of Ba/F3 JAK2V617F cellsdriven leukemic disease. Supplementary Figure S2. Modulation of aberrant JAK2/STAT5 signaling in vitro and in vivo following treatment with ruxolitinib (RUX) and panobinostat (PAN), alone and in combination. Supplementary Figure S3. Efficacy and tolerability of ruxolitinib (RUX) in a mouse model of JAK2V617F-driven MPN-like disease. Supplementary Figure S4. Modulation of STAT5 phosphorylation in spleen and bone marrow following treatment with ruxolitinib (RUX) in a mouse model of JAK2V617F-driven MPN-like disease. Supplementary Figure S5. Histological analysis following treatment with ruxolitinib (RUX) in a mouse model of JAK2V617F-driven MPN-like disease. Supplementary Figure S6. Efficacy and tolerability of panobinostat (PAN) in a mouse model of JAK2V617F-driven MPN-like disease Supplementary Figure S7. Modulation of protein acetylation and acetylated histone H3 in spleen and bone marrow, respectively, following treatment with 21 panobinostat (PAN) in a mouse model of JAK2V617F-driven MPN-like disease. Supplementary Figure S8. Histological analysis following treatment with panobinostat (PAN) in a mouse model of JAK2V617F-driven MPN-like disease. Supplementary Figure S9. Efficacy and tolerability of ruxolitinib (RUX) and panobinostat (PAN), alone and in combination, in a mouse model of JAK2V617Fdriven MPN-like disease. Supplementary Figure S10. Impact of ruxolitinib (RUX) and panobinostat (PAN), alone and in combination, on relative levels of retrovirally transduced JAK2V617F in a mouse model of JAK2V617F-driven MPN-like disease. Supplementary Figure S11. Histological analysis following treatment with ruxolitinib (RUX) and panobinostat (PAN), alone and in combination, in a mouse model of JAK2V617F-driven MPN-like disease

Supplementary Figure Legends 1-3 from Effects of the Dual Phosphatidylinositol 3-Kinase/Mammalian Target of Rapamycin Inhibitor NVP-BEZ235 on the Tumor Vasculature: Implications for Clinical Imaging

Supplementary Figures 1-3 from Effects of the Dual Phosphatidylinositol 3-Kinase/Mammalian Target of Rapamycin Inhibitor NVP-BEZ235 on the Tumor Vasculature: Implications for Clinical Imaging

Background & aims: \ud Obesity prevalence in people with phenylketonuria (PKU) is comparable to that of the general population but the underlying aetiology remains unknown. To assess body composition, dietary intake, moderate physical activity duration (MPAD) and energy expenditure (MPAEE), resting metabolic rate (RMR), diet-induced thermogenesis (DIT), fasting and postprandial fat (FOx) and carbohydrate oxidation (CHOOx), in PKU people and healthy Controls.\ud \ud Methods: \ud Participants were PKU people (n = 16) and healthy controls (n = 15). Body composition was measured with stable isotopes using deuterium as tracer, dietary intake from 4-day food diaries, MPAD and MPAEE from 7-day activity counts measured by triaxial accelerometers, calibrated against individual rates of oxygen consumption and carbon dioxide production, RMR, DIT, FOx and CHOOx by indirect calorimetry.\ud \ud Results: \ud Body composition, DIT, FOx, CHOOx and RMR did not differ between the PKU and the Control groups. MPAD (PKU, 73 ± 26 min/week; Control, 152 ± 43 min/week) and MPAEE (PKU, 404 ± 127 kcal/week; Control, 741 ± 153 kcal/week) were lower (P < 0.05) in the PKU than the Control group. Raised phenylalanine levels were inversely related with MPAD and MPAEE. Energy intake and energy provided by protein did not differ between the groups, while energy proportion obtained from carbohydrate was higher (PKU, 60 ± 2%; Control, 51 ± 2%; P < 0.05) and from fat lower (PKU, 24 ± 2%; Control, 35 ± 3%; P < 0.05) in the PKU than in the Control group.\ud \ud Conclusion: \ud People with PKU spent less time and expend less energy in moderate physical activity and have a higher intake of energy from CHO which may be involved in the underlying mechanisms of obesity in PKU.

Purpose African Swine Fever (ASF) is a serious disease that impacts on domestic pig production. It has no approved vaccines or effective treatments. The potential costs of ASF entering Australia include the costs of eradication, costs to industry until the disease is eradicated, and long-term costs of ASF’s presence should eradication prove economically or technically infeasible. The analysis in this report estimated the costs to the economy of 2 plausible scenarios for an ASF incursion into Australia - incursions into either the feral pig population or the domestic production system - as well as the costs of ASF becoming endemic. Key findings • A small-scale outbreak of ASF in feral pigs followed by eradication of the disease was estimated to cost 101 million to 127 million Dollar. • A small-scale outbreak of ASF in domestic pigs followed by eradication of the disease was estimated to cost 117 million to 263 million Dollar. • Larger outbreaks would cost more and make eradication more difficult, particularly outbreaks in the feral population. • Endemic ASF was estimated to cost between about 0.4 billion and 2.5 billion Dollar. • The estimated cost of endemic ASF was influenced by the rate at which it spreads through the country, which in turn influences the extent to which enhanced biosecurity practices across the Australian pork supply chain and heightened surveillance activities are implemented, the frequency of spillover into the production system, and the cost of spillover events.

A scoping review covering approaches to gender affirming care being taught in undergraduate physician assistant (PA), medical doctor (MD), and doctor of osteopathy (DO) programs in the United States.

Investigations into the textures in the ironstones, relationships between Au-Bi-Cu mineralisation and associated chlorite, metal zonation and distribution and paragenesis have been made on the White Devil Deposit in the Proterozoic Tennant Creek district of the Northern Territory. The textures within the ironstone revealed that they consist of intergrown and interlocking masses of magnetite laths pseudomorphed after hematite. The consistent textures and overgrowth of primary magnetite on the laths and in fractures indicates the ironstones existed in entirety and were deformed prior to magnetite alteration. The open porous lath texture of the ironstones has implications for ore deposition. The mineralising stage was accompanied by the deposition of chlorite. The gold, bismuth sulphosalts and chalcopyrite are intergrown with coarse grained chlorite in the spaces between the magnetite laths and around magnetite stringers. The chlorite associated with ore and altered sediments ranges from low temperature green Mg-rich pycnochlorite to high temperature blue Fe-rich ripidolite. Geothermometry has shown that the chlorite has a temperature range from 230 to 330°C. Talc alteration of Mg chlorite occurred during the waning stage of the hydrothermal event. The metal zonation within the deposit is not clear, even within the orebodies studied. The initial hematite ironstone formed in chloride sediments within the shear. The fluids became more reduced and interacted with the hematite, altering it to magnetite. The mineralising stage saw the introduction of higher temperature fluids which interacted with the magnetite causing the deposition of gold and chlorite followed by bismuth and copper with associated chlorite at higher temperatures. As the fluids cooled, alteration of Mg chlorite to talc, martitisation and quartz flooding of the Pinter Ironstone occurred.