DATAFILE_HAN_DINGEMANSE_2017_PRSB

DATAFILE_HAN_DINGEMANSE_2017_PRS

Additional file 1: of You are what you eat: diet shapes body composition, personality and behavioural stability

Supplementary material. Figure S1. Schematic representations of hypothesized population-level average behavioural responses and associated among- and within-individual variances across environments. Figure S2. The experimental set-up. Figure S3. The effect of diet and sex on the expression of behavioural and morphological traits. Table S1. Diet effects on variance components and repeatability of phenotypes. Table S2. Variance components (with standard errors in parentheses) for each unique combination of treatment for sex, and a range of phenotypic traits. Table S3. Additional linear mixed models to test the effect of social partner treatment on aggression and mating activity. (DOCX 479 kb

RSPB-2018-0951_Tuni et al_DATA

RSPB-2018-0951_Tuni et al_DAT

Image_1_Real-world landscape transition of death causes in the immunotherapy era for metastatic non-small cell lung cancer.jpeg

BackgroundWith approval of anti-PD-1/PD-L1, metastatic non-small cell lung cancer (NSCLC) has entered the era of immunotherapy. Since immune-related adverse events (irAEs) occur commonly in patients receiving anti-PD-1/PD-L1, the landscape of death causes may have changed in metastatic NSCLC. We aim to compare patterns of death causes in metastatic NSCLC between the pre-immunotherapy and immunotherapy era to identify the consequent landscape transition of death causes.MethodsIn this cohort study, 298,485 patients with metastatic NSCLC diagnosed between 2000 and 2018 were identified from the Surveillance, Epidemiology, and End Results Program. Unsupervised clustering with Bayesian inference method was performed for all patients’ death causes, which separated them into two death patterns: the pre-immunotherapy era group and the immunotherapy era group. Relative risk (RR) of each death cause between two groups was estimated using Poisson regression. Reduced death risk as survival time was calculated with locally weighted scatterplot smooth (Lowess) regression.ResultsTwo patterns of death causes were identified by unsupervised clustering for all patients. Thus, we separated them into two groups, the immunotherapy era (2015-2017, N=40,172) and the pre-immunotherapy era (2000-2011, N=166,321), in consideration of obscure availability to immunotherapy for patients diagnosed in 2012-2014, when the follow-up cutoff was set as three years. Although all-cause death risk had reduced (29.2%, 13.7% and 27.8% for death risks of lung cancer, non-cancer and other cancers), non-cancer deaths in the immunotherapy era (N=2,100, 5.2%; RR=1.155, 95%CI: 1.101-1.211, PConclusionsThe real-world landscape of death causes has changed in metastatic NSCLC when entering the immunotherapy era, and the increased non-cancer diseases may contribute to the changes that may be associated with commonly occurring irAEs.</p

miR-150 Deficiency Protects against FAS-Induced Acute Liver Injury in Mice through Regulation of AKT

<div><p>Although miR-150 is implicated in the regulation of immune cell differentiation and activation, it remains unknown whether miR-150 is involved in liver biology and disease. This study was performed to explore the potential role of miR-150 in LPS/D-GalN and Fas-induced liver injuries by using wild type and miR-150 knockout (KO) mice. Whereas knockout of miR-150 did not significantly alter LPS/D-GalN-induced animal death and liver injury, it protected against Fas-induced liver injury and mortality. The Jo2-induced increase in serum transaminases, apoptotic hepatocytes, PARP cleavage, as well as caspase-3/7, caspase-8, and caspase-9 activities were significantly attenuated in miR-150 KO mice. The liver tissues from Jo2-treated miR-150 KO mice expressed higher levels of Akt1, Akt2, total Akt, as well as p-Akt(Ser473) compared to the wild type livers. Pretreatment with the Akt inhibitor V reversed Jo2-induced liver injury in miR-150 KO mice. The primary hepatocytes isolated from miR-150 KO mice also showed protection against Fas-induced apoptosis <i>in vitro</i> (characterized by less prominent PARP cleavage, less nuclear fragmentation and less caspase activation) in comparison to hepatocytes from wild type mice. Luciferase reporter assays in hepatocytes transfected with the <i>Akt1</i> or <i>Akt2</i> 3’-UTR reporter constructs (with or without mutation of miR-150 binding site) established <i>Akt1</i> and <i>Akt2</i> as direct targets of <i>miR-150</i>. Tail vein injection of lentiviral particles containing pre-<i>miR-150</i> enhanced Jo2-induced liver injury in miR-150 KO mice. These findings demonstrate that miR-150 deficiency prevents Fas-induced hepatocyte apoptosis and liver injury through regulation of the Akt pathway.</p></div

Table_1_Real-world landscape transition of death causes in the immunotherapy era for metastatic non-small cell lung cancer.xlsx

BackgroundWith approval of anti-PD-1/PD-L1, metastatic non-small cell lung cancer (NSCLC) has entered the era of immunotherapy. Since immune-related adverse events (irAEs) occur commonly in patients receiving anti-PD-1/PD-L1, the landscape of death causes may have changed in metastatic NSCLC. We aim to compare patterns of death causes in metastatic NSCLC between the pre-immunotherapy and immunotherapy era to identify the consequent landscape transition of death causes.MethodsIn this cohort study, 298,485 patients with metastatic NSCLC diagnosed between 2000 and 2018 were identified from the Surveillance, Epidemiology, and End Results Program. Unsupervised clustering with Bayesian inference method was performed for all patients’ death causes, which separated them into two death patterns: the pre-immunotherapy era group and the immunotherapy era group. Relative risk (RR) of each death cause between two groups was estimated using Poisson regression. Reduced death risk as survival time was calculated with locally weighted scatterplot smooth (Lowess) regression.ResultsTwo patterns of death causes were identified by unsupervised clustering for all patients. Thus, we separated them into two groups, the immunotherapy era (2015-2017, N=40,172) and the pre-immunotherapy era (2000-2011, N=166,321), in consideration of obscure availability to immunotherapy for patients diagnosed in 2012-2014, when the follow-up cutoff was set as three years. Although all-cause death risk had reduced (29.2%, 13.7% and 27.8% for death risks of lung cancer, non-cancer and other cancers), non-cancer deaths in the immunotherapy era (N=2,100, 5.2%; RR=1.155, 95%CI: 1.101-1.211, PConclusionsThe real-world landscape of death causes has changed in metastatic NSCLC when entering the immunotherapy era, and the increased non-cancer diseases may contribute to the changes that may be associated with commonly occurring irAEs.</p

The levels of miR-150 and Akt in WT and miR-150 KO mice.

<p>(A) Mature miR-150 sequence was aligned among mouse, human, rat, dog and gorilla. (B) miR-150 expression in different tissues from WT and miR-150 KO mice. Total RNA was isolated from eight-week-old male WT and miR-150 KO mice. 1μg of total RNA was used for reverse transcription followed by qRT-PCR analysis. Relative expression of miR-150 was normalized to U6. Data are expressed as mean ± SD. (C) The levels of miR-150 and miR-221 in WT and miR-150 KO livers with or without Jo2 treatment. WT and miR-150 KO mice were intraperitoneally injected with or without 0.5 μg/g of body weight Jo2 (n = 6–8 per group). The livers were harvested 4 hours after Jo2 injection. The levels of miR-150 and miR-221 in the livers were determined by qRT-PCR (Data are expressed as mean ± SD, *<i>p</i><0.05). (D) Western blot for Akt1, Akt 2, total Akt, p-Akt(Ser473), 4E-BP1 and p-4E-BP1(Thr37/46) (with GAPDH as the loading control). The blots shown in this figure were from two individual mice for each group. Quantifications of relative protein levels are shown at the right panel (the data are expressed as mean ± SD, *<i>p</i><0.05).</p

Effect of Akt inhibitor V on Fas-induced liver injury and hepatocyte apoptosis.

<p>(A) WT and miR-150 KO mice were injected intraperitoneally with vehicle or Akt inhibitor V (1mg/kg of body weight) 30 minutes before PBS administration (n = 3 each group). Mice were sacrificed 3 hours after PBS administration. Western blot showed that Akt inhibitor V treatment inhibited the phosphorylation of Akt. (B) WT and miR-150 KO mice were injected intraperitoneally with vehicle or Akt inhibitor V (1mg/kg of body weight) 30 minutes before Jo2 (0.5μg/g body weight) administration (n = 6 each group). Mice were sacrificed 3 hours after Jo2 administration. H&E staining (100×, scale bar 20μm) of formalin-fixed, paraffin-embedded liver tissues. (C) Serum levels of ALT and AST in mice with or without Akt inhibitor V pretreatment (data are expressed as mean ± SD **<i>p</i><0.01). (D) TUNEL staining of the liver tissues from WT and miR-150 KO mice 3 hours after Jo2 injection (with or without Akt inhibitor V pretreatment). Representative photographs (100×, scale bar 20μm) are shown at the left panel; quantification of TUNEL-positive cells is shown at the right panel (the data are expressed as mean ± SD; *<i>p</i><0.05, **<i>p</i><0.01). (E) Immunostaining for cleaved caspase-3 in liver tissues from mice with or without Akt inhibitor V pretreatment (100×, scale bar 20μm). Quantification of cleaved caspase-3 positive cells is shown at the right panel (the data are expressed as mean ± SD; *<i>p</i><0.05, **<i>p</i><0.01). (F) Caspase-3/7, caspase-8, and caspase-9 activities in liver tissue from mice with or without Akt inhibitor V pretreatment (the data are expressed as mean ± SD; *<i>p</i><0.05, **<i>p</i><0.01).</p

The levels of miR-150 and Akt in WT and miR-150 KO mice.

<p>(A) Mature miR-150 sequence was aligned among mouse, human, rat, dog and gorilla. (B) miR-150 expression in different tissues from WT and miR-150 KO mice. Total RNA was isolated from eight-week-old male WT and miR-150 KO mice. 1μg of total RNA was used for reverse transcription followed by qRT-PCR analysis. Relative expression of miR-150 was normalized to U6. Data are expressed as mean ± SD. (C) The levels of miR-150 and miR-221 in WT and miR-150 KO livers with or without Jo2 treatment. WT and miR-150 KO mice were intraperitoneally injected with or without 0.5 μg/g of body weight Jo2 (n = 6–8 per group). The livers were harvested 4 hours after Jo2 injection. The levels of miR-150 and miR-221 in the livers were determined by qRT-PCR (Data are expressed as mean ± SD, *<i>p</i><0.05). (D) Western blot for Akt1, Akt 2, total Akt, p-Akt(Ser473), 4E-BP1 and p-4E-BP1(Thr37/46) (with GAPDH as the loading control). The blots shown in this figure were from two individual mice for each group. Quantifications of relative protein levels are shown at the right panel (the data are expressed as mean ± SD, *<i>p</i><0.05).</p

miR-150 deficiency protects against Fas-induced caspase and PARP cleavage.

<p>WT and miR-150 KO mice were intraperitoneally injected with 0.5μg/g of body weight Jo2 (n = 6 or 8 per group, respectively). The animals were sacrificed 0, 2 and 4 hours after Jo2 injection. (A) Caspase activity in liver tissue 4 hours after Jo2 injection. The levels of caspase-3/7, caspase-8, and caspase-9 activities in miR-150 KO livers were significantly lower compared to the WT livers (the data are expressed as mean ± SD) **<i>p</i><0.01. (B) Western blotting analysis to detect PARP and caspases cleavage. (C) Formalin-fixed and paraffin-embedded sections (5 μm thick) were stained with antibody against cleaved caspase-3 (100×, scale bar 20μm). miR-150 KO livers showed fewer numbers of caspase- 3-positive hepatocytes compared to the WT livers (4 hours after Jo2 treatment). Quantification of cleaved caspase-3 positive hepatocytes is shown at the right panel (data are expressed as mean ± SD, **<i>p</i><0.01).</p