KAP1 targets actively transcribed genomic loci to exert pleomorphic effects on RNA polymerase II activity

KAP1 (KRAB-associated protein 1) is best known as a co-repressor responsible for inducing heterochromatin formation, notably at transposable elements. However, it has also been observed to bind the transcription start site of actively expressed genes. To address this paradox, we characterized the protein interactome of KAP1 in the human K562 erythro-leukaemia cell line. We found that the regulator can associate with a wide range of nucleic acid binding proteins, nucleosome remodellers, chromatin modifiers and other transcription modulators. We further determined that KAP1 is recruited at actively transcribed polymerase II promoters, where its depletion resulted in pleomorphic effects, whether expression of these genes was normally constitutive or inducible, consistent with the breadth of possible KAP1 interactors. This article is part of a discussion meeting issue 'Crossroads between transposons and gene regulation'

Bispecific PD1-IL2v and anti-PD-L1 break tumor immunity resistance by enhancing stem-like tumor-reactive CD8⁺ T cells and reprogramming macrophages.

Immunotherapies have shown remarkable, albeit tumor-selective, therapeutic benefits in the clinic. Most patients respond transiently at best, highlighting the importance of understanding mechanisms underlying resistance. Herein, we evaluated the effects of the engineered immunocytokine PD1-IL2v in a mouse model of de novo pancreatic neuroendocrine cancer that is resistant to checkpoint and other immunotherapies. PD1-IL2v utilizes anti-PD-1 as a targeting moiety fused to an immuno-stimulatory IL-2 cytokine variant (IL2v) to precisely deliver IL2v to PD-1 <sup>+</sup> T cells in the tumor microenvironment. PD1-IL2v elicited substantial infiltration by stem-like CD8 <sup>+</sup> T cells, resulting in tumor regression and enhanced survival in mice. Combining anti-PD-L1 with PD1-IL2v sustained the response phase, improving therapeutic efficacy both by reprogramming immunosuppressive tumor-associated macrophages and enhancing T cell receptor (TCR) immune repertoire diversity. These data provide a rationale for clinical trials to evaluate the combination therapy of PD1-IL2v and anti-PD-L1, particularly in immunotherapy-resistant tumors infiltrated with PD-1 <sup>+</sup> stem-like T cells

A snow and glacier hydrological model for large catchments – case study for the Naryn River, central Asia

In this paper we implement a degree day snowmelt and glacier melt model in the Dynamic fluxEs and ConnectIvity for Predictions of HydRology (DECIPHeR) model. The purpose is to develop a hydrological model that can be applied to large glaciated and snow-fed catchments yet is computationally efficient enough to include model uncertainty in streamflow predictions. The model is evaluated by simulating monthly discharge at six gauging stations in the Naryn River catchment (57 833 km2) in central Asia over the period 1951 to a variable end date between 1980 and 1995 depending on the availability of discharge observations. The spatial distribution of simulated snow cover is validated against MODIS weekly snow extent for the years 2001–2007. Discharge is calibrated by selecting parameter sets using Latin hypercube sampling and assessing the model performance using six evaluation metrics. The model shows good performance in simulating monthly discharge for the calibration period (NSE is 0.74&lt;NSE&lt;0.87) and validation period (0.7&lt;NSE&lt;0.9), where the range of NSE values represents the 5th–95th percentile prediction limits across the gauging stations. The exception is the Uch-Kurgan station, which exhibits a reduction in model performance during the validation period attributed to commissioning of the Toktogul reservoir in 1975 which impacted the observations. The model reproduces the spatial extent in seasonal snow cover well when evaluated against MODIS snow extent; 86 % of the snow extent is captured (mean 2001–2007) for the median ensemble member of the best 0.5 % calibration simulations. We establish the present-day contributions of glacier melt, snowmelt and rainfall to the total annual runoff and the timing of when these components dominate river flow. The model predicts well the observed increase in discharge during the spring (April–May) associated with the onset of snow melting and peak discharge during the summer (June, July and August) associated with glacier melting. Snow melting is the largest component of the annual runoff (89 %), followed by the rainfall (9 %) and the glacier melt component (2 %), where the values refer to the 50th percentile estimates at the catchment outlet gauging station Uch-Kurgan. In August, glacier melting can contribute up to 66 % of the total runoff at the highly glacierized Naryn headwater sub-catchment. The glaciated area predicted by the best 0.5 % calibration simulations overlaps the Landsat observations for the late 1990s and mid-2000s. Despite good predictions for discharge, the model produces a large range of estimates for the glaciated area (680–1196 km2) (5th–95th percentile limits) at the end of the simulation period. To constrain these estimates further, additional observations such as glacier mass balance, snow depth or snow extent should be used directly to constrain model simulations.</p

Rehydrating efficacy of maple water after exercise-induced dehydration

Abstract Dehydration impairs physiological function and physical performance, thus understanding effective rehydration strategies is paramount. Despite growing interest in natural rehydrating beverages, no study has examined maple water (MW). Purpose To investigate the rehydrating efficacy of MW after exercise-induced dehydration. Methods Using a single-blind, counterbalanced, crossover design, we compared the rehydrating efficacy of MW vs. maple-flavored bottled water (control) in 26 young healthy (22 ± 4 yrs., 24 ± 4 kg/m2) males (n = 13) and females (n = 13) after exercise-induced dehydration (~ 2.0%ΔBody Weight [BW]) in the heat (30 °C, 50% relative humidity [RH]). Hydration indicators (BW, salivary and urine osmolality [SOsm/UOsm], urine specific gravity [USG], urine volume [UV], urine color [UC]), thirst, fatigue, and recovery (heart rate [HR)], and HR variability [HRV]) were taken at baseline, post-exercise, 0.5, 1, and 2 h post-consumption of 1 L of MW or control. Results Following similar dehydration (~ 2%ΔBW), MW had no differential (p > 0.05) impact on any measure of rehydration. Likely due to greater beverage osmolality (81 ± 1.4 vs. 11 ± 0.7 mOsmol/kg), thirst sensation remained 12% higher with MW (p <  0.05). When sex was considered, females had lower UV, elevated UOsm (p < 0.05), trends for higher ΔBW, USG, but similar SOsm. Analysis of beverages and urine for antioxidant potential (AP) revealed a four-fold greater AP in MW, which increased peak urine AP (9.4 ± 0.7 vs. 7.6 ± 1.0 mmol, MW vs. control, p <  0.05). Conclusion Electrolyte-containing MW, was similar in effectiveness to water, but has antioxidant properties. Furthermore, trends for sex differences were discovered in urinary, but not salivary, hydration markers, with discrepancies in kinetics between fluid compartments both warranting further study

Transposable Elements and Their KRAB-ZFP Controllers Regulate Gene Expression in Adult Tissues.

KRAB-containing zinc finger proteins (KRAB-ZFPs) are early embryonic controllers of transposable elements (TEs), which they repress with their cofactor KAP1 through histone and DNA methylation, a process thought to result in irreversible silencing. Using a target-centered functional screen, we matched murine TEs with their cognate KRAB-ZFP. We found the paralogs ZFP932 and Gm15446 to bind overlapping but distinguishable subsets of ERVK (endogenous retrovirus K), repress these elements in embryonic stem cells, and regulate secondarily the expression of neighboring genes. Most importantly, we uncovered that these KRAB-ZFPs and KAP1 control TEs in adult tissues, in cell culture and in vivo, where they partner up to modulate cellular genes. Therefore, TEs and KRAB-ZFPs establish transcriptional networks that likely regulate not only development but also many physiological events. Given the high degree of species specificity of TEs and KRAB-ZFPs, these results have important implications for understanding the biology of higher vertebrates, including humans

Lymph node dendritic cells harbor inducible replication-competent HIV despite years of suppressive ART.

Although gut and lymph node (LN) memory CD4 T cells represent major HIV and simian immunodeficiency virus (SIV) tissue reservoirs, the study of the role of dendritic cells (DCs) in HIV persistence has long been limited to the blood due to difficulties to access lymphoid tissue samples. In this study, we show that LN migratory and resident DC subpopulations harbor distinct phenotypic and transcriptomic profiles. Interestingly, both LN DC subpopulations contain HIV intact provirus and inducible replication-competent HIV despite the expression of the antiviral restriction factor SAMHD1. Notably, LN DC subpopulations isolated from HIV-infected individuals treated for up to 14 years are transcriptionally silent but harbor replication-competent virus that can be induced upon TLR7/8 stimulation. Taken together, these results uncover a potential important contribution of LN DCs to HIV infection in the presence of ART

A gene-rich, transcriptionally active environment and the pre-deposition of repressive marks are predictive of susceptibility to KRAB/KAP1-mediated silencing.

AbstractBACKGROUND: KRAB-ZFPs (Krüppel-associated box domain-zinc finger proteins) are vertebrate-restricted transcriptional repressors encoded in the hundreds by the mouse and human genomes. They act via an essential cofactor, KAP1, which recruits effectors responsible for the formation of facultative heterochromatin. We have recently shown that KRAB/KAP1 can mediate long-range transcriptional repression through heterochromatin spreading, but also demonstrated that this process is at times countered by endogenous influences.METHOD: To investigate this issue further we used an ectopic KRAB-based repressor. This system allowed us to tether KRAB/KAP1 to hundreds of euchromatic sites within genes, and to record its impact on gene expression. We then correlated this KRAB/KAP1-mediated transcriptional effect to pre-existing genomic and chromatin structures to identify specific characteristics making a gene susceptible to repression.RESULTS: We found that genes that were susceptible to KRAB/KAP1-mediated silencing carried higher levels of repressive histone marks both at the promoter and over the transcribed region than genes that were insensitive. In parallel, we found a high enrichment in euchromatic marks within both the close and more distant environment of these genes.CONCLUSION: Together, these data indicate that high levels of gene activity in the genomic environment and the pre-deposition of repressive histone marks within a gene increase its susceptibility to KRAB/KAP1-mediated repression