The rockefeller university

Regulatory T (Treg) cells, expressing the transcription factor Foxp3, are obligatory gatekeepers of immune responsiveness, yet the mechanisms by which Foxp3 governs the Treg transcriptional network remain incompletely understood. Using a novel chemogenetic system of inducible Foxp3 protein degradation in vivo, we found that while Foxp3 was indispensable for the establishment of transcriptional and functional programs of newly generated Treg cells, Foxp3 loss in mature Treg cells resulted in minimal functional and transcriptional changes under steady state. This resilience of the Foxp3-dependent program in mature Treg cells was acquired over an unexpectedly long timescale; however, in settings of severe inflammation, Foxp3 loss led to a pronounced perturbation of Treg cell transcriptome and fitness. Furthermore, tumoral Treg cells were uniquely sensitive to Foxp3 degradation, which led to impairment in their suppressive function and tumor shrinkage in the absence of pronounced adverse effects. These studies demonstrate a context-dependent differential requirement for Foxp3 for Treg transcriptional and functional programs.

QuestionCan an education and communication-based intervention improve knowledge and caregiving self-efficacy among home health aides and attendants (HHAs) caring for adults with heart failure (HF)?FindingsIn this pilot randomized clinical trial including 102 agency-employed HHAs randomized to training alone or in addition to an application that allowed HHAs to exchange text messages with nurse supervisors, training improved HHAs' HF knowledge and HF caregiving self-efficacy. The addition of the application did not improve these primary outcomes, but it significantly reduced HHAs' self-reported preventable 911 calls, a secondary outcome.MeaningThese results support a future RCT that will test the effectiveness of the intervention on outcomes for both HHAs and their patients with HF. This pilot randomized clinical trial examines the effectiveness of an education- and communication-based intervention among home health aides caring for patients with heart failure. ImportanceHome health aides (HHAs) frequently care for adults with heart failure (HF), but many lack HF training, confidence with HF caregiving, and cannot reach their nurse supervisors by telephone when they need guidance. This may have negative consequences for HHAs and patients.ObjectiveTo examine the effectiveness of an education- and communication-based intervention among HHAs caring for patients with HF.Design, Setting, and ParticipantsThis 2-group pilot randomized clinical trial was conducted in partnership with a large home care agency in New York, New York, from May 2022 to May 2024. HHAs caring for a patient with HF participated. Outcomes were ascertained on an intent-to-treat basis at baseline, mid-study (45 days after the training course), and 90 days.InterventionsThe enhanced usual care (EUC) group received HF training, and the intervention group received HF training plus a mobile health application that allowed HHAs to message nurses.Main Outcomes and MeasuresCo-primary outcomes were HF knowledge (assessed using the Dutch HF Knowledge Scale [DHFKS]; range 0-15; higher score indicates greater knowledge) and HF caregiver self-efficacy (assessed using the Caregiver Contribution to Self-Care in HF Index; range, 0-100; higher score indicates greater efficacy). The secondary outcome was self-reported preventable 911 calls. Exploratory outcomes included patient emergency department (ED) visits and hospitalizations. Mixed-effects models were used to compare trajectories of outcomes between and within study groups.ResultsA total of 102 HHAs (mean [SD] age, 54 [10.5] years; 98 [96.1%] female) were assessed, including 50 in the EUC group and 52 in the intervention group. Overall, 62 HHAs (62.0%) were Black, 1 HHA (1.0%) was American Indian or Alaska Native, 7 HHAs (7.0%) were Asian, 9 HHAs (9.0%) were White, and 21 HHAs (21.0%) identified as other race; 27 HHAs (27.0%) were Hispanic. Within the intervention group, DHFKS scores improved at 90 days, from a median (IQR) score of 6.1 (5.5-6.7) points at baseline to 7.7 (7.0-8.4) points at 90 days (P = .02); however the change did not differ between groups. Across both groups, HHAs with the lowest baseline DHFKS and self-efficacy had the greatest increases at 90 days (median [IQR] change: DHFKS, 1.45 [0.84-2.04] points; self-efficacy, 8.06 [4.42-11.71] points). At 90 days, there were no statistically significant within-group differences in the proportion of HHAs reporting preventable 911 calls group (intervention: 0.51 [95% CI, 0.37-0.64] at baseline vs. 0.34 [95% CI, 0.2-0.49] at 90 days; P = .06; EUC: 0.42 [95% CI, 0.28-0.56] at baseline vs 0.54 [95% CI, 0.38-0.70] at 90 days; P = .21), but the difference between groups was statistically significant (P = .04). This pilot study was not powered for patient-level outcomes, so the risk of ED visits for patients of intervention HHAs (incidence rate ratio, 0.56 [95% CI, 0.25-1.28]; P = .17) should be considered exploratory.Conclusions and RelevanceIn this randomized clinical trial of HHAs caring for patients with HF, HF training improved HHAs' knowledge and self-efficacy, with greatest gains among those with the lowest baseline scores. The ability to message nurses was associated with fewer preventable 911 calls among HHAs in the intervention group. These findings can inform the design of a large-scale trial to better support and integrate HHAs providing HF care.Trial RegistrationClinicalTrials.gov Identifier: NCT04239911

BACKGROUND & AIMS: Hepatic encephalopathy (HE) is a common complication following transjugular intrahepatic portosystemic shunt (TIPS) insertion. However, the prognostic significance of overt HE post-TIPS remains controversial. METHODS: We screened 2137 patients who underwent TIPS insertion at 8 German tertiary care centers between 2004 and 2021. Patients with pre-emptive TIPS placement, hepatocellular carcinoma, missing data, and non-PTFE covered stents were excluded. Competing risk analysis was performed, considering liver transplantation as a competing event. To correct for immortal time bias, landmark analyses were conducted, with the landmark being set at 30 and 90 days post-TIPS. Outcome data were assessed for up to 30 months post-TIPS insertion. RESULTS: A total of 1356 patients (median Model for End-stage Liver Disease [MELD], 13 [interquartile range (IQR), 10-17]; age, 60 years [IQR, 54-67 years]; 64% male; 12% HE before TIPS), were included. Overall, HE post-TIPS was linked to impaired survival (P < .001; subdistribution hazard ratio [sHR], 1.41; 95% confidence interval [CI],1.15-1.73). However, this was only confirmed if HE occurred within the first 30 days post-TIPS (early HE; P < .001; sHR, 2.02; 95% CI, 1.59-2.57). Additionally, patients with a history of HE (P < .001; sHR, 1.59; 95% CI, 1.21-2.07) and history of HE and early HE post-TIPS (P < .001; sHR, 3.44; 95% CI, 2.34-5.04) showed impaired survival. These findings were confirmed in the landmark and multivariable analyses. CONCLUSIONS: Early HE post-TIPS is associated with significantly reduced survival. Therefore, patients who experience early HE or have a history of HE should be closely monitored by physicians, as they constitute a particularly vulnerable group with impaired survival.

Solute carriers (SLCs) regulate cellular and organismal metabolism by transporting small molecules and ions across membranes, yet the physiological substrates of similar to 20% remain elusive. To address this, we developed a machine-learning platform to predict gene-metabolite associations. This approach identifies UNC93A and SLC45A4 as candidate plasma membrane transporters for acetylglucosamine and polyamines, respectively. Additionally, we uncover SLC25A45 as a mitochondrial transporter linked to serum levels of methylated basic amino acids, products of protein catabolism. Mechanistically, SLC25A45 is necessary for the mitochondrial import of methylated basic amino acids, including ADMA and TML, the latter serving as a precursor for carnitine synthesis. In line with this observation, SLC25A45 loss impairs carnitine synthesis and blunts upregulation of carnitine-containing metabolites under fasted conditions. By facilitating mitochondrial TML import, SLC25A45 connects protein catabolism to carnitine production, sustaining beta-oxidation during fasting. Altogether, our study identifies putative substrates for three SLCs and provides a resource for transporter deorphanization.

The receptor-binding domain (RBD) of the SARS-CoV-2 spike (S) protein continues to evolve, facilitating antibody evasion. It remains unclear whether any conserved RBD epitopes persist across SARS-CoV-2 variants and whether vaccination and/or breakthrough infection (BTI) can elicit antibodies capable of targeting these conserved regions to counter future variants. Here, using a heterogeneous double-bait single B-cell sorting strategy, we identify a subset of antibodies with broad-spectrum RBD binding, including recognition of SARS-CoV-1 and emerging variants such as EG.5.1, BA.2.86, JN.1, and KP.2/3. These broadly binding antibodies (bbAbs) exhibit elevated levels of somatic hypermutation but are infrequently derived from clonally expanded B lymphocytes. Passive transfer of representative bbAbs reduces viral infection in a male hamster model. Structural analyses reveals that these bbAbs primarily target three distinct, highly conserved RBD epitopes, suggesting potential regions of future mutational pressure and highlighting the presence of conserved and immunogenic RBD conformations that may serve as a foundation for the development of broadly protective vaccines.

Membrane protein homo-oligomers named higher-order transient structures (HOTS) are formed through cohesive self-interactions in the range of a few kBT. The small magnitude of these interactions underlies the rapid reversibility of HOTS on the timescale of membrane signaling processes, permitting the dynamic modulation of signals. At the same time, weak interactions present an apparent paradox: HOTS should form only if the concentration of a particular protein is sufficiently high to produce oligomerization by mass action. And yet, HOTS are observed experimentally with membrane proteins present in cell membranes at concentrations of only a few per mu m2. In this study, we employ principles of statistical thermodynamics to explain how cells can alter the configurational entropy of the oligomerization reaction, thereby achieving HOTS formation at low concentrations of the protein in the membrane. We propose that this modification of the configurational entropy, a process we call configurational length scaling, is an important aspect of HOTS formation in cell membranes and possibly other cellular compartments.

Current methods for variant effect prediction do not differentiate between pathogenic variants resulting in different disease outcomes and are restricted in application due to a focus on variants with a single molecular consequence. We have developed Variant-to-Phenotype (V2P), a multi-task, multi-output machine learning model to predict variant pathogenicity conditioned on top-level Human Phenotype Ontology disease phenotypes (n = 23) for single nucleotide variants and insertions/deletions throughout the human genome. V2P leverages a unique approach for the modeling of variant effect that incorporates resultant disease phenotypes as output and during training to improve the quality of variant disease phenotype and effect predictions, simultaneously. We describe the architecture, training strategy, and biological features contributing to V2P's output, revealing initial characteristics underlying the relationship between disease genotype and phenotype. Moreover, we demonstrate the benefit of incorporating disease phenotypes for variant effect predictions by comparing V2P with several variant effect predictors across various high-quality evaluation datasets from manually curated databases and functional assays. Finally, we examine how V2P's predictions result in the successful identification of pathogenic variants in real and simulated patient sequencing data, outperforming other tested methods in initial comparisons. V2P offers a complete mapping of human genetic variants to disease-phenotypes, offering a uniquely conditioned set of variant effect characterizations.

Generation of in vitro human induced pluripotent cell (hiPSC)-derived skeletal muscle progenitor cells (SMPCs) holds great promise for regenerative medicine for skeletal muscle wasting diseases, for example Duchenne muscular dystrophy (DMD). While multiple approaches have been described to obtain SMPCs in vitro, hiPSC-derived SMPCs generated using transgene-free protocols are usually obtained in a low amount and resemble a more embryonal/fetal stage of differentiation. Here, we demonstrate that modulation of the JAK2/STAT3 signaling pathway during an in vitro skeletal muscle differentiation protocol increases the yield of PAX7+and CD54+human SMPCs (hSMPCs) and drives them to a higher maturation stage, in both human embryonic stem (ES) and patient-derived induced pluripotent cells (iPSCs). Importantly, the obtained SMPCs are able to differentiate into multinucleated myotubes in vitro and engraft in vivo. These findings reveal that modulation of the JAK2/STAT3 signaling pathway is a potential therapeutic avenue to generate SMPCs in vitro with potential for cell therapy approaches.

Although adenosine was identified in the brain many decades ago, our understanding of when, where, and how it functions has expanded rapidly in recent years, driven in part by innovative technological advances. Adenosine is now increasingly recognized as a key neuromodulator that dynamically regulates brain circuits important for sleep/wakefulness, movement, cognition, and homeostasis. In addition, growing attention has been directed toward the molecular mechanisms governing adenosine production and its downstream signaling pathways, both of which hold great promise as therapeutic targets for neuropsychiatric disorders and neurodegenerative diseases. This review highlights recent progress in detecting adenosine, unraveling its signaling pathways in vitro and in vivo, and understanding how it regulates brain function under physiological and pathological conditions.