Translational Therapeutics (United States)

Cytokines dimerize two receptor chains to activate Janus kinases and STAT transcription factors that regulate immune cells but have therapeutic liabilities. We engineered “Trikines” to compel cis formation of three-chain cytokine receptor complexes at the cell surface that induce bespoke STAT transcriptional signaling programs optimized for therapeutic efficacy. Designed Trikines co-activated pSTAT5 and pSTAT3 signatures distinct from any natural cytokines, by assembling trimeric combinations of Interleukin-2 (IL-2), Interleukin-10 (IL-10), and Interleukin-21 (IL-21) receptors. An IL-2-based-Trikine restrained terminal differentiation of T cells, promoted stemness, and enhanced durability of tumor control without toxicity. Unexpectedly, an IL-10-based Trikine induced immune infiltration into poorly immunogenic tumors, showing striking efficacy in small cell lung cancer and pancreatic cancer models. Trikines obviate the need for cell engineering to customize STAT signatures for immunotherapy.

TBCK syndrome is a severe degenerative leukoencephalopathy with multisystem involvement. Neurodevelopmental, craniofacial, and pulmonary challenges are among the topmost effects on these children. TBCK has been implicated in endo-lysosomal regulation, RNA transport, and mTOR-associated pathways, all of which are critical for the development of mineralized tissue. Although craniofacial abnormalities can be clinically apparent, conventional imaging approaches may overlook subtle defects in mineral quality. Here, we apply our multimodal framework to investigate the mineralization of enamel, dentin, and alveolar bone in a Tbck knockout mouse model. This is the first time our multimodal framework will be applied to a genetic condition. Using micro-computed tomography (microCT), histology, nanoindentation, energy-dispersive spectroscopy, and Raman spectroscopy, we identify tissue- and stage-dependent mineral effects undetected by microCT alone. Tbck loss resulted in differences in enamel and dentin element compositions as early as secretory and transition stages, while mechanical properties remained undetected until maturation stage. Notably, Tbck knockout enamel exhibited reduced calcium and phosphorus content, along with increased carbon content during early mineralization, consistent with the retained organic matrix. Additionally, marked and opposing alterations in magnesium and iron levels began at the secretory stage. Together, these findings define a previously unrecognized mineralization signature associated with TBCK deficiency and establish multimodal hard-tissue analysis as a sensitive approach for detecting early craniofacial phenotypes in rare genetic disorders.

Summary Photo-activatable drugs (PDs) are rapidly emerging as precision therapeutics across biomedical research, yet their potential in epilepsy treatment has remained understudied. Given that 30% of epilepsies are medically refractory, and anti-seizure medications often cause multi-organ side-effects, PDs could break new ground. Here, we evaluate light-switchable ion-channel blockers QAQ and CQAQ, and a newly developed caged propofol (CaP), in murine brain slices, and postsurgical brain tissue from patients with epilepsy or brain tumors. In mice, we show that QAQ/CQAQ reversibly suppress neuronal firing, while photo-activated CaP prolonged inhibitory post-synaptic currents and enhanced leak current. While QAQ caused identical effects in human tissue, CQAQ unexpectedly increased firing. Activated CaP robustly suppressed epileptiform activity across species. This work establishes CaP as a tool for neuroscience and disease-related studies. Our results highlight the necessity for early human model testing in biomedical research, and showcase photopharmacology as a potential powerful approach to control human epilepsy.

Purpose:Germline deletions affecting the Y-chromosomal gr/gr region were reported in 2005 as associated with susceptibility to testicular germ cell tumor (TGCT), a highly heritable tumor type that is the most common cancer type affecting adult men under the age of 45. Attempts to replicate this association have been equivocal, primarily due to limited power.Methods:Here, we compare and validate two computational approaches to gr/gr deletion calling in high-, low- and ultra-low-coverage whole genome sequencing data, applying these to two datasets from UK Biobank and the TECAC consortium. We generate dataset-specific effect size estimates for the gr/gr deletion-TGCT association using Firth’s bias-reduced logistic regression across a total of 198,306 men of European-like ancestry (2231 with and 196,075 without TGCT).Results:Upon random-effects meta-analysis of estimated effect sizes in the two datasets, we found no significant association between gr/gr deletion status and TGCT risk (combined odds ratio=1.24, 95% CI=0.74–2.07, p=0.42), nor upon stratification of seminoma and non-seminoma/mixed histological subtypes.Conclusion:Our analysis suggests gr/gr deletion status alone is likely not predictive of TGCT risk in population-scale analyses of European-like individuals; however, the importance of other proposed determinants of gr/gr deletion impact, including Y-haplogroups and semen phenotype, remains unexplored at scale.

ABSTRACTIntroductionPersistent infection with high-risk Human Papillomavirus (hr-HPV) in women is a leading cause of cervical cancer, and its co-infection among people living with HIV (PLHIV) increases the risk of HPV-associated cancer, including oropharyngeal and anogenital cancers. In sub-Saharan Africa, awareness of HPV is limited, screening and vaccine adoption are critically low, undermining efforts toward sexually transmitted infection (STI) elimination.MethodsFromAwareness to ACTion (FACT) study assessed HPV knowledge, screening, and vaccine uptake in adolescent-mother pairs participating in the HOMINY (HPV, Human Immunodeficiency Virus, andOralMicrobiotaInterplay inNigerianYouths) prospective cohort study. Participants were enrolled, including adolescents aged 9-18 years (N=636) and mothers aged 29-59 years (N=385). FACT was conducted at the University of Benin Teaching Hospital, Nigeria, with adolescent participants grouped as perinatally acquired HIV, HIV-exposed without acquisition and HIV-unexposed, and mothers by HIV serostatus. In line with the national immunisation programme protecting girls in Nigeria, sensitisation programmes were integrated into the research study to promote awareness and adoption of screening and vaccination practices. Knowledge and attitudes regarding HPV and its vaccination benefits were assessed through thematic discussions, field notes, interactive sessions, and questionnaires administered over the study period.ResultsAt baseline, HPV awareness was low, with 34.5% of mothers and 1.4% of adolescents being aware of HPV. Post-sensitisation, awareness increased significantly to 64.4% and 19% in mothers and adolescents, respectively. Vaccination uptake rose from 0% to 3.4% in adolescents, and the proportion of mothers who underwent HPV-related screening (Visual Inspection with Acetic Acid and/or Papanicolaou test) increased from 38.7 % at baseline to 52.4 % after a year (p < 0.0001). Barriers to the adoption of preventive services included misconceptions, healthcare provider gaps, myths, misinformation, mistrust, skepticism, and limited access.ConclusionsHPV awareness programmes significantly improved knowledge, vaccination uptake, and screening practices in this vulnerable population. As part of comprehensive STI elimination strategies, integrating HPV education and vaccination initiatives into HIV care and research will enhance prevention and address significant barriers. Lessons from a unique programmatic science framework provide critical insights for scaling vaccine delivery, and the design of future vaccine programmes.

Working memory (WM) deficits are common in psychiatric disorders that are associated with decreased prefrontal cortex activity. As WM is essential for cognitive functions, deficits interfere with daily life and treatment. The lateralization of WM components remains unclear, but stimuli matching, often assessed using the n-back task, has been associated with right-hemispheric dominance. This study examines whether anodal transcranial direct current stimulation (tDCS) targeting the right dorsolateral prefrontal cortex (DLPFC) could enhance WM performance in alcohol-dependent patients (AD), tobacco users (TU) and healthy controls (HC). In a double-blind, sham-controlled study, tDCS was applied to upregulate right DLPFC activity. A total of 46 participants received anodal tDCS with a current intensity of 1.5 mA for 20 min or sham stimulation. In addition to the 7x5cm anode, a large reference electrode (10x10cm) was situated over the contralateral supraorbital area. While being stimulated, participants performed the n-back task as a measure of WM performance. Results revealed no significant differences in WM performance between active and sham stimulated participants, nor between groups, and no significant interaction between stimulation condition and group. Bayesian analysis supported the null effects. These findings do not provide evidence that single-session right DLPFC stimulation reliably enhances working memory across all stimulus types. The outcomes may have been influenced by task–stimulation mismatch, sample heterogeneity, small sample size, and stimulation parameters, which could limit the ability to detect subtle tDCS effects in both clinical and healthy populations.

Adrenocortical carcinoma (ACC) is an under-studied, aggressive cancer of the adrenal glands where surgical resection is currently the only effective curative option. However, after surgery the majority of patients experience tumor progression. The median overall survival of 12 months has not improved since approval of mitotane in 1970. The lack of effective therapies in ACC is partially due to the lack of preclinical models that accurately represent human ACC. Most attempts to generate ACC cell lines or animal models have been unsuccessful. The few existing models do not adequately reflect the oncogenic signaling pathways or intratumoral heterogeneity of human ACC. These limited model systems have hindered identification of drivers of tumor progression and immune escape mechanisms, thereby limiting development and testing of novel therapeutic approaches such as targeted therapies or immunotherapies. We developed human ACC patient-derived tumor constructs (PTCs) encapsulated in synthetic extracellular matrix. Our ACC PTCs exhibit hallmarks of ACC: proliferation, expression of key ACC biomarkers, such as SF1, and production of cortisol. We provide characterization in the form of immunofluorescence staining for ACC biomarkers, and confirmation of PTC proliferation and cortisol production - a hallmark of ACC. We then demonstrate the utility of ACC PTCs for evaluation of chemotherapies currently used clinically (mitotane) with and without experimental cocktails of etoposide, doxorubicin, and cisplatin, experimental targeted therapies, and cellular immunotherapies, primarily in the form of natural killer (NK) cell therapy. In particular, the latter - cellular immunotherapies - are cutting edge studies demonstrating potential to evaluate immunotherapies in ACC clinical scenarios. Together these data provide evidence that patient-derived ACC models can serve as an important tool for identification of future points of intervention and testing of novel therapeutic strategies to improve ACC clinical care.

BACKGROUND:Large clinical trials compared distinct nonsteroidal anti-inflammatory drugs in terms of their risk of adverse cardiovascular events. However, whether pharmacologically equipotent doses were used, that is, whether a similar degree of COX (cyclooxygenase)-2 inhibition was achieved, was not considered. We compared drug target inhibition and blood pressure (BP) response to celecoxib and naproxen.METHODS:Sixteen healthy participants were treated with celecoxib (200 mg/d), naproxen (500 mg/d), or placebo for 7 days in a double-blind, crossover design. The degree of COX inhibition was assessed ex vivo using established whole blood assays and in vivo by quantifying urinary metabolites of thromboxane A2 (COX-1) and prostacyclin (COX-2). Ambulatory BP was measured throughout the final dosing interval.RESULTS:Both nonsteroidal anti-inflammatory drugs inhibited COX-2 activity relative to placebo, but naproxen inhibited COX-2 activity to a greater degree (62.9±21.7%) than celecoxib (35.7±25.2%; P<0.05). Similarly, naproxen treatment inhibited prostacyclin formation in vivo (48.0±24.9%) to a greater degree than celecoxib (26.7±24.6%; P<0.05). Naproxen significantly increased BP compared with celecoxib (mean arterial pressure, +2.5 [95% CI, 1.5–3.5] mm Hg; systolic BP, +4.0 [95% CI, 2.9–5.1] mm Hg; and diastolic BP, +1.8 [95% CI, 0.8–2.8] mm Hg; P<0.05 for all). The difference in systolic BP relative to placebo was associated with the degree of COX-2 inhibition (P<0.05).CONCLUSIONS:Future studies should consider pharmacokinetic and pharmacodynamic properties, as well as patient-specific factors that may modulate the cardiovascular risk of nonsteroidal anti-inflammatory drug use.REGISTRATION:URL: https://www.clinicaltrials.gov; Unique identifier: NCT02502006.

Background:Understanding how variation in brain structure and function contributes to psychiatric and behavioral phenotypes remains a key challenge. The absence of neuroimaging data in many study samples limits this effort.Methods:We used genome-wide association study (GWAS) summary statistics from the UK Biobank to impute 301 brain imaging-derived phenotype (IDP) genetic scores (IGS) in the Yale-Penn cohort, which is enriched for substance use disorders (n = 10,275; 52.8% European-like [EUR] and 47.2% African-like [AFR] genetic ancestry). The brain IDPs include white matter microstructure, regional volume, and resting-state functional connectivity measures, for which we generated IGS in the Yale-Penn participants. We then conducted a brain-wide phenome-wide association study (pheWAS) of the 301 IGS across 692 behavioral, psychiatric, and environmental traits.Results:Among EUR individuals, we identified 19 IGS with significant associations that survived within-trait corrections for multiple testing. These included links between genetically predicted white matter integrity and sedative abuse, tobacco withdrawal, attention deficit hyperactivity disorder (ADHD); structural brain volumes and cocaine dependence, ADHD, and conduct disorder; and functional connectivity with substance-related symptoms and social phobia. Among AFR individuals, we identified 15 IDPs with significant associations, including associations between genetically predicted white matter integrity and stimulant use disorder, regional brain volumes and opioid withdrawal/dependence, and functional connectivity and cocaine craving.Conclusions:Genetically imputed brain features capture biological variation associated with psychiatric traits. This work provides a framework for leveraging genetic data to link neuroimaging measures to substance use and mental health outcomes in samples that lack imaging data.

Clinical progression during psychosis has been closely associated with grey matter abnormalities resulting from atypical brain development. However, the complex interplay between psychopathology and heterogeneous maturational trajectories challenges the identification of neuroanatomical features that anticipate symptomatic decline. To investigate cortical volume longitudinal deviations in first-episode psychosis (FEP) using normative modelling, exploring their relationship with long-term cognitive and symptomatic outcomes, as well as their cytoarchitectural and neurobiological underpinnings. We collected magnetic resonance imaging (MRI), cognitive and symptomatic data from 195 healthy controls and 357 drug-naïve or minimally medicated FEP individuals that were followed up 1, 3, 5 and 10 years following the first episode (1209 MRI scans and assessments in total). Using normative modelling, we derived subject-specific centile scores for cortical volume to investigate atypical deviations in FEP and their relationship to long-term cognitive and symptomatic deterioration. The resulting centile association maps were further characterised by examining their cytoarchitectural and neurobiological attributes using normative atlases. FEP centiles demonstrated a widespread reduction at treatment initiation, with longitudinal analysis showing an increase during treatment time, indicating convergence towards normal maturation trajectories. Interestingly, this effect was reduced in highly medicated individuals. Additionally, we found that cognitive impairments experienced during early FEP stages worsened under long-term medication. Positive symptomatology was negatively associated with regional centiles, and individuals with higher centiles benefited most from treatment. Cytoarchitectural and neurobiological analyses revealed that regional centiles related to FEP, as well as to symptomatology, were associated with specific molecular features, such as regional serotonin and dopamine receptor densities. Collectively, these findings underscore the potential use of centile-based normative modelling for a better understanding of how atypical cortical development contributes to the long-term clinical progression of neurodevelopmental conditions.