Biology subjects

Weller, A.

Publications and source records attributed to Weller, A..

4 recordsLinked to original sources

bioRxiv2025

Optimization and evaluation of complementary degrader discovery assays for application in screening

Targeted protein degradation (TPD) mediated by molecular glues is an innovative pharmaceutical paradigm. By binding to and modulating the surface of an E3-ligase component, molecular glue degraders can facilitate the recruitment of a specific target protein (or vice versa) and, ultimately, invoke target degradation. This mode of action results in specific challenges for the development of rational discovery strategies, and complex hit validation workflows may be required to reliably eliminate compounds that elicit non-specific effects. With the aim to guide screening efforts, we optimized two orthogonal cell-based, target-centric assays for degrader discovery: (1) a time-resolved FRET assay directly quantifying levels of target protein and its degradation (signal inhibition), and (2) an assay coupling TPD to cell growth (signal rescue). To enable a deeper understanding of the individual assays strengths and limitations, we compared their statistical performance as well as respective hit populations by screening a specifically designed collection of about 1000 compounds containing well annotated reference compounds and known frequent hitters. We found that the signal rescue format reliably and specifically captured active target degraders while it efficiently filtered out interfering or frequent hitter compounds. Importantly, this format achieved to retrieve lower potency hits, which might be desirable in order to confidently include as many diverse chemical starting points as possible at the start of a drug discovery project.

pharmacology and toxicology↗

bioRxiv2025

PRDM16 controls smooth muscle cell fate in atherosclerosis

Vascular smooth muscle cells (SMCs) normally exist in a contractile state but can undergo fate switching to produce various cell phenotypes in response to pathologic stimuli1-3. In atherosclerosis, these phenotypically modulated SMCs regulate plaque composition and influence the risk of major adverse cardiovascular events4,5. We found that PRDM16, a transcription factor that is genetically associated with cardiovascular disease, is highly expressed in arterial SMCs and downregulated during SMC fate switching in human and mouse atherosclerosis. Loss of Prdm16 in SMCs of mice activates a synthetic modulation program under homeostatic conditions. Single cell analyses show that loss of Prdm16 drives a synthetic program in all SMC populations. Upon exposure to atherogenic stimuli, SMC-selective Prdm16 deficient mice develop SMC-rich, fibroproliferative plaques that contain few foam cells. Acute loss of Prdm16 results in the formation of collagen-rich lesions with thick fibrous caps. Reciprocally, increasing PRDM16 expression in SMCs blocks synthetic processes, including migration, proliferation, and fibrosis. Mechanistically, PRDM16 binds to chromatin and decreases activating histone marks at synthetic genes. Altogether, these results define PRDM16 as a critical determinant of SMC identity and atherosclerotic lesion composition.

molecular biology↗

bioRxiv2024

Temporary suspension of mineral phosphorus reduces mobilizable bone zinc in adult laying hens irrespective of the dietary zinc supply

This study examined the effects of short-term dietary zinc (Zn) and phosphorus (P) variations on the mobilizable bone Zn pool and overall Zn status in adult laying hens. Forty-eight hens (50% Lohmann Brown Classic, 50% Lohmann LSL Classic) were housed in pairs (one hen per breed per pen) across 24 pens. The pens were randomly assigned to one of two dietary P levels (0.37% or 0.84% in DM) using a high-protein corn-soybean diet (11.4 MJ AME/kg, 21.5% CP) during a 14-day acclimatization period. Following acclimatization, pens from both P groups were further randomized into four dietary treatments in a 2 x 2 factorial design, varying in P levels (low vs. high) and Zn supplementation (28 vs. 131 mg/kg) over an 8-day experimental feeding phase. Performance metrics, egg production and quality, and tissue mineral concentrations (plasma, liver, bone, and eggs) were measured. Statistical analyses were performed using linear mixed models in SAS 9.4, incorporating random effects of pen nested within treatment group and fixed effects of dietary P, dietary Zn, breed, and their interactions. Tukey-corrected 95% confidence intervals were used to estimate effect differences, with significance set at P < 0.05. Performance metrics, including egg production and body weight, were unaffected by dietary treatments (P > 0.1), indicating no clinical symptoms of Zn deficiency. However, hens on low-Zn diets exhibited significant reductions in plasma Zn concentration (-0.83 mg/L; P = 0.0008) and liver Zn concentration (-6.78 mg/kg DM; P = 0.01), confirming subclinical Zn deficiency. Low-Zn diets also increased the femoral molar Ca:P ratio by 0.15 (P = 0.01), irrespective of dietary P supply. Interestingly, low-P diets led to a significant reduction in femur Zn content (-0.46 mg; P = 0.0009), regardless of Zn supplementation, following 21 days of reduced P feeding. These findings highlight the higher susceptibility of laying hens to phytate antagonism compared to broilers, as evidenced by measurable subclinical Zn deficiency under short-term Zn deprivation. Additionally, a temporary suspension of mineral P supply appeared to impair the mobilizable bone Zn pool. The underlying functional mechanisms driving these interactions remain unclear and warrant further investigation.

physiology↗

bioRxiv2024

Helping behavior is associated with increased affiliative behavior, activation of the prosocial brain network and elevated oxytocin receptor expression in the nucleus accumbens

A prosocial response to others in distress is increasingly recognized as a natural behavior for many social species, from humans to rodents. While prosocial behavior is more frequently observed towards familiar conspecifics, even within the same social context some individuals are more prone to help than others. For instance, in a rat helping behavior test, rats can release a distressed conspecific trapped inside a restrainer by opening the restrainer door. Typically, rats are motivated to release a trapped cagemate, and consistently release the trapped rat ( openers), yet around 30% do not open the restrainer ( non-openers). To characterize the difference between these populations, behavioral and neural activity were compared between opener and non-opener rats tested with a trapped cagemate in the helping test. Behaviorally, openers showed significantly more social affiliative behavior both before and after door-opening compared to non-openers. Analysis of brain-wide neural activity based on the immediate early gene c-Fos revealed increased activity in openers in the previously identified prosocial neural network compared to non-openers. The network includes regions associated with empathy in humans (somatosensory cortex, insula, cingulate cortex and frontal cortex), and motivation and reward regions such as the nucleus accumbens. Oxytocin receptor mRNA expression levels were higher in the accumbens but not the anterior insula. Several transcription control pathways were also significantly upregulated in openers accumbens. These findings indicate that prosocial behavior may be predicted by affiliative behavior and activity in the prosocial neural network and provide targets for the investigation of causal mechanisms underlying prosocial behavior. Significance StatementProsocial behavior is observed in many social species, including rodents, yet the determinants underlying why some animals help and others do not is poorly understood. Here, we show behavioral and neural differences between prosocial and non-prosocial pairs in a rat helping behavior test, with increased social interaction and nucleus accumbens oxytocin receptor gene expression in animals that helped.

neuroscience↗