Biology subjects

Wahl, L.

Publications and source records attributed to Wahl, L..

4 recordsLinked to original sources

bioRxiv2024

Synthesis and evaluation of chemical linchpins for highly selective CK2α targeting

Casein kinase-2 (CK2) are serine/threonine kinases with dual co-factor (ATP and GTP) specificity, that are involved in the regulation of a wide variety of cellular functions. Small molecules targeting CK2 have been described in the literature targeting different binding pockets of the kinase with a focus on type I inhibitors such as the recently published chemical probe SGC-CK2-1. In this study, we investigated whether known allosteric inhibitors binding to a pocket adjacent to helix D could be combined with ATP mimetic moieties defining a novel class of ATP competitive compounds with a unique binding mode. Linking both binding sites requires a chemical linking moiety that would introduce a 90-degree angle between the ATP mimetic ring system and the D targeting moiety, which was realized using a sulfonamide. The synthesized inhibitors were highly selective for CK2 with binding constants in the nM range and low micromolar activity. While these inhibitors need to be further improved, the present work provides a structure-based design strategy for highly selective CK2 inhibitors.

biochemistry↗

bioRxiv2024

Multi-parametric assays capture sex- and environment-dependent modifiers of behavioral phenotypes in autism mouse models

Current phenotyping approaches for murine autism models often focus on one selected behavioral feature, making the translation onto a spectrum of autistic characteristics in humans challenging. Furthermore, sex and environmental factors are rarely considered. Here, we aimed to capture the full spectrum of behavioral manifestations in three autism mouse models to develop a "behavioral fingerprint" that takes environmental and sex influences under consideration. To this end, we employed a wide range of classical standardized behavioral tests; and two multi-parametric behavioral assays: the Live Mouse Tracker and Motion Sequencing (MoSeq), on male and female Shank2, Tsc1 and Purkinje cell specific-Tsc1 mutant mice raised in standard or enriched environments. Our aim was to integrate our high dimensional data into one single platform to classify differences in all experimental groups along dimensions with maximum discriminative power. Multi-parametric behavioral assays enabled far more accurate classification of experimental groups compared to classical tests, and dimensionality reduction analysis demonstrated significant additional gains in classification accuracy, highlighting the presence of sex, environmental and genotype differences in our experimental groups. Together, our results provide a complete phenotypic description of all tested groups, suggesting multi-parametric assays can capture the entire spectrum of the heterogenous phenotype in autism mouse models.

neuroscience↗

bioRxiv2023

Detecting ataxia using automated analysis of motor coordination and balance of mice on the balance beam

BackgroundThe balance beam assay is a well-known paradigm to assess motor coordination in mouse models of neurodegenerative diseases. Classically, these experiments are quantified using manual annotation, which is time-consuming and prone to inter-experimenter variability. MethodsWe present an open-source analysis pipeline that allows for the automated quantification of motor function in mice crossing the balance beam. ResultsUsing an established Pcp2-Ppp3r1 ataxia mouse model, we validated the analysis pipeline by comparing the motor performance of Pcp2-Ppp3r1 animals to their wildtype littermates. Pcp2-Ppp3r1 animals showed a significant increase in the number of missteps and increased time to traverse the beam. Moreover, we compared the results of the automated classification of missteps and stops to that of 3 independent observers, which showed no significant differences between the classifier and the cumulative observer score for missteps. ConclusionWe show that our pipeline can reliably report crossing time and missteps, offering a high-throughput, automated option for the analysis of balance beam data. Method summaryOur method consists of an easy-to-follow and low-cost manual for building a balance beam setup and an analysis pipeline of mouse movement while crossing the beam. We present a MATLAB script combining FFMPEG and ImageJ (using the MIJ package) to pre-process the videos and extract the time for the mouse to cross the beam. Mice movements on the beam were then tracked using motr and stops and missteps were extracted using a trained JAABA classifier. The trained classifier, the code and all accompanying files were deposited in an open source repository

animal behavior and cognition↗

bioRxiv2021

Activated PI3Kδ syndrome, an immunodeficiency disorder, leads to sensorimotor deficits recapitulated in a murine model.

The phosphoinositide-3-kinase (PI3K) family plays a major role in cell signalling and is predominant in leukocytes. Gain-of-function (GOF) mutations in the PIK3CD gene lead to the development of activated PI3K{delta} syndrome (APDS), a rare primary immunodeficiency disorder. A subset of APDS patients also displays neurodevelopmental delay symptoms, suggesting a potential role of PIK3CD in cognitive and behavioural function. However, the extent and nature of the neurodevelopmental deficits has not been previously quantified. Here, we assessed the cognitive functions of two APDS patients, and investigated the causal role of the PIK3CD GOF mutation in neurological deficits using a murine model of this disease. We used E1020K knock-in mice, harbouring the most common APDS mutation in patients. We found that APDS patients present with visuomotor deficits, exacerbated by autism spectrum disorder comorbidity, whereas p110{delta}E1020K mice exhibited impairments in motor behaviour, learning and repetitive behaviour patterning. Our data indicate that PIK3CD GOF mutations increase the risk for neurodevelopmental deficits, supporting previous findings on the interplay between the nervous and the immune system. Further, our results validate the knock-in mouse model, and offer an objective assessment tool for patients that could be incorporated in diagnosis and in the evaluation of treatments.

neuroscience↗