Moritz J Paha, Arshi Mustafa, Lyvin Tat, Sandy Richter, Linnaeus Bundalian, Sworadip Chowdhury, Akhil Velluva, Robin-Tobias Jauss, Knut Krohn, Kathleen Schön, Max Holzer, Anna S Kirstein, Meena Balasubramanian, María Juliana Ballesta-Martínez, Lynne M Bird, Kirsty Bradshaw, Ange-Line Bruel, Alfredo Brusco, Yanmin Chen, Aurore Garde, Silvia Guarducci, Yoel Gofin, Rachel E Harrison, Alexis Heidlebaugh, Anna Hurst, Julian A Martinez-Agosto, Jessica L Mester, Alistair T Pagnamenta, Marilena Pantaleo, Andrea Paras, Angela Peron, Tiziana Pisano, Ahna M Rabani, Marcello Scala, Elena Shukarova-Angelovska, Sui Yang, Slavica Trajkova, Sheila Umayam, Tianyun Wang, Rami Abou Jamra, Johannes R Lemke, Wieland Kiess, Konstantinos S Zarbalis, Antje Garten, Diana Le Duc
Current evidence is dominated by conventional AR animal models, whereas disease-syndrome integrated TCM models represent a relatively small but rapidly expanding subset of the literature. Standardization of naturally relevant allergen models, development of objective syndrome-reflective biomarkers, and establishment of a "symptom-immunology-omics" multidimensional evaluation system are urgently needed to enhance the reliability and translational value of TCM syndrome animal models in AR research.
We provide a comprehensive phenotypic characterization of loss-of-function (LoF) variants in WDFY3 based on the largest cohort reported to date (n = 32). Our findings define a monogenic disorder marked by neuropsychiatric features (including autism and ADHD), mild to moderate neurodevelopmental delay, and variable brain growth - most commonly macrocephaly and in one case reduced head circumference. To investigate the effect of WDFY3 LoF we performed knockdown (KD) in the human neuroblastoma cell line SH-SY5Y and used a Wdfy3-haploinsufficient mouse model. WDFY3 KD impaired clearance of protein aggregates and decreased protein levels of the ubiquitin-binding protein p62. Transcriptomic analyses identified dysregulation in gene networks that are primarily involved in nervous system development, neuron projection, MAPK/ERK1/2, and WNT/β-catenin signaling. This was confirmed at protein level. Neuronal cell proliferation was increased, as evidenced by elevated cell counts and proliferation markers Ki-67 and PCNA. Conversely, neuronal cell differentiation was impaired, as indicated by reduced neurite outgrowth. We show that β-catenin is downregulated, while GSK3B is upregulated. LiCl activates WNT/β-catenin signaling by inhibiting GSK3B. We thus tested LiCl treatment, which significantly increased WDFY3 expression. Moreover, in vitro treatment improved the neurite branching capacity of Wdfy3-haploinsufficient neurons above the level of the wild type, although this specific effect appeared independent of WNT/β-catenin signaling. In sum, we expand on the phenotype related to WDFY3 LoF and provide insights into the pathophysiology using in vitro human and murine neuronal models. Our findings suggest that LiCl may serve as a potential therapeutic strategy for patients with WDFY3 LoF variants.