Pérez Baca María Del Rocío, Palomares-Bralo María, Vanhooydonck Michiel, Hamerlinck Lisa, D'haene Eva, Leimbacher Sebastian, Jacobs Eva Z, De Cock Laurenz, D'haenens Erika, Dheedene Annelies, Malfait Zoë, Vantomme Lies, Silva Ananilia, Rooney Kathleen, Zhao Xiaonan, Saeidian Amir Hossein, Owen Nichole Marie, Santos-Simarro Fernando, Lleuger-Pujol Roser, García-Miñaúr Sixto, Losantos-García Itsaso, Menten Björn, Gestri Gaia, Ragge Nicola, Sadikovic Bekim, Bogaert Elke, Vleminckx Kris, Naert Thomas, Syx Delfien, Callewaert Bert, Vergult Sarah
Center for Medical Genetics Ghent, Ghent University Hospital, Ghent, Belgium; Department of Biomolecular Medicine, Ghent University, Ghent, Belgium.
CIBERER-ISCIII and INGEMM, Institute of Medical and Molecular Genetics, Hospital Universitario La Paz, Madrid, Spain; ITHACA-European Reference Network, Madrid, Spain.
Am J Hum Genet. 2025 Jun 5;112(6):1388-1414. doi: 10.1016/j.ajhg.2025.04.008. Epub 2025 May 13.
8q21.11 microdeletions involving ZFHX4 have previously been associated with a syndromic form of intellectual disability, hypotonia, unstable gait, and hearing loss. We report on 63 individuals-57 probands and 6 affected family members-with protein-truncating variants (n = 41), (micro)deletions (n = 21), or an inversion (n = 1) affecting ZFHX4. Probands display variable developmental delay and intellectual disability, distinctive facial characteristics, morphological abnormalities of the central nervous system, behavioral alterations, short stature, hypotonia, and occasionally cleft palate and anterior segment dysgenesis. The phenotypes associated with 8q21.11 microdeletions and ZFHX4 intragenic loss-of-function (LoF) variants largely overlap, although leukocyte-derived DNA shows a mild common methylation profile for (micro)deletions. ZFHX4 shows increased expression during human brain development and neuronal differentiation. Furthermore, ZFHX4-interacting factors identified via immunoprecipitation followed by mass spectrometry (IP-MS) suggest an important role for ZFHX4 in cellular pathways, especially during histone modifications, protein trafficking, signal transduction, cytosolic transport, and development. Additionally, using CUT&RUN, we observed that ZFHX4 binds the promoter of genes with crucial roles in embryonic, neuronal, and axonal development. Moreover, we investigated whether the disruption of zfhx4 causes craniofacial abnormalities in zebrafish. First-generation (F0) zfhx4 crispant zebrafish, a (mosaic) mutant for zfhx4 LoF variants, have significantly shorter Meckel's cartilage and smaller ethmoid plates compared to control zebrafish. Behavioral assays showed a decreased movement frequency in the zfhx4 crispant zebrafish in comparison with controls. Furthermore, structural abnormalities were found in the zebrafish hindbrain. In conclusion, our findings delineate a ZFHX4-associated neurodevelopmental disorder and suggest a role for zfhx4 in facial skeleton patterning, palatal development, and behavior.
涉及ZFHX4的8q21.11微缺失先前已与一种伴有智力残疾、肌张力减退、步态不稳和听力丧失的综合征形式相关联。我们报告了63名个体——57名先证者和6名受影响的家庭成员，他们携带影响ZFHX4的蛋白质截短变体（n = 41）、（微）缺失（n = 21）或倒位（n = 1）。先证者表现出不同程度的发育迟缓、智力残疾、独特的面部特征、中枢神经系统形态异常、行为改变、身材矮小、肌张力减退，偶尔还伴有腭裂和眼前节发育异常。与8q21.11微缺失和ZFHX4基因内功能丧失（LoF）变体相关的表型在很大程度上重叠，尽管白细胞衍生的DNA显示（微）缺失有轻度的共同甲基化谱。ZFHX4在人类大脑发育和神经元分化过程中表达增加。此外，通过免疫沉淀后质谱分析（IP-MS）鉴定出的ZFHX4相互作用因子表明ZFHX4在细胞途径中起重要作用，尤其是在组蛋白修饰、蛋白质运输、信号转导、胞质运输和发育过程中。此外，使用CUT&RUN技术，我们观察到ZFHX4结合在胚胎、神经元和轴突发育中起关键作用的基因的启动子。此外，我们研究了zfhx4的破坏是否会导致斑马鱼出现颅面异常。第一代（F0）zfhx4基因敲降斑马鱼是zfhx4 LoF变体的（嵌合）突变体，与对照斑马鱼相比，其Meckel软骨明显更短，筛骨板更小。行为分析表明，与对照相比，zfhx4基因敲降斑马鱼的运动频率降低。此外，在斑马鱼后脑发现了结构异常。总之，我们的研究结果描绘了一种与ZFHX4相关的神经发育障碍，并表明zfhx4在面部骨骼模式形成、腭部发育和行为中起作用。
