Department of Pediatrics, Section of Neurology and Developmental Neuroscience, Baylor College of Medicine, Houston, TX, USA; Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, Houston, TX, USA; Cain Pediatric Neurology Research Foundation Laboratories, Jan and Dan Duncan Neurological Research Institute, Houston, TX, USA.
Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, Houston, TX, USA; Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.
Am J Hum Genet. 2024 Jan 4;111(1):96-118. doi: 10.1016/j.ajhg.2023.12.004.
PPFIA3 encodes the protein-tyrosine phosphatase, receptor-type, F-polypeptide-interacting-protein-alpha-3 (PPFIA3), which is a member of the LAR-protein-tyrosine phosphatase-interacting-protein (liprin) family involved in synapse formation and function, synaptic vesicle transport, and presynaptic active zone assembly. The protein structure and function are evolutionarily well conserved, but human diseases related to PPFIA3 dysfunction are not yet reported in OMIM. Here, we report 20 individuals with rare PPFIA3 variants (19 heterozygous and 1 compound heterozygous) presenting with developmental delay, intellectual disability, hypotonia, dysmorphisms, microcephaly or macrocephaly, autistic features, and epilepsy with reduced penetrance. Seventeen unique PPFIA3 variants were detected in 18 families. To determine the pathogenicity of PPFIA3 variants in vivo, we generated transgenic fruit flies producing either human wild-type (WT) PPFIA3 or five missense variants using GAL4-UAS targeted gene expression systems. In the fly overexpression assays, we found that the PPFIA3 variants in the region encoding the N-terminal coiled-coil domain exhibited stronger phenotypes compared to those affecting the C-terminal region. In the loss-of-function fly assay, we show that the homozygous loss of fly Liprin-α leads to embryonic lethality. This lethality is partially rescued by the expression of human PPFIA3 WT, suggesting human PPFIA3 function is partially conserved in the fly. However, two of the tested variants failed to rescue the lethality at the larval stage and one variant failed to rescue lethality at the adult stage. Altogether, the human and fruit fly data reveal that the rare PPFIA3 variants are dominant-negative loss-of-function alleles that perturb multiple developmental processes and synapse formation.
PPFIA3 编码蛋白酪氨酸磷酸酶,受体型,F-多肤-相互作用蛋白-α-3(PPFIA3),它是 LAR-蛋白酪氨酸磷酸酶相互作用蛋白(liprin)家族的成员,参与突触形成和功能、突触小泡运输以及突触前活性区组装。该蛋白结构和功能在进化上高度保守,但 OMIM 中尚未报道与 PPFIA3 功能障碍相关的人类疾病。在这里,我们报告了 20 名个体携带罕见的 PPFIA3 变异(19 个杂合子和 1 个复合杂合子),表现为发育迟缓、智力残疾、张力减退、畸形、小头或大头、自闭症特征和癫痫,外显率降低。在 18 个家庭中检测到 17 个独特的 PPFIA3 变异。为了确定 PPFIA3 变异在体内的致病性,我们使用 GAL4-UAS 靶向基因表达系统生成了产生人野生型(WT)PPFIA3 或五种错义变异的转基因果蝇。在果蝇过表达实验中,我们发现编码 N 端卷曲螺旋结构域的 PPFIA3 变异表现出比影响 C 端区域的变异更强的表型。在功能丧失果蝇实验中,我们表明果蝇 Liprin-α 的纯合缺失导致胚胎致死。人 PPFIA3 WT 的表达部分挽救了这种致死性,表明人 PPFIA3 功能在果蝇中部分保守。然而,两种测试的变异在幼虫阶段未能挽救致死性,一种变异在成虫阶段未能挽救致死性。总之,人类和果蝇的数据表明,罕见的 PPFIA3 变异是显性负性功能丧失等位基因,扰乱了多个发育过程和突触形成。