PDF(Pubmed)
Abstract：
Helsmoortel-Van der Aa syndrome (HVDAS) is a rare genetic disorder caused by variants in the activity-dependent neuroprotector homeobox (ADNP) gene; hence, it is also called ADNP syndrome. ADNP is a multitasking protein with the function as a transcription factor, playing a critical role in brain development. Furthermore, ADNP variants have been identified as one of the most common single-gene causes of autism spectrum disorder (ASD) and intellectual disability.
We assembled a cohort of 15 Chinese pediatric patients, identified 13 variants in the coding region of ADNP gene, and evaluated their clinical phenotypes. Additionally, we constructed the corresponding ADNP variants and performed western blotting and immunofluorescence analysis to examine their protein expression and subcellular localization in human HEK293T and SH-SY5Y cells.
Our study conducted a thorough characterization of the clinical manifestations in 15 children with ADNP variants, and revealed a broad spectrum of symptoms including global developmental delay, intellectual disability, ASD, facial abnormalities, and other features. In vitro studies were carried out to check the expression of ADNP with identified variants. Two cases presented missense variants, while the remainder exhibited nonsense or frameshift variants, leading to truncated mutants in in vitro overexpression systems. Both overexpressed wildtype ADNP and all the different mutants were found to be confined to the nuclei in HEK293T cells; however, the distinctive pattern of nuclear bodies formed by the wildtype ADNP was either partially or entirely disrupted by the mutant proteins. Moreover, two variants of p.Y719* on the nuclear localization signal (NLS) of ADNP disrupted the nuclear expression pattern, predominantly manifesting in the cytoplasm in SH-SY5Y cells.
Our study was limited by a relatively small sample size and the absence of a longitudinal framework to monitor the progression of patient conditions over time. Additionally, we lacked in vivo evidence to further indicate the causal implications of the identified ADNP variants.
Our study reported the first cohort of HVDAS patients in the Chinese population and provided systematic clinical presentations and laboratory examinations. Furthermore, we identified multiple genetic variants and validated them in vitro. Our findings offered valuable insights into the diverse genetic variants associated with HVDAS.
We assembled a cohort of 15 Chinese pediatric patients, identified 13 variants in the coding region of ADNP gene, and evaluated their clinical phenotypes. Additionally, we constructed the corresponding ADNP variants and performed western blotting and immunofluorescence analysis to examine their protein expression and subcellular localization in human HEK293T and SH-SY5Y cells.
Our study conducted a thorough characterization of the clinical manifestations in 15 children with ADNP variants, and revealed a broad spectrum of symptoms including global developmental delay, intellectual disability, ASD, facial abnormalities, and other features. In vitro studies were carried out to check the expression of ADNP with identified variants. Two cases presented missense variants, while the remainder exhibited nonsense or frameshift variants, leading to truncated mutants in in vitro overexpression systems. Both overexpressed wildtype ADNP and all the different mutants were found to be confined to the nuclei in HEK293T cells; however, the distinctive pattern of nuclear bodies formed by the wildtype ADNP was either partially or entirely disrupted by the mutant proteins. Moreover, two variants of p.Y719* on the nuclear localization signal (NLS) of ADNP disrupted the nuclear expression pattern, predominantly manifesting in the cytoplasm in SH-SY5Y cells.
Our study was limited by a relatively small sample size and the absence of a longitudinal framework to monitor the progression of patient conditions over time. Additionally, we lacked in vivo evidence to further indicate the causal implications of the identified ADNP variants.
Our study reported the first cohort of HVDAS patients in the Chinese population and provided systematic clinical presentations and laboratory examinations. Furthermore, we identified multiple genetic variants and validated them in vitro. Our findings offered valuable insights into the diverse genetic variants associated with HVDAS.
摘要：
背景：Helsmoortel-VanderAa综合征（HVDAS）是一种罕见的遗传性疾病，由活动依赖性神经保护者同源盒（ADNP）基因变异引起；因此，它也被称为ADNP综合征。ADNP是一种多任务蛋白，具有转录因子的功能，在大脑发育中起关键作用。此外，ADNP变体已被确定为自闭症谱系障碍（ASD）和智力障碍的最常见单基因原因之一。
方法：我们收集了一个由15名中国儿科患者组成的队列，在ADNP基因的编码区鉴定出13个变异体，并评估其临床表型。此外,我们构建了相应的ADNP变体,并进行了蛋白质印迹和免疫荧光分析,以检测它们在人HEK293T和SH-SY5Y细胞中的蛋白表达和亚细胞定位.
结果：我们的研究对15例ADNP变异患儿的临床表现进行了彻底的表征，并揭示了广泛的症状，包括全球发育迟缓，智力残疾,ASD,面部异常,和其他功能。进行体外研究以检查具有鉴定的变体的ADNP的表达。两个案例呈现错义变体，而其余的则表现出无义或移码变体，在体外过表达系统中导致截短的突变体。发现过表达的野生型ADNP和所有不同的突变体都被限制在HEK293T细胞的细胞核内；然而，由野生型ADNP形成的核体的独特模式被突变蛋白部分或全部破坏。此外，在ADNP的核定位信号（NLS）上p.Y719*的两个变体破坏了核表达模式，主要表现在SH-SY5Y细胞的细胞质中。
结论：我们的研究受限于相对较小的样本量和缺乏纵向框架来监测患者病情随时间的进展。此外,我们缺乏体内证据来进一步表明已鉴定的ADNP变体的因果关系.
结论：我们的研究报告了中国人群中第一个HVDAS患者队列，并提供了系统的临床表现和实验室检查。此外，我们鉴定了多种遗传变异并在体外进行了验证.我们的发现为与HVDAS相关的各种遗传变异提供了有价值的见解。
方法：我们收集了一个由15名中国儿科患者组成的队列，在ADNP基因的编码区鉴定出13个变异体，并评估其临床表型。此外,我们构建了相应的ADNP变体,并进行了蛋白质印迹和免疫荧光分析,以检测它们在人HEK293T和SH-SY5Y细胞中的蛋白表达和亚细胞定位.
结果：我们的研究对15例ADNP变异患儿的临床表现进行了彻底的表征，并揭示了广泛的症状，包括全球发育迟缓，智力残疾,ASD,面部异常,和其他功能。进行体外研究以检查具有鉴定的变体的ADNP的表达。两个案例呈现错义变体，而其余的则表现出无义或移码变体，在体外过表达系统中导致截短的突变体。发现过表达的野生型ADNP和所有不同的突变体都被限制在HEK293T细胞的细胞核内；然而，由野生型ADNP形成的核体的独特模式被突变蛋白部分或全部破坏。此外，在ADNP的核定位信号（NLS）上p.Y719*的两个变体破坏了核表达模式，主要表现在SH-SY5Y细胞的细胞质中。
结论：我们的研究受限于相对较小的样本量和缺乏纵向框架来监测患者病情随时间的进展。此外,我们缺乏体内证据来进一步表明已鉴定的ADNP变体的因果关系.
结论：我们的研究报告了中国人群中第一个HVDAS患者队列，并提供了系统的临床表现和实验室检查。此外，我们鉴定了多种遗传变异并在体外进行了验证.我们的发现为与HVDAS相关的各种遗传变异提供了有价值的见解。
