Adnp-mutant mice with cognitive inflexibility, CaMKIIα hyperactivity, and synaptic plasticity deficits

Cho, Heejin; Yoo, Taesun; Moon, Hyang Mi; Kang, Hyojin; Yang, Yeji; Kang, MinSoung; Yang, Esther; Lee, Dowoon; Hwang, Daehee; Kim, H.; Kim, Doyoun; Kim, Jin Young; Kim, Eunjoon

doi:10.1038/s41380-023-02129-5

Cited by 6 publications

(7 citation statements)

References 96 publications

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“…We also uncovered the nuclear localization and transcription factor activities of ADNP, as it regulates the chromatin structure and is implicated in the expression of hundreds of essential genes [ 4 , 7 , 8 , 9 , 10 , 11 , 12 ]. These findings have been substantiated and extended by others [ 6 , 13 , 14 , 15 , 16 , 17 , 18 , 19 ]. Most recent clinical findings include the association of de novo ADNP mutations in congenital cranial anomalies [ 20 ].…”

Section: Introductionsupporting

confidence: 79%

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NAP (Davunetide): The Neuroprotective ADNP Drug Candidate Penetrates Cell Nuclei Explaining Pleiotropic Mechanisms

Ganaiem,

Gildor,

Shazman

et al. 2023

Cells

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(1) Background: Recently, we showed aberrant nuclear/cytoplasmic boundaries/activity-dependent neuroprotective protein (ADNP) distribution in ADNP-mutated cells. This malformation was corrected upon neuronal differentiation by the ADNP-derived fragment drug candidate NAP (davunetide). Here, we investigated the mechanism of NAP nuclear protection. (2) Methods: CRISPR/Cas9 DNA-editing established N1E-115 neuroblastoma cell lines that express two different green fluorescent proteins (GFPs)—labeled mutated ADNP variants (p.Tyr718* and p.Ser403*). Cells were exposed to NAP conjugated to Cy5, followed by live imaging. Cells were further characterized using quantitative morphology/immunocytochemistry/RNA and protein quantifications. (3) Results: NAP rapidly distributed in the cytoplasm and was also seen in the nucleus. Furthermore, reduced microtubule content was observed in the ADNP-mutated cell lines. In parallel, disrupting microtubules by zinc or nocodazole intoxication mimicked ADNP mutation phenotypes and resulted in aberrant nuclear–cytoplasmic boundaries, which were rapidly corrected by NAP treatment. No NAP effects were noted on ADNP levels. Ketamine, used as a control, was ineffective, but both NAP and ketamine exhibited direct interactions with ADNP, as observed via in silico docking. (4) Conclusions: Through a microtubule-linked mechanism, NAP rapidly localized to the cytoplasmic and nuclear compartments, ameliorating mutated ADNP-related deficiencies. These novel findings explain previously published gene expression results and broaden NAP (davunetide) utilization in research and clinical development.

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Section: Introductionsupporting

confidence: 79%

“…We also uncovered the nuclear localization and transcription factor activities of ADNP, as it regulates the chromatin structure and is implicated in the expression of hundreds of essential genes [4,[7][8][9][10][11][12]. These findings have been substantiated and extended by others [6,[13][14][15][16][17][18][19].…”

Section: Introductionmentioning

confidence: 64%

NAP (Davunetide): The Neuroprotective ADNP Drug Candidate Penetrates Cell Nuclei Explaining Pleiotropic Mechanisms

Ganaiem,

Gildor,

Shazman

et al. 2023

Cells

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“…In the literature, the mutation of p.Y719* has been extensively studied [ 8 , 18 , 24 , 26 , 39 , 54 ], which was detected in 2 out of 15 cases (p.Y719*, c.2157C>G; p.Y719*, c.2157C>A) in current study. This variant, located on NLS, is a nonsense variant that results in a truncated protein lacking the homeobox domain and HP1-binding motif.…”

Section: Discussionmentioning

confidence: 49%

Clinical impact and in vitro characterization of ADNP variants in pediatric patients

Ge,

Tian,

et al. 2024

Molecular Autism

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Background 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. Methods 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. Results 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. Limitations 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. Conclusions 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.

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“…To characterize functions for the ChAHP complex, previous investigations have focused on ADNP mutant models based on the argument that ADNP is an exclusive and essential component of ChAHP 2,15 . However, ADNP mutants arrest at the onset of neurodevelopment 2,20,29,30,49,50 , precluding genetic analysis of later processes. To bypass early lethality, we generated telencephalon-specific Adnp and Chd4 cKOs.…”

Section: Discussionmentioning

confidence: 99%

“…An additional limitation is that we focused our genomic and transcriptomic profiling on embryonic stages of development. Indeed, a recent study performed transcriptomic profiling of brains from germline Adnp heterozygotes at juvenile and adult stages 49 . Few if any DEGs appear to be common between the two studies.…”

Section: The Chahp Complex and Neurodevelopmental Gene Regulationmentioning

confidence: 99%

The ChAHP chromatin remodelling complex regulates neurodevelopmental disorder risk genes to scale the production of neocortical layers

Clémot-Dupont,

Lourenço Fernandes,

Larrigan

et al. 2024

Preprint

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Although chromatin remodellers are among the most important risk genes associated with neurodevelopmental disorders (NDDs), the roles of these complexes during brain development are in many cases unclear. Here, we focused on the recently discovered ChAHP chromatin remodelling complex. The zinc finger and homeodomain transcription factor ADNP is a core subunit of this complex, and de novo ADNP mutations lead to intellectual disability and autism spectrum disorder. However, germline Adnp knockout mice were previously shown to exhibit early embryonic lethality, obscuring subsequent roles for the ChAHP complex in neurogenesis. Here, we employed single cell transcriptomics, cut&run-seq, and histological approaches to characterize mice conditionally ablated for the ChAHP subunits Adnp and Chd4. We show that during neocortical development, Adnp and Chd4 orchestrate the production of late-born, upper-layer neurons through a two-step process. First, Adnp is required to sustain progenitor proliferation specifically during the developmental window for upper-layer cortical neurogenesis. Accordingly, we found that Adnp recruits Chd4 to genes associated with progenitor proliferation. Second, in postmitotic differentiated neurons, we define a network of risk genes linked to NDDs that are regulated by Adnp and Chd4. Taken together, these data demonstrate that ChAHP is critical for driving the expansion upper-layer cortical neurons, and for regulating neuronal gene expression programs, suggesting that these processes may potentially contribute to NDD etiology.

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Adnp-mutant mice with cognitive inflexibility, CaMKIIα hyperactivity, and synaptic plasticity deficits

Cited by 6 publications

References 96 publications

NAP (Davunetide): The Neuroprotective ADNP Drug Candidate Penetrates Cell Nuclei Explaining Pleiotropic Mechanisms

NAP (Davunetide): The Neuroprotective ADNP Drug Candidate Penetrates Cell Nuclei Explaining Pleiotropic Mechanisms

Clinical impact and in vitro characterization of ADNP variants in pediatric patients

The ChAHP chromatin remodelling complex regulates neurodevelopmental disorder risk genes to scale the production of neocortical layers

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