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<!DOCTYPE html> <html lang="en" class="grade-c"> <head> <title>The autism-associated gene chromodomain helicase DNA-binding protein 8 (CHD8) regulates noncoding RNAs and autism-related genes | Translational Psychiatry</title> <link rel="alternate" type="application/rss+xml" href="https://www.nature.com/tp.rss"/> <link rel="preconnect" href="https://cmp.nature.com" crossorigin> <meta http-equiv="X-UA-Compatible" content="IE=edge"> <meta name="applicable-device" content="pc,mobile"> <meta name="viewport" content="width=device-width,initial-scale=1.0,maximum-scale=5,user-scalable=yes"> <meta name="360-site-verification" content="5a2dc4ab3fcb9b0393241ffbbb490480" /> <script data-test="dataLayer"> window.dataLayer = [{"content":{"category":{"contentType":"original article","legacy":{"webtrendsPrimaryArticleType":"research","webtrendsSubjectTerms":"autism-spectrum-disorders","webtrendsContentCategory":null,"webtrendsContentCollection":null,"webtrendsContentGroup":"Translational 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name="dc.title" content="The autism-associated gene chromodomain helicase DNA-binding protein 8 (CHD8) regulates noncoding RNAs and autism-related genes"/> <meta name="dc.source" content="Translational Psychiatry 2015 5:5"/> <meta name="dc.format" content="text/html"/> <meta name="dc.publisher" content="Nature Publishing Group"/> <meta name="dc.date" content="2015-05-19"/> <meta name="dc.type" content="OriginalPaper"/> <meta name="dc.language" content="En"/> <meta name="dc.copyright" content="2015 The Author(s)"/> <meta name="dc.rights" content="2015 The Author(s)"/> <meta name="dc.rightsAgent" content="journalpermissions@springernature.com"/> <meta name="dc.description" content="Chromodomain helicase DNA-binding protein 8 (CHD8) was identified as a leading autism spectrum disorder (ASD) candidate gene by whole-exome sequencing and subsequent targeted-sequencing studies. 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data-test="article-identifier"> <li class="c-article-identifiers__item" data-test="article-category">Original Article</li> <li class="c-article-identifiers__item"> <a href="https://www.springernature.com/gp/open-research/about/the-fundamentals-of-open-access-and-open-research" data-track="click" data-track-action="open access" data-track-label="link" class="u-color-open-access" data-test="open-access">Open access</a> </li> <li class="c-article-identifiers__item">Published: <time datetime="2015-05-19">19 May 2015</time></li> </ul> <h1 class="c-article-title" data-test="article-title" data-article-title="">The autism-associated gene chromodomain helicase DNA-binding protein 8 (<i>CHD8</i>) regulates noncoding RNAs and autism-related genes</h1> <ul class="c-article-author-list c-article-author-list--short" data-test="authors-list" data-component-authors-activator="authors-list"><li class="c-article-author-list__item"><a data-test="author-name" data-track="click" data-track-action="open author" data-track-label="link" href="#auth-B-Wilkinson-Aff1" data-author-popup="auth-B-Wilkinson-Aff1" data-author-search="Wilkinson, B">B Wilkinson</a><sup class="u-js-hide"><a href="#Aff1">1</a></sup>, </li><li class="c-article-author-list__item"><a data-test="author-name" data-track="click" data-track-action="open author" data-track-label="link" href="#auth-N-Grepo-Aff2" data-author-popup="auth-N-Grepo-Aff2" data-author-search="Grepo, N">N Grepo</a><sup class="u-js-hide"><a href="#Aff2">2</a></sup>, </li><li class="c-article-author-list__item c-article-author-list__item--hide-small-screen"><a data-test="author-name" data-track="click" data-track-action="open author" data-track-label="link" href="#auth-B_L-Thompson-Aff3" data-author-popup="auth-B_L-Thompson-Aff3" data-author-search="Thompson, B L">B L Thompson</a><sup class="u-js-hide"><a href="#Aff3">3</a></sup>, </li><li class="c-article-author-list__item c-article-author-list__item--hide-small-screen"><a data-test="author-name" data-track="click" data-track-action="open author" data-track-label="link" href="#auth-J-Kim-Aff2-Aff4" data-author-popup="auth-J-Kim-Aff2-Aff4" data-author-search="Kim, J">J Kim</a><sup class="u-js-hide"><a href="#Aff2">2</a>,<a href="#Aff4">4</a></sup>, </li><li class="c-article-author-list__item c-article-author-list__item--hide-small-screen"><a data-test="author-name" data-track="click" data-track-action="open author" data-track-label="link" href="#auth-K-Wang-Aff2-Aff4" data-author-popup="auth-K-Wang-Aff2-Aff4" data-author-search="Wang, K">K Wang</a><sup class="u-js-hide"><a href="#Aff2">2</a>,<a href="#Aff4">4</a></sup>, </li><li class="c-article-author-list__item c-article-author-list__item--hide-small-screen"><a data-test="author-name" data-track="click" data-track-action="open author" data-track-label="link" href="#auth-O_V-Evgrafov-Aff2-Aff4" data-author-popup="auth-O_V-Evgrafov-Aff2-Aff4" data-author-search="Evgrafov, O V">O V Evgrafov</a><sup class="u-js-hide"><a href="#Aff2">2</a>,<a href="#Aff4">4</a></sup>, </li><li class="c-article-author-list__item c-article-author-list__item--hide-small-screen"><a data-test="author-name" data-track="click" data-track-action="open author" data-track-label="link" href="#auth-W-Lu-Aff5-Aff6" data-author-popup="auth-W-Lu-Aff5-Aff6" data-author-search="Lu, W">W Lu</a><sup class="u-js-hide"><a href="#Aff5">5</a>,<a href="#Aff6">6</a></sup>, </li><li class="c-article-author-list__item c-article-author-list__item--hide-small-screen"><a data-test="author-name" data-track="click" data-track-action="open author" data-track-label="link" href="#auth-J_A-Knowles-Aff2-Aff4" data-author-popup="auth-J_A-Knowles-Aff2-Aff4" data-author-search="Knowles, J A">J A Knowles</a><sup class="u-js-hide"><a href="#Aff2">2</a>,<a href="#Aff4">4</a></sup> & </li><li class="c-article-author-list__show-more" aria-label="Show all 9 authors for this article" title="Show all 9 authors for this article">…</li><li class="c-article-author-list__item"><a data-test="author-name" data-track="click" data-track-action="open author" data-track-label="link" href="#auth-D_B-Campbell-Aff2-Aff4" data-author-popup="auth-D_B-Campbell-Aff2-Aff4" data-author-search="Campbell, D B" data-corresp-id="c1">D B Campbell<svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-mail-medium"></use></svg></a><sup class="u-js-hide"><a href="#Aff2">2</a>,<a href="#Aff4">4</a></sup> </li></ul><button aria-expanded="false" class="c-article-author-list__button"><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-down-medium"></use></svg><span>Show authors</span></button> <p class="c-article-info-details" data-container-section="info"> <a data-test="journal-link" href="/tp" data-track="click" data-track-action="journal homepage" data-track-category="article body" data-track-label="link"><i data-test="journal-title">Translational Psychiatry</i></a> <b data-test="journal-volume"><span class="u-visually-hidden">volume</span> 5</b>, <span class="u-visually-hidden">page </span>e568 (<span data-test="article-publication-year">2015</span>)<a href="#citeas" class="c-article-info-details__cite-as u-hide-print" data-track="click" data-track-action="cite this article" data-track-label="link">Cite this article</a> </p> <div class="c-article-metrics-bar__wrapper u-clear-both"> <ul class="c-article-metrics-bar u-list-reset"> <li class=" c-article-metrics-bar__item" data-test="access-count"> <p class="c-article-metrics-bar__count">8113 <span class="c-article-metrics-bar__label">Accesses</span></p> </li> <li class="c-article-metrics-bar__item" data-test="altmetric-score"> <p class="c-article-metrics-bar__count">6 <span class="c-article-metrics-bar__label">Altmetric</span></p> </li> <li class="c-article-metrics-bar__item"> <p class="c-article-metrics-bar__details"><a href="/articles/tp201562/metrics" data-track="click" data-track-action="view metrics" data-track-label="link" rel="nofollow">Metrics <span class="u-visually-hidden">details</span></a></p> </li> </ul> </div> </header> <div class="u-js-hide" data-component="article-subject-links"> <h3 class="c-article__sub-heading">Subjects</h3> <ul class="c-article-subject-list"> <li class="c-article-subject-list__subject"><a href="/subjects/autism-spectrum-disorders" data-track="click" data-track-action="view subject" data-track-label="link">Autism spectrum disorders</a></li> </ul> </div> </div> <div class="c-article-body"> <section aria-labelledby="Abs1" data-title="Abstract" lang="en"><div class="c-article-section" id="Abs1-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Abs1">Abstract</h2><div class="c-article-section__content" id="Abs1-content"><p>Chromodomain helicase DNA-binding protein 8 (<i>CHD8</i>) was identified as a leading autism spectrum disorder (ASD) candidate gene by whole-exome sequencing and subsequent targeted-sequencing studies. <i>De novo</i> loss-of-function mutations were identified in 12 individuals with ASD and zero controls, accounting for a highly significant association. Small interfering RNA-mediated knockdown of <i>CHD8</i> in human neural progenitor cells followed by RNA sequencing revealed that CHD8 insufficiency results in altered expression of 1715 genes, including both protein-coding and noncoding RNAs. Among the 10 most changed transcripts, 4 (40%) were noncoding RNAs. The transcriptional changes among protein-coding genes involved a highly interconnected network of genes that are enriched in neuronal development and in previously identified ASD candidate genes. These results suggest that CHD8 insufficiency may be a central hub in neuronal development and ASD risk.</p></div></div></section> <section aria-labelledby="inline-recommendations" data-title="Inline Recommendations" class="c-article-recommendations" data-track-component="inline-recommendations"> <h3 class="c-article-recommendations-title" id="inline-recommendations">Similar content being viewed by others</h3> <div class="c-article-recommendations-list"> <div class="c-article-recommendations-list__item"> <article class="c-article-recommendations-card" itemscope itemtype="http://schema.org/ScholarlyArticle"> <div class="c-article-recommendations-card__img"><img src="https://media.springernature.com/w215h120/springer-static/image/art%3A10.1038%2Fs41588-022-01104-0/MediaObjects/41588_2022_1104_Fig1_HTML.png" loading="lazy" alt=""></div> <div class="c-article-recommendations-card__main"> <h3 class="c-article-recommendations-card__heading" itemprop="name headline"> <a class="c-article-recommendations-card__link" itemprop="url" href="https://www.nature.com/articles/s41588-022-01104-0?fromPaywallRec=false" data-track="select_recommendations_1" data-track-context="inline recommendations" data-track-action="click recommendations inline - 1" data-track-label="10.1038/s41588-022-01104-0">Rare coding variation provides insight into the genetic architecture and phenotypic context of autism </a> </h3> <div class="c-article-meta-recommendations" data-test="recommendation-info"> <span class="c-article-meta-recommendations__item-type">Article</span> <span class="c-article-meta-recommendations__date">18 August 2022</span> </div> </div> </article> </div> <div class="c-article-recommendations-list__item"> <article class="c-article-recommendations-card" itemscope itemtype="http://schema.org/ScholarlyArticle"> <div class="c-article-recommendations-card__img"><img src="https://media.springernature.com/w215h120/springer-static/image/art%3A10.1038%2Fs41598-020-70656-0/MediaObjects/41598_2020_70656_Fig1_HTML.png" loading="lazy" alt=""></div> <div class="c-article-recommendations-card__main"> <h3 class="c-article-recommendations-card__heading" itemprop="name headline"> <a class="c-article-recommendations-card__link" itemprop="url" href="https://www.nature.com/articles/s41598-020-70656-0?fromPaywallRec=false" data-track="select_recommendations_2" data-track-context="inline recommendations" data-track-action="click recommendations inline - 2" data-track-label="10.1038/s41598-020-70656-0">Homozygous deletions implicate non-coding epigenetic marks in Autism spectrum disorder </a> </h3> <div class="c-article-meta-recommendations" data-test="recommendation-info"> <span class="c-article-meta-recommendations__item-type">Article</span> <span class="c-article-meta-recommendations__access-type">Open access</span> <span class="c-article-meta-recommendations__date">20 August 2020</span> </div> </div> </article> </div> <div class="c-article-recommendations-list__item"> <article class="c-article-recommendations-card" itemscope itemtype="http://schema.org/ScholarlyArticle"> <div class="c-article-recommendations-card__img"><img src="https://media.springernature.com/w215h120/springer-static/image/art%3A10.1038%2Fs41380-024-02806-z/MediaObjects/41380_2024_2806_Fig1_HTML.png" loading="lazy" alt=""></div> <div class="c-article-recommendations-card__main"> <h3 class="c-article-recommendations-card__heading" itemprop="name headline"> <a class="c-article-recommendations-card__link" itemprop="url" href="https://www.nature.com/articles/s41380-024-02806-z?fromPaywallRec=false" data-track="select_recommendations_3" data-track-context="inline recommendations" data-track-action="click recommendations inline - 3" data-track-label="10.1038/s41380-024-02806-z">Monoallelic loss-of-function variants in <i>GSK3B</i> lead to autism and developmental delay </a> </h3> <div class="c-article-meta-recommendations" data-test="recommendation-info"> <span class="c-article-meta-recommendations__item-type">Article</span> <span class="c-article-meta-recommendations__date">29 October 2024</span> </div> </div> </article> </div> </div> </section> <script> window.dataLayer = window.dataLayer || []; window.dataLayer.push({ recommendations: { recommender: 'semantic', model: 'specter', policy_id: 'NA', timestamp: 1739784045, embedded_user: 'null' } }); </script> <div class="main-content"> <section data-title="Introduction"><div class="c-article-section" id="Sec1-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec1">Introduction</h2><div class="c-article-section__content" id="Sec1-content"><p>Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder characterized by impairments in social interaction, communication and behavioral flexibility.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 1" title="American Psychiatric Association and American Psychiatric Association DSM-5 Task Force Diagnostic and Statistical Manual of Mental Disorders: DSM-5. American Psychiatric Association: Arlington, VA, USA, 2013." href="/articles/tp201562#ref-CR1" id="ref-link-section-d446063898e563">1</a></sup> Due to the vast clinical and genetic heterogeneity of ASD, the identification of causal genetic determinants has proven challenging.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 2" title="Krumm N, O'Roak BJ, Shendure J, Eichler EE . A de novo convergence of autism genetics and molecular neuroscience. Trends Neurosci 2014; 37: 95–105." href="/articles/tp201562#ref-CR2" id="ref-link-section-d446063898e567">2</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 3" title="Jeste SS, Geschwind DH . Disentangling the heterogeneity of autism spectrum disorder through genetic findings. Nat Rev Neurol 2014; 10: 74–81." href="/articles/tp201562#ref-CR3" id="ref-link-section-d446063898e570">3</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 4" title="Levitt P, Campbell DB . The genetic and neurobiologic compass points toward common signaling dysfunctions in autism spectrum disorders. J Clin Invest 2009; 119: 747–754." href="/articles/tp201562#ref-CR4" id="ref-link-section-d446063898e573">4</a></sup> However, multiple independent studies have now provided substantial evidence for the contribution of <i>de novo</i> loss-of-function (LoF) mutations in chromodomain helicase DNA-binding protein 8 (<i>CHD8</i>) to ASD.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 5" title="Sanders SJ, Murtha MT, Gupta AR, Murdoch JD, Raubeson MJ, Willsey AJ et al. De novo mutations revealed by whole-exome sequencing are strongly associated with autism. Nature 2012; 485: 237–241." href="/articles/tp201562#ref-CR5" id="ref-link-section-d446063898e583">5</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 6" title="O'Roak BJ, Vives L, Girirajan S, Karakoc E, Krumm N, Coe BP et al. Sporadic autism exomes reveal a highly interconnected protein network of de novo mutations. Nature 2012; 485: 246–250." href="/articles/tp201562#ref-CR6" id="ref-link-section-d446063898e586">6</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 7" title="Neale BM, Kou Y, Liu L, Ma'ayan A, Samocha KE, Sabo A et al. Patterns and rates of exonic de novo mutations in autism spectrum disorders. Nature 2012; 485: 242–245." href="/articles/tp201562#ref-CR7" id="ref-link-section-d446063898e589">7</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 8" title="O'Roak BJ, Vives L, Fu W, Egertson JD, Stanaway IB, Phelps IG et al. Multiplex targeted sequencing identifies recurrently mutated genes in autism spectrum disorders. Science 2012; 338: 1619–1622." href="/articles/tp201562#ref-CR8" id="ref-link-section-d446063898e592">8</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 9" title="Bernier R, Golzio C, Xiong B, Stessman HA, Coe BP, Penn O et al. Disruptive CHD8 mutations define a subtype of autism early in development. Cell 2014; 158: 263–276." href="/articles/tp201562#ref-CR9" id="ref-link-section-d446063898e595">9</a></sup> Initially, exome sequencing on 209 ASD probands and their unaffected family members discovered two <i>de novo</i> LoF mutations in <i>CHD8</i>.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 6" title="O'Roak BJ, Vives L, Girirajan S, Karakoc E, Krumm N, Coe BP et al. Sporadic autism exomes reveal a highly interconnected protein network of de novo mutations. Nature 2012; 485: 246–250." href="/articles/tp201562#ref-CR6" id="ref-link-section-d446063898e606">6</a></sup> Subsequent targeted sequencing of 44 candidate genes in 2446 ASD probands identified an additional seven <i>de novo</i> LoF mutations in <i>CHD8</i>, establishing a frequency of mutation that is the highest of all genes screened.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 7" title="Neale BM, Kou Y, Liu L, Ma'ayan A, Samocha KE, Sabo A et al. Patterns and rates of exonic de novo mutations in autism spectrum disorders. Nature 2012; 485: 242–245." href="/articles/tp201562#ref-CR7" id="ref-link-section-d446063898e616">7</a></sup> Furthermore, a second targeted-sequencing study of an additional 3370 children with ASD or developmental delay identified three <i>de novo</i> LoF mutations in <i>CHD8</i>.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 8" title="O'Roak BJ, Vives L, Fu W, Egertson JD, Stanaway IB, Phelps IG et al. Multiplex targeted sequencing identifies recurrently mutated genes in autism spectrum disorders. Science 2012; 338: 1619–1622." href="/articles/tp201562#ref-CR8" id="ref-link-section-d446063898e627">8</a></sup> Altogether, these highly significant findings have placed <i>CHD8</i> as a genuine ASD risk factor and account for 0.2% (12/6,176) of ASD cases. The <i>CHD8</i> LoF mutations have been found throughout the coding region of the gene, with truncating mutations as early as amino acid 62 of the 2581 amino acid CHD8 protein. Truncating mutations were found in the chromodomain, the dex domain and the helicase domain. A detailed map of all the identified <i>CHD8</i> LoF mutations was published recently.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 9" title="Bernier R, Golzio C, Xiong B, Stessman HA, Coe BP, Penn O et al. Disruptive CHD8 mutations define a subtype of autism early in development. Cell 2014; 158: 263–276." href="/articles/tp201562#ref-CR9" id="ref-link-section-d446063898e641">9</a></sup> In addition to <i>de novo</i> mutations in <i>CHD8</i>, six potentially disrupting mutations with unknown or inherited modes of transmission have also been identified<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 8" title="O'Roak BJ, Vives L, Fu W, Egertson JD, Stanaway IB, Phelps IG et al. Multiplex targeted sequencing identifies recurrently mutated genes in autism spectrum disorders. Science 2012; 338: 1619–1622." href="/articles/tp201562#ref-CR8" id="ref-link-section-d446063898e651">8</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 9" title="Bernier R, Golzio C, Xiong B, Stessman HA, Coe BP, Penn O et al. Disruptive CHD8 mutations define a subtype of autism early in development. Cell 2014; 158: 263–276." href="/articles/tp201562#ref-CR9" id="ref-link-section-d446063898e654">9</a></sup> along with a balanced chromosomal abnormality leading to disruption of <i>CHD8</i>.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 10" title="Talkowski ME, Rosenfeld JA, Blumenthal I, Pillalamarri V, Chiang C, Heilbut A et al. Sequencing chromosomal abnormalities reveals neurodevelopmental loci that confer risk across diagnostic boundaries. Cell 2012; 149: 525–537." href="/articles/tp201562#ref-CR10" id="ref-link-section-d446063898e661">10</a></sup> <i>CHD8</i> LoF mutations have not been found in any of the 8792 controls included in these analyses, emphasizing the impact of <i>CHD8</i> LoF mutations on ASD risk.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 9" title="Bernier R, Golzio C, Xiong B, Stessman HA, Coe BP, Penn O et al. Disruptive CHD8 mutations define a subtype of autism early in development. Cell 2014; 158: 263–276." href="/articles/tp201562#ref-CR9" id="ref-link-section-d446063898e672">9</a></sup> Phenotypic characterization of individuals with <i>CHD8</i> disrupting mutations indicate a subset of ASD that includes macrocephaly, distinct facial features and gastrointestinal difficulties.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 8" title="O'Roak BJ, Vives L, Fu W, Egertson JD, Stanaway IB, Phelps IG et al. Multiplex targeted sequencing identifies recurrently mutated genes in autism spectrum disorders. Science 2012; 338: 1619–1622." href="/articles/tp201562#ref-CR8" id="ref-link-section-d446063898e679">8</a></sup></p><p>Although a critical role of CHD8 in development is revealed by the embryonic lethality of <i>Chd8</i> knockout mice,<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 11" title="Nishiyama M, Oshikawa K, Tsukada Y, Nakagawa T, Iemura S, Natsume T et al. CHD8 suppresses p53-mediated apoptosis through histone H1 recruitment during early embryogenesis. Nat Cell Biol 2009; 11: 172–182." href="/articles/tp201562#ref-CR11" id="ref-link-section-d446063898e688">11</a></sup> the function of CHD8 in neural cell lineages has been largely unexplored. As CHD8 actively associates with core transcriptional machinery,<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 12" title="Rodríguez-Paredes M, Ceballos-Chávez M, Esteller M, García-Domínguez M, Reyes JC . The chromatin remodeling factor CHD8 interacts with elongating RNA polymerase II and controls expression of the cyclin E2 gene. Nucleic Acids Res 2009; 37: 2449–2460." href="/articles/tp201562#ref-CR12" id="ref-link-section-d446063898e692">12</a></sup> transcription factors<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 13" title="Ishihara K, Oshimura M, Nakao M . CTCF-dependent chromatin insulator is linked to epigenetic remodeling. Mol Cell 2006; 23: 733–742." href="/articles/tp201562#ref-CR13" id="ref-link-section-d446063898e696">13</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 14" title="Subtil-Rodríguez A, Vázquez-Chávez E, Ceballos-Chávez M, Rodríguez-Paredes M, Martín-Subero JI, Esteller M et al. The chromatin remodeller CHD8 is required for E2F-dependent transcription activation of S-phase genes. Nucleic Acids Res 2014; 42: 2185–2196." href="/articles/tp201562#ref-CR14" id="ref-link-section-d446063898e699">14</a></sup> and histone-modifying complexes,<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 15" title="Yates JA, Menon T, Thompson BA, Bochar DA . Regulation of HOXA2 gene expression by the ATP-dependent chromatin remodeling enzyme CHD8. FEBS Lett 2010; 584: 689–693." href="/articles/tp201562#ref-CR15" id="ref-link-section-d446063898e703">15</a></sup> transcriptional dysregulation conferred by CHD8 insufficiency may provide evidence for the neurodevelopmental phenotypes observed in ASD. To emulate the potential effects of the identified LoF mutations, we performed small interfering RNA (siRNA)-mediated knockdown of <i>CHD8</i> followed by genome-wide transcriptional profiling through RNA sequencing (RNA-seq). Here, we show that knockdown of <i>CHD8</i> in SK-N-SH human neural progenitor cells results in altered expression of a highly interconnected network of genes, which are enriched in several processes essential for neuronal development. Remarkably, numerous previously identified ASD candidate genes are also differentially expressed in response to knockdown of <i>CHD8</i>, suggesting a convergence of multiple genes contributing to ASD onset. The data presented here point toward a role of <i>CHD8</i> in maintaining the active transcription of neural-specific genes and begins to elucidate the potential contributions of decreased functional CHD8 to the pathogenesis of ASD.</p></div></div></section><section data-title="Materials and methods"><div class="c-article-section" id="Sec2-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec2">Materials and methods</h2><div class="c-article-section__content" id="Sec2-content"><h3 class="c-article__sub-heading" id="Sec3">Cell culture</h3><p>To measure gene expression in human neural progenitor cells, SK-N-SH cells (American Type Culture Collection; Manassas, VA, USA) were maintained in minimal essential medium supplemented with 10% heat-inactivated fetal bovine serum, 1% penicillin/streptomycin, non-essential amino acids, and 1.5 g l<sup>−1</sup> sodium bicarbonate in 183-cm flasks at 37 °C and 5% CO<sub>2</sub>.</p><h3 class="c-article__sub-heading" id="Sec4">siRNA transfection</h3><p>To determine the impact of CHD8 knockdown on gene expression in human neural progenitor cells, SK-N-SH cells were seeded into six-well 10-cm plates and grown for 24 h (~70% confluency) before transfection. Transfections were carried out with either siRNA silencer select negative control No. 1 (catalog no. 4390843, Ambion/Life Technologies; Carlsbad, CA, USA) or siRNA targeting <i>CHD8</i> (catalog no. 33582, Ambion/Life Technologies)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 16" title="Caldon CE, Sergio CM, Schütte J, Boersma MN, Sutherland RL, Carroll JS et al. Estrogen regulation of cyclin E2 requires cyclin D1 but not c-Myc. Mol Cell Biol 2009; 29: 4623–4639." href="/articles/tp201562#ref-CR16" id="ref-link-section-d446063898e747">16</a></sup> at a concentration of 20 n<span class="u-small-caps">M</span> using Lipofectamine RNAiMAX Reagent (Invitrogen/Life Technologies; Carlsbad, CA, USA) according to the manufacturer's protocol. Cells were then collected 72 h post siRNA transfection and processed for downstream applications. Experiments were performed in quadruplicate.</p><h3 class="c-article__sub-heading" id="Sec5">Western blot analyses</h3><p>To determine the degree to which siRNA knockdown of <i>CHD8</i> transcript results in decreased CHD8 protein, total protein was isolated using the Illustra triplePrep kit (GE Healthcare; Waukesha, WI, USA) and protein concentration was determined using the DC protein assay (Bio-Rad; Hercules, CA, USA). Total protein (10 μg) was then separated on a 4–20% gradient criterion TGX gel (Bio-Rad) and transferred to a nitrocellulose membrane by capillary transfer at 80 V for 3 h using a Bio-Rad Criterion blotter system. Blots were incubated overnight at 4 °C with anti-CHD8 (catalog no. 7656, Cell Signaling Technology; Danvers, MA, USA) and anti-GAPDH (catalog no. 1228, Cell Signaling Technology) primary antibodies diluted 1:1000 in blocking buffer. Anti-rabbit immunoglobulin G, horseradish-peroxidase-linked secondary antibody (catalog no. 7074, Cell Signaling Technology) was diluted 1:2000 and incubated with the membrane for 1.5 h at room temperature. Signal was developed using SuperSignal West Dura Extended Duration Substrate (Thermo Fisher Scientific; Waltham, MA, USA) and captured on a BioSpectrum Imaging System (UVP; Upland, CA, USA). The total density ratio of CHD8 to GAPDH was determined using the VisionWorks LS version 6.8 software (UVP) and analyzed by Student's paired <i>t</i>-test for comparison (<i>n</i>=4).</p><h3 class="c-article__sub-heading" id="Sec6">RNA-seq</h3><p>To determine genes that are differentially expressed in response to knockdown of <i>CHD8</i> on a genome-wide scale, cells were homogenized with QIAshredder spin columns (Qiagen; Valencia, CA, USA) followed by total cellular RNA extraction using the Qiagen RNeasy kit according to the manufacturer's protocol. RNeasy extraction typically excludes fragments <200 bp. Therefore, the data presented here focuses on long noncoding RNAs (ncRNAs). Quality (RNA integrity number) was assessed using an Agilent Technologies 2200 TapeStation Instrument (Agilent Technologies; Santa Clara, CA, USA) and 260/280 absorbance ratios were obtained using the NanoDrop ND-1000 Spectrophotometer (Thermo Fisher Scientific). cDNA library preparation was carried out using the Illumina Truseq Stranded Total RNA Sample Preparation kit and sequencing was performed on the HiSeq2000 sequencer to generate 101 bp single-end reads (Illumina; San Diego, CA, USA). An approximate average of 45 million reads per replicate was obtained for samples treated with siRNA targeting <i>CHD8</i> and an average of 40 million reads per replicate was obtained for samples treated with negative control siRNA (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 1</a>).</p><h3 class="c-article__sub-heading" id="Sec7">RNA-seq analysis</h3><p>Quality control pre-processing of raw reads was carried out using Cutadapt.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 17" title="Martin M . Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet J 2011; 17, pp 10–12." href="/articles/tp201562#ref-CR17" id="ref-link-section-d446063898e797">17</a></sup> Reads were first mapped to the transcriptome of either UCSC Hg19 or ENSEMBL GrCH37 release 75 and the remaining reads were then mapped to the unannotated regions of the respective human genome build using Tophat2.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 18" title="Kim D, Pertea G, Trapnell C, Pimentel H, Kelley R, Salzberg SL . TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol 2013; 14: R36." href="/articles/tp201562#ref-CR18" id="ref-link-section-d446063898e801">18</a></sup> The percentage of reads that were aligned to the genome for each replicate along with their respective quality control metrics are outlined in <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 1</a>. Following alignment, differential gene expression analysis was carried out using Cuffdiff<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 19" title="Trapnell C, Roberts A, Goff L, Pertea G, Kim D, Kelley DR et al. Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks. Nat Protoc 2012; 7: 562–578." href="/articles/tp201562#ref-CR19" id="ref-link-section-d446063898e808">19</a></sup> along with a supplied GTF file of the corresponding transcriptome to be analyzed. Cuffdiff analysis was performed with default settings with the exception of the masking of all annotated ribosomal RNAs and the no-effective-length-correction option, which was used to reduce overestimation of short transcript expression. Genes with a false discovery rate (<i>Q</i>-value) of <0.05 were considered significant. All gene ontology (GO) and protein–protein interaction analyses were carried out using the list of significantly differentially expressed genes identified using the UCSC Hg19 human genome annotation. To explore changes in genes transcribing ncRNAs, we also carried out the RNA-seq analysis using the ENSEMBL GrCH37, release 75, gene annotation, which contains a much larger quantity of ncRNA transcripts compared with the UCSC Hg19 annotated transcriptome.</p><h3 class="c-article__sub-heading" id="Sec8">Quantitative PCR</h3><p>To measure the degree to which siRNA knockdown of <i>CHD8</i> results in decreased transcript levels and verify the quality of RNA-seq data analysis, cell pellets were processed for RNA isolation using the Qiagen RNeasy kit (Qiagen). Superscipt III (Life Technologies; Grand Island, NY, USA) was used for reverse transcription of cDNA libraries. Quantitative PCR (qPCR) was performed using Life Technologies Taqman Gene Expression qPCR kits on a Life Technologies OneStepPlus real-time PCR machine. Each sample was run in triplicate along with the housekeeping gene, <i>GAPDH</i>. Assays used for validation (Life Technologies) include <i>ABCA1</i> (Hs01059118_m1), <i>CHD8</i> (Hs00394229_m1), <i>BDNF</i> (Hs02718934_s1), <i>CUL1</i> (Hs01117001_m1), <i>GAPDH</i> (Hs9999905_m1), <i>GJA1</i> (Hs00748445_s1), <i>HMGA2</i> (Hs00171569_m1), <i>HSPB3</i> (Hs00937412_s1), <i>IGF2</i> (Hs04188276_m1), <i>IGFBP3</i> (Hs00365742_g1), <i>IL1R1</i> (Hs00991002_m1), <i>IL8</i> (Hs00174103_m1), <i>LINC00473</i> (Hs00293257_m1), <i>LINC01021</i> (Hs01388536_m1), <i>MMP3</i> (Hs00968305_m1), <i>PITX3</i> (Hs01013935_g1), <i>SCN2A</i> (Hs01109877_m1), <i>SERPINE1</i> (Hs01126606_m1), <i>SEMA3D</i> (Hs00380877_m1), <i>SYTL2</i> (Hs00909223_m1), <i>TGM2</i> (Hs00190278_m1) and <i>TNS3</i> (Hs00994933_m1). Relative quantity of the assayed transcript was determined by the 2<sup>−ΔΔCt</sup> method using <i>GAPDH</i> as a reference.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 20" title="Schmittgen TD, Livak KJ . Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 2008; 3: 1101–1108." href="/articles/tp201562#ref-CR20" id="ref-link-section-d446063898e904">20</a></sup> For RNA-seq validation, the observed qPCR changes were compared with those obtained in RNA-seq data analysis through the use of Spearman and Pearson correlation tests carried out in the R statistical environment using the (cor.test) package.</p><h3 class="c-article__sub-heading" id="Sec9">Functional annotation and network construction</h3><p>To begin to functionally characterize and cluster the differentially expressed transcripts, the Database for Annotation, Visualization and Integrated Discovery (DAVID)<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 21" title="Dennis G, Sherman BT, Hosack DA, Yang J, Gao W, Lane HC et al. DAVID: Database for Annotation, Visualization, and Integrated Discovery. Genome Biol 2003; 4: P3." href="/articles/tp201562#ref-CR21" id="ref-link-section-d446063898e916">21</a></sup> was used to analyze for the enrichment of GO terms and UP_TISSUE (Uniprot tissue annotation database) categories. All analyses were carried out with the default options. Terms that were redundant or had a corrected <i>P-</i>value >0.05 were eliminated. Disease Association Protein–Protein Link Evaluator<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 22" title="Rossin EJ, Lage K, Raychaudhuri S, Xavier RJ, Tatar D, Benita Y et al. Proteins encoded in genomic regions associated with immune-mediated disease physically interact and suggest underlying biology. PLoS Genet 2011; 7: e1001273." href="/articles/tp201562#ref-CR22" id="ref-link-section-d446063898e923">22</a></sup> was used to determine direct interactions and connectivity among the protein products of the differentially expressed genes using the default settings. Network construction of the resulting interactions was carried out using Cytoscape version 3.1.1 software.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 23" title="Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res 2003; 13: 2498–2504." href="/articles/tp201562#ref-CR23" id="ref-link-section-d446063898e927">23</a></sup></p><h3 class="c-article__sub-heading" id="Sec10">ASD candidate genes statistical analysis</h3><p>To analyze whether or not a lack of functional <i>CHD8</i> results in the differential expression of other ASD candidate genes, a list of ASD candidate genes was obtained from the Simons Foundation Autism Research Initiative (SFARI) AutDB database.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 24" title="Abrahams BS, Arking DE, Campbell DB, Mefford HC, Morrow EM, Weiss LA et al. SFARI Gene 2.0: a community-driven knowledgebase for the autism spectrum disorders (ASDs). Mol Autism 2013; 4: 36." href="/articles/tp201562#ref-CR24" id="ref-link-section-d446063898e941">24</a></sup> A restricted list of ASD candidate genes was then curated by using only those genes with SFARI gene rankings 1–4 (strong evidence–minimal evidence) and S (syndromatic) as previously reported.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 25" title="Parikshak NN, Luo R, Zhang A, Won H, Lowe JK, Chandran V et al. Integrative functional genomic analyses implicate specific molecular pathways and circuits in autism. Cell 2013; 155: 1008–1021." href="/articles/tp201562#ref-CR25" id="ref-link-section-d446063898e945">25</a></sup> Genes above the threshold of reliable detection in Cuffdiff (a total of 13 020 genes) served as the background for enrichment tests, which were measured by the two-tailed Fisher's exact test using the (fisher.test) package in R.</p></div></div></section><section data-title="Results"><div class="c-article-section" id="Sec11-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec11">Results</h2><div class="c-article-section__content" id="Sec11-content"><h3 class="c-article__sub-heading" id="Sec12">Quantitative modeling of <i>CHD8 de novo</i> loss-of-function mutations</h3><p>The ASD-associated <i>CHD8 de novo</i> LoF mutations likely result in an ~50% reduction in <i>CHD8</i> transcript and CHD8 protein in the developing brains of individuals with ASD. To quantitatively model these dosage-dependent consequences of <i>CHD8</i> LoF mutations, siRNA-mediated knockdown of <i>CHD8</i> was performed in the human neural progenitor cell line, SK-N-SH. Following this, the knockdown efficiency of both <i>CHD8</i> transcript and CHD8 protein levels were confirmed via qPCR and western blotting, respectively (<a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/tp201562#Fig1">Figure 1</a>).</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-test="figure" data-container-section="figure" id="figure-1" data-title="Figure 1"><figure><figcaption><b id="Fig1" class="c-article-section__figure-caption" data-test="figure-caption-text">Figure 1</b></figcaption><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><a class="c-article-section__figure-link" data-test="img-link" data-track="click" data-track-label="image" data-track-action="view figure" href="/articles/tp201562/figures/1" rel="nofollow"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_Article_BFtp201562_Fig1_HTML.jpg?as=webp"><img aria-describedby="Fig1" src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_Article_BFtp201562_Fig1_HTML.jpg" alt="figure 1" loading="lazy" width="685" height="155"></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-1-desc"><p>Quantitative modeling of disruptive mutations in CHD8. (<b>a</b>) Quantitative PCR showed that a 20 n<span class="u-small-caps">M</span> concentration of <i>CHD8</i> siRNA resulted in an ~50% reduction of <i>CHD8</i> transcript. (<b>b</b>) Representative western blot indicated decreased CHD8 protein following <i>CHD8</i> siRNA-mediated knockdown. (<b>c</b>) Quantitative analysis of western blot data shows a significant reduction in CHD8 protein following <i>CHD8</i> siRNA treatment. Error bars are s.e.m., **<i>P</i><0.01, *<i>P</i><0.05. CHD8, chromodomain helicase DNA-binding protein 8; siRNA, small interfering RNA.</p></div></div><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="article-link" data-track="click" data-track-label="button" data-track-action="view figure" href="/articles/tp201562/figures/1" data-track-dest="link:Figure1 Full size image" aria-label="Full size image figure 1" rel="nofollow"><span>Full size image</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><h3 class="c-article__sub-heading" id="Sec13">CHD8 insufficiency results in genome-wide transcriptional changes</h3><p>As transcriptional changes due to CHD8 insufficiency may be able to inform the mechanisms by which a lack of <i>CHD8</i> influences ASD pathogenesis, we performed genome-wide transcriptome profiling of <i>CHD8</i>-depleted human neuronal progenitor cells, SK-N-SH, via RNA-seq. Differential gene expression analysis using the UCSC Hg19 genome annotation revealed 1715 differentially expressed genes (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Tables 2 and 3</a>), with a majority of protein-coding genes downregulated (~63%). Subsequent qPCR validation was performed for genes categorized as either having large fold changes in expression or possessing functions related to neurodevelopmental processes identified by GO analysis. Comparison of gene expression levels measured by RNA-Seq and qPCR indicated a highly significant Pearson correlation coefficient (<a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/tp201562#Fig2">Figure 2</a>), validating the magnitude of gene expression changes identified by RNA-seq. Both protein-coding and ncRNAs with altered expression are distributed across the genome in a pattern consistent with length of the chromosome; there does not appear to be any clustering of the genes with altered expression (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Figures 1 and 2</a>).</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-test="figure" data-container-section="figure" id="figure-2" data-title="Figure 2"><figure><figcaption><b id="Fig2" class="c-article-section__figure-caption" data-test="figure-caption-text">Figure 2</b></figcaption><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><a class="c-article-section__figure-link" data-test="img-link" data-track="click" data-track-label="image" data-track-action="view figure" href="/articles/tp201562/figures/2" rel="nofollow"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_Article_BFtp201562_Fig2_HTML.jpg?as=webp"><img aria-describedby="Fig2" src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_Article_BFtp201562_Fig2_HTML.jpg" alt="figure 2" loading="lazy" width="625" height="314"></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-2-desc"><p>qPCR validation of differentially expressed genes identified by RNA-Seq. Fifteen genes excluding <i>CHD8</i> were assayed via qPCR to establish the validity of the differential expression analysis based on the obtained RNA-seq data. qPCR and RNA-seq values showed a statistically significant correlation with both Pearson and Spearman methods. qPCR, quantitative PCR.</p></div></div><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="article-link" data-track="click" data-track-label="button" data-track-action="view figure" href="/articles/tp201562/figures/2" data-track-dest="link:Figure2 Full size image" aria-label="Full size image figure 2" rel="nofollow"><span>Full size image</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><h3 class="c-article__sub-heading" id="Sec14">CHD8 influences the expression of ncRNA</h3><p>NcRNAs represent a relatively unexplored source of functional transcripts that may contribute to neurodevelopmental phenotypes.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 26" title="Wilkinson B, Campbell DB . Contribution of long noncoding RNAs to autism spectrum disorder risk. Int Rev Neurobiol 2013; 113: 35–59." href="/articles/tp201562#ref-CR26" id="ref-link-section-d446063898e1087">26</a></sup> Using the ENSEMBL GrCH37, release 75, gene annotation, we identified 657 differentially expressed genes and found that ~12.5% of these were ncRNAs (<a data-track="click" data-track-label="link" data-track-action="table anchor" href="/articles/tp201562#Tab1">Table 1</a>, <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Tables 4 and 5</a>). The identified ncRNA account for 40% of the top 10 differentially expressed genes (<a data-track="click" data-track-label="link" data-track-action="table anchor" href="/articles/tp201562#Tab1">Table 1</a>). In contrast to protein-coding genes, the majority of ncRNAs are upregulated (~60%) instead of downregulated following <i>CHD8</i> knockdown.</p><div class="c-article-table" data-test="inline-table" data-container-section="table" id="table-1"><figure><figcaption class="c-article-table__figcaption"><b id="Tab1" data-test="table-caption">Table 1 Top 10 differentially expressed genes following knockdown of <i>CHD8</i></b></figcaption><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="table-link" data-track="click" data-track-action="view table" data-track-label="button" rel="nofollow" href="/articles/tp201562/tables/1" aria-label="Full size table 1"><span>Full size table</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><h3 class="c-article__sub-heading" id="Sec15">Loss of functional CHD8 affects numerous ASD susceptibility genes</h3><p>Comparing the list of differentially expressed genes to those genes contained in the SFARI AutDB gene database,<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 24" title="Abrahams BS, Arking DE, Campbell DB, Mefford HC, Morrow EM, Weiss LA et al. SFARI Gene 2.0: a community-driven knowledgebase for the autism spectrum disorders (ASDs). Mol Autism 2013; 4: 36." href="/articles/tp201562#ref-CR24" id="ref-link-section-d446063898e1488">24</a></sup> there is an overlap of 65 genes excluding <i>CHD8</i> (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 6</a>). Of the 631 genes in the AutDB database, 409 were expressed in SK-N-SH cells. Among the 1715 genes differentially expressed following <i>CHD8</i> knockdown in SK-N-SH cells, 65 were overlapping (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 7</a>). Although the entire list of differentially expressed genes does not reach statistical significance for enrichment (<i>P</i>=0.059, OR (odds ratio)=1.307, Fisher’s exact test), considering only those genes that are downregulated significantly strengthens this enrichment (<i>P</i>=4.793 × 10<sup>−3</sup>, OR=1.59). To focus on genes with substantial evidence for a role in ASD, a restricted list of 250 genes (159 of which are expressed in SK-N-SH cells) was curated from the SFARI database taking into account SFARI gene rankings as previously described.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 25" title="Parikshak NN, Luo R, Zhang A, Won H, Lowe JK, Chandran V et al. Integrative functional genomic analyses implicate specific molecular pathways and circuits in autism. Cell 2013; 155: 1008–1021." href="/articles/tp201562#ref-CR25" id="ref-link-section-d446063898e1513">25</a></sup> Following this narrowing, the list of differentially expressed genes due to knockdown of <i>CHD8</i> contained a total of 27 overlapping genes excluding <i>CHD8</i> (<a data-track="click" data-track-label="link" data-track-action="table anchor" href="/articles/tp201562#Tab2">Table 2</a>), which did not reach statistical significance for enrichment (<i>P</i>=0.153, OR=1.378). Again, considering only those genes that are downregulated significantly strengthened the enrichment (<i>P</i>=0.038, OR=1.673). In comparing all genes from the SFARI database and the narrow list of ASD genes, upregulated genes fail to show significant enrichment in both the cases. These results suggest no advantage of narrowing the list of genes by evidence-based scoring. These results further suggest that CHD8 insufficiency may regulate a network of genes that contribute to ASD risk. Although previously identified ASD candidate genes are enriched among the 1715 differentially expressed genes following <i>CHD8</i> knockdown, the magnitude of gene expression changes is similar between ASD candidate genes and genes not previously implicated in ASD. These results suggest that, in addition to altering expression of genes implicated in ASD, <i>CHD8</i> knockdown also impacts expression of genes that are either unrelated to ASD or not yet identified as contributors to ASD risk.</p><div class="c-article-table" data-test="inline-table" data-container-section="table" id="table-2"><figure><figcaption class="c-article-table__figcaption"><b id="Tab2" data-test="table-caption">Table 2 ASD-associated genes differentially expressed following knockdown of <i>CHD8</i></b></figcaption><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="table-link" data-track="click" data-track-action="view table" data-track-label="button" rel="nofollow" href="/articles/tp201562/tables/2" aria-label="Full size table 2"><span>Full size table</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><h3 class="c-article__sub-heading" id="Sec16">Transcriptional consequences of CHD8 insufficiency are enriched in processes essential for neuronal development</h3><p>GO analysis using the DAVID web server for all differentially expressed genes shows an enrichment of genes within GO categories describing several functions that have previously been associated with CHD8 including regulation of proliferation, apoptosis and transcription (<a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/tp201562#Fig3">Figure 3a</a>, <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 8</a>).<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 11" title="Nishiyama M, Oshikawa K, Tsukada Y, Nakagawa T, Iemura S, Natsume T et al. CHD8 suppresses p53-mediated apoptosis through histone H1 recruitment during early embryogenesis. Nat Cell Biol 2009; 11: 172–182." href="/articles/tp201562#ref-CR11" id="ref-link-section-d446063898e2271">11</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 12" title="Rodríguez-Paredes M, Ceballos-Chávez M, Esteller M, García-Domínguez M, Reyes JC . The chromatin remodeling factor CHD8 interacts with elongating RNA polymerase II and controls expression of the cyclin E2 gene. Nucleic Acids Res 2009; 37: 2449–2460." href="/articles/tp201562#ref-CR12" id="ref-link-section-d446063898e2274">12</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 27" title="Menon T, Yates JA, Bochar DA . Regulation of androgen-responsive transcription by the chromatin remodeling factor CHD8. Mol Endocrinol 2010; 24: 1165–1174." href="/articles/tp201562#ref-CR27" id="ref-link-section-d446063898e2277">27</a></sup> We also performed GO analysis grouping the differentially expressed genes by their direction of differential expression. Although upregulated genes show GO enrichment in mitochondrial membrane-related terms (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 9</a>), GO analysis of downregulated genes shows an enrichment in several neuron-specific processes in addition to those categories observed for all differentially expressed genes. These include GO categories such as regulation of neuron differentiation, neurogenesis and neuron projection development (<a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/tp201562#Fig3">Figure 3b</a>, <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 10</a>), all of which may be processes disrupted during ASD pathogenesis. In addition, utilizing the Uniprot tissue (UP_TISSUE) annotation database, we found the downregulated genes to be highly enriched in brain tissue (<i>P</i>=2.01 × 10<sup>−15</sup>, Benjamini–Hotchberg correction), whereas upregulated genes were not enriched in any specific tissue (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 11</a>). Overall, these results suggest that CHD8 retains conserved functions and also has an essential role in the maintenance of tissue-specific genes involved in neuron development.</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-test="figure" data-container-section="figure" id="figure-3" data-title="Figure 3"><figure><figcaption><b id="Fig3" class="c-article-section__figure-caption" data-test="figure-caption-text">Figure 3</b></figcaption><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><a class="c-article-section__figure-link" data-test="img-link" data-track="click" data-track-label="image" data-track-action="view figure" href="/articles/tp201562/figures/3" rel="nofollow"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_Article_BFtp201562_Fig3_HTML.jpg?as=webp"><img aria-describedby="Fig3" src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_Article_BFtp201562_Fig3_HTML.jpg" alt="figure 3" loading="lazy" width="685" height="184"></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-3-desc"><p>Representative enrichment of gene ontology among differentially expressed genes. (<b>a</b>) Gene ontology (GO) enrichment analysis on all differentially expressed genes due to knockdown of <i>CHD8</i>. (<b>b</b>) GO enrichment analysis on genes downregulated in response to knockdown of <i>CHD8</i>. Upregulated genes subjected to GO analysis indicated enrichment of mitochondrial membrane genes only (not shown). <i>P</i>-value represents Benjamini–Hotchberg correction for multiple tests. CHD8, chromodomain helicase DNA-binding protein 8.</p></div></div><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="article-link" data-track="click" data-track-label="button" data-track-action="view figure" href="/articles/tp201562/figures/3" data-track-dest="link:Figure3 Full size image" aria-label="Full size image figure 3" rel="nofollow"><span>Full size image</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div><h3 class="c-article__sub-heading" id="Sec17">CHD8 regulates a highly interconnected network of genes</h3><p>To assess the connectivity among the differentially expressed genes following knockdown of <i>CHD8</i>, we utilized the Disease Association Protein–Protein Link Evaluator.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 22" title="Rossin EJ, Lage K, Raychaudhuri S, Xavier RJ, Tatar D, Benita Y et al. Proteins encoded in genomic regions associated with immune-mediated disease physically interact and suggest underlying biology. PLoS Genet 2011; 7: e1001273." href="/articles/tp201562#ref-CR22" id="ref-link-section-d446063898e2343">22</a></sup> Analysis of all differentially expressed genes shows significant enrichment in both direct and indirect connectivity (<i>P</i>=3.00 × 10<sup>−3</sup> for both), identifying 2022 direct interactions (<a data-track="click" data-track-label="link" data-track-action="figure anchor" href="/articles/tp201562#Fig4">Figure 4</a>, <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 12</a>). Analysis of the downregulated genes indicated a significant increase in indirect connectivity (<i>P</i>=3.00 × 10<sup>−3</sup> for direct connectivity and <i>P</i>=9.99 × 10<sup>−4</sup> for indirect connectivity). Although upregulated genes showed no enrichment in ASD candidate genes and weak enrichment in GO categories, Disease Association Protein–Protein Link Evaluator showed an unexpected enrichment in direct and indirect connectivity among upregulated genes (<i>P</i>=2.00 × 10<sup>−3</sup> and <i>P</i>=3.20 × 10<sup>−2</sup>, respectively). These results suggest a network of connected genes upregulated by <i>CHD8</i>.</p><div class="c-article-section__figure js-c-reading-companion-figures-item" data-test="figure" data-container-section="figure" id="figure-4" data-title="Figure 4"><figure><figcaption><b id="Fig4" class="c-article-section__figure-caption" data-test="figure-caption-text">Figure 4</b></figcaption><div class="c-article-section__figure-content"><div class="c-article-section__figure-item"><a class="c-article-section__figure-link" data-test="img-link" data-track="click" data-track-label="image" data-track-action="view figure" href="/articles/tp201562/figures/4" rel="nofollow"><picture><source type="image/webp" srcset="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_Article_BFtp201562_Fig4_HTML.jpg?as=webp"><img aria-describedby="Fig4" src="//media.springernature.com/lw685/springer-static/image/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_Article_BFtp201562_Fig4_HTML.jpg" alt="figure 4" loading="lazy" width="685" height="402"></picture></a></div><div class="c-article-section__figure-description" data-test="bottom-caption" id="figure-4-desc"><p>Protein–protein interaction analysis of differentially expressed genes—Disease Association Protein–Protein Link Evaluator (DAPPLE) was used to determine direct relationships among the protein products of the genes that were differentially expressed due to knockdown of <i>CHD8</i>. A subnetwork of genes with associated corrected <i>P</i>-values <0.05, indicating seed proteins that are more likely to be connected to other seed proteins within the network than by chance, was generated and visualized using Cytoscape software. Increasing significance corresponds to increasing node size. Differential expression of the associated gene product is shown with increasing densities of blue and red indicating the degree of down- and upregulation conferred by knockdown of <i>CHD8</i>, respectively. CHD8, chromodomain helicase DNA-binding protein 8.</p></div></div><div class="u-text-right u-hide-print"><a class="c-article__pill-button" data-test="article-link" data-track="click" data-track-label="button" data-track-action="view figure" href="/articles/tp201562/figures/4" data-track-dest="link:Figure4 Full size image" aria-label="Full size image figure 4" rel="nofollow"><span>Full size image</span><svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-chevron-right-small"></use></svg></a></div></figure></div></div></div></section><section data-title="Discussion"><div class="c-article-section" id="Sec18-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec18">Discussion</h2><div class="c-article-section__content" id="Sec18-content"><p>From the initial discovery through exome sequencing to confirmation by targeted sequencing of additional ASD probands, <i>de novo</i> LoF mutations in <i>CHD8</i> are highly implicated in contributing to ASD.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 5" title="Sanders SJ, Murtha MT, Gupta AR, Murdoch JD, Raubeson MJ, Willsey AJ et al. De novo mutations revealed by whole-exome sequencing are strongly associated with autism. Nature 2012; 485: 237–241." href="/articles/tp201562#ref-CR5" id="ref-link-section-d446063898e2425">5</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 6" title="O'Roak BJ, Vives L, Girirajan S, Karakoc E, Krumm N, Coe BP et al. Sporadic autism exomes reveal a highly interconnected protein network of de novo mutations. Nature 2012; 485: 246–250." href="/articles/tp201562#ref-CR6" id="ref-link-section-d446063898e2428">6</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 7" title="Neale BM, Kou Y, Liu L, Ma'ayan A, Samocha KE, Sabo A et al. Patterns and rates of exonic de novo mutations in autism spectrum disorders. Nature 2012; 485: 242–245." href="/articles/tp201562#ref-CR7" id="ref-link-section-d446063898e2431">7</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 8" title="O'Roak BJ, Vives L, Fu W, Egertson JD, Stanaway IB, Phelps IG et al. Multiplex targeted sequencing identifies recurrently mutated genes in autism spectrum disorders. Science 2012; 338: 1619–1622." href="/articles/tp201562#ref-CR8" id="ref-link-section-d446063898e2434">8</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 9" title="Bernier R, Golzio C, Xiong B, Stessman HA, Coe BP, Penn O et al. Disruptive CHD8 mutations define a subtype of autism early in development. Cell 2014; 158: 263–276." href="/articles/tp201562#ref-CR9" id="ref-link-section-d446063898e2437">9</a></sup> Due to the heterogeneity of ASD, it is likely to be influenced by the combinatorial effects of hundreds of genes under abnormal regulation. Although genetic influences may differ among ASD cases, strong evidence suggests that the functional contribution of these various factors may converge on similar pathways.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 4" title="Levitt P, Campbell DB . The genetic and neurobiologic compass points toward common signaling dysfunctions in autism spectrum disorders. J Clin Invest 2009; 119: 747–754." href="/articles/tp201562#ref-CR4" id="ref-link-section-d446063898e2441">4</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 28" title="Berg JM, Geschwind DH . Autism genetics: searching for specificity and convergence. Genome Biol 2012; 13: 247." href="/articles/tp201562#ref-CR28" id="ref-link-section-d446063898e2444">28</a></sup> Our data suggest that <i>CHD8</i> may be a central node in one of these pathways as CHD8 insufficiency in human neural progenitor cells results in altered expression of previously identified ASD candidate genes. Among recently identified genes disrupted in autism,<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 29" title="De Rubeis S, He X, Goldberg AP, Poultney CS, Samocha K, Cicek AE et al. Synaptic, transcriptional and chromatin genes disrupted in autism. Nature 2014; 515: 209–215." href="/articles/tp201562#ref-CR29" id="ref-link-section-d446063898e2452">29</a></sup> <i>CACNA2D3</i> and <i>ETFB</i> were among the 1715 genes with altered expression following CHD8 knockdown, suggesting potential biological pathways that contribute to ASD.</p><p>Considering the enrichment of ASD candidate genes among the differentially expressed protein-coding genes, the ncRNA genes identified in our analysis may also serve as contributing factors to ASD phenotypes. The ncRNA represents a relatively unexplored category of genes that participate in several regulatory functions and may potentially contribute to the neurodevelopmental phenotypes observed in ASD.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 26" title="Wilkinson B, Campbell DB . Contribution of long noncoding RNAs to autism spectrum disorder risk. Int Rev Neurobiol 2013; 113: 35–59." href="/articles/tp201562#ref-CR26" id="ref-link-section-d446063898e2465">26</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 30" title="Kerin T, Ramanathan A, Rivas K, Grepo N, Coetzee GA, Campbell DB . A noncoding RNA antisense to moesin at 5p14.1 in autism. Sci Transl Med 2012; 4: 128ra40." href="/articles/tp201562#ref-CR30" id="ref-link-section-d446063898e2468">30</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 31" title="Ziats MN, Rennert OM . Aberrant expression of long noncoding RNAs in autistic brain. J Mol Neurosci 2013; 49: 589–593." href="/articles/tp201562#ref-CR31" id="ref-link-section-d446063898e2471">31</a></sup> For example, the ncRNAs colorectal neoplasia differentially expressed (CRNDE) and telomerase RNA component, have both been shown to undergo differential expression during neuronal differentiation of pluripotent stem cells suggesting their participation in neurogenesis.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 32" title="Lin M, Pedrosa E, Shah A, Hrabovsky A, Maqbool S, Zheng D et al. RNA-Seq of human neurons derived from iPS cells reveals candidate long non-coding RNAs involved in neurogenesis and neuropsychiatric disorders. PLoS One 2011; 6: e23356." href="/articles/tp201562#ref-CR32" id="ref-link-section-d446063898e2475">32</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 33" title="Ng SY, Johnson R, Stanton LW . Human long non-coding RNAs promote pluripotency and neuronal differentiation by association with chromatin modifiers and transcription factors. EMBO J 2012; 31: 522–533." href="/articles/tp201562#ref-CR33" id="ref-link-section-d446063898e2478">33</a></sup> In particular, CRNDE is intimately linked with the insulin/IGF signaling pathway, as treatment with IGF1 or IGF2 downregulates nuclear-retained CRNDE transcripts.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 34" title="Ellis BC, Graham LD, Molloy PL . CRNDE, a long non-coding RNA responsive to insulin/IGF signaling, regulates genes involved in central metabolism. Biochim Biophys Acta 2014; 1843: 372–386." href="/articles/tp201562#ref-CR34" id="ref-link-section-d446063898e2482">34</a></sup> This is of importance to ASD as administration of IGF1 has been shown to rescue phenotypes associated with Rett Syndrome<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 35" title="Marchetto MC, Carromeu C, Acab A, Yu D, Yeo GW, Mu Y et al. A model for neural development and treatment of Rett syndrome using human induced pluripotent stem cells. Cell 2010; 143: 527–539." href="/articles/tp201562#ref-CR35" id="ref-link-section-d446063898e2486">35</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 36" title="Tropea D, Giacometti E, Wilson NR, Beard C, McCurry C, Fu DD et al. Partial reversal of Rett Syndrome-like symptoms in MeCP2 mutant mice. Proc Natl Acad Sci USA 2009; 106: 2029–2034." href="/articles/tp201562#ref-CR36" id="ref-link-section-d446063898e2489">36</a></sup> and is now in clinical trials.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 37" title="Khwaja OS, Ho E, Barnes KV, O'Leary HM, Pereira LM, Finkelstein Y et al. Safety, pharmacokinetics, and preliminary assessment of efficacy of mecasermin (recombinant human IGF-1) for the treatment of Rett syndrome. Proc Natl Acad Sci USA 2014; 111: 4596–4601." href="/articles/tp201562#ref-CR37" id="ref-link-section-d446063898e2493">37</a></sup> Our findings of upregulation of CRNDE following decreased expression of <i>CHD8</i> suggest that this ncRNA may represent a novel therapeutic target. In addition, our data shows downregulation of the ncRNA TUG1, which has been shown to directly associate with the chromatin modifying complex, PRC2, and to be regulated by p53.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 38" title="Khalil AM, Guttman M, Huarte M, Garber M, Raj A, Rivea Morales D et al. Many human large intergenic noncoding RNAs associate with chromatin-modifying complexes and affect gene expression. Proc Natl Acad Sci USA 2009; 106: 11667–11672." href="/articles/tp201562#ref-CR38" id="ref-link-section-d446063898e2501">38</a></sup> This may represent a novel ncRNA target of CHD8, as CHD8 is known to interact with p53 and colocalize to promoters of p53-regulated genes.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 11" title="Nishiyama M, Oshikawa K, Tsukada Y, Nakagawa T, Iemura S, Natsume T et al. CHD8 suppresses p53-mediated apoptosis through histone H1 recruitment during early embryogenesis. Nat Cell Biol 2009; 11: 172–182." href="/articles/tp201562#ref-CR11" id="ref-link-section-d446063898e2505">11</a></sup></p><p>Consistent with previous studies showing the participation of CHD8 in large protein complexes involving members of the mixed lineage leukemia (MLL) family,<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 39" title="Dou Y, Milne TA, Tackett AJ, Smith ER, Fukuda A, Wysocka J et al. Physical association and coordinate function of the H3 K4 methyltransferase MLL1 and the H4 K16 acetyltransferase MOF. Cell 2005; 121: 873–885." href="/articles/tp201562#ref-CR39" id="ref-link-section-d446063898e2511">39</a></sup> depletion of endogenous CHD8 in neural cells results in decreased expression of the MLL genes, <i>MLLT1</i> and <i>MLLT3</i>, along with <i>MN1</i>, which associates with MLL to regulate transcription.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 40" title="Milne TA, Hughes CM, Lloyd R, Yang Z, Rozenblatt-Rosen O, Dou Y et al. Menin and MLL cooperatively regulate expression of cyclin-dependent kinase inhibitors. Proc Natl Acad Sci USA 2005; 102: 749–754." href="/articles/tp201562#ref-CR40" id="ref-link-section-d446063898e2524">40</a></sup> In addition, our data show the conservation of specific changes in gene expression including genes involved in Wnt signaling (<i>DKK1</i> and <i>NKD2</i>) and the p53 target, <i>CDKN1A</i>, observed in <i>CHD8</i>-depleted HCT116 colorectal cancer cells and U2OS osteosarcoma cells, respectively.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 11" title="Nishiyama M, Oshikawa K, Tsukada Y, Nakagawa T, Iemura S, Natsume T et al. CHD8 suppresses p53-mediated apoptosis through histone H1 recruitment during early embryogenesis. Nat Cell Biol 2009; 11: 172–182." href="/articles/tp201562#ref-CR11" id="ref-link-section-d446063898e2541">11</a>, <a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 41" title="Thompson BA, Tremblay V, Lin G, Bochar DA . CHD8 is an ATP-dependent chromatin remodeling factor that regulates beta-catenin target genes. Mol Cell Biol 2008; 28: 3894–3904." href="/articles/tp201562#ref-CR41" id="ref-link-section-d446063898e2544">41</a></sup> Approximately 13% of differentially expressed genes in our analysis of human neural progenitor cells were shown to have CHD8 bound to their promoters in previous work characterizing the binding sites of CHD8 in human C33A cervical carcinoma cells (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 13</a>).<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 14" title="Subtil-Rodríguez A, Vázquez-Chávez E, Ceballos-Chávez M, Rodríguez-Paredes M, Martín-Subero JI, Esteller M et al. The chromatin remodeller CHD8 is required for E2F-dependent transcription activation of S-phase genes. Nucleic Acids Res 2014; 42: 2185–2196." href="/articles/tp201562#ref-CR14" id="ref-link-section-d446063898e2552">14</a></sup> Although these data point toward the conservation of specific regulatory mechanisms involving CHD8, our analyses also suggest a tissue-specific function of CHD8 in maintaining the active transcription of genes related to neuronal development. This is illustrated by the significant enrichment of neuron-development GO categories, brain tissue specificity and the significant connectivity of genes downregulated by CHD8 insufficiency. In contrast, upregulated genes lack tissue enrichment, suggesting that they are not specific to any one particular tissue type.</p><p>Sugathan <i>et al.</i><sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 42" title="Sugathan A, Biagioli M, Golzio C, Erdin S, Blumenthal I, Manavalan P et al. CHD8 regulates neurodevelopmental pathways associated with autism spectrum disorder in neural progenitors. Proc Natl Acad Sci USA 2014; 111: E4468–E4477." href="/articles/tp201562#ref-CR42" id="ref-link-section-d446063898e2561">42</a></sup> recently described the binding sites of CHD8 and changes in transcription resulting from knockdown of <i>CHD8</i> in iPSC-derived neural progenitor cells where they found <i>CHD8</i> to be localized to 5568 promoters and 1756 differentially expressed genes, respectively. Comparing the Sugathan <i>et al.</i><sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 42" title="Sugathan A, Biagioli M, Golzio C, Erdin S, Blumenthal I, Manavalan P et al. CHD8 regulates neurodevelopmental pathways associated with autism spectrum disorder in neural progenitors. Proc Natl Acad Sci USA 2014; 111: E4468–E4477." href="/articles/tp201562#ref-CR42" id="ref-link-section-d446063898e2573">42</a></sup> list of differentially expressed genes with the list of 1715 genes observed in the current study, there is an overlap of 159 genes (9.37%, <a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 14</a>). There were 633 (37.30%) differentially expressed genes in our data set that Sugathan <i>et al.</i><sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 42" title="Sugathan A, Biagioli M, Golzio C, Erdin S, Blumenthal I, Manavalan P et al. CHD8 regulates neurodevelopmental pathways associated with autism spectrum disorder in neural progenitors. Proc Natl Acad Sci USA 2014; 111: E4468–E4477." href="/articles/tp201562#ref-CR42" id="ref-link-section-d446063898e2583">42</a></sup> reported to have CHD8 bound in close proximity to the transcription start site (<a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Table 15</a>), indicating that there is a substantial proportion of genes directly regulated by CHD8. In addition, the differentially expressed genes that are shared between our data and that of Sugathan <i>et al.</i><sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 42" title="Sugathan A, Biagioli M, Golzio C, Erdin S, Blumenthal I, Manavalan P et al. CHD8 regulates neurodevelopmental pathways associated with autism spectrum disorder in neural progenitors. Proc Natl Acad Sci USA 2014; 111: E4468–E4477." href="/articles/tp201562#ref-CR42" id="ref-link-section-d446063898e2592">42</a></sup> include <i>H1F0</i>, <i>TP53INP1</i> and <i>MLLT3</i>, suggesting conserved functions of CHD8 to participate in protein complexes containing members of the MLL family<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 39" title="Dou Y, Milne TA, Tackett AJ, Smith ER, Fukuda A, Wysocka J et al. Physical association and coordinate function of the H3 K4 methyltransferase MLL1 and the H4 K16 acetyltransferase MOF. Cell 2005; 121: 873–885." href="/articles/tp201562#ref-CR39" id="ref-link-section-d446063898e2606">39</a></sup> and regulate p53 signaling through interactions with Histone H1.<sup><a data-track="click" data-track-action="reference anchor" data-track-label="link" data-test="citation-ref" aria-label="Reference 11" title="Nishiyama M, Oshikawa K, Tsukada Y, Nakagawa T, Iemura S, Natsume T et al. CHD8 suppresses p53-mediated apoptosis through histone H1 recruitment during early embryogenesis. Nat Cell Biol 2009; 11: 172–182." href="/articles/tp201562#ref-CR11" id="ref-link-section-d446063898e2610">11</a></sup> As we also find <i>CHD8</i> knockdown results in an enrichment of differentially expressed ASD-associated genes, this illustrates the involvement of <i>CHD8</i> in regulating neurodevelopmental genes, which may differ between cell types and emphasizes the need for these transcriptional networks to be analyzed in several cellular models.</p><p>Our data indicate that a decrease of functional <i>CHD8</i> in human neural progenitor cells induces numerous transcriptional changes that potentially contribute to the pathogenesis of ASD. As the goal of this study was to uncover the consequences of CHD8 insufficiency as a whole rather than each of the particular <i>CHD8</i> LoF mutations identified in individual patients, the modeling of the exact <i>CHD8</i> LoF mutations through genome-editing techniques or induced pluripotent stem cell generation should be considered a priority. With the function of CHD8 in neurons just beginning to be explored, this work lays a foundation for further functional studies and identifies numerous molecular targets, including targetable ncRNAs, that could potentially be modulated for the purposes of therapeutic intervention.</p></div></div></section> </div> <div class="u-mt-32"> <div id="MagazineFulltextArticleBodySuffix"><section aria-labelledby="Bib1" data-title="References"><div class="c-article-section" id="Bib1-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Bib1">References</h2><div class="c-article-section__content" id="Bib1-content"><div data-container-section="references"><ol class="c-article-references" data-track-component="outbound reference" data-track-context="references section"><li class="c-article-references__item js-c-reading-companion-references-item" data-counter="1"><p class="c-article-references__text" id="ref-CR1">American Psychiatric Association and American Psychiatric Association DSM-5 Task Force <i>Diagnostic and Statistical Manual of Mental Disorders: DSM-5</i>. 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id="Aff1"><p class="c-article-author-affiliation__address">Development, Stem Cells, and Regeneration Ph.D. Program, University of Southern California, Los Angeles, CA, USA</p><p class="c-article-author-affiliation__authors-list">B Wilkinson</p></li><li id="Aff2"><p class="c-article-author-affiliation__address">Zilkha Neurogenetic Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA</p><p class="c-article-author-affiliation__authors-list">N Grepo, J Kim, K Wang, O V Evgrafov, J A Knowles & D B Campbell</p></li><li id="Aff3"><p class="c-article-author-affiliation__address">Division of Occupational Science and Occupational Therapy, Herman Ostrow School of Dentistry, University of Southern California, Los Angeles, CA, USA</p><p class="c-article-author-affiliation__authors-list">B L Thompson</p></li><li id="Aff4"><p class="c-article-author-affiliation__address">Department of Psychiatry and the Behavioral Sciences, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA</p><p class="c-article-author-affiliation__authors-list">J Kim, K Wang, O V Evgrafov, J A Knowles & D B Campbell</p></li><li id="Aff5"><p class="c-article-author-affiliation__address">Broad Center for Regenerative Medicine and Stem Cell Research, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA</p><p class="c-article-author-affiliation__authors-list">W Lu</p></li><li id="Aff6"><p class="c-article-author-affiliation__address">Department of Biochemistry and Molecular Biology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA</p><p class="c-article-author-affiliation__authors-list">W Lu</p></li></ol><div class="u-js-hide u-hide-print" data-test="author-info"><span class="c-article__sub-heading">Authors</span><ol class="c-article-authors-search u-list-reset"><li id="auth-B-Wilkinson-Aff1"><span class="c-article-authors-search__title u-h3 js-search-name">B Wilkinson</span><div 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id="additional-information-content"><p><a data-track="click" data-track-label="link" data-track-action="supplementary material anchor" href="/articles/tp201562#MOESM225">Supplementary Information</a> accompanies the paper on the Translational Psychiatry website</p></div></div></section><section data-title="Supplementary information"><div class="c-article-section" id="Sec19-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="Sec19">Supplementary information</h2><div class="c-article-section__content" id="Sec19-content"><div data-test="supplementary-info"><div id="figshareContainer" class="c-article-figshare-container" data-test="figshare-container"></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM225"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary figure 1 (xls 50 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM225_ESM.xls" data-supp-info-image="">Supplementary Figure 1 (XLS 50 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM226"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary figure 2 (xls 50 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM226_ESM.xls" data-supp-info-image="">Supplementary Figure 2 (XLS 50 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM227"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 1 (xls 21 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM227_ESM.xls" data-supp-info-image="">Supplementary Table 1 (XLS 21 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM228"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 2 (xls 244 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM228_ESM.xls" data-supp-info-image="">Supplementary Table 2 (XLS 244 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM229"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 3 (xls 29 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM229_ESM.xls" data-supp-info-image="">Supplementary Table 3 (XLS 29 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM230"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 4 (xls 115 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM230_ESM.xls" data-supp-info-image="">Supplementary Table 4 (XLS 115 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM231"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 5 (xls 33 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM231_ESM.xls" data-supp-info-image="">Supplementary Table 5 (XLS 33 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM232"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 6 (xls 24 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM232_ESM.xls" data-supp-info-image="">Supplementary Table 6 (XLS 24 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM233"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 7 (xls 20 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM233_ESM.xls" data-supp-info-image="">Supplementary Table 7 (XLS 20 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM234"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 8 (xls 45 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM234_ESM.xls" data-supp-info-image="">Supplementary Table 8 (XLS 45 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM235"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 9 (xls 23 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM235_ESM.xls" data-supp-info-image="">Supplementary Table 9 (XLS 23 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM236"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 10 (xls 56 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM236_ESM.xls" data-supp-info-image="">Supplementary Table 10 (XLS 56 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM237"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 11 (xls 31 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM237_ESM.xls" data-supp-info-image="">Supplementary Table 11 (XLS 31 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM238"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 12 (xls 126 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM238_ESM.xls" data-supp-info-image="">Supplementary Table 12 (XLS 126 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM239"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 13 (xls 29 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM239_ESM.xls" data-supp-info-image="">Supplementary Table 13 (XLS 29 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM240"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 14 (xls 32 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM240_ESM.xls" data-supp-info-image="">Supplementary Table 14 (XLS 32 kb)</a></h3></div><div class="c-article-supplementary__item" data-test="supp-item" id="MOESM241"><h3 class="c-article-supplementary__title u-h3"><a class="print-link" data-track="click" data-track-action="view supplementary info" data-test="supp-info-link" data-track-label="supplementary table 15 (xls 47 kb)" href="https://static-content.springer.com/esm/art%3A10.1038%2Ftp.2015.62/MediaObjects/41398_2015_BFtp201562_MOESM241_ESM.xls" data-supp-info-image="">Supplementary Table 15 (XLS 47 kb)</a></h3></div></div></div></div></section><section data-title="Rights and permissions"><div class="c-article-section" id="rightslink-section"><h2 class="c-article-section__title js-section-title js-c-reading-companion-sections-item" id="rightslink">Rights and permissions</h2><div class="c-article-section__content" id="rightslink-content"> <p>This work is licensed under a Creative Commons Attribution 4.0 International License. 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id="citeas">Cite this article</h3><p class="c-bibliographic-information__citation">Wilkinson, B., Grepo, N., Thompson, B. <i>et al.</i> The autism-associated gene chromodomain helicase DNA-binding protein 8 (<i>CHD8</i>) regulates noncoding RNAs and autism-related genes. <i>Transl Psychiatry</i> <b>5</b>, e568 (2015). https://doi.org/10.1038/tp.2015.62</p><p class="c-bibliographic-information__download-citation u-hide-print"><a data-test="citation-link" data-track="click" data-track-action="download article citation" data-track-label="link" data-track-external="" rel="nofollow" href="https://citation-needed.springer.com/v2/references/10.1038/tp.2015.62?format=refman&flavour=citation">Download citation<svg width="16" height="16" focusable="false" role="img" aria-hidden="true" class="u-icon"><use xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="#icon-eds-i-download-medium"></use></svg></a></p><ul class="c-bibliographic-information__list" data-test="publication-history"><li 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