Two de novo loss-of-function (LoF) variants in the ASH1L gene were identified in ASD probands from the Simons Simplex Collection (PMIDs 24267886, 25363768), while a third de novo LoF variant in this gene was identified in one ASD proband from 2,270 trios screened by the Autism Sequencing Consortium in De Rubeis et al., 2014 (PMID 25363760). Analysis of rare coding variation in 3,871 ASD cases and 9,937 ancestry-matched or paternal controls from the Autism Sequencing Consortium (ASC) in this report identified ASH1L as a gene meeting high statistical significance with a 0.05 < FDR 0.1, meaning that this gene had a 90% chance of being a true autism gene. A fourth de novo LoF variant in the ASH1L gene was identified in an ASD proband in Tammimies et al., 2015 (PMID 26325558). This gene was identified in Iossifov et al. 2015 as a strong candidate to be an ASD risk gene based on a combination of de novo mutational evidence and the absence or very low frequency of mutations in controls (PMID 26401017). An additional de novo LoF variant in ASH1L was identifed in a proband from the Pediatric Cardiac Genetics Consortium who presented with ASD, developmental delay, and intellectual disability in addition to congenital heart disease (Homsy et al., 2015). De novo and inherited missense variants that were predicted to be deleterious were identified in ASD probands from the Autism Clinical and Genetic Resources in China (ACGC) cohort in (PMID 27824329). De novo LoF variants in ASH1L have also been identified in individuals with intellectual disability in Stessman et al., 2017 (PMID 28191889) and Okamoto et al., 2017 (PMID 28394464). A two-stage analysis of rare de novo and inherited coding variants in 42,607 ASD cases, including 35,130 new cases from the SPARK cohort, in Zhou et al., 2022 identified ASH1L as a gene reaching exome-wide significance (P < 2.5E-06).
Molecular Function
This gene encodes a member of the trithorax group of transcriptional activators. The encoded product functions as a histone methyltransferase specifically methylating 'Lys-36' of histone H3 (H3K36me).
External Links
References
Type
Title
Type of Disorder
Associated Disorders
Author, Year
Primary
Coexpression networks implicate human midfetal deep cortical projection neurons in the pathogenesis of autism.
271 Tourette syndrome nuclear family trios and 337 control subjects of Han Chinese descent from the Affiliated Hospital of Qingdao University; all TS patients diagnosed according to DSM-V criteria.
272 Tourette syndrome nuclear family trios and 337 control subjects of Han Chinese descent from the Affiliated Hospital of Qingdao University; all TS patients diagnosed according to DSM-V criteria.
Ash1l null mice exhibit Increased premature lethality starts after P7 and growth retardation. Surviving mutants have skeletal abnormalities, blepharitis, and infertility caused by developmental defects in both male and female reproductive organs. The malformed reproductive organs observed in Ash1l null mice might result from the decreased expression of Hoxa10 in uterin or Hoxa11 and Hoxd10 in epididymis.
References
Type
Title
Author, Year
Additional
Histone methyltransferase Ash1l suppresses interleukin-6 production and inflammatory autoimmune diseases by inducing the ubiquitin-editing enzyme A20.
Model Type:
Genetic
Model Genotype:
Homozygous
Mutation:
A transposon- mediated (piggyBac) DNA insertion of actin-promoter-driven RFP gene between exon 15 and 16 of Ash1l allele, which results in reduced expression of Ash1l mRNA.
Allele Type: Transgenic
Strain of Origin: FVB
Genetic Background: C57BL/6J
ES Cell Line: Mutant ES Cell Line: Model Source:
Model Type:
Genetic
Model Genotype:
Homozygous
Mutation:
A gene trap vector, pGT01xf, containing the beta-geo reporter gene (beta-galactosidase and neomycin resistance fusion gene) was inserted into intron 1 of the Ash1l gene, which results in mis-splicing leading to a truncated product missing AT Hook domains, the SET associated domains, the bromodomain, and the PHD domain.
Allele Type: Gene trapped
Strain of Origin: Not specified
Genetic Background: C57BL/6J and DBA/2J
ES Cell Line: 129P2/OlaHsd
Mutant ES Cell Line: Not specified
Model Source: Not specified
Model Type:
Genetic
Model Genotype:
Heterozygous
Mutation:
A gene trap vector, pGT01xf, containing the beta-geo reporter gene (beta-galactosidase and neomycin resistance fusion gene) was inserted into intron 1 of the Ash1l gene, which results in mis-splicing leading to a truncated product missing AT Hook domains, the SET associated domains, the bromodomain, and the PHD domain.
Allele Type: Gene trapped
Strain of Origin: Not specified
Genetic Background: C57BL/6J and DBA/2J
ES Cell Line: 129P2/OlaHsd
Mutant ES Cell Line: Not specified
Model Source: Not specified
Model Type:
Genetic
Model Genotype:
Homozygous
Mutation:
Deletion of 11 bp in the exon 2 of Ash1l gene was generated by pronuclei injection of a targeting sgRNA and Cas9-coding mRNA (CRISPR/Cas9 system), which resulted in a null allelle of Ash1L.
Allele Type: Targeted (knockout)
Strain of Origin: Genetic Background: C57BL/6
ES Cell Line: Mutant ES Cell Line: Model Source:
Model Type:
Genetic
Model Genotype:
Heterozygous
Mutation:
Deletion of 11 bp in the exon 2 of Ash1l gene was generated by pronuclei injection of a targeting sgRNA and Cas9-coding mRNA (CRISPR/Cas9 system), which resulted in a null allelle of Ash1L.
Allele Type: Targeted (knockout)
Strain of Origin: Genetic Background: C57BL/6
ES Cell Line: Mutant ES Cell Line: Model Source:
Model Type:
Genetic LOF
Model Genotype:
Homozygous
Mutation:
Ash1l conditional knockout (cko) mouse line with a cre recombinase-mediated deletion of exon 4 that results in altered splicing of mrna that creates a premature stop codon before the sequences encoding the first functional aws (associated with set), set, bromo, bah and phd domains. the truncated ash1l protein contains the n-terminal 1,694 amino acids but no functional domains, thus mimicking the disruptive mutations found in patients. (ash1l+/2f)
Allele Type: Knockout
Strain of Origin: Not reported
Genetic Background: C57BL/6
ES Cell Line: NA
Mutant ES Cell Line: NA
Model Source: 34145365
Model Type:
Genetic LOF
Model Genotype:
Heterozygous
Mutation:
Ash1l conditional knockout (cko) mouse line with a cre recombinase-mediated deletion of exon 4 that results in altered splicing of mrna that creates a premature stop codon before the sequences encoding the first functional aws (associated with set), set, bromo, bah and phd domains. the truncated ash1l protein contains the n-terminal 1,694 amino acids but no functional domains, thus mimicking the disruptive mutations found in patients. heterozygous ash1l-ko mice (ash1l+/1f) were obtained by crossing the wildtype ash1l+/2f mice with cmv-cre mice, through which one allele of ash1l gene was deleted in both germlines and somatic cells in progenies.
Allele Type: Knockout
Strain of Origin: Not reported
Genetic Background: C57BL/6
ES Cell Line: NA
Mutant ES Cell Line: NA
Model Source: 34145365
Model Type:
Genetic LOF
Model Genotype:
Homozygous
Mutation:
Mice with ash1l deleted in the npcs and npc-derived neuronal and glial lineages in the developing mouse brain by crossing the ash1l-conditional ready mice bearing floxed exon 4 with a neural progenitor cell (npc)-specific cre (nestin-cre) mouse line. homozygous ash1l-cko (ash1l-nes-cko, ash1l2f/2f;nestin-cre+/â??)
Allele Type: Conditional knockout
Strain of Origin: Not reported
Genetic Background: C57BL/6
ES Cell Line: NA
Mutant ES Cell Line: NA
Model Source: 34145365
Model Type:
Genetic LOF
Model Genotype:
Heterozygous
Mutation:
Mice with ash1l deleted in the npcs and npc-derived neuronal and glial lineages in the developing mouse brain by crossing the ash1l-conditional ready mice bearing floxed exon 4 with a neural progenitor cell (npc)-specific cre (nestin-cre) mouse line. heterozygous (ash1l2f/+;nestin-cre+/â??)
Allele Type: Conditional knockout
Strain of Origin: Not reported
Genetic Background: C57BL/6
ES Cell Line: NA
Mutant ES Cell Line: NA
Model Source: 34145365
Model Type:
Genetic LOF
Model Genotype:
Homozygous
Mutation:
Tamoxifen-inducible ash1l-cko mouse line (ash1l2f/2f;rosa26-creert2+/+) generated by crossing the ash1l-cko mice bearing floxed exon 4 with a rosa26-creert2 line.
Allele Type: Conditional knockout
Strain of Origin: Not reported
Genetic Background: C57BL/6
ES Cell Line: NA
Mutant ES Cell Line: NA
Model Source: 34145365
Model Type:
Genetic
Model Genotype:
Homozygous
Mutation:
Exon 2 of Ash1l was flanked with one pair of loxP sites. Ash1l fl/fl females were crossed with Emx1-Cre then Ash1lfl/+ (Ash1lEmx1-het) males were crossed to obtain homozygous mice with conditional deletion of Ash1l in neurons of forebrain, especially neocortex and hippocampus.
Allele Type: conditional knockout
Strain of Origin: Genetic Background: C57BL/6
ES Cell Line: Mutant ES Cell Line: Model Source:
Description: Peritoneal macrophages from ash1l-silenced mice produce more il-6 and tnf in response to lps or poly(i:c) than those from wildtype controls; the serum of ash1l-silenced mice contains increased levels of il-6 and tnf in response to injected e. coli (10e8) , lps, or poly(i:c) relative to wildtype controls
Exp Paradigm: Immunoassay: cytometric bead array (cba): il-6, tnf; thioglycolate-elicited mouse peritoneal macrophages are treated with lps (100 ng/ml) or poly(i:c) (10 microgram/ml); mice are injected intraperitoneally with e. coli (10e8) , lps (15 mg/kg), or poly(i:c) (20 mg/kg)
Description: Ash1l-silenced aged mice have higher immunoglobulins (igm, igg1, igg2a, igg2b, and iga) and il-6 in serum, have more severe infiltration of mononuclear cells in various organs, exhibit increased susceptibility to membranous glomerulonephritis, and have more deposition of immune complexes of autoantibodies in the glomerulis; higher incidence of ash1l-silenced mice are subject to collagen ii-induced arthritis (i.e. increased thickness in hind ankle joints and more severe swelling in the paws, increased concentration of il-6 in serum, and more severe cartilage and bone destruction and mononuclear cells infiltration in joint capsule) relative to wildtype controls
Exp Paradigm: Histology; elisa; collagen ii-induced arthritis (cia)
Description: Ash1l-silenced mice have severe inflammatory hyperemia in the lungs with increased infiltration of mononuclear cells and red blood cells 6 hr after e. coli or lps injection; ash1l-silenced mice have higher bacterial load in the blood and are more susceptible to e. coli-induced sepsis
Exp Paradigm: Histology
Description: Ash1l-silenced macrophages exhibit reduced a20 expression upon lps and poly(i:c) stimulation as compared to wildtype controls
Exp Paradigm: Lps or poly(i:c) induced peritoneal macrophages; nuclear run-on assay: tnfaip3; western blot: a20 (tnfaip3 protein)- western blot
Description: The chromatin accessibility and the concentration of h3k4me3 modification of the tnfaip3 promoter is significantly decreased in ash1l-silenced macrophages upon tlr stimulation as compared to wildtype controls
Exp Paradigm: Dnase i hypersensitive sites sequencing (dnase-seq); chromatin immunoprecipitation (chip)
Dnase i hypersensitive sites sequencing (dnase-seq)
Description: Enhanced nf-kappab and mapk signalings in ash1l-silenced macrophages in response to lps or tnf compared to wildtype controls
Exp Paradigm: Western blot: phosphorylation of ikkalpha, ikkbeta, ikappabalpha, and p65 (the nuclear translocation of p65 is also assayed) as well as phosphorlation of erk, jnk, and p38; k63-ubiquitination on nemo and traf6
Description: The uterine function in supporting implantation and decidualization were impaired in the mutant mice.
Exp Paradigm: Histology: chicago sky blue 6b staining
Description: Increased apoptotic cells in the caudal epididymis of the mutant mice compared to heterozygous controls
Exp Paradigm: Activated caspase 3 immunostaining
Description: Female knockout mice were infertile with no change in the birth rate of the pups that developed from the mutant blastocysts in foster mothers; there is no change in the in vitro fertilization rate by using ash1l null eggs; no change in the estrous cycle in ash1l nulls
Exp Paradigm: Female only; embryo transfer
Description: Decreased litters produced per male mutant mouse; decreased count of motile sperm collected from the mutant vas deferens; no significant change in the sperm count at epididymis; no change in the in vitro fertilization rate
Exp Paradigm: Male only; motile sperm count; h&e histochemistry
Description: 4 out of 7 knockout mice have an abnormal expansion of the corneal epithelium, disorganization of the basal lamina, and an apparent reduction in sebaceous glands
Exp Paradigm: General observations-general observations
Reproductive system development: uterus morphology1
Decreased
Description: Uteri are smaller and twisted in the mutants. the stromal layer of the uterus was thinner in mutants and the uterine glands were 4-fold less than in the littermate controls
Exp Paradigm: General observations
Description: 4 out of 7 knockout mice have an abnormal expansion of the corneal epithelium, disorganization of the basal lamina, and an apparent reduction in sebaceous glands
Exp Paradigm: General observations- histology
Reproductive system development: epididymis morphology1
Decreased
Description: 17 out of 20 epididymides from mutants had a abnormal morphology from corpus to caput, a widening of the cauda, and a twisting of the ductus deferens
Exp Paradigm: The gross structure of the isolated tissue was visualized by x-gal staining
Description: Knockout embryos (60%) had an eighth rib attached to the sternum on either the right side (25%) left side (25%), or both sides (10%) as well as the ribs were attached in a staggered manner to the sternum
Exp Paradigm: Alcian blue/alizarin red staining
Description: Decreased body weight was observed at 2 weeks or 3 weeks old knockout mice (compared to littermate controls)
Exp Paradigm: Body weight measurement
Description: Decreased protein levels of hoxd10 in initial segment, caput, and ductus deferens; decreased protein levels of hoxa11 in the initial segment of the mutant epididymis
Exp Paradigm: Quantitative pcr (qrt-pcr)
Description: Decreased protein levels of either foxa2 or hoxa10 at the mutant uterine glands or the stroma, respectively
Exp Paradigm: Immunostaining: foxa2, hoxa10
Description: Decreased ash1l protein expression at the cells lining epididymal tubules in mutant mice
Exp Paradigm: Immunostaining; rt-qpcr-immunostaining
Description: Ash1l null hippocampal neurons exhibit no activity-dependent repression of nrxn1alpha relative to wildtype controls
Exp Paradigm: Quantitative pcr (qrt-pcr): nrxn1alpha; qrt-pcrs are conducted 24 hours after high potassium stimulation or by optical stimulation (primary cultured neurons tansfected by lentiviral vector, lv: camkii-chr2)
Description: No homozygous ash1l nulls are observed at 1 week of age from intercrosses of ash1l hets
Exp Paradigm: Genotypic ratio of progeny from heterozygous parents
Genotypic ratio of progeny from heterozygous parents
Description: elimination rates of boutons at 24 h after conditioning were significantly lower in Ash1l het mice than that in WT mice, while the bouton formation rates remained unchanged; tone-dependent fear conditioning learning induced an increase in total bouton number in Ash1l het mice than in WT mice at 24 h after conditioning; in WT but not in Ash1l het mice, density of axonal boutons in activated AUD-LA projections (mCherry+) was significantly lower than that in non-activated AUD-LA projections after tone-dependent fear conditioning
Exp Paradigm: axons labeled by AAV-EF1a-DIO-EYFP in Rbp4-Cre mice; Ash1l crossed to FosTRAP mice
Chromatin modification: histone modification: open chromatin2
decreased
Description: Learning-induced significant accumulation of the repressive marker H3K27me3 on EphA7 gene; learning also induced a higher level of accumulation of EZH2 on EphA7 promoter region in Ash1l het mice compared with the WT mice; H3K27me3 levels in the EphA7 promoter region and adjacent gene body regions were increased in the Ash1l mutant mice
Exp Paradigm: tone dependent Go-No-go task; ephrin-A5 Fc was bilaterally infused in the auditory cortex immediately after the training phase
Chromatin modification: histone modification: open chromatin2
decreased
Description: H3K36me2, the direct target of Ash1l, showed no difference around EphA7 gene loci between WT and Ash1l het mice in auditory cortex; H3K27me3 signal showed significant increase in EphA7 gene loci of Ash1l het mice compared with WT mice; H3K27 acetylation was decreased near the EphA7 promoter region in Ash1l het mice compared with the WT mice
Cued or contextual fear conditioning: memory of cue2
abnormal
Description: Ash1l het mice showed comparable freezing with WT for the conditioned CS+ stimulus but showed significantly increased freezing with the CS- stimulus; Ash1l het mice showed impaired discrimination ability
Cued or contextual fear conditioning: memory of context2
decreased
Description: Ash1l het mice showed reduced freezing on day 3 during learning; difference in freezing was not observed between distinct contexts indicating impairment of discrimination ability
Description: Ash1l het mice had performance similar to WT mice; Ash1l het mice showed higher level of licking probability in many untrained auditory signals compared to WT mice
Exp Paradigm: tone dependent Go-No-go task
Description: transcriptome analysis showed 563 differentially expressed genes in the auditory cortex and 155 in the dorsal striatum; Ash1l downregulated genes involved in regulation of synaptic transmission, synapse structure, axon part, and synaptic membrane in the auditory cortex and dorsal striatum
Description: activated neurons in Ash1l het mice showed a much lower expression level of EphA7 than that in WT mice and did not show elevated expression of EphA7 compared with silent neurons; After tone dependent fear conditioning training for 12h, the expression of EphA7 was significantly reduced in Ash1l het mice compared to WT mice in the task-activated neurons
Exp Paradigm: mice crossed with EGR1-EGFP transgenic line; fluorescence-activated cell sorting (FACS)
Description: transcriptome analysis showed 1691 differentially expressed genes in the auditory cortex and 861 in the dorsal striatum; Ash1l downregulated genes involved in regulation of synaptic transmission, synapse structure, axon part, and synaptic membrane in the auditory cortex and dorsal striatum
Description: Ash1l hets exhibit decreased h3k36me2 at nrxn1alpha promoter region relative to wildtype controls in hippocampus; overall epigenetic histone modification in the hippocampus of ash1l hets is not changed
Exp Paradigm: Chromatin immunoprecipitation (chip): h3k36me2, h3k4me3
Description: Decreased ash1l expression at the hippocampus of ash1l hets relative to wildtype controls
Exp Paradigm: Western blot: ash1l; quantitative pcr (qrt-pcr): ash1l-western blot
Description: Some satb2+ neurons were not properly located on the upper layers and scattered in the bottom layers suggesting delayed lamination of neuronal cells during embryonic cortical layer formation
Description: Without maternal uterine support, all ash1l-ko newborns died within 24 h after birth suggesting ash1l might be critical for establishing and maintaining a stable physiological condition for neonatal survival
Description: Both male and female mutant mice had significantly more locomotor activity and longer running distances.
Exp Paradigm: TSE PhenoMaster/LabMaster System
Description: Levels of myelination in the cortices were significantly lower at p21 but reached to a comparable level around postnatal two months, indicating delayed myelination
Description: Male and female mutant mice displayed much more severe epileptic behaviors including heavy myoclonic jerks, lying on belly with rapid body twitches, and clonic-tonic spasm. Mutant mice had spike-wave electrical discharges with increased amplitude, which was consistent with the severe epileptic behaviors observed in these mice.
Exp Paradigm: intraperitoneal injection of pentylenetetrazole
Description: Mutant mice show markedly reduced subcutaneous and visceral adipose tissue depots. Change appeared to mainly affect white adipose tissues but not brown adipose tissues.
Cued or contextual fear conditioning: memory of cue1
abnormal
Description: Ash1l cko mice showed comparable freezing to WT in response to the CS+ stimulus but CS- provoked higher level of freezing in the Ash1l cko mice; Ash1l cko showed significantly impaired discrimination ability
Description: compared with normal neurons, the elimination rate of axonal boutons in Ash1l-deficient neurons was significantly lower at 24 h after conditioning; no difference was detected for newly emerged boutons between Ash1l-deficient and Ash1l-normal neurons
Exp Paradigm: injected AAV-hSyn-Cre and AAVEF1a-DIO-EYFP into the right side of AUD
