ABSTRACT
The evolution of reproductive barriers is the first step in the formation of new species and can help us understand the diversification of life on Earth. These reproductive barriers often take the form of hybrid incompatibilities, in which alleles derived from two different species no longer interact properly in hybrids1-3. Theory predicts that hybrid incompatibilities may be more likely to arise at rapidly evolving genes4-6 and that incompatibilities involving multiple genes should be common7,8, but there has been sparse empirical data to evaluate these predictions. Here we describe a mitonuclear incompatibility involving three genes whose protein products are in physical contact within respiratory complex I of naturally hybridizing swordtail fish species. Individuals homozygous for mismatched protein combinations do not complete embryonic development or die as juveniles, whereas those heterozygous for the incompatibility have reduced complex I function and unbalanced representation of parental alleles in the mitochondrial proteome. We find that the effects of different genetic interactions on survival are non-additive, highlighting subtle complexity in the genetic architecture of hybrid incompatibilities. Finally, we document the evolutionary history of the genes involved, showing signals of accelerated evolution and evidence that an incompatibility has been transferred between species via hybridization.
Subject(s)
Cell Nucleus , Electron Transport Complex I , Fishes , Genes, Lethal , Genetic Speciation , Hybridization, Genetic , Mitochondrial Proteins , Animals , Alleles , Electron Transport Complex I/genetics , Fishes/classification , Fishes/embryology , Fishes/genetics , Fishes/growth & development , Homozygote , Genes, Lethal/genetics , Species Specificity , Embryonic Development/genetics , Mitochondrial Proteins/genetics , Cell Nucleus/genetics , Heterozygote , Evolution, MolecularABSTRACT
The concept of synthetic lethality has great potential for anticancer therapy as a new strategy to specifically kill cancer cells while sparing normal cells. To further understand the potential molecular interactions and gene characteristics involved in synthetic lethality, we performed a comprehensive analysis of predicted cancer-specific genetic interactions. Many genes were identified as cancer-associated genes that contributed to multiple biological processes and pathways, and the gene features were not random, indicating their potential roles in human carcinogenesis. Some relevant genes detected in multiple cancers were prone to be enriched in specific biological progresses and pathways, especially processes associated with DNA damage, chromosome-related functions and cancer pathways. These findings strongly implicated potential roles for these genes in cancer pathophysiology and functional relationships, as well as applications for future anticancer drug discovery. Further experimental validation indicated that the synthetic lethal interaction of APC and GFER may provide a potential anticancer strategy for patients with APC-mutant colon cancer. These results will contribute to further exploration of synthetic lethal interactions and broader application of the concept of synthetic lethality in anticancer therapeutics.
Subject(s)
Antineoplastic Agents , Genes, Lethal , Neoplasms , Humans , Antineoplastic Agents/pharmacology , Antineoplastic Agents/therapeutic use , Carcinogenesis/genetics , Colonic Neoplasms/drug therapy , Colonic Neoplasms/genetics , DNA Damage , Genes, Lethal/genetics , Genes, Lethal/physiology , Neoplasms/drug therapy , Neoplasms/genetics , OncogenesABSTRACT
Postzygotic isolation by genomic conflict is a major cause for the formation of species. Despite its importance, the molecular mechanisms that result in the lethality of interspecies hybrids are still largely unclear. The genus Drosophila, which contains over 1600 different species, is one of the best characterized model systems to study these questions. We showed in the past that the expression levels of the two hybrid incompatibility factors Hmr and Lhr diverged in the two closely related Drosophila species, D. melanogaster and D. simulans, resulting in an increased level of both proteins in interspecies hybrids. The overexpression of the two proteins also leads to mitotic defects, a misregulation in the expression of transposable elements and decreased fertility in pure species. In this work, we describe a distinct six subunit protein complex containing HMR and LHR and analyse the effect of Hmr mutations on complex integrity and function. Our experiments suggest that HMR needs to bring together components of centromeric and pericentromeric chromatin to fulfil its physiological function and to cause hybrid male lethality.
Subject(s)
Drosophila Proteins/genetics , Reproductive Isolation , Animals , Centromere/metabolism , DNA Transposable Elements/genetics , Drosophila Proteins/metabolism , Drosophila melanogaster/genetics , Drosophila melanogaster/metabolism , Drosophila simulans/genetics , Drosophila simulans/metabolism , Genes, Lethal/genetics , Genetic Speciation , Hybridization, Genetic/genetics , Reproduction/geneticsABSTRACT
Gangliosides are expressed on plasma membranes throughout the body and enriched in the nervous system. A critical role for complex a- and b-series gangliosides in central and peripheral nervous system ageing has been established through transgenic manipulation of enzymes in ganglioside biosynthesis. Disrupting GalNAc-transferase (GalNAc-T), thus eliminating all a- and b-series complex gangliosides (with consequent over-expression of GM3 and GD3) leads to an age-dependent neurodegeneration. Mice that express only GM3 ganglioside (double knockout produced by crossing GalNAc-T-/- and GD3 synthase-/- mice, Dbl KO) display markedly accelerated neurodegeneration with reduced survival. Degenerating axons and disrupted node of Ranvier architecture are key features of complex ganglioside-deficient mice. Previously, we have shown that reintroduction of both a- and b-series gangliosides into neurons on a global GalNAcT-/- background is sufficient to rescue this age-dependent neurodegenerative phenotype. To determine the relative roles of a- and b-series gangliosides in this rescue paradigm, we herein reintroduced GalNAc-T into neurons of Dbl KO mice, thereby reconstituting a-series but not b-series complex gangliosides. We assessed survival, axon degeneration, axo-glial integrity, inflammatory markers and lipid-raft formation in these Rescue mice compared to wild-type and Dbl KO mice. We found that this neuronal reconstitution of a-series complex gangliosides abrogated the adult lethal phenotype in Dbl KO mice, and partially attenuated the neurodegenerative features. This suggests that whilst neuronal expression of a-series gangliosides is critical for survival during ageing, it is not entirely sufficient to restore complete nervous system integrity in the absence of either b-series or glial a-series gangliosides.
Subject(s)
G(M3) Ganglioside/metabolism , Gangliosides/metabolism , Genes, Lethal/genetics , Neurons/metabolism , Animals , Axons/pathology , Heredodegenerative Disorders, Nervous System/genetics , Heredodegenerative Disorders, Nervous System/pathology , Inflammation/metabolism , Membrane Microdomains/metabolism , Mice , Mice, Inbred C57BL , Mice, Knockout , Mice, Transgenic , N-Acetylgalactosaminyltransferases/genetics , Phenotype , Ranvier's Nodes/pathology , Sialyltransferases/genetics , Survival Analysis , Polypeptide N-acetylgalactosaminyltransferaseABSTRACT
Whole-genome doubling (WGD) is common in human cancers, occurring early in tumorigenesis and generating genetically unstable tetraploid cells that fuel tumour development1,2. Cells that undergo WGD (WGD+ cells) must adapt to accommodate their abnormal tetraploid state; however, the nature of these adaptations, and whether they confer vulnerabilities that can be exploited therapeutically, is unclear. Here, using sequencing data from roughly 10,000 primary human cancer samples and essentiality data from approximately 600 cancer cell lines, we show that WGD gives rise to common genetic traits that are accompanied by unique vulnerabilities. We reveal that WGD+ cells are more dependent than WGD- cells on signalling from the spindle-assembly checkpoint, DNA-replication factors and proteasome function. We also identify KIF18A, which encodes a mitotic kinesin protein, as being specifically required for the viability of WGD+ cells. Although KIF18A is largely dispensable for accurate chromosome segregation during mitosis in WGD- cells, its loss induces notable mitotic errors in WGD+ cells, ultimately impairing cell viability. Collectively, our results suggest new strategies for specifically targeting WGD+ cancer cells while sparing the normal, non-transformed WGD- cells that comprise human tissue.
Subject(s)
Genome, Human/genetics , Mitosis/drug effects , Neoplasms/genetics , Neoplasms/pathology , Tetraploidy , Abnormal Karyotype/drug effects , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Cell Line, Tumor , Female , Genes, Lethal/genetics , Humans , Kinesins/deficiency , Kinesins/genetics , Kinesins/metabolism , M Phase Cell Cycle Checkpoints/drug effects , Male , Mitosis/genetics , Proteasome Endopeptidase Complex/metabolism , Reproducibility of Results , Spindle Apparatus/drug effectsABSTRACT
Cenani Lenz syndrome is a rare autosomal recessive disorder associated with variable degree of limb malformations, dysmorphism, and renal agenesis. It is caused due to pathogenic variants in the LRP4 gene, which plays an important role in limb and renal development. Mutations in the APC gene have also been occasionally associated with CLS. The phenotypic spectrum ranges from mild to very severe perinatal lethal type depending on the type of variant. We report a pathogenic variant, c.2710 del T (p.Trp904GlyfsTer5) in theLRP4 gene, in a fetus with lethal Cenani Lenz syndrome with antenatal presentation of tetraphocomelia and symmetrical involvement of hands and feet.
Subject(s)
Congenital Abnormalities/genetics , Kidney Diseases/congenital , Kidney/abnormalities , LDL-Receptor Related Proteins/genetics , Limb Deformities, Congenital/genetics , Syndactyly/genetics , Aborted Fetus/pathology , Adenomatous Polyposis Coli Protein/genetics , Congenital Abnormalities/mortality , Congenital Abnormalities/pathology , Female , Genes, Lethal/genetics , Genetic Predisposition to Disease , Homozygote , Humans , Indian Ocean Islands/epidemiology , Kidney/pathology , Kidney Diseases/genetics , Kidney Diseases/mortality , Kidney Diseases/pathology , Limb Deformities, Congenital/mortality , Limb Deformities, Congenital/pathology , Male , Mutation/genetics , Pedigree , Phenotype , Pregnancy , Syndactyly/mortality , Syndactyly/pathologyABSTRACT
The CRISPR/Cas system offers new opportunities for targeted gene modifications in a wide range of organisms. In medaka (Oryzias latipes), a vertebrate model organism, a wild-type Cas9-based approach is commonly used to establish desired strains, however, its use in lethal genes is still challenging due to excess gene disruptions triggered by DNA double strand breaks (DSBs). To overcome this problem, we aimed to develop a new knock-in system using Cas9 nickase (Cas9n) that can reduce DNA DSBs. We revealed that Cas9n allowed reduction of the DSB-induced unwanted mutagenesis via non-homologous end-joining at both on- and off- target sites. Further, with a new donor plasmid (p2BaitD) that provides a linear template through Cas9n-mediated nicks, we successfully integrated reporter cassettes via homology-directed repair (HDR) into all three loci tested, including a lethal gene. In the experiment targeting the lethal gene, the combination of p2BaitD and Cas9n achieved higher survival rates than the Cas9-based approach, which enabled the desired knock-in founders. Additionally, through a technical blend of our knock-in system with a recently developed One-step mating protocol, we successfully established a homozygous knock-in strain in one generation period. This study presents evidence of an effective method to generate an HDR-mediated gene knock-in in medaka and other organisms, which is useful for establishing screening platforms for genes or drugs toxicity or other applications.
Subject(s)
CRISPR-Cas Systems/genetics , Deoxyribonuclease I/genetics , Genes, Lethal/genetics , Animals , DNA Breaks, Double-Stranded , DNA Repair , Deoxyribonuclease I/metabolism , Oryzias/geneticsABSTRACT
Catalytic-inactivating mutations within the Drosophila enhancer H3K4 mono-methyltransferase Trr and its mammalian homologs, MLL3/4, cause only minor changes in gene expression compared with whole-gene deletions for these COMPASS members. To identify essential histone methyltransferase-independent functions of Trr, we screened to identify a minimal Trr domain sufficient to rescue Trr-null lethality and demonstrate that this domain binds and stabilizes Utx in vivo. Using the homologous MLL3/MLL4 human sequences, we mapped a short â¼80-amino-acid UTX stabilization domain (USD) that promotes UTX stability in the absence of the rest of MLL3/4. Nuclear UTX stability is enhanced when the USD is fused with the MLL4 HMG-box. Thus, COMPASS-dependent UTX stabilization is an essential noncatalytic function of Trr/MLL3/MLL4, suggesting that stabilizing UTX could be a therapeutic strategy for cancers with MLL3/4 loss-of-function mutations.
Subject(s)
Conserved Sequence/genetics , DNA-Binding Proteins/genetics , Drosophila Proteins/genetics , Drosophila melanogaster/genetics , Genes, Lethal/genetics , Histone-Lysine N-Methyltransferase/genetics , Oxidoreductases, N-Demethylating/genetics , Animals , Gene Deletion , Gene Expression Regulation/genetics , HCT116 Cells , Humans , Protein Domains , Protein StabilityABSTRACT
The theory of population genetics leads to the expectation that in very large populations the frequencies of recessive lethal mutations are close to the square root of the mutation rate, corresponding to mutation-selection balance. There are numerous examples where the frequencies of such alleles are orders of magnitude larger than this result. In this work we theoretically investigate the role of temporal fluctuations in the heterozygous effect (h) for lethal mutations in very large populations. For fluctuations of h, around a mean value of [Formula: see text], we find a biased outcome that is described by an effective dominance coefficient, heff, that is generally less than the mean dominance coefficient, i.e., [Formula: see text]. In the case where the mean dominance coefficient is zero, the effective dominance coefficient is negative: heff < 0, corresponding to the lethal allele behaving as though overdominant and having an elevated mean frequency. This case plausibly explains mean allele frequencies that are an order of magnitude larger than the equilibrium frequency of a recessive allele with a constant dominance coefficient. Our analysis may be relevant to explaining lethal disorders with anomalously high frequencies, such as cystic fibrosis and Tay-Sachs, and may open the door to further investigations into the statistics of fluctuations of the heterozygous effect.
Subject(s)
Genes, Lethal/genetics , Genetics, Population , Models, Theoretical , Selection, Genetic/genetics , Alleles , Gene Frequency/genetics , Genes, Dominant/genetics , Genes, Recessive/genetics , Heterozygote , Humans , Mutation/genetics , Mutation RateABSTRACT
Congenital disorders of glycosylation (CDG) are a group of genetic, mostly multisystem disorders, which often involve the central nervous system. ALG3-CDG is one the some 130 known CDG. Here we report two siblings with a severe phenotype and intrauterine death. Whole-exome sequencing revealed two novel variants in ALG3: NM_005787.6:c.512G>T (p.Arg171Leu) inherited from the mother and NM_005787.6:c.511C>T (p.Arg171Trp) inherited from the father.
Subject(s)
Central Nervous System/metabolism , Congenital Disorders of Glycosylation/genetics , Genes, Lethal/genetics , Mannosyltransferases/genetics , Aborted Fetus/pathology , Central Nervous System/pathology , Congenital Disorders of Glycosylation/metabolism , Congenital Disorders of Glycosylation/pathology , Female , Humans , Male , Mothers , Mutation/genetics , Phenotype , Siblings , Exome SequencingABSTRACT
Roan (Rn) horses show a typical seasonal change of color. Their body is covered with colored and white hair. We performed a descriptive statistical analysis of breeding records of Icelandic horses to challenge the hypothesis of roan being lethal in utero under homozygous condition. The roan to non-roan ratio of foals from roan × roan matings revealed homozygous roan Icelandic horses to be viable. Even though roan is known to be inherited in a dominant mode and epistatic to other coat colors, the causative mutation is still unknown. Nevertheless, an association between roan phenotype and the KIT gene was shown for different horse breeds. In the present study, we identified KIT variants by Sanger sequencing, and show that KIT is also associated with roan in the Icelandic horse breed.
Subject(s)
Hair Color/genetics , Horse Diseases/genetics , Horses/genetics , Proto-Oncogene Proteins c-kit/genetics , Animals , Breeding , Color , Genes, Lethal/genetics , Horse Diseases/pathology , Mutation/genetics , PhenotypeABSTRACT
Chromatin modifiers play critical roles in epidermal development, but the functions of histone deacetylases in this context are poorly understood. The class I HDAC, HDAC3, is of particular interest because it plays divergent roles in different tissues by partnering with tissue-specific transcription factors. We found that HDAC3 is expressed broadly in embryonic epidermis and is required for its orderly stepwise stratification. HDAC3 protein stability in vivo relies on NCoR and SMRT, which function redundantly in epidermal development. However, point mutations in the NCoR and SMRT deacetylase-activating domains, which are required for HDAC3's enzymatic function, permit normal stratification, indicating that HDAC3's roles in this context are largely independent of its histone deacetylase activity. HDAC3-bound sites are significantly enriched for predicted binding motifs for critical epidermal transcription factors including AP1, GRHL, and KLF family members. Our results suggest that among these, HDAC3 operates in conjunction with KLF4 to repress inappropriate expression of Tgm1, Krt16, and Aqp3 In parallel, HDAC3 suppresses expression of inflammatory cytokines through a Rela-dependent mechanism. These data identify HDAC3 as a hub coordinating multiple aspects of epidermal barrier acquisition.
Subject(s)
Cell Differentiation/genetics , Epidermal Cells/cytology , Epidermis/embryology , Histone Deacetylases/genetics , Histone Deacetylases/metabolism , Animals , Embryo, Mammalian , Gene Deletion , Gene Expression Regulation, Developmental , Genes, Lethal/genetics , Kruppel-Like Factor 4 , Kruppel-Like Transcription Factors/genetics , Mice , Mice, Inbred C57BL , Mutation , Nuclear Receptor Co-Repressor 1/genetics , Nuclear Receptor Co-Repressor 1/metabolism , Nuclear Receptor Co-Repressor 2/genetics , Nuclear Receptor Co-Repressor 2/metabolism , Protein Interaction Domains and Motifs/genetics , Transcription Factor RelA/genetics , Transcription Factor RelA/metabolismABSTRACT
Genetic interactions, including synthetic lethal effects, can now be systematically identified in cancer cell lines using high-throughput genetic perturbation screens. Despite this advance, few genetic interactions have been reproduced across multiple studies and many appear highly context-specific. Here, by developing a new computational approach, we identified 220 robust driver-gene associated genetic interactions that can be reproduced across independent experiments and across non-overlapping cell line panels. Analysis of these interactions demonstrated that: (i) oncogene addiction effects are more robust than oncogene-related synthetic lethal effects; and (ii) robust genetic interactions are enriched among gene pairs whose protein products physically interact. Exploiting the latter observation, we used a protein-protein interaction network to identify robust synthetic lethal effects associated with passenger gene alterations and validated two new synthetic lethal effects. Our results suggest that protein-protein interaction networks can be used to prioritise therapeutic targets that will be more robust to tumour heterogeneity.
Subject(s)
Epistasis, Genetic/genetics , Gene Expression Regulation, Neoplastic/genetics , Loss of Function Mutation/genetics , Neoplasms/genetics , Protein Interaction Maps/genetics , Cell Line, Tumor , Computational Biology , Genes, Lethal/genetics , Humans , Oncogenes/geneticsABSTRACT
Neu-Laxova syndrome (NLS) is a lethal genetic multiple congenital anomaly syndrome of unknown prevalence representing the severe spectrum of serine biosynthesis defects associated with PHGDH, PSAT1, or PSP gene mutations. The purpose of this study was to describe clinical/molecular and pathologic features of a NLS case caused by novel heterozygous missense variant in PHGDH gene identified in his consanguineous parents.
Subject(s)
Abnormalities, Multiple/genetics , Brain Diseases/genetics , Congenital Abnormalities/genetics , Fetal Growth Retardation/genetics , Ichthyosis/genetics , Limb Deformities, Congenital/genetics , Microcephaly/genetics , Phosphoglycerate Dehydrogenase/genetics , Stillbirth/genetics , Abnormalities, Multiple/mortality , Abnormalities, Multiple/pathology , Brain Diseases/mortality , Brain Diseases/pathology , Brazil/epidemiology , Congenital Abnormalities/mortality , Congenital Abnormalities/pathology , Consanguinity , Female , Fetal Growth Retardation/mortality , Fetal Growth Retardation/pathology , Genes, Lethal/genetics , Genetic Predisposition to Disease , Humans , Ichthyosis/mortality , Ichthyosis/pathology , Limb Deformities, Congenital/mortality , Limb Deformities, Congenital/pathology , Microcephaly/mortality , Microcephaly/pathology , Mutation, Missense/genetics , Pregnancy , Stillbirth/epidemiologyABSTRACT
We present a case of lethal fetal akinesia deformation sequence (FADS) caused by a frameshift variant in trans with a 148 kbp deletion encompassing 3-36 exons of AGRN. Pathogenic variants in AGRN have been described in families with a form of congenital myasthenic syndrome (CMS), manifesting in the early childhood with variable fatigable muscle weakness. To the best of our knowledge, this is the first case of FADS caused by defects in AGRN gene. FADS has been reported to be caused by pathogenic variants in genes previously associated with CMS including these involved in endplate development and maintenance: MuSK, DOK7, and RAPSN. FADS seems to be the most severe form of CMS. None of the reported in the literature CMS cases associated with AGRN had two null variants, like the case presented herein. This indicates a strong genotype-phenotype correlation.
Subject(s)
Agrin/genetics , Arthrogryposis/genetics , Genes, Lethal/genetics , Myasthenic Syndromes, Congenital/genetics , Adult , Arthrogryposis/diagnostic imaging , Arthrogryposis/pathology , Child , Female , Fetus/diagnostic imaging , Fetus/pathology , Humans , Male , Mutation/genetics , Myasthenic Syndromes, Congenital/pathology , Pedigree , PregnancyABSTRACT
Deriving cell populations from human embryonic stem cells (hESCs) for cell-based therapy is considered a promising strategy to achieve functional cells, yet its translation to clinical practice depends on achieving fully defined differentiated cells. In this work, we generated a miRNA-responsive lethal mRNA construct that selectively induces rapid apoptosis in hESCs by expressing a mutant (S184del) Bax variant. Insertion of miR-499 target sites in the construct enabled to enrich hESC-derived cardiomyocytes (CMs) in culture. A deterministic non-linear model was developed and validated with experimental data, to predict the outcome for each treatment cycle and the number of treatment cycle repetitions required to achieve completely purified cTNT-positive cells. The enriched hESC-CMs displayed physiological sarcomere orientation, functional calcium handling and after transplantation into SCID-NOD mice did not form teratomas. The modular miRNA responsive lethal mRNA construct could be employed in additional directed differentiation protocols, by adjusting the miRNA to the specific cells of choice.
Subject(s)
Cell Differentiation/genetics , MicroRNAs/genetics , Organogenesis/genetics , bcl-2-Associated X Protein/genetics , Animals , Apoptosis/genetics , Cell Line , Gene Expression Regulation, Developmental , Genes, Lethal/genetics , Genetic Vectors/genetics , Human Embryonic Stem Cells/metabolism , Humans , Mice , MicroRNAs/antagonists & inhibitors , Myocytes, Cardiac/metabolism , Myocytes, Cardiac/transplantation , RNA, Messenger/geneticsABSTRACT
Brain morphogenesis is an important process contributing to higher-order cognition, however our knowledge about its biological basis is largely incomplete. Here we analyze 118 neuroanatomical parameters in 1,566 mutant mouse lines and identify 198 genes whose disruptions yield NeuroAnatomical Phenotypes (NAPs), mostly affecting structures implicated in brain connectivity. Groups of functionally similar NAP genes participate in pathways involving the cytoskeleton, the cell cycle and the synapse, display distinct fetal and postnatal brain expression dynamics and importantly, their disruption can yield convergent phenotypic patterns. 17% of human unique orthologues of mouse NAP genes are known loci for cognitive dysfunction. The remaining 83% constitute a vast pool of genes newly implicated in brain architecture, providing the largest study of mouse NAP genes and pathways. This offers a complementary resource to human genetic studies and predict that many more genes could be involved in mammalian brain morphogenesis.
Subject(s)
Brain , Genetic Association Studies , Morphogenesis/genetics , Neuroanatomy , Neurogenesis/genetics , Animals , Brain/metabolism , Cell Cycle , Cognition , Cytoskeleton , Gene Regulatory Networks , Genes, Lethal/genetics , Humans , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Models, Animal , Mutation , Phenotype , SynapsesABSTRACT
Structural maintenance of chromosomes flexible hinge domain-containing 1 (SMCHD1) is an architectural factor critical for X-chromosome inactivation (XCI) and the repression of select autosomal gene clusters. In mice, homozygous nonsense mutations in Smchd1 cause female-specific embryonic lethality due to an XCI defect. However, although human mutations in SMCHD1 are associated with congenital arhinia and facioscapulohumeral muscular dystrophy type 2 (FSHD2), the diseases do not show a sex-specific bias, despite the essential nature of XCI in humans. To investigate whether there is a dosage imbalance for the sex chromosomes, we here analyze transcriptomic data from arhinia and FSHD2 patient blood and muscle cells. We find that X-linked dosage compensation is maintained in these patients. In mice, SMCHD1 controls not only protocadherin (Pcdh) gene clusters, but also Hox genes critical for craniofacial development. Ablating Smchd1 results in aberrant expression of these genes, coinciding with altered chromatin states and three-dimensional (3D) topological organization. In a subset of FSHD2 and arhinia patients, we also found dysregulation of clustered PCDH, but not HOX genes. Overall, our study demonstrates preservation of XCI in arhinia and FSHD2, and implicates SMCHD1 in the regulation of the 3D organization of select autosomal gene clusters.