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1.
Nat Commun ; 13(1): 7695, 2022 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-36509783

RESUMO

Mutations in PRPF31 cause autosomal dominant retinitis pigmentosa, an untreatable form of blindness. Gene therapy is a promising treatment for PRPF31-retinitis pigmentosa, however, there are currently no suitable animal models in which to develop AAV-mediated gene augmentation. Here we establish Prpf31 mutant mouse models using AAV-mediated CRISPR/Cas9 knockout, and characterize the resulting retinal degeneration phenotype. Mouse models with early-onset morphological and functional impairments like those in patients were established, providing new platforms in which to investigate pathogenetic mechanisms and develop therapeutic methods. AAV-mediated PRPF31 gene augmentation restored the retinal structure and function in a rapidly degenerating mouse model, demonstrating the first in vivo proof-of-concept for AAV-mediated gene therapy to treat PRPF31-retinitis pigmentosa. AAV-CRISPR/Cas9-PRPF31 knockout constructs also mediated efficient PRPF31 knockout in human and non-human primate retinal explants, laying a foundation for establishing non-human primate models using the method developed here.


Assuntos
Degeneração Retiniana , Retinite Pigmentosa , Camundongos , Animais , Humanos , Proteínas do Olho/metabolismo , Degeneração Retiniana/genética , Sistemas CRISPR-Cas/genética , Retinite Pigmentosa/genética , Retinite Pigmentosa/terapia , Genes Dominantes , Mutação , Modelos Animais de Doenças
2.
Mol Vis ; 28: 359-368, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36338669

RESUMO

Purpose: To identify the molecular mechanisms of the development of autosomal dominant retinitis pigmentosa (adRP) with incomplete penetrance in an Israeli Muslim Arab family. Methods: Two patients with adRP underwent a detailed ophthalmic evaluation, including funduscopic examination, visual field testing, optical coherence tomography, and electroretinography. Genetic analysis was performed using a combination of whole exome sequencing (WES) and Sanger sequencing. The pathogenicity of the identified intronic variant was evaluated in silico using several web-based tools, in vitro using a minigene-based assay, and in vivo using reverse transcription PCR analysis of lymphocyte-derived RNA. The relative abundance of alternatively spliced transcripts was evaluated using amplicon-based next-generation sequencing. The relative expression levels of PRPF31 and CNOT3 were measured using quantitative PCR (qPCR) analysis. Results: The two patients recruited in this study had childhood-onset RP, with night blindness as the initial symptom, followed by concentric restriction of the visual field. The funduscopic findings included narrowed retinal blood vessels and peripheral bone spicule pigmentation. By the third decade of life, the full-field electroretinography findings had been remarkably attenuated. In these patients, we identified a novel heterozygous intronic variant at position +5 of PRPF31 intron 11 (c.1146+5G>T). The same variant was also detected in one asymptomatic family member. Through in silico analysis, the variant was predicted to alter the splicing of intron 11. An in vitro splicing assay and a reverse transcription PCR analysis of lymphocyte-derived RNA revealed that the mutant allele yielded mainly a shorter transcript in which exon 11 was skipped. The skipping of exon 11 was expected to cause a frameshift and an aberrant truncated protein (p.Tyr359Serfs*29). The qPCR analysis revealed reduced PRPF31 expression levels in the mutation carriers, without a significant difference between the affected patient and his asymptomatic brother. We evaluated several factors that have been suggested to correlate with non-penetrance of PRPF31 mutations, including the number of cis-acting MSR1 elements adjacent to the PRPF31 core promoter, CNOT3 expression level, and CNOT3 rs4806718 single-nucleotide polymorphism. None of these factors correlated with non-penetrance in the family in this study. Conclusions: We report a novel intronic mutation in PRPF31 underlying adRP. This report expands the spectrum of pathogenic mutations in PRPF31 and further demonstrates the importance of intronic mutations. Moreover, it demonstrates the phenomenon of incomplete penetrance previously associated with PRPF31 mutations. The fact that the non-penetrance in the family in this study could not be explained by any of the known mechanisms suggests the possible contribution of a novel modifier of PRPF31 penetrance.


Assuntos
Proteínas do Olho , Retinite Pigmentosa , Masculino , Humanos , Criança , Proteínas do Olho/genética , Proteínas do Olho/metabolismo , Linhagem , Retinite Pigmentosa/diagnóstico , Mutação/genética , RNA , Genes Dominantes , Fatores de Transcrição/genética
3.
Genes (Basel) ; 13(11)2022 10 28.
Artigo em Inglês | MEDLINE | ID: mdl-36360203

RESUMO

Osteopetrosis (from the Greek "osteo": bone; "petrosis": stone) is a clinically and genetically heterogeneous group of rare diseases of the skeleton, sharing the same main characteristic of an abnormally increased bone density. Dense bones in radiological studies are considered the hallmark of these diseases, and the reason for the common term used: "Marble bone disease". Interestingly, a radiologist, Dr. Albers-Schonberg, described this disease for the first time in Germany in 1904. Indeed, radiology has a key role in the clinical diagnosis of osteopetrosis and is fundamental in assessing the disease severity and complications, as well as in follow-up controls and the evaluation of the response to treatment. Osteopetrosis includes a broad spectrum of genetic mutations with very different clinical symptoms, age onset, and prognosis (from mild to severe). This diversity translates into different imaging patterns related to specific mutations, and different disease severity. The main recognized types of osteopetrosis are the infantile malignant forms with autosomal recessive transmission (ARO-including the rarer X-linked recessive form); the intermediate autosomal recessive form (IAO); and the autosomal dominant ones ADO, type I, and type II, the latter being called 'Albers-Schonberg' disease. Imaging features may change among those distinct types with different patterns, severities, skeletal segment involvement, and speeds of progression. There are several classical and well-recognized radiological features related to osteopetrosis: increased bone density (all types with different degrees of severity assuming a 'Marble Bone Appearance' especially in the ARO type), different metaphyseal alterations/enlargement including the so-called 'Erlenmeyer flask deformity' (particularly of femoral bones, more frequent in ADO type 2, and less frequent in ARO and IAO), 'bone in bone' appearance (more frequent in ADO type 2, less frequent in ARO and IAO), and 'rugger-jersey spine' appearance (typical of ADO type 2). After conducting an overview of the epidemiological and clinical characteristic of the disease, this review article aims at summarizing the main radiological features found in different forms of osteopetrosis together with their inheritance pattern.


Assuntos
Osteopetrose , Radiologia , Humanos , Osteopetrose/diagnóstico por imagem , Osteopetrose/genética , Osteopetrose/patologia , Genes Dominantes , Padrões de Herança , Carbonato de Cálcio
4.
Int J Mol Sci ; 23(20)2022 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-36293299

RESUMO

In plants, the accumulation of carotenoids can maintain the balance of the photosystem and improve crop nutritional quality. Therefore, the molecular mechanisms underlying carotenoid synthesis and accumulation should be further explored. In this study, carotenoid accumulation differed significantly among parental Brassica rapa. Genetic analysis was carried out using the golden inner leaf '1900264' line and the light-yellow inner leaf '1900262' line, showing that the golden inner leaf phenotype was controlled by a single dominant gene. Using bulked-segregant analysis sequencing, BraA09g007080.3C encoding the ORANGE protein was selected as a candidate gene. Sequence alignment revealed that a 4.67 kb long terminal repeat insertion in the third exon of the BrGOLDEN resulted in three alternatively spliced transcripts. The spatiotemporal expression results indicated that BrGOLDEN might regulate the expression levels of carotenoid-synthesis-related genes. After transforming BrGOLDEN into Arabidopsis thaliana, the seed-derived callus showed that BrGOLDENIns and BrGOLDENDel lines presented a yellow color and the BrGOLDENLdel line presented a transparent phenotype. In addition, using the yeast two-hybrid assay, BrGOLDENIns, BrGOLDENLdel, and Brgoldenwt exhibited strong interactions with BrPSY1, but BrGOLDENDel did not interact with BrPSY1 in the split-ubiquitin membrane system. In the secondary and 3D structure analysis, BrGOLDENDel was shown to have lost the PNFPSFIPFLPPL sequences at the 125 amino acid position, which resulted in the α-helices of BrGOLDENDel being disrupted, restricting the formation of the 3D structure and affecting the functions of the protein. These findings may provide new insights into the regulation of carotenoid synthesis in B. rapa.


Assuntos
Arabidopsis , Brassica rapa , Brassica rapa/genética , Brassica rapa/metabolismo , Genes Dominantes , Carotenoides/metabolismo , Arabidopsis/genética , Aminoácidos/genética , Ubiquitinas/genética
5.
Int J Mol Sci ; 23(20)2022 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-36293404

RESUMO

Cucurbits powdery mildew (CPM) is one of the main limiting factors of melon cultivation worldwide. Resistance to races 1, 2, and 5 has been reported in the African accession TGR-1551, whose resistance is controlled by a dominant-recessive epistasis. The dominant and recessive quantitative trail loci (QTL) have previously been located in chromosomes 5 and 12, respectively. We used several densely genotyped BC3 families derived from the cross between TGR-1551 and the susceptible cultivar 'Bola de Oro' to finely map these resistance regions. The further phenotyping and genotyping of the selected BC5, BC5S1, BC5S2, BC4S1, BC4xPS, and (BC4xPS) S1 offspring allowed for the narrowing of the candidate intervals to a 250 and 381 kb region in chromosomes 5 and 12, respectively. Moreover, the temperature effect over the resistance provided by the dominant gene has been confirmed. High resolution melting markers (HRM) were tightly linked to both resistance regions and will be useful in marker-assisted selection programs. Candidate R genes with variants between parents that caused a potential modifier impact on the protein function were identified within both intervals. These candidate genes provide targets for future functional analyses to better understand the resistance to powdery mildew in melons.


Assuntos
Ascomicetos , Humanos , Mapeamento Cromossômico , Ascomicetos/genética , Doenças das Plantas/genética , Erysiphe , Genes Dominantes , Resistência à Doença/genética
7.
Proc Natl Acad Sci U S A ; 119(32): e2113795119, 2022 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-35917346

RESUMO

KIF1A is a kinesin superfamily motor protein that transports synaptic vesicle precursors in axons. Cargo binding stimulates the dimerization of KIF1A molecules to induce processive movement along microtubules. Mutations in human Kif1a lead to a group of neurodegenerative diseases called KIF1A-associated neuronal disorder (KAND). KAND mutations are mostly de novo and autosomal dominant; however, it is unknown if the function of wild-type KIF1A motors is inhibited by heterodimerization with mutated KIF1A. Here, we have established Caenorhabditis elegans models for KAND using CRISPR-Cas9 technology and analyzed the effects of human KIF1A mutation on axonal transport. In our C. elegans models, both heterozygotes and homozygotes exhibited reduced axonal transport. Suppressor screening using the disease model identified a mutation that recovers the motor activity of mutated human KIF1A. In addition, we developed in vitro assays to analyze the motility of heterodimeric motors composed of wild-type and mutant KIF1A. We find that mutant KIF1A significantly impaired the motility of heterodimeric motors. Our data provide insight into the molecular mechanism underlying the dominant nature of de novo KAND mutations.


Assuntos
Transporte Axonal , Caenorhabditis elegans , Cinesinas , Doenças Neurodegenerativas , Vesículas Sinápticas , Animais , Transporte Axonal/genética , Caenorhabditis elegans/genética , Modelos Animais de Doenças , Genes Dominantes , Humanos , Cinesinas/genética , Atividade Motora/genética , Mutação , Doenças Neurodegenerativas/genética , Vesículas Sinápticas/genética , Vesículas Sinápticas/metabolismo
8.
Theor Appl Genet ; 135(10): 3563-3570, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36030437

RESUMO

KEY MESSAGE: Identification and mapping of an inhibitor of Ndhrl1 mediating nitrogen-dependent hypersensitive reaction-like phenotype in wheat. Hypersensitive reaction-like (HRL) traits are characteristic of spontaneous lesions including yellowish spots, brown spots or white-stripe that appeared randomly and dispersedly on all the leaves in the absence of plant pathogens. Our previous studies have shown that the wheat line P7001 showed an HRL trait at low nitrogen supply, and this trait was controlled by the gene Ndhrl1 (Nitrogen-dependent hypersensitive reaction-like 1). In order to investigate the robustness of the trait expression mediated by Ndhrl1 under different genetic backgrounds, seven genetic populations, with P7001 being the common female parent, were constructed and analyzed. F1 plants from six of the seven combinations showed HRL trait and Ndhrl1 segregated in a dominant way of HRL: non-HRL = 3:1 in the six populations (F2). Exceptionally, the F1 plants of P7001/Fielder combination showed non-HRL trait and HRL trait in the F2 population showed a contrasting recessive segregation ratio of HRL: non-HRL = 1:3, suggesting Fielder may have another HRL-related gene. Using 55 K SNP array and PCR-based markers, the HRL-related gene in Fielder was mapped to an interval of 5.63-12.91 Mb on the short arm of chromosome 2B with the flanking markers Yzu660R075552 and Yzu660F075941. A recombinant with genomic region of Fielder at Ndhrl1 locus showing HRL trait demonstrated that Fielder also harbored Ndhrl1 same as P7001. Thus, Fielder carries a single dominant suppressor of Ndhrl1, designated as Ihrl1 (Inhibitor of hypersensitive reaction-like). Interestingly, Ihrl1 is tightly linked to Ndhrl1 and may be also involved in nitrogen metabolic and (or) signaling pathways.


Assuntos
Nitrogênio , Triticum , Mapeamento Cromossômico , Genes Dominantes , Genes de Plantas , Ligação Genética , Fenótipo , Doenças das Plantas/genética , Triticum/genética
9.
Planta ; 256(4): 66, 2022 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-36036325

RESUMO

MAIN CONCLUSION: By constructing an F2 population, a new potential dominant resistance gene to TuMV in Brassica rapa was mapped and identified. Brassica rapa is the most widely grown vegetable crop in China, and turnip mosaic virus (TuMV) is a great threat to its production. Hence, it is a very important work to excavate more and novel resistance genes in B. rapa. In this study, the resistant line B80124 and the susceptible line B80450 were used to construct the F2 populations, and through genetic analysis, the resistance to TuMV was found to be controlled by a dominant gene. Bulked segregant analysis sequence (BSA-seq) was used for the primary mapping, and an intersection (22.25-25.03 Mb) was obtained. After fine mapping using single nucleotide polymorphisms (SNP) markers, the candidate region was narrowed to 330 kb between the SNP markers A06S11 and A06S14, including eight genes relating to disease resistance. Using the transcriptome analysis and sequence identification, BraA06g035130.3C was screened as the final candidate gene, and it contained two deletion mutations, leading to frameshift in the susceptible line B80450. In addition, the phylogenetic analysis, hydrophilia and hydrophobicity analysis, subcellular location prediction analysis, amino acid bias analysis, and 3D modeling structures of BraA06g035130.3C were conducted to predict its functions. This study was conducive to the identification of a new TuMV resistance gene in B. rapa, which is of important scientific significance and application value for the improvement of TuMV resistance traits and molecular design breeding for Brassica crops.


Assuntos
Brassica rapa , Genes Dominantes , Filogenia , Doenças das Plantas , Potyvirus
10.
Sci Transl Med ; 14(654): eabn0449, 2022 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-35857824

RESUMO

Programmable RNA editing tools enable the reversible correction of mutant transcripts, reducing the potential risk associated with permanent genetic changes associated with the use of DNA editing tools. However, the potential of these RNA tools to treat disease remains unknown. Here, we evaluated RNA correction therapy with Cas13-based RNA base editors in the myosin VI p.C442Y heterozygous mutation (Myo6C442Y/+) mouse model that recapitulated the phenotypes of human dominant-inherited deafness. We first screened several variants of Cas13-based RNA base editors and guide RNAs (gRNAs) targeting Myo6C442Y in cultured cells and found that mini dCas13X.1-based adenosine base editor (mxABE), composed of truncated Cas13X.1 and the RNA editing enzyme adenosine deaminase acting on RNA 2 deaminase domain variant (ADAR2ddE488Q), exhibited both high efficiency of A > G conversion and low frequency of off-target edits. Single adeno-associated virus (AAV)-mediated delivery of mxABE in the cochlea corrected the mutated Myo6C442Y to Myo6WT allele in homozygous Myo6C442Y/C442Y mice and resulted in increased Myo6WT allele in the injected cochlea of Myo6C442Y/+ mice. The treatment rescued auditory function, including auditory brainstem response and distortion product otoacoustic emission up to 3 months after AAV-mxABE-Myo6 injection in Myo6C442Y/+ mice. We also observed increased survival rate of hair cells and decreased degeneration of hair bundle morphology in the treated compared to untreated control ears. These findings provide a proof-of-concept study for RNA editing tools as a therapeutic treatment for various semidominant forms of hearing loss and other diseases.


Assuntos
Surdez , Perda Auditiva , Animais , Artrogripose , Genes Dominantes , Células Ciliadas Auditivas , Perda Auditiva/genética , Perda Auditiva/terapia , Humanos , Camundongos , RNA
11.
Genes (Basel) ; 13(8)2022 07 26.
Artigo em Inglês | MEDLINE | ID: mdl-35893064

RESUMO

Retinitis pigmentosa (RP) is the most common group of inherited retinal degenerations and pathogenic variants in the Rhodopsin (RHO) gene are major cause for autosomal dominant RP (adRP). Despite extensive attempts to treat RHO-associated adRP, standardized curative treatment is still lacking. Recently developed base editors offer an exciting opportunity to correct pathogenic single nucleotide variants and are currently able to correct all transition variants and some transversion variants. In this study, we analyzed previously reported pathogenic RHO variants (n = 247) for suitable PAM sites for currently available base editors utilizing the Streptococcus pyogenes Cas9 (SpCas9), Staphylococcus aureus Cas9 (SaCas9) or the KKH variant of SaCas9 (KKH-SaCas9) to assess DNA base editing as a treatment option for RHO-associated adRP. As a result, 55% of all the analyzed variants could, in theory, be corrected with base editors, however, PAM sites were available for only 32% of them and unwanted bystander edits were predicted for the majority of the designed guide RNAs. As a conclusion, base editing offers exciting possibilities to treat RHO-associated adRP in the future, but further research is needed to develop base editing constructs that will provide available PAM sites for more variants and that will not introduce potentially harmful bystander edits.


Assuntos
Edição de Genes , Retinite Pigmentosa , Rodopsina , DNA/genética , Genes Dominantes , Humanos , Mutação , Retinite Pigmentosa/genética , Retinite Pigmentosa/terapia , Rodopsina/genética , Staphylococcus aureus/genética
12.
Theor Appl Genet ; 135(7): 2187-2196, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35668203

RESUMO

KEY MESSAGE: A candidate non photosensitive gene S m F TS H10 was identified by combining bulked segregant analysis and map­based cloning. Low light condition often leads to poor coloration of photosensitive eggplant. Here, we obtained a non-photosensitive eggplant that can synthesize large amount of anthocyanin under shading conditions. Genetic analysis of F1 and F2 populations revealed that the phenotype of non-photosensitivity was regulated by a single dominant nuclear gene, herein temporarily designated SmFTSH10. Through Bulked segregant analysis (BSA), SNP haplotyping and fine genetic mapping delimited SmFTSH10 to a 290 kb region of eggplant chromosome 10 flanking by markers dCAPS21 and dCAPS32. Sequence analysis revealed C-base deletion in the fourth exon of SmFTSH10 resulted in premature termination of translation. The expression level of SmFTSH10 decreased significantly in anthocyanin-rich parts of mutant '145' compared with the wild-type 'LSHX'. Sequencing of 10 recombinants revealed that the C-base deletion in the 4th exon of SmFTSH10 was co-segregated with the non-photosensitive phenotype, and the sequencing analysis of the natural population of eggplant also showed that the Indel in SmFTSH10 had a high accuracy in the identification of the photosensitivity of eggplant. Light-responsive expression patterns analysis suggests that it has the same expression trend as the genes involved in eggplant anthocyanin biosynthesis, which supports SmFTSH10 as the most possible candidate gene of non-photosensitivity. These findings provide a new insight into understanding the molecular mechanisms of anthocyanin biosynthesis in non-photosensitive eggplant.


Assuntos
Solanum melongena , Antocianinas , Mapeamento Cromossômico , Genes Dominantes , Mutação INDEL , Solanum melongena/genética , Solanum melongena/metabolismo
13.
Invest Ophthalmol Vis Sci ; 63(5): 19, 2022 05 02.
Artigo em Inglês | MEDLINE | ID: mdl-35579903

RESUMO

Purpose: Heterozygous truncating variants of TOPORS have been reported to cause autosomal dominant retinitis pigmentosa (adRP). The purpose of this study was to investigate whether all heterozygous truncating variants, including copy number variants (CNVs), are pathogenic. Methods: TOPORS truncating variants were collected and reviewed through an in-house dataset and existing databases. Individuals with truncating variants underwent ophthalmological evaluation. Results: Six truncating variants were detected in seven families. Three N-terminus truncating variants were detected in three families without RP, and the other three were identified in four unrelated families with typical RP. Based on the in-house dataset and published literature, 17 truncating variants were identified in 47 families with RP. All RP-associated truncating alleles, except one, were distributed in the last exon of TOPORS and clustered in amino acid residues 807 to 867 (46/47, 97.9%). Conversely, in the gnomAD database, only one truncating allele (1/27, 3.7%) was in this region, and the others were outside (26/27, 96.3%), suggesting that the pathogenic truncating variants were significantly clustered in residues 807 to 867 (χ2 = 65.6, P = 1.1 × 10-17). Additionally, three CNVs involving the N-terminus of TOPORS were recorded in control populations but were absent in affected patients. Conclusions: This study suggests that all pathogenic truncating variants of TOPORS were clustered in residues 807 to 867, whereas the truncating variants outside this region and the CNVs involving the N-terminus were not associated with RP. A dominant-negative effect, rather than haploinsufficiency, is speculated to be the underlying pathogenesis. These findings provide valuable information for interpreting variation in TOPORS and other genes in similar situations, especially for CNVs.


Assuntos
Aminoácidos , Proteínas de Neoplasias , Proteínas Nucleares , Retinite Pigmentosa , Ubiquitina-Proteína Ligases , Aminoácidos/genética , Análise Mutacional de DNA , Genes Dominantes , Humanos , Proteínas Mutantes/genética , Mutação , Proteínas de Neoplasias/genética , Proteínas Nucleares/genética , Linhagem , Retinite Pigmentosa/genética , Ubiquitina-Proteína Ligases/genética
14.
Int J Mol Sci ; 23(8)2022 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-35457072

RESUMO

Inherited diseases caused by connexin mutations are found in multiple organs and include hereditary deafness, congenital cataract, congenital heart diseases, hereditary skin diseases, and X-linked Charcot-Marie-Tooth disease (CMT1X). A large number of knockout and knock-in animal models have been used to study the pathology and pathogenesis of diseases of different organs. Because the structures of different connexins are highly homologous and the functions of gap junctions formed by these connexins are similar, connexin-related hereditary diseases may share the same pathogenic mechanism. Here, we analyze the similarities and differences of the pathology and pathogenesis in animal models and find that connexin mutations in gap junction genes expressed in the ear, eye, heart, skin, and peripheral nerves can affect cellular proliferation and differentiation of corresponding organs. Additionally, some dominant mutations (e.g., Cx43 p.Gly60Ser, Cx32 p.Arg75Trp, Cx32 p.Asn175Asp, and Cx32 p.Arg142Trp) are identified as gain-of-function variants in vivo, which may play a vital role in the onset of dominant inherited diseases. Specifically, patients with these dominant mutations receive no benefits from gene therapy. Finally, the complete loss of gap junctional function or altered channel function including permeability (ions, adenosine triphosphate (ATP), Inositol 1,4,5-trisphosphate (IP3), Ca2+, glucose, miRNA) and electric activity are also identified in vivo or in vitro.


Assuntos
Doença de Charcot-Marie-Tooth , Conexinas , Animais , Doença de Charcot-Marie-Tooth/genética , Conexina 26/genética , Conexinas/genética , Junções Comunicantes/genética , Junções Comunicantes/patologia , Genes Dominantes , Humanos , Mutação
15.
Proc Natl Acad Sci U S A ; 119(11): e2115202119, 2022 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-35271391

RESUMO

SignificanceIn humans, genetic mutations in the retinal pigment epithelium (RPE) 65 are associated with blinding diseases, for which there is no effective therapy alleviating progressive retinal degeneration in affected patients. Our findings uncovered that the increased free opsin caused by enhancing the ambient light intensity increased retinal activation, and when compounded with the RPE visual cycle dysfunction caused by the heterozygous D477G mutation and aggregation, led to the onset of retinal degeneration.


Assuntos
Proteínas do Olho , Genes Dominantes , Distrofias Retinianas , cis-trans-Isomerases , Animais , Proteínas do Olho/genética , Camundongos , Camundongos Knockout , Mutação , Retina/enzimologia , Retina/patologia , Distrofias Retinianas/genética , Visão Ocular , cis-trans-Isomerases/genética
16.
Hum Genet ; 141(3-4): 445-453, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35254497

RESUMO

Novel hearing loss (HL) genes are constantly being discovered, and evidence from independent studies is essential to strengthen their position as causes of hereditary HL. To address this issue, we searched our genetic data of families with autosomal dominant HL (ADHL) who had been tested with high-throughput DNA sequencing methods. For CD164, only one pathogenic variant in one family has so far been reported. For LMX1A, just two previous studies have revealed its involvement in ADHL. In this study we found two families with the same pathogenic variant in CD164 and one family with a novel variant in LMX1A (c.686C>A; p.(Ala229Asp)) that impairs its transcriptional activity. Our data show recurrence of the same CD164 variant in two HL families of different geographic origin, which strongly suggests it is a mutational hotspot. We also provide further evidence for haploinsufficiency as the pathogenic mechanism underlying LMX1A-related ADHL.


Assuntos
Surdez , Endolina , Perda Auditiva Neurossensorial , Perda Auditiva , Proteínas com Homeodomínio LIM , Fatores de Transcrição , Artrogripose , Surdez/genética , Endolina/genética , Genes Dominantes , Perda Auditiva/genética , Perda Auditiva Neurossensorial/genética , Perda Auditiva Neurossensorial/patologia , Humanos , Proteínas com Homeodomínio LIM/genética , Mutação , Linhagem , Fatores de Transcrição/genética
17.
J Hum Genet ; 67(9): 553-556, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35338243

RESUMO

Heterozygous mutations in Bicaudal D2 Drosophila homolog 2 (BICD2) gene, encodes a vesicle transport protein involved in dynein-mediated movement along microtubules, are responsible for an exceedingly rare autosomal dominant spinal muscular atrophy type 2A which starts in the childhood and predominantly effects lower extremities. Recently, a more severe form, type 2B, has also been described. Here, we present a patient born to a consanguineous union and who suffered from intellectual disability, speech delay, epilepsy, happy facial expression, truncal obesity with tappering fingers, and joint hypermobility. Whole-exome sequencing analysis revealed a rare, homozygous missense mutation (c.731T>C; p.Leu244Pro) in BICD2 gene. This finding presents the first report in the literature for homozygous BICD2 mutations and its association with a Cohen-Like syndrome. Patients presenting with Cohen-Like phenotypes should be further interrogated for mutations in BICD2.


Assuntos
Deficiência Intelectual , Atrofia Muscular Espinal , Genes Dominantes , Humanos , Deficiência Intelectual/genética , Proteínas Associadas aos Microtúbulos/genética , Atrofia Muscular Espinal/genética , Mutação , Mutação de Sentido Incorreto
19.
Ann Hum Genet ; 86(4): 207-217, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-35292975

RESUMO

AIM: Autosomal dominant non-syndromic hearing loss is a common sensorineural disorder with extremely high genetic heterogeneity. CEA antigen-related cell adhesion molecule 16(CEACAM16)is a secreted glycoprotein encoded by the CEACAM16 gene. Mutations in CEACAM16 lead to autosomal dominant non-syndromic hearing loss in humans, due defects in the tectorial membrane of the inner ear. Here we reported a novel missense variant in CEACAM16 gene causes autosomal dominant non-syndromic hearing loss. MATERIAL AND METHODS: A four-generation Chinese family affected by late-onset and progressive hearing loss was enrolled in this study. The proband was analyzed by targeted next-generation sequencing and bioinformatic analysis. And in vitro experiments were performed in overexpressed transfected HEK293T cells to investigate the pathogenesis of the mutant protein. RESULTS: We identified a novel missense variant in the CEACAM16 gene c.763A>G; (p.Arg255Gly) as causing autosomal dominant non-syndromic hearing loss in the Chinese family. Using Western blot analysis, ELISA, and immunofluorescence we found increased expression level of the secreted mutant CEACAM16 protein, both intracellularly and extracellularly, compared with wild type CEACAM16 protein. CONCLUSION: Our study showed that the p.Arg255Gly variant leads to increased secretion of mutant CEACAM16 protein, with potential deleterious effect to the function of the protein. Our findings expand the mutation spectrum of CEACAM16, and further the understanding CEACAM16 function and implications in disease.


Assuntos
Artrogripose , Surdez , Perda Auditiva Neurossensorial , Artrogripose/genética , Moléculas de Adesão Celular/genética , Surdez/genética , Genes Dominantes , Perda Auditiva Neurossensorial/genética , Humanos , Mutação de Sentido Incorreto , Linhagem
20.
STAR Protoc ; 3(1): 101150, 2022 03 18.
Artigo em Inglês | MEDLINE | ID: mdl-35146449

RESUMO

Identifying a disease gene and determining its causality in patients can be challenging. Here, we present an approach to predicting the pathogenicity of deletions and missense variants for an autosomal dominant gene. We provide online resources for identifying patients and determining constraint metrics to isolate the causal gene among several candidates encompassed in a shared region of deletion. We also provide instructions for optimizing functional annotation programs that may be otherwise inaccessible to a nonexpert or novice in computational approaches. For complete details on the use and execution of this protocol, please refer to Gennarino et al. (2018).


Assuntos
Artrogripose , Genes Dominantes , Humanos , Mutação de Sentido Incorreto/genética , Virulência
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