Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 50
Filtrar
1.
Cancer Med ; 2020 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-33159431

RESUMO

This study aimed to estimate the rates of biopsy undersampling and progression for four prostate cancer (PCa) active surveillance (AS) cohorts within the Movember Foundation's Global Action Plan Prostate Cancer Active Surveillance (GAP3) consortium. We used a hidden Markov model (HMM) to estimate factors that define PCa dynamics for men on AS including biopsy under-sampling and progression that are implied by longitudinal data in four large cohorts included in the GAP3 database. The HMM was subsequently used as the basis for a simulation model to evaluate the biopsy strategies previously proposed for each of these cohorts. For the four AS cohorts, the estimated annual progression rate was between 6%-13%. The estimated probability of a biopsy successfully sampling undiagnosed non-favorable risk cancer (biopsy sensitivity) was between 71% and 80%. In the simulation study of patients diagnosed with favorable risk cancer at age 50, the mean number of biopsies performed before age 75 was between 4.11 and 12.60, depending on the biopsy strategy. The mean delay time to detection of non-favorable risk cancer was between 0.38 and 2.17 years. Biopsy undersampling and progression varied considerably across study cohorts. There was no single best biopsy protocol that is optimal for all cohorts, because of the variation in biopsy under-sampling error and annual progression rates across cohorts. All strategies demonstrated diminishing benefits from additional biopsies.

2.
Arch Virol ; 165(12): 2817-2828, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32990841

RESUMO

Enterovirus 71 (EV-A71) and coxsackievirus A16 (CV-A16) are the major pathogens responsible for hand, foot and mouth disease (HFMD), but the mechanism by which these viruses cause disease remains unclear. In this study, we used transcriptome sequencing technology to investigate changes in the transcriptome profiles after infection with EV-A71 and CV-A16 in human bronchial epithelial (16HBE) cells. Using systematic bioinformatics analysis, we then searched for useful clues regarding the pathogenesis of HFMD. As a result, a total of 111 common differentially expressed genes were present in both EV-A71- and CV-A16-infected cells. A trend analysis of these 111 genes showed that 91 of them displayed the same trend in EV-A71 and CV-A16 infection, including 49 upregulated genes and 42 downregulated genes. These 91 genes were further used to conduct GO, pathway, and coexpression network analysis. It was discovered that enriched GO terms (such as histone acetylation and positive regulation of phosphorylation) and pathways (such as glycosylphosphatidylinositol (GPI)-anchor biosynthesis and DNA replication) might be closely associated with the pathogenic mechanism of these two viruses, and key genes (such as TBCK and GPC) might be involved in the progression of HFMD. Finally, we randomly selected 10 differentially expressed genes for qRT-PCR to validate the transcriptome sequencing data. The experimental qRT-PCR results were roughly in agreement with the results of transcriptome sequencing. Collectively, our results provide clues to the mechanism of pathogenesis of HFMD induced by EV-A71 and CV-A16.


Assuntos
Enterovirus Humano A/patogenicidade , Células Epiteliais/virologia , Perfilação da Expressão Gênica , Doença de Mão, Pé e Boca/patologia , Linhagem Celular , Replicação do DNA , Células Epiteliais/fisiologia , Regulação da Expressão Gênica , Doença de Mão, Pé e Boca/virologia , Humanos
3.
J Med Genet ; 2020 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-32381727

RESUMO

BACKGROUND: Early-onset scoliosis (EOS), defined by an onset age of scoliosis less than 10 years, conveys significant health risk to affected children. Identification of the molecular aetiology underlying patients with EOS could provide valuable information for both clinical management and prenatal screening. METHODS: In this study, we consecutively recruited a cohort of 447 Chinese patients with operative EOS. We performed exome sequencing (ES) screening on these individuals and their available family members (totaling 670 subjects). Another cohort of 13 patients with idiopathic early-onset scoliosis (IEOS) from the USA who underwent ES was also recruited. RESULTS: After ES data processing and variant interpretation, we detected molecular diagnostic variants in 92 out of 447 (20.6%) Chinese patients with EOS, including 8 patients with molecular confirmation of their clinical diagnosis and 84 patients with molecular diagnoses of previously unrecognised diseases underlying scoliosis. One out of 13 patients with IEOS from the US cohort was molecularly diagnosed. The age at presentation, the number of organ systems involved and the Cobb angle were the three top features predictive of a molecular diagnosis. CONCLUSION: ES enabled the molecular diagnosis/classification of patients with EOS. Specific clinical features/feature pairs are able to indicate the likelihood of gaining a molecular diagnosis through ES.

4.
Biosci Biotechnol Biochem ; 84(7): 1394-1400, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32180505

RESUMO

This study investigated the antioxidant defense system involved in the tolerance of soybean (Glycine max) to aluminum (Al) stress. Physiological assays showed that the amount of superoxide free radicals (O2 -), hydrogen peroxide (H2O2), and malondialdehyde (MDA) were significantly lower in an Al-resistant soybean cultivar (cv. PI416937) than in an Al-sensitive soybean cultivar (cv. Huachun18). Comparative analysis of microarray data from both genotypes following Al-stress treatment revealed that the expression of a series of antioxidant enzymes genes was induced in the Al-resistant cultivar. The quantitative real time-PCR (qRT-PCR) assay showed that the transcript levels of genes encoding antioxidant enzymes, including GmCAT1, GmPOD1, GmGST1, GmAPX, GmGSH1, and GmSOD, were higher in the Al-resistant cultivar than in the Al-sensitive cultivar in Al-stress conditions. Furthermore, GmCAT1-overexpressing Arabidopsis plants had improved tolerance to Al-stress and lower O2 -, H2O2, and MDA contents than wild-type plants. Therefore, providing evidence that the antioxidant defense system is essential for Al tolerance in soybean. ABBREVIATIONS: Al: aluminum; O2 -: superoxide free radicals; ROS: reactive oxygen species; H2O2: hydrogen peroxide; MDA: malondialdehyde; qRT-PCR: quantitative reverse transcription polymerase chain reaction; GO: gene ontology; WT: wild type; MS medium: Murashige and Skoog medium.

5.
J Assist Reprod Genet ; 37(4): 829-840, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32124190

RESUMO

PURPOSE: To investigate the relation between mutations in ciliopathy-related SPAG6 and RSPH3 and male infertility with severe asthenoteratospermia characterized by multiple flagellar malformations and reveal the intracytoplasmic sperm injection (ICSI) outcomes of those primary ciliary dyskinesia (PCD) patients. METHODS: Whole-exome sequencing was applied to identify the pathogenic genes for the five PCD patients. The ICSI outcomes of those patients were compared with eight DNAH1-mutated patients and 215 oligo-asthenospermia (OAT) patients. RESULTS: We identified, for the first time, the compound heterozygous SPAG6 mutations (c.143_145del: p.48_49del, c.585delA: p.Lys196Serfs*6) in a sporadic PCD patient. Further, a novel homozygous nonsynonymous RSPH3 mutation (c.C799T: p.Arg267Cys) was identified in another PCD patient with consanguineous parents. The pathogenicity of these mutations in the assembly of sperm flagella was confirmed by flagellar ultrastructure analysis, immunofluorescence, and quantitative real-time PCR. All five patients underwent six ICSI cycles. The fertilization rate, blastocyst development rate, and clinical pregnancy rate were 69.3%, 50.0%, and 66.7%, respectively. Four of the five couples, including the subjects carrying mutations in SPAG6 or RSPH3, got healthy children born after ICSI. Additionally, the ICSI outcomes of the five PCD couples were statistically comparable with those of the eight DNAH1-mutated couples and the 215 OAT couples. CONCLUSIONS: Mutations in ciliopathy-related SPAG6 and RSPH3 cause severe asthenoteratospermia characterized by multiple flagellar malformations, resulting in sterility. ICSI is an optimal management with a positive pregnancy outcome.

6.
J Med Genet ; 57(7): 445-453, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32051257

RESUMO

BACKGROUND: Asthenoteratospermia, one of the most common causes for male infertility, often presents with defective sperm heads and/or flagella. Multiple morphological abnormalities of the sperm flagella (MMAF) is one of the common clinical manifestations of asthenoteratospermia. Variants in several genes including DNAH1, CEP135, CATSPER2 and SUN5 are involved in the genetic pathogenesis of asthenoteratospermia. However, more than half of the asthenoteratospermia cases cannot be explained by the known pathogenic genes. METHODS AND RESULTS: Two asthenoteratospermia-affected men with severe MMAF (absent flagella in >90% spermatozoa) from consanguineous families were subjected to whole-exome sequencing. The first proband had a homozygous missense mutation c.188G>A (p.Arg63Gln) of DZIP1 and the second proband had a homozygous stop-gain mutation c.690T>G (p.Tyr230*). Both of the mutations were neither detected in the human population genome data (1000 Genomes Project, Exome Aggregation Consortium) nor in our own data of a cohort of 875 Han Chinese control populations. DZIP1 encodes a DAZ (a protein deleted in azoospermia) interacting protein, which was associated with centrosomes in mammalian cells. Immunofluorescence staining of the centriolar protein Centrin1 indicated that the spermatozoa of the proband presented with abnormal centrosomes, including no concentrated centriolar dot or more than two centriolar dots. HEK293T cells transfected with two DZIP1-mutated constructs showed reduced DZIP1 level or truncated DZIP1. The Dzip1-knockout mice, generated by the CRSIPR-Cas9, revealed consistent phenotypes of severe MMAF. CONCLUSION: Our study strongly suggests that homozygous DZIP1 mutations can induce asthenoteratospermia with severe MMAF. The deficiency of DZIP1 induces sperm centrioles dysfunction and causes the absence of flagella.

7.
Plant Physiol Biochem ; 147: 215-222, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31869734

RESUMO

Ethylene-response factor (ERF) proteins are members of a transcription factor family involved in plant growth and environmental stress responses, but the biological functions of ERF members in adzuki bean (Vigna angularis var. angularis) remain unknown. In addition, it is unclear whether these proteins have a role in regulating responses to abiotic stressors. Here, we identified 47 ERF genes by analyzing the adzuki bean genome. Whole-transcriptome analyses of plants under saline-alkaline stress suggested that the expression of 13 ERF genes was induced in response to saline-alkaline stress. Analysis of the cis-acting elements showed that the promoters of these saline-alkaline stress-inducible ERF genes contained LTRs, DREs, MYBs, ABREs, MYCs, CGTCA-, and TGACG-motifs, which are involved in abiotic stress responses. The expression of VaERF3 was induced by NaHCO3, polyethylene glycol 6000, NaCl, and ABA (abscisic acid), as determined by qRT-PCR. Overexpression of VaERF3 in transgenic Arabidopsis resulted in higher levels of proline accumulation and lower malondialdehyde and reactive oxygen species contents in plants grown under saline-alkaline stress conditions. Moreover, VaERF3 encoded a nuclear-localized transcriptional activator that promoted the expression of stress-responsive genes. Collectively, these results are of great significance in elucidating the mechanisms of saline-alkaline stress responses in adzuki bean.


Assuntos
Fatores de Terminação de Peptídeos , Proteínas de Plantas , Estresse Fisiológico , Vigna , Ácido Abscísico/farmacologia , Arabidopsis/genética , Etilenos/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Estudo de Associação Genômica Ampla , Fatores de Terminação de Peptídeos/genética , Fatores de Terminação de Peptídeos/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Polietilenoglicóis/farmacologia , Bicarbonato de Sódio/farmacologia , Cloreto de Sódio/farmacologia , Estresse Fisiológico/genética , Vigna/genética , Vigna/metabolismo
8.
J Med Genet ; 57(2): 89-95, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31501240

RESUMO

BACKGROUND: Male infertility is a prevalent issue worldwide, mostly due to the impaired sperm motility. Multiple morphological abnormalities of the sperm flagella (MMAF) present aberrant spermatozoa with absent, short, coiled, bent and irregular-calibre flagella resulting in severely decreased motility. Previous studies reported several MMAF-associated genes accounting for approximately half of MMAF cases. METHODS AND RESULT: We conducted genetic analysis using whole-exome sequencing in 88 Han Chinese MMAF probands. CFAP65 homozygous mutations were identified in four unrelated consanguineous families, and CFAP65 compound heterozygous mutations were found in two unrelated cases with MMAF. All these CFAP65 mutations were null, including four frameshift mutations (c.1775delC [p.Pro592Leufs*8], c.3072_3079dup [p.Arg1027Profs*41], c.1946delC [p.Pro649Argfs*5] and c.1580delT [p.Leu527Argfs*31]) and three stop-gain mutations (c.4855C>T [p.Arg1619*], c.5270T>A [p.Leu1757*] and c.5341G>T [p.Glu1781*]). Additionally, two homozygous CFAP65 variants likely affecting splicing were identified in two MMAF-affected men of Tunisian and Iranian ancestries, respectively. These biallelic variants of CFAP65 were verified by Sanger sequencing and were absent or very rare in large data sets aggregating sequence information from various human populations. CFAP65, encoding the cilia and flagella associated protein 65, is highly and preferentially expressed in the testis. Here we also generated a frameshift mutation in mouse orthologue Cfap65 using CRISPR-Cas9 technology. Remarkably, the phenotypes of Cfap65-mutated male mice were consistent with human MMAF. CONCLUSIONS: Our experimental observations performed on both human subjects and on Cfap65-mutated mice demonstrate that the presence of biallelic mutations in CFAP65 causes the MMAF phenotype and impairs sperm motility.

9.
J Med Genet ; 57(1): 31-37, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31048344

RESUMO

BACKGROUND: Male infertility due to multiple morphological abnormalities of the sperm flagella (MMAF) is a genetically heterogeneous disorder. Previous studies revealed several MMAF-associated genes, which account for approximately 60% of human MMAF cases. The pathogenic mechanisms of MMAF remain to be illuminated. METHODS AND RESULTS: We conducted genetic analyses using whole-exome sequencing in 50 Han Chinese probands with MMAF. Two homozygous stop-gain variants (c.910C>T (p.Arg304*) and c.3400delA (p.Ile1134Serfs*13)) of the SPEF2 (sperm flagellar 2) gene were identified in two unrelated consanguineous families. Consistently, an Iranian subject from another cohort also carried a homozygous SPEF2 stop-gain variant (c.3240delT (p.Phe1080Leufs*2)). All these variants affected the long SPEF2 transcripts that are expressed in the testis and encode the IFT20 (intraflagellar transport 20) binding domain, important for sperm tail development. Notably, previous animal studies reported spontaneous mutations of SPEF2 causing sperm tail defects in bulls and pigs. Our further functional studies using immunofluorescence assays showed the absence or a remarkably reduced staining of SPEF2 and of the MMAF-associated CFAP69 protein in the spermatozoa from SPEF2-affected subjects. CONCLUSIONS: We identified SPEF2 as a novel gene for human MMAF across the populations. Functional analyses suggested that the deficiency of SPEF2 in the mutated subjects could alter the localisation of other axonemal proteins.


Assuntos
Proteínas de Ciclo Celular/genética , Homozigoto , Infertilidade Masculina/genética , Mutação , Cauda do Espermatozoide/metabolismo , China , Análise Mutacional de DNA , Humanos , Infertilidade Masculina/metabolismo , Infertilidade Masculina/patologia , Irã (Geográfico) , Masculino , Linhagem , Cauda do Espermatozoide/patologia , Cauda do Espermatozoide/ultraestrutura , Sequenciamento Completo do Exoma
12.
J Cancer ; 10(27): 6858-6864, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31839820

RESUMO

Background: Inflammation might play an important role in promoting cancer growth partly by affecting tumor angiogenesis. We explored the role of Glasgow prognostic score (GPS) in metastatic colorectal cancer patients receiving first-linebevacizumab. Methods: All consecutive metastatic colorectal cancer patients treated with first-line chemotherapy plus or not plus bevacizumab were eligible. Pre-treatment GPS were collected for all cases. Results: In the chemotherapy group for patients with GPS of 0, 1 and 2, median progression-free survival (PFS) was 8.67, 8.10, and 8.27months, respectively (P = 0.17). Median overall survival (OS) was 24.87, 23.30, and 17.93months, respectively (P = 0.04). In the bevacizumab group, median PFS was 11.83, 8.10, and 6.87 months, respectively (P = 0.01), and median OS was 30.80, 19.47, and 18.67 months, respectively (P = 0.03).In whole group patients with a GPS of 0, both PFS and OS were in favor of patients treated with bevacizumab plus chemotherapy compared with who treated with chemotherapy alone (PFS 11.83 vs. 8.67 months, p=0.03; OS 30.80 vs. 24.87 months, p=0.04). Conclusion: GPS of 0 was correlated with good prognosis. Bevacizumab added a survival advantage only in metastatic colorectal cancer patients with a GPS of 0.

13.
Am J Hum Genet ; 105(6): 1168-1181, 2019 12 05.
Artigo em Inglês | MEDLINE | ID: mdl-31735294

RESUMO

As a type of severe asthenoteratospermia, multiple morphological abnormalities of the flagella (MMAF) are characterized by the presence of immotile spermatozoa with severe flagellar malformations. MMAF is a genetically heterogeneous disorder, and the known MMAF-associated genes can only account for approximately 60% of human MMAF cases. Here we conducted whole-exome sequencing and identified bi-allelic truncating mutations of the TTC29 (tetratricopeptide repeat domain 29) gene in three (3.8%) unrelated cases from a cohort of 80 MMAF-affected Han Chinese men. TTC29 is preferentially expressed in the testis, and TTC29 protein contains the tetratricopeptide repeat domains that play an important role in cilia- and flagella-associated functions. All of the men harboring TTC29 mutations presented a typical MMAF phenotype and dramatic disorganization in axonemal and/or other peri-axonemal structures. Immunofluorescence assays of spermatozoa from men harboring TTC29 mutations showed deficiency of TTC29 and remarkably reduced staining of intraflagellar-transport-complex-B-associated proteins (TTC30A and IFT52). We also generated a Ttc29-mutated mouse model through the use of CRISPR-Cas9 technology. Remarkably, Ttc29-mutated male mice also presented reduced sperm motility, abnormal flagellar ultrastructure, and male subfertility. Furthermore, intracytoplasmic sperm injections performed for Ttc29-mutated mice and men harboring TTC29 mutations consistently acquired satisfactory outcomes. Collectively, our experimental observations in humans and mice suggest that bi-allelic mutations in TTC29, as an important genetic pathogeny, can induce MMAF-related asthenoteratospermia. Our study also provided effective guidance for clinical diagnosis and assisted reproduction treatments.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Biomarcadores Tumorais , Estudos de Casos e Controles , Terapia Combinada , Feminino , Seguimentos , Regulação Neoplásica da Expressão Gênica , Humanos , Metástase Linfática , Masculino , Camundongos , Pessoa de Meia-Idade , Invasividade Neoplásica , Recidiva Local de Neoplasia/tratamento farmacológico , Recidiva Local de Neoplasia/patologia , Neoplasias/patologia , Prognóstico , Estudos Retrospectivos , Taxa de Sobrevida
14.
Infect Genet Evol ; 73: 401-410, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31176031

RESUMO

Hand, foot and mouth disease (HFMD) is mainly caused by human enterovirus 71 (EV71) and coxsackievirus A16 (CA16), which circulate alternatively or together in epidemic areas. Although the two viruses exhibit genetic homology, their clinical manifestations have some discrepancies. However, the factors underlying these differences remain unclear. Herein, we mainly focused on the alterations and roles of putative novel miRNAs in human umbilical vein endothelial cells (HUVECs) following EV71 and CA16 infections using high-throughput sequencing. The results identified 247 putative novel, differentially expressed miRNAs, of which only 11 miRNAs presented an opposite trend between the EV71- and CA16-infected samples and were used for target prediction. Gene ontology (GO) and pathway enrichment analysis of the predicted targets displayed the top 15 significant biological processes, molecular functions, cell components and pathways. Subsequently, regulatory miRNA-predicted targets and miRNA-GO and miRNA-pathway networks were constructed to further reveal the complex regulatory mechanisms of the miRNAs during infection. Therefore, our data provide useful insights that will help elucidate the different host-pathogen interactions following EV71 and CA16 infections and may offer novel therapeutic targets for these infections.


Assuntos
Infecções por Coxsackievirus/genética , Enterovirus Humano A/patogenicidade , Infecções por Enterovirus/genética , Enterovirus/patogenicidade , MicroRNAs/genética , Células Cultivadas , Biologia Computacional , Infecções por Coxsackievirus/virologia , Infecções por Enterovirus/virologia , Perfilação da Expressão Gênica/métodos , Ontologia Genética , Redes Reguladoras de Genes/genética , Doença de Mão, Pé e Boca/genética , Doença de Mão, Pé e Boca/virologia , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Interações Hospedeiro-Patógeno/genética , Células Endoteliais da Veia Umbilical Humana , Humanos
16.
Physiol Mol Biol Plants ; 25(2): 523-532, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30956433

RESUMO

To produce high quality, glyphosate-resistant soybeans, we crossed Jinda 73 and glyphosate-resistant RR1 (Roundup Ready First Generation) (RR1) resulting in 34 hybrid strains. To determine the effects of glyphosate on soybean metabolism, we grew the two parents upto the seedling stage, and measured chlorophyll, soluble sugar, malondialdehyde (MDA), relative conductivity and proline. Then, we treated the plants with glyphosate and measured the same factors again. Results showed that the chlorophyll content of Jinda 73 and RR1 decreased after spraying glyphosate. Glyphosate increased the level of soluble sugar, MDA, relative conductivity and proline in Jinda 73, but had no significant effect on RR1. We determined glyphosate resistance of the parents and the 34 hybrid, offspring strains by documenting the growth response in the field after treatment with glyphosate. Results showed that 29 hybrid, offspring strains have complete glyphosate resistance. Polymerase chain reaction (PCR) shows that the strains which have complete resistance to glyphosate have imported the CP4 5-enolpyhruvylshikimate-3- phosphate synthase (CP4 EPSPS) gene successfully. We selected three high quality, glyphosate-resistant strains (F7-3, F7-16 and F7-21), which had higher protein and oil levels as compared with Jinda 73.

17.
Am J Hum Genet ; 104(4): 738-748, 2019 04 04.
Artigo em Inglês | MEDLINE | ID: mdl-30929735

RESUMO

Male infertility is a major concern affecting human reproductive health. Asthenoteratospermia can cause male infertility through reduced motility and abnormal morphology of spermatozoa. Several genes, including DNAH1 and some CFAP family members, are involved in multiple morphological abnormalities of the sperm flagella (MMAF). However, these known genes only account for approximately 60% of human MMAF cases. Here, we conducted further genetic analyses by using whole-exome sequencing in a cohort of 65 Han Chinese men with MMAF. Intriguingly, bi-allelic mutations of TTC21A (tetratricopeptide repeat domain 21A) were identified in three (5%) unrelated, MMAF-affected men, including two with homozygous stop-gain mutations and one with compound heterozygous mutations of TTC21A. Notably, these men consistently presented with MMAF and additional abnormalities of sperm head-tail conjunction. Furthermore, a homozygous TTC21A splicing mutation was identified in two Tunisian cases from an independent MMAF cohort. TTC21A is preferentially expressed in the testis and encodes an intraflagellar transport (IFT)-associated protein that possesses several tetratricopeptide repeat domains that perform functions crucial for ciliary function. To further investigate the potential roles of TTC21A in spermatogenesis, we generated Ttc21a mutant mice by using CRISPR-Cas9 technology and revealed sperm structural defects of the flagella and the connecting piece. Our consistent observations across human populations and in the mouse model strongly support the notion that bi-allelic mutations in TTC21A can induce asthenoteratospermia with defects of the sperm flagella and head-tail conjunction.


Assuntos
Infertilidade Masculina/genética , Proteínas Associadas aos Microtúbulos/genética , Mutação , Espermatozoides/anormalidades , Alelos , Processamento Alternativo , Animais , Sistemas CRISPR-Cas , China , Exoma , Flagelos/patologia , Homozigoto , Humanos , Masculino , Camundongos , Fenótipo , Motilidade Espermática , Sequenciamento Completo do Exoma
18.
Reprod Biomed Online ; 38(5): 769-778, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30904354

RESUMO

RESEARCH QUESTION: Multiple morphological abnormalities of the sperm flagella (MMAF) comprise a rare congenital disease that can cause primary male infertility. Several pathogenic genes (e.g. AKAP4, DNAH1, CFAP43 and CFAP44) are associated with MMAF but the pathogenic mechanisms have not been elucidated. DESIGN: Whole-exome sequencing (WES) was applied to identify the pathogenic genes in 13 Chinese patients with MMAF; the patients were unrelated but all had consanguineous parents (usually first cousins). Real-time polymerase chain reaction and immunofluorescence staining were employed to assess the pathogenicity of these mutations. RESULTS: Four novel homozygous CFAP43 mutations in four (30.8%) MMAF patients and one novel homozygous CFAP44 mutation in one (7.7%) other case were identified. The four novel homozygous CFAP43 mutations included one frameshift mutation (c.1140_1143del: p.Asn380Lysfs*3), one nonsense mutation (c.739A>T: p.Lys247*) and two missense mutations (c.1474G>C: p.Gln492Arg; c.4600C>G: p.Leu1534Val). The novel mutation in CFAP44 was a homozygous nonsense mutation (c.4963C>T: p.Arg1655*). Co-segregation of the mutations was verified by Sanger sequencing of the families. The relative mRNA expression levels of CFAP43 in patients 1 and 9 and the levels of CFAP44 in patient 5 were significantly lower than those in control sperm samples. Immunofluorescence analysis of CFAP43 showed the protein was absent in the sperm flagella of patients 1 and 9. Furthermore, two previously reported mutations of DNAH1 were also identified in another four (30.8%) patients. CONCLUSIONS: This study demonstrated that CFAP43 and CFAP44 mutations are important causes of MMAF in the Chinese population. These novel mutations broaden the spectrum of CFAP43 and CFAP44 mutations.


Assuntos
Proteínas do Citoesqueleto/genética , Infertilidade Masculina/genética , Proteínas dos Microtúbulos/genética , Peptídeo Hidrolases/genética , Espermatozoides/anormalidades , Grupo com Ancestrais do Continente Asiático , Proteínas do Citoesqueleto/metabolismo , Humanos , Infertilidade Masculina/metabolismo , Masculino , Proteínas dos Microtúbulos/metabolismo , Mutação , Peptídeo Hidrolases/metabolismo , Cauda do Espermatozoide , Espermatozoides/metabolismo
20.
Nat Commun ; 10(1): 433, 2019 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-30683861

RESUMO

Aberrant sperm flagella impair sperm motility and cause male infertility, yet the genes which have been identified in multiple morphological abnormalities of the flagella (MMAF) can only explain the pathogenic mechanisms of MMAF in a small number of cases. Here, we identify and functionally characterize homozygous loss-of-function mutations of QRICH2 in two infertile males with MMAF from two consanguineous families. Remarkably, Qrich2 knock-out (KO) male mice constructed by CRISPR-Cas9 technology present MMAF phenotypes and sterility. To elucidate the mechanisms of Qrich2 functioning in sperm flagellar formation, we perform proteomic analysis on the testes of KO and wild-type mice. Furthermore, in vitro experiments indicate that QRICH2 is involved in sperm flagellar development through stabilizing and enhancing the expression of proteins related to flagellar development. Our findings strongly suggest that the genetic mutations of human QRICH2 can lead to male infertility with MMAF and that QRICH2 is essential for sperm flagellar formation.


Assuntos
Infertilidade Masculina/genética , Mutação com Perda de Função , Proteínas dos Microtúbulos/genética , Cauda do Espermatozoide/metabolismo , Proteínas de Ancoragem à Quinase A/deficiência , Proteínas de Ancoragem à Quinase A/genética , Adulto , Animais , Proteínas de Ligação ao Cálcio/deficiência , Proteínas de Ligação ao Cálcio/genética , Consanguinidade , Expressão Gênica , Perfilação da Expressão Gênica , Proteínas de Choque Térmico/deficiência , Proteínas de Choque Térmico/genética , Humanos , Infertilidade Masculina/metabolismo , Infertilidade Masculina/patologia , Masculino , Camundongos , Camundongos Knockout , Linhagem , Fosfoproteínas/deficiência , Fosfoproteínas/genética , Motilidade Espermática , Cauda do Espermatozoide/patologia , Cauda do Espermatozoide/ultraestrutura , Testículo/química , Testículo/metabolismo , Sequenciamento Completo do Genoma
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...