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1.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 39(8): 819-823, 2022 Aug 10.
Artigo em Chinês | MEDLINE | ID: mdl-35929929

RESUMO

OBJECTIVE: To explore the genetic basis for fetuses with renal anomalies. METHODS: Genomic DNA of four fetuses and their parents was extracted from amniotic fluid and peripheral blood samples and subjected to whole genome sequencing. Candidate variants were predicted according to the American College of Medical Genetics and Genomics (ACMG) guidelines and validated by SNP-array and Sanger sequencing. RESULTS: Two fetuses were found to carry a 1.45 Mb pathogenic microdeletion in 17q12 and a pathogenic 1.85 Mb microduplication at 1q21.1-21.2, respectively. One fetus was found to harbor compound heterozygous variants c.8301del (p.Asn2768Thrfs*18) and c.4481del (p.Asn1494Thrfs*6) of the PKHD1 gene, which were predicted to be pathogenic. And one fetus has harbored homozygous c.1372dup (p.Thr458Asnfs*5) variants of the BBS12 gene, which was predicted to be likely pathogenic. All variants were validated by Sanger sequencing. CONCLUSION: Whole genome sequencing can enable efficient prenatal diagnosis for fetuses with renal anomalies with high accuracy.


Assuntos
Feto , Diagnóstico Pré-Natal , Feminino , Feto/anormalidades , Humanos , Gravidez , Sequenciamento Completo do Exoma , Sequenciamento Completo do Genoma
2.
Genome Med ; 14(1): 82, 2022 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-35922826

RESUMO

BACKGROUND: Small intestinal neuroendocrine tumors (SI-NETs) are the most common neoplasms of the small bowel. The majority of tumors are located in the distal ileum with a high incidence of multiple synchronous primary tumors. Even though up to 50% of SI-NET patients are diagnosed with multifocal disease, the mechanisms underlying multiple synchronous lesions remain elusive. METHODS: We performed whole genome sequencing of 75 de-identified synchronous primary tumors, 15 metastases, and corresponding normal samples from 13 patients with multifocal ileal NETs to identify recurrent somatic genomic alterations, frequently affected signaling pathways, and shared mutation signatures among multifocal SI-NETs. Additionally, we carried out chromosome mapping of the most recurrent copy-number alterations identified to determine which parental allele had been affected in each tumor and assessed the clonal relationships of the tumors within each patient. RESULTS: Absence of shared somatic variation between the synchronous primary tumors within each patient was observed, indicating that these tumors develop independently. Although recurrent copy-number alterations were identified, additional chromosome mapping revealed that tumors from the same patient can gain or lose different parental alleles. In addition to the previously reported CDKN1B loss-of-function mutations, we observed potential loss-of-function gene alterations in TNRC6B, a candidate tumor suppressor gene in a small subset of ileal NETs. Furthermore, we show that multiple metastases in the same patient can originate from either one or several primary tumors. CONCLUSIONS: Our study demonstrates major genomic diversity among multifocal ileal NETs, highlighting the need to identify and remove all primary tumors, which have the potential to metastasize, and the need for optimized targeted treatments.


Assuntos
Neoplasias Intestinais , Tumores Neuroendócrinos , Neoplasias Pancreáticas , Humanos , Neoplasias Intestinais/genética , Neoplasias Intestinais/patologia , Mutação , Tumores Neuroendócrinos/genética , Proteínas de Ligação a RNA/genética , Neoplasias Gástricas , Sequenciamento Completo do Genoma
3.
Sci Rep ; 12(1): 10830, 2022 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-35790751

RESUMO

The potential germline effects of radiation exposure to military veterans present at British nuclear tests in Australia and the South Pacific is of considerable interest. We analyzed germline mutations in 60 families of UK military personnel comprising 30 control and 30 nuclear test veterans (NTV). Using whole-genome sequencing we studied the frequency and spectra of de novo mutations to investigate the transgenerational effect of veterans' (potential) exposure to radiation at nuclear bomb test sites. We find no elevation in total de novo single nucleotide variants, small insertion-deletions, structural variants or clustered mutations among the offspring of nuclear test veterans compared to those of control personnel. We did observe an elevated occurrence of single base substitution mutations within mutation signature SBS16, due to a subset of NTV offspring. The relevance of this elevation to potential exposure of veteran fathers and, future health risks, require further investigation. Overall, we find no evidence of increased mutations in the germline of a group of British nuclear test veterans. ISRCTN Registry 17461668.


Assuntos
Veteranos , Células Germinativas , Mutação em Linhagem Germinativa , Humanos , Mutação , Sequenciamento Completo do Genoma
4.
Commun Biol ; 5(1): 667, 2022 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-35790849

RESUMO

Efficient, accurate molecular characterization of genetically modified (GM) organisms is challenging, especially for those transgenic events transferred with genes/elements of recipient species. Herein, we decipher the comprehensive molecular characterization of one novel GM rice event G281 which was transferred with native promoters and an RNA interference (RNAi) expression cassette using paired-end whole genome sequencing (PE-WGS) and modified TranSeq approach. Our results show that transgenes integrate at rice chromosome 3 locus 16,439,674 included a 36 bp deletion of rice genomic DNA, and the whole integration contains two copies of the complete transfer DNA (T-DNA) in a head-to-head arrangement. No unintended insertion or backbone sequence of the transformed plasmid is observed at the whole genome level. Molecular characterization of the G281 event will assist risk assessment and application for a commercial license. In addition, we speculate that our approach could be further used for identifying the transgene integration of cisgenesis/intragenesis crops since both ends of T-DNA in G281 rice were from native gene or elements which is similar with that of cisgenesis/intrasgenesis. Our results from the in silico mimicking cisgenesis event confirm that the mimic rice Gt1 gene insertion and its flanking sequences are successfully identified, demonstrating the applicability of PE-WGS for molecular characterization of cisgenesis/intragenesis crops.


Assuntos
Oryza , Produtos Agrícolas/genética , DNA/metabolismo , Oryza/genética , Oryza/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Transgenes , Sequenciamento Completo do Genoma
5.
Methods Mol Biol ; 2509: 233-250, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35796967

RESUMO

Whole genome bisulfite sequencing (WGBS) is a high-throughput DNA sequencing-based technique that is used to determine genome-wide DNA methylation patterns at base resolution. Library construction by post-bisulfite adaptor tagging (PBAT ) extends the application of WGBS to several hundred cells and minimizes the required number of library amplification cycles. We herein describe a PBAT protocol to prepare WGBS libraries from 200 cells and introduce the outline of a downstream bioinformatic analysis. The prepared library can typically generate 800 million sequencing reads, which is sufficient to cover the human and mouse genomes approximately 15 times, using the Illumina NovaSeq 6000 sequencing system.


Assuntos
Metilação de DNA , Sulfitos , Animais , DNA/genética , Biblioteca Gênica , Genoma , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Camundongos , Análise de Sequência de DNA/métodos , Sequenciamento Completo do Genoma/métodos
7.
Nature ; 607(7920): 732-740, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-35859178

RESUMO

Detailed knowledge of how diversity in the sequence of the human genome affects phenotypic diversity depends on a comprehensive and reliable characterization of both sequences and phenotypic variation. Over the past decade, insights into this relationship have been obtained from whole-exome sequencing or whole-genome sequencing of large cohorts with rich phenotypic data1,2. Here we describe the analysis of whole-genome sequencing of 150,119 individuals from the UK Biobank3. This constitutes a set of high-quality variants, including 585,040,410 single-nucleotide polymorphisms, representing 7.0% of all possible human single-nucleotide polymorphisms, and 58,707,036 indels. This large set of variants allows us to characterize selection based on sequence variation within a population through a depletion rank score of windows along the genome. Depletion rank analysis shows that coding exons represent a small fraction of regions in the genome subject to strong sequence conservation. We define three cohorts within the UK Biobank: a large British Irish cohort, a smaller African cohort and a South Asian cohort. A haplotype reference panel is provided that allows reliable imputation of most variants carried by three or more sequenced individuals. We identified 895,055 structural variants and 2,536,688 microsatellites, groups of variants typically excluded from large-scale whole-genome sequencing studies. Using this formidable new resource, we provide several examples of trait associations for rare variants with large effects not found previously through studies based on whole-exome sequencing and/or imputation.


Assuntos
Bancos de Espécimes Biológicos , Bases de Dados Genéticas , Variação Genética , Genoma Humano , Genômica , Sequenciamento Completo do Genoma , África/etnologia , Ásia/etnologia , Estudos de Coortes , Sequência Conservada , Éxons/genética , Genoma Humano/genética , Haplótipos/genética , Humanos , Mutação INDEL , Irlanda/etnologia , Repetições de Microssatélites , Polimorfismo de Nucleotídeo Único/genética , Reino Unido
8.
BMC Microbiol ; 22(1): 186, 2022 07 29.
Artigo em Inglês | MEDLINE | ID: mdl-35906551

RESUMO

BACKGROUND: Cellulolytic microorganisms are considered a key player in the degradation of feed fiber. These microorganisms can be isolated from various resources, such as animal gut, plant surfaces, soil and oceans. A new strain of Bacillus amyloliquefaciens, TL106, was isolated from faeces of a healthy Tibetan pigs. This strain can produce cellulase and shows strong antimicrobial activity in mice. Thus, in this study, to better understand the strain of B. amyloliquefaciens TL106 on degradation of cellulose, the genome of the strain TL106 was completely sequenced and analyzed. In addition, we also explored the cellulose degradation ability of strain TL106 in vitro. RESULTS: TL106 was completely sequenced with the third generation high-throughput DNA sequencing. In vitro analysis with enzymatic hydrolysis identified the activity of cellulose degradation. TL106 consisted of one circular chromosome with 3,980,960 bp and one plasmid with 16,916 bp, the genome total length was 3.99 Mb and total of 4,130 genes were predicted. Several genes of cellulases and hemicellulase were blasted in Genbank, including ß-glucosidase, endoglucanase, ß-glucanase and xylanase genes. Additionally, the activities of amylase (20.25 U/mL), cellulase (20.86 U/mL), xylanase (39.71 U/mL) and ß-glucanase (36.13 U/mL) in the fermentation supernatant of strain TL106 were higher. In the study of degradation characteristics, we found that strain TL106 had a better degradation effect on crude fiber, neutral detergent fiber, acid detergent fiber, starch, arabinoxylan and ß-glucan of wheat and highland barley . CONCLUSIONS: The genome of B. amyloliquefaciens TL106 contained several genes of cellulases and hemicellulases, can produce carbohydrate-active enzymes, amylase, cellulase, xylanase and ß-glucanase. The supernatant of fermented had activities of strain TL106. It could degrade the fiber fraction and non-starch polysaccharides (arabinoxylans and ß-glucan) of wheat and highland barley. The present study demonstrated that the degradation activity of TL106 to crude fiber which can potentially be applied as a feed additive to potentiate the digestion of plant feed by monogastric animals.


Assuntos
Bacillus amyloliquefaciens , Celulase , Hordeum , beta-Glucanas , Amilases , Animais , Bacillus amyloliquefaciens/genética , Bacillus amyloliquefaciens/metabolismo , Celulase/genética , Celulase/metabolismo , Celulose/metabolismo , Detergentes , Fibras na Dieta , Camundongos , Suínos , Tibet , Triticum , Sequenciamento Completo do Genoma , beta-Glucosidase/genética
9.
Int J Mol Sci ; 23(14)2022 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-35886856

RESUMO

Some prevention strategies, including vaccines and antibiotic alternatives, have been developed to reduce enterotoxigenic Escherichia coli proliferation in animal production. In this study, a wild-type strain of BE311 with a virulent heat-stable enterotoxin gene identical to E. coli K99 was isolated for its high potential for gene expression ability. The whole genome of E. coli BE311 was sequenced for gene analyses and editing. Subsequently, the fluorescent gene mCherry was successfully knocked into the genome of E. coli BE311 by CRISPR/Cas9. The E. coli BE311-mCherry strain was precisely quantified through the fluorescence intensity and red colony counting. The inflammatory factors in different intestinal tissues all increased significantly after an E. coli BE311-mCherry challenge in Sprague-Dawley rats (p < 0.05). The heat-stable enterotoxin gene of E. coli BE311 was knocked out, and an attenuated vaccine host E. coli BE311-STKO was constructed. Flow cytometry showed apoptotic cell numbers were lower following a challenge of IPEC-J2 cells with E. coli BE311-STKO than with E. coli BE311. Therefore, the E. coli BE311-mCherry and E. coli BE311-STKO strains that were successfully constructed based on the gene knock-in and knock-out technology could be used as ideal candidates in ETEC challenge models and for the development of attenuated vaccines.


Assuntos
Escherichia coli Enterotoxigênica , Infecções por Escherichia coli , Proteínas de Escherichia coli , Animais , Sistemas CRISPR-Cas/genética , Escherichia coli Enterotoxigênica/genética , Enterotoxinas/química , Enterotoxinas/genética , Infecções por Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Edição de Genes/métodos , Imagem Óptica/métodos , Ratos , Ratos Sprague-Dawley , Sequenciamento Completo do Genoma
10.
Virulence ; 13(1): 1242-1251, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-35891618

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern have been emerging. However, knowledge of temporal and spatial dynamics of SARS-CoV-2 is limited. This study characterized SARS-CoV-2 evolution in immunosuppressed patients with long-term SARS-CoV-2 shedding for 73-250 days, without specific treatment. We conducted whole-genome sequencing of 27 serial samples, including 26 serial samples collected from various anatomic sites of two patients and the first positive sample from patient 2's mother. We analysed the intrahost temporal dynamics and genomic diversity of the viral population within different sample types. Intrahost variants emerging during infection showed diversity between individual hosts. Remarkably, N501Y, P681R, and E484K, key substitutions within spike protein, emerged in vivo during infection and became the dominant population. P681R, which had not yet been detected in the publicly available genome in Korea, appeared within patient 1 during infection. Mutually exclusive substitutions at residues R346 (R346S and R346I) and E484 (E484K and E484A) of spike protein and continuous turnover of these substitutions occurred. Unique genetic changes were observed in urine samples. A household transmission from patient 2 to his mother, at least 38 days after the diagnosis, was characterized. Viruses may differently mutate and adjust to the host selective pressure, which could enable the virus to replicate efficiently for fitness in each host. Intrahost variants could be candidate variants likely to spread to the population eventually. Our findings may provide new insights into the dynamics of SARS-CoV-2 in response to interactions between the virus and host.


Assuntos
COVID-19 , Hospedeiro Imunocomprometido , SARS-CoV-2 , Eliminação de Partículas Virais , COVID-19/transmissão , Humanos , Mutação , SARS-CoV-2/genética , Glicoproteína da Espícula de Coronavírus/genética , Sequenciamento Completo do Genoma
11.
Nat Commun ; 13(1): 4057, 2022 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-35882841

RESUMO

While many genetic diseases have effective treatments, they frequently progress rapidly to severe morbidity or mortality if those treatments are not implemented immediately. Since front-line physicians frequently lack familiarity with these diseases, timely molecular diagnosis may not improve outcomes. Herein we describe Genome-to-Treatment, an automated, virtual system for genetic disease diagnosis and acute management guidance. Diagnosis is achieved in 13.5 h by expedited whole genome sequencing, with superior analytic performance for structural and copy number variants. An expert panel adjudicated the indications, contraindications, efficacy, and evidence-of-efficacy of 9911 drug, device, dietary, and surgical interventions for 563 severe, childhood, genetic diseases. The 421 (75%) diseases and 1527 (15%) effective interventions retained are integrated with 13 genetic disease information resources and appended to diagnostic reports ( https://gtrx.radygenomiclab.com ). This system provided correct diagnoses in four retrospectively and two prospectively tested infants. The Genome-to-Treatment system facilitates optimal outcomes in children with rapidly progressive genetic diseases.


Assuntos
Variações do Número de Cópias de DNA , Criança , Humanos , Lactente , Estudos Retrospectivos , Sequenciamento Completo do Genoma
12.
BMC Genomics ; 23(1): 537, 2022 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-35870884

RESUMO

BACKGROUND: The implementation of whole genome sequencing (WGS) by PulseNet, the molecular subtyping network for foodborne diseases, has transformed surveillance, outbreak detection, and public health laboratory practices in the United States. In 2017, the New Hampshire Public Health Laboratories, a member of PulseNet, commenced the use of WGS in tracking foodborne pathogens across the state. We present some of the initial results of New Hampshire's initiative to transition to WGS in tracking Salmonella enterica, a bacterial pathogen that is responsible for non-typhoidal foodborne infections and enteric fever. We characterize the population structure and evolutionary history of 394 genomes of isolates recovered from human clinical cases in New Hampshire from 2017 to 2020. RESULTS: The New Hampshire S. enterica population is phylogenetically diverse, consisting of 78 sequence types (ST) and 67 serotypes. Six lineages dominate the population: ST 11 serotype Enteritidis, ST 19 Typhimurium, ST 32 Infantis, ST 118 Newport, ST 22 Braenderup, and ST 26 Thompson. Each lineage is derived from long ancestral branches in the phylogeny, suggesting their extended presence in the region and recent clonal expansion. We detected 61 genes associated with resistance to 14 antimicrobial classes. Of these, unique genes of five antimicrobial classes (aminocoumarins, aminoglycosides, fluoroquinolones, nitroimidazoles, and peptides) were detected in all genomes. Rather than a single clone carrying multiple resistance genes expanding in the state, we found multiple lineages carrying different combinations of independently acquired resistance determinants. We estimate the time to the most recent common ancestor of the predominant lineage ST 11 serotype Enteritidis (126 genomes) to be 1965 (95% highest posterior density intervals: 1927-1982). Its population size expanded until 1978, followed by a population decline until 1990. This lineage has been expanding since then. Comparison with genomes from other states reveal lack of geographical clustering indicative of long-distance dissemination. CONCLUSIONS: WGS studies of standing pathogen diversity provide critical insights into the population and evolutionary dynamics of lineages and antimicrobial resistance, which can be translated to effective public health action and decision-making. We highlight the need to strengthen efforts to implement WGS-based surveillance and genomic data analyses in state public health laboratories.


Assuntos
Salmonella enterica , Febre Tifoide , Animais , Antibacterianos/farmacologia , Genoma Bacteriano , Humanos , Laboratórios , New Hampshire , Filogenia , Saúde Pública , Estados Unidos , Sequenciamento Completo do Genoma/métodos
13.
PLoS One ; 17(7): e0270897, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35877652

RESUMO

The unintentional movement of agronomic pests and pathogens is steadily increasing due to the intensification of global trade. Being able to identify accurately and rapidly early stages of an invasion is critical for developing successful eradication or management strategies. For most invasive organisms, molecular diagnostics is today the method of choice for species identification. However, the currently implemented tools are often developed for certain taxa and need to be adapted for new species, making them ill-suited to cope with the current constant increase in new invasive species. To alleviate this impediment, we developed a fast and accurate sequencing tool allowing to modularly obtain genetic information at different taxonomical levels. Using whole genome amplification (WGA) followed by Oxford nanopore MinION sequencing, our workflow does not require any a priori knowledge on the investigated species and its classification. While mainly focusing on harmful plant pathogenic insects, we also demonstrate the suitability of our workflow for the molecular identification of bacteria (Erwinia amylovora and Escherichia coli), fungi (Cladosporium herbarum, Colletotrichum salicis, Neofabraea alba) and nematodes (Globodera rostochiensis). On average, the pairwise identity between the generated consensus sequences and best GenBank BLAST matches was 99.6 ± 0.6%. Additionally, assessing the generated insect genomic dataset, the potential power of the workflow to detect pesticide resistance genes, as well as arthropod-infecting viruses and endosymbiotic bacteria is demonstrated.


Assuntos
Ascomicetos , Sequenciamento por Nanoporos , Nanoporos , Ascomicetos/genética , Bactérias/genética , Biosseguridade , Sequenciamento de Nucleotídeos em Larga Escala , Análise de Sequência de DNA , Sequenciamento Completo do Genoma
14.
15.
World J Microbiol Biotechnol ; 38(9): 164, 2022 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-35842870

RESUMO

Bacillus velezensis FTL7 which exhibited potent antimicrobial peptide producing capacity was isolated from a marine sediment sample of the West Coast region, South India, and characterized through experimental and genomic analysis approaches. FTL7 showed potential antimicrobial activity against a broad range of foodborne pathogenic bacteria like Listeria monocytogenes Scott A, Bacillus cereus (ATCC 11778), Salmonella Typhimurium (MTCC 1251), Staphylococcus aureus (ATCC 25923), and Escherichia coli (MTCC 443). It also exhibited strong inhibitory activity against Kocuria rhyzophila (ATCC 934) and Bacillus subtilis subsp. spizizenii (ATCC 6633). Phylogenetic analysis by 16S rRNA gene sequence showed that Bacillus velezensis FTL7 was closely related to B. velezensis LBUM288 (GenBank accession number MG461457) with 100% identity. Whole-genome sequencing of the strain FTL7 was carried out using Illumina sequencing technology to get a better insight into the mechanisms of controlling pathogens by FTL7. The strain FTL7 has a chromosome size of 3849,077 bp with a GC content of 46.56%. The genome consists of 3635 coding sequences, 64 RNA, 59 tRNAs, 5 ncRNAs, and 69 pseudogenes. The presence of genes responsible for the synthesis of non-ribosomal peptides and bacteriocins was identified through genome annotation. Thus, many Bacillus strains, including B. velezensis, have been demonstrated as excellent producers of antimicrobial substances.


Assuntos
Anti-Infecciosos , Bacillus , Anti-Infecciosos/farmacologia , Bacillus cereus/genética , Filogenia , RNA Ribossômico 16S/genética , Sequenciamento Completo do Genoma
16.
Genome Biol Evol ; 14(7)2022 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-35881514

RESUMO

Herein, we provide the first whole-genome sequence of the purple butter clam (Saxidomus purpuratus), an economically important bivalve shellfish. Specifically, we sequenced and de novo assembled the genome of Sa. purpuratus based on PromethION long reads and Hi-C data. The 978-Mb genome of Sa. purpuratus comprises 19 chromosomes with 36,591 predicted protein-coding genes. The N50 length of Sa. purpuratus genome is 52 Mb, showing the highest continuous assembly among bivalve genomes. The Benchmarking by Universal Single-Copy Orthologs assessment indicated that 95.07% of complete metazoan universal single-copy orthologs (n = 954) were present in the assembly. Approximately 51% of Sa. purpuratus genome comprises repetitive sequences. Based on the high-quality Sa. purpuratus genome, we resolved half of the immune-associated genes, namely, scavenger receptor (SR) proteins, which are collinear to those in the closely related Cyclina sinensis genome. This finding suggested a high degree of conservation among immune-associated genes. Twenty-two (19%) SR proteins are tandemly duplicated in Sa. purpuratus genome, suggesting putative convergence evolution. Overall, Sa. purpuratus genome provides a new resource for the discovery of economically important traits and immune-response genes.


Assuntos
Bivalves , Cromossomos , Animais , Bivalves/genética , Cromossomos/genética , Genoma , Anotação de Sequência Molecular , Sequências Repetitivas de Ácido Nucleico , Sequenciamento Completo do Genoma
17.
PLoS One ; 17(7): e0272129, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35901010

RESUMO

The accurate identification of genetic variants contributing to therapeutic drug response or adverse effects is the first step in implementation of precision drug therapy. Targeted sequencing has recently become a common methodology for large-scale studies of genetic variation thanks to its favorable balance between low cost, high throughput, and deep coverage. Here, we present ClinPharmSeq, a targeted sequencing panel of 59 genes with associations to pharmacogenetic (PGx) phenotypes, as a platform to explore the relationship between drug response and genetic variation, both common and rare. For validation, we sequenced DNA from 64 ethnically diverse Coriell samples with ClinPharmSeq to call star alleles (haplotype patterns) in 27 genes using the bioinformatics tool PyPGx. These reference samples were extensively characterized by multiple laboratories using PGx testing assays and, more recently, whole genome sequencing. We found that ClinPharmSeq can consistently generate deep-coverage data (mean = 274x) with high uniformity (30x or above = 94.8%). Our genotype analysis identified a total of 185 unique star alleles from sequencing data, and showed that diplotype calls from ClinPharmSeq are highly concordant with that from previous publications (97.6%) and whole genome sequencing (97.9%). Notably, all 19 star alleles with complex structural variation including gene deletions, duplications, and hybrids were recalled with 100% accuracy. Altogether, these results demonstrate that the ClinPharmSeq platform offers a feasible path for broad implementation of PGx testing and optimization of individual drug treatments.


Assuntos
Sequenciamento de Nucleotídeos em Larga Escala , Farmacogenética , Alelos , Genótipo , Haplótipos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Farmacogenética/métodos , Sequenciamento Completo do Genoma/métodos
18.
Science ; 377(6605): 511-517, 2022 07 29.
Artigo em Inglês | MEDLINE | ID: mdl-35901164

RESUMO

We analyzed 131 human brains (44 neurotypical, 19 with Tourette syndrome, 9 with schizophrenia, and 59 with autism) for somatic mutations after whole genome sequencing to a depth of more than 200×. Typically, brains had 20 to 60 detectable single-nucleotide mutations, but ~6% of brains harbored hundreds of somatic mutations. Hypermutability was associated with age and damaging mutations in genes implicated in cancers and, in some brains, reflected in vivo clonal expansions. Somatic duplications, likely arising during development, were found in ~5% of normal and diseased brains, reflecting background mutagenesis. Brains with autism were associated with mutations creating putative transcription factor binding motifs in enhancer-like regions in the developing brain. The top-ranked affected motifs corresponded to MEIS (myeloid ectopic viral integration site) transcription factors, suggesting a potential link between their involvement in gene regulation and autism.


Assuntos
Envelhecimento , Encéfalo , Envelhecimento/genética , Humanos , Mutagênese , Mutação , Sequenciamento Completo do Genoma
19.
Elife ; 112022 07 26.
Artigo em Inglês | MEDLINE | ID: mdl-35880398

RESUMO

Transmission is a driver of tuberculosis (TB) epidemics in high-burden regions, with assumed negligible impact in low-burden areas. However, we still lack a full characterization of transmission dynamics in settings with similar and different burdens. Genomic epidemiology can greatly help to quantify transmission, but the lack of whole genome sequencing population-based studies has hampered its application. Here, we generate a population-based dataset from Valencia region and compare it with available datasets from different TB-burden settings to reveal transmission dynamics heterogeneity and its public health implications. We sequenced the whole genome of 785 Mycobacterium tuberculosis strains and linked genomes to patient epidemiological data. We use a pairwise distance clustering approach and phylodynamic methods to characterize transmission events over the last 150 years, in different TB-burden regions. Our results underscore significant differences in transmission between low-burden TB settings, i.e., clustering in Valencia region is higher (47.4%) than in Oxfordshire (27%), and similar to a high-burden area as Malawi (49.8%). By modeling times of the transmission links, we observed that settings with high transmission rate are associated with decades of uninterrupted transmission, irrespective of burden. Together, our results reveal that burden and transmission are not necessarily linked due to the role of past epidemics in the ongoing TB incidence, and highlight the need for in-depth characterization of transmission dynamics and specifically tailored TB control strategies.


Assuntos
Epidemias , Mycobacterium tuberculosis , Tuberculose , Humanos , Mycobacterium tuberculosis/genética , Dinâmica Populacional , Tuberculose/epidemiologia , Sequenciamento Completo do Genoma
20.
Viruses ; 14(7)2022 07 04.
Artigo em Inglês | MEDLINE | ID: mdl-35891452

RESUMO

Orf virus (ORFV) is distributed worldwide and is the causative agent of contagious ecthyma that mainly occurs in sheep and goats. This disease was reported for the first time at the end of 18th century in Europe but very little is currently known about the temporal and geographic origins of this virus. In the present study, the use of new Italian whole genomes allowed for better inference on the evolutionary history of ORFV. In accordance with previous studies, two genome types (S and G) were described for infection of sheep and goats, respectively. These two well-differentiated groups of genomes originated for evolutive convergence in the late 1800s in two different areas of the world (Europe for S type and Asia for G type), but it was only in the early 1900s that the effective size of ORFV increased among hosts and the virus spread across the whole European continent. The Italian strains which were sequenced in the present study were isolated on the Mediterranean island of Sardinian and showed to be exclusive to this geographic area. One of them is likely representative of the early European forms of ORFV which infected sheep and became extinct about one century ago. Such an ancient Sardinian strain may have reached the island simple by chance, where it quickly adapted to the new habitat.


Assuntos
Ectima Contagioso , Vírus do Orf , Animais , Cabras , Vírus do Orf/genética , Filogenia , Ovinos , Sequenciamento Completo do Genoma
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