A Solve-RD ClinVar-based reanalysis of 1522 index cases from ERN-ITHACA reveals common pitfalls and misinterpretations in exome sequencing

Purpose Within the Solve-RD project (https://solve-rd.eu/), the European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies aimed to investigate whether a reanalysis of exomes from unsolved cases based on ClinVar annotations could establish additional diagnoses. We present the results of the “ClinVar low-hanging fruit” reanalysis, reasons for the failure of previous analyses, and lessons learned. Methods Data from the first 3576 exomes (1522 probands and 2054 relatives) collected from European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies was reanalyzed by the Solve-RD consortium by evaluating for the presence of single-nucleotide variant, and small insertions and deletions already reported as (likely) pathogenic in ClinVar. Variants were filtered according to frequency, genotype, and mode of inheritance and reinterpreted. Results We identified causal variants in 59 cases (3.9%), 50 of them also raised by other approaches and 9 leading to new diagnoses, highlighting interpretation challenges: variants in genes not known to be involved in human disease at the time of the first analysis, misleading genotypes, or variants undetected by local pipelines (variants in off-target regions, low quality filters, low allelic balance, or high frequency). Conclusion The “ClinVar low-hanging fruit” analysis represents an effective, fast, and easy approach to recover causal variants from exome sequencing data, herewith contributing to the reduction of the diagnostic deadlock

Dynamic dans evolution of two notable LTR retrotransposons lineages in Coffea genus (Rubiaceae family)

Les éléments transposables (ET) sont des portions d’ADN capables de se déplacer et d’augmenter le nombre de leurs copies dans les génomes. Deux grands types de transposition, correspondant à deux grandes classes d’ET, sont retrouvés chez la quasi-totalité des génomes étudiés à ce jour. Les rétrotransposons à LTR (Long Terminal Repeats, LTR-RT), appartenant à la Classe 1, sont les composants majoritaires des génomes des plantes. Leur prolifération peut avoir un impact important sur l’organisation, la variation de taille, l’évolution des génomes et l’activité des gènes.Le café, largement consommé dans le monde et produit uniquement par des pays du Sud, est issu de deux espèces cultivées d’origine africaine : Coffea arabica et C. canephora. Le genre Coffea est constitué de 139 espèces occupant des habitats très variés en Afrique, dans les îles de l’ouest de l’océan Indien, l’Inde, l’Asie tropicale et du sud-est et au nord de l’Australie. Toutes les espèces son diploïdes, à l’exception notable de C. arabica, allotétraploïde, issu d’une hybridation interspécifique récente entre les deux espèces diploïdes : C. canephora et C. eugenioides. Pour autant, la taille des génomes des espèces diploïdes varie du simple au double. Les nombreuses données génomiques aujourd’hui disponibles au sein du genre Coffea permettent d’étudier la dynamique des LTR-RT constituant au minimum 42% du génome de C. canephora, l’espèce séquencée et disponible dans les bases de données publiques.Dans ce travail, deux lignées remarquables de LTR-RT, Bianca et SIRE, ont été étudiées par des approches bio- informatiques. Bianca sensu stricto, présente uniquement chez les monocotylédones, est représentée chez les dicotylédones par la famille Divo, très peu étudiée à ce jour. L’activation récente de Divo sans induire sa propre structuration, est étroitement associée à la différenciation génétique de C. canephora. Par contre, tout en étant présente dans toutes les espèces de caféiers étudiées, l’activation semble sporadique. À l’opposé, les éléments SIRE, la seule lignée de LTR-RT de la superfamille des Copia contenant un domaine enveloppe comme les rétrovirus, montre des variations structurales importantes entre les accessions des espèces diploïdes à l’origine de C arabica et plus globalement, et en parallèle de l’évolution du genre.Nos travaux montrent que la compréhension de la dynamique des LTR-RT dans un genre peut permettre de mieux appréhender son histoire évolutive, chaque famille de LTR-RT pouvant apporter un éclairage différent. Nos résultats indiquent qu’à la fois les clades biogéographiques (phylogénie moléculaire des caféiers) mais aussi certaines accessions d’espèces diploïdes ont des histoires particulières. Celles-ci seraient vraisemblablement liées à la colonisation de nouvelles niches et à la dynamique des LTR-RT composant les génomes des Coffea.Transposable elements (TEs) are DNA fragments that are able to move and to increase their copy numbers. Two transposition mechanisms corresponding to the two main TE classes are found in almost all organisms. LTR retrotransposons (Long Terminal Repeats, LTR-RTs), belonging to Class 1, are the main components of plant genomes. Genome organisation, size variation, evolution and gene activity can be strongly impacted by their proliferation.Worldwide consumed and produced by South countries, coffee is obtained from two African cultivated species: Coffea arabica and C. canephora. The Coffea genus includes 139 species occurring in diverse habitats in Africa, Madagascar, Mascarene Islands, Comoros, India, Southeast and Tropical Asia and North Australia. All the species are diploids, except the noteworthy allotetraploid C. arabica, originated from a recent inter-specific hybridisation between two diploids: C. canephora and C. eugenioides. However, genome size of diploid species can vary for up to two folds. Today, the numerous genomic data available for Coffea allows the study of LTR- RTs, constituting at least 42% of C. canephora genome, the sequenced species available in public databases.In this work, two notable LTR-RT lineages, Bianca and SIRE, have been studied by bioinformatics approaches. Bianca s.s., is present only in Monocots and it is represented in Dicots by the Divo family, poorly studied nowadays. The recent activation of Divo, without leading to its own structuring, is closely associated to the genetic differentiation of C. canephora. However, this activation seems sporadic as being present in all the coffee-trees species studied here. On the opposite, SIRE elements, which are the only Copia LTR-RTs carrying an envelope-like gene as retroviruses, show an important structuring variation between accessions among C. arabica progenitors, and in parallel to the genus evolution.Our work shows that understanding the LTR-RTs dynamics in a genus allows a better perception of its evolutionary history, with the possibility of different evolutionary timing given by different LTR-RTs families. Our results also indicate that both the biogeographic clades (coffee molecular phylogeny) and also some diploid accessions have peculiar histories, probably related to the colonisation of new ecological niches and to the LTR- RTs dynamics

Dynamique et évolution de deux lignées remarquables de rétrotransposons à LTR dans le genre Coffea (famille des Rubiacées)

Transposable elements (TEs) are DNA fragments that are able to move and to increase their copy numbers. Two transposition mechanisms corresponding to the two main TE classes are found in almost all organisms. LTR retrotransposons (Long Terminal Repeats, LTR-RTs), belonging to Class 1, are the main components of plant genomes. Genome organisation, size variation, evolution and gene activity can be strongly impacted by their proliferation.Worldwide consumed and produced by South countries, coffee is obtained from two African cultivated species: Coffea arabica and C. canephora. The Coffea genus includes 139 species occurring in diverse habitats in Africa, Madagascar, Mascarene Islands, Comoros, India, Southeast and Tropical Asia and North Australia. All the species are diploids, except the noteworthy allotetraploid C. arabica, originated from a recent inter-specific hybridisation between two diploids: C. canephora and C. eugenioides. However, genome size of diploid species can vary for up to two folds. Today, the numerous genomic data available for Coffea allows the study of LTR- RTs, constituting at least 42% of C. canephora genome, the sequenced species available in public databases.In this work, two notable LTR-RT lineages, Bianca and SIRE, have been studied by bioinformatics approaches. Bianca s.s., is present only in Monocots and it is represented in Dicots by the Divo family, poorly studied nowadays. The recent activation of Divo, without leading to its own structuring, is closely associated to the genetic differentiation of C. canephora. However, this activation seems sporadic as being present in all the coffee-trees species studied here. On the opposite, SIRE elements, which are the only Copia LTR-RTs carrying an envelope-like gene as retroviruses, show an important structuring variation between accessions among C. arabica progenitors, and in parallel to the genus evolution.Our work shows that understanding the LTR-RTs dynamics in a genus allows a better perception of its evolutionary history, with the possibility of different evolutionary timing given by different LTR-RTs families. Our results also indicate that both the biogeographic clades (coffee molecular phylogeny) and also some diploid accessions have peculiar histories, probably related to the colonisation of new ecological niches and to the LTR- RTs dynamics.Les éléments transposables (ET) sont des portions d’ADN capables de se déplacer et d’augmenter le nombre de leurs copies dans les génomes. Deux grands types de transposition, correspondant à deux grandes classes d’ET, sont retrouvés chez la quasi-totalité des génomes étudiés à ce jour. Les rétrotransposons à LTR (Long Terminal Repeats, LTR-RT), appartenant à la Classe 1, sont les composants majoritaires des génomes des plantes. Leur prolifération peut avoir un impact important sur l’organisation, la variation de taille, l’évolution des génomes et l’activité des gènes.Le café, largement consommé dans le monde et produit uniquement par des pays du Sud, est issu de deux espèces cultivées d’origine africaine : Coffea arabica et C. canephora. Le genre Coffea est constitué de 139 espèces occupant des habitats très variés en Afrique, dans les îles de l’ouest de l’océan Indien, l’Inde, l’Asie tropicale et du sud-est et au nord de l’Australie. Toutes les espèces son diploïdes, à l’exception notable de C. arabica, allotétraploïde, issu d’une hybridation interspécifique récente entre les deux espèces diploïdes : C. canephora et C. eugenioides. Pour autant, la taille des génomes des espèces diploïdes varie du simple au double. Les nombreuses données génomiques aujourd’hui disponibles au sein du genre Coffea permettent d’étudier la dynamique des LTR-RT constituant au minimum 42% du génome de C. canephora, l’espèce séquencée et disponible dans les bases de données publiques.Dans ce travail, deux lignées remarquables de LTR-RT, Bianca et SIRE, ont été étudiées par des approches bio- informatiques. Bianca sensu stricto, présente uniquement chez les monocotylédones, est représentée chez les dicotylédones par la famille Divo, très peu étudiée à ce jour. L’activation récente de Divo sans induire sa propre structuration, est étroitement associée à la différenciation génétique de C. canephora. Par contre, tout en étant présente dans toutes les espèces de caféiers étudiées, l’activation semble sporadique. À l’opposé, les éléments SIRE, la seule lignée de LTR-RT de la superfamille des Copia contenant un domaine enveloppe comme les rétrovirus, montre des variations structurales importantes entre les accessions des espèces diploïdes à l’origine de C arabica et plus globalement, et en parallèle de l’évolution du genre.Nos travaux montrent que la compréhension de la dynamique des LTR-RT dans un genre peut permettre de mieux appréhender son histoire évolutive, chaque famille de LTR-RT pouvant apporter un éclairage différent. Nos résultats indiquent qu’à la fois les clades biogéographiques (phylogénie moléculaire des caféiers) mais aussi certaines accessions d’espèces diploïdes ont des histoires particulières. Celles-ci seraient vraisemblablement liées à la colonisation de nouvelles niches et à la dynamique des LTR-RT composant les génomes des Coffea

Horizontal transfer of transposons between and within crustaceans and insects

BACKGROUND: Horizontal transfer of transposable elements (HTT) is increasingly appreciated as an important source of genome and species evolution in eukaryotes. However, our understanding of HTT dynamics is still poor in eukaryotes because the diversity of species for which whole genome sequences are available is biased and does not reflect the global eukaryote diversity. RESULTS: In this study we characterized two Mariner transposable elements (TEs) in the genome of several terrestrial crustacean isopods, a group of animals particularly underrepresented in genome databases. The two elements have a patchy distribution in the arthropod tree and they are highly similar (>93% over the entire length of the element) to insect TEs (Diptera and Hymenoptera), some of which were previously described in Ceratitis rosa (Crmar2) and Drosophila biarmipes (Mariner-5_Dbi). In addition, phylogenetic analyses and comparisons of TE versus orthologous gene distances at various phylogenetic levels revealed that the taxonomic distribution of the two elements is incompatible with vertical inheritance. CONCLUSIONS: We conclude that the two Mariner TEs each underwent at least three HTT events. Both elements were transferred once between isopod crustaceans and insects and at least once between isopod crustacean species. Crmar2 was also transferred between tephritid and drosophilid flies and Mariner-5 underwent HT between hymenopterans and dipterans. We demonstrate that these various HTTs took place recently (most likely within the last 3 million years), and propose iridoviruses and/or Wolbachia endosymbionts as potential vectors of these transfers

Distribution of Divo in Coffea genomes, a poorly described family of angiosperm LTR-Retrotransposons

International audienceCoffea arabica (the Arabica coffee) is an allotetraploid species originating from a recent hybridization between two diploid species: C. canephora and C. eugenioides. Transposable elements can drive structural and functional variation during the process of hybridization and allopolyploid formation in plants. To learn more about the evolution of the C. arabica genome, we characterized and studied a new Copia LTR-Retrotransposon (LTR-RT) family in diploid and allotetraploid Coffea genomes called Divo. It is a complete and relatively compact LTR-RT element (similar to 5 kb), carrying typical Gag and Pol Copia type domains. Reverse Trancriptase (RT) domain-based phylogeny demonstrated that Divo is a new and well-supported family in the Bianca lineage, but strictly restricted to dicotyledonous species. In C. canephora, Divo is expressed and showed a genomic distribution along gene rich and gene poor regions. The copy number, the molecular estimation of insertion time and the analysis at orthologous locations of insertions in diploid and allotetraploid coffee genomes suggest that Divo underwent a different and recent transposition activity in C. arabica and C. canephora when compared to C. eugenioides. The analysis of this novel LTR-RT family represents an important step toward uncovering the genome structure and evolution of C. arabica allotetraploid genome

The evolutionary history of three Baracoffea species from western Madagascar revealed by chloroplast and nuclear genomes

International audienceThe wild species of the Coffea genus present a very wide morphological, genetic, and biochemical diversity. Wild species are recognized more resistant to diseases, pests, and environmental variations than the two species currently cultivated worldwide: C. arabica (Arabica) and C. canephora (Robusta). Consequently, wild species are now considered as a crucial resource for adapting cultivated coffee trees to climate change. Within the Coffea genus, 79 wild species are native to the Indian Ocean islands of Comoros, Mayotte, Mauritius, Re ´union and Madagascar, out of a total of 141 taxa worldwide. Among them, a group of 9 species called "Baracoffea" are particularly atypical in their morphology and adaptation to the sandy soils of the dry deciduous forests of western Madagascar. Here, we have attempted to shed light on the evolutionary history of three Baracoffea species: C. ambongensis, C. boinensis and C. bissetiae by analyzing their chloroplast and nuclear genomes. We assembled the complete chloroplast genomes de novo and extracted 28,800 SNP (Single Nucleotide Polymorphism) markers from the nuclear genomes. These data were used for phylogenetic analysis of Baracoffea with Coffea species from Madagascar and Africa. Our new data support the monophyletic origin of Baracoffea within the Coffea of Madagascar, but also reveal a divergence with a sister clade of four species: C. augagneurii, C. ratsimamangae, C. pervilleana and C. Mcphersonii (also called C. vohemarensis), belonging to the Subterminal botanical series and living in dry or humid forests of northern Madagascar. Based on a bioclimatic analysis, our work suggests that Baracoffea may have diverged from a group of Malagasy Coffea from northern Madagascar and adapted to the specific dry climate and low rainfall of western Madagascar. The genomic data generated in the course of this work will contribute to the understanding of the adaptation mechanisms of these particularly singular species

The evolutionary history of three Baracoffea species from western Madagascar revealed by chloroplast and nuclear genomes.

The wild species of the Coffea genus present a very wide morphological, genetic, and biochemical diversity. Wild species are recognized more resistant to diseases, pests, and environmental variations than the two species currently cultivated worldwide: C. arabica (Arabica) and C. canephora (Robusta). Consequently, wild species are now considered as a crucial resource for adapting cultivated coffee trees to climate change. Within the Coffea genus, 79 wild species are native to the Indian Ocean islands of Comoros, Mayotte, Mauritius, Réunion and Madagascar, out of a total of 141 taxa worldwide. Among them, a group of 9 species called "Baracoffea" are particularly atypical in their morphology and adaptation to the sandy soils of the dry deciduous forests of western Madagascar. Here, we have attempted to shed light on the evolutionary history of three Baracoffea species: C. ambongensis, C. boinensis and C. bissetiae by analyzing their chloroplast and nuclear genomes. We assembled the complete chloroplast genomes de novo and extracted 28,800 SNP (Single Nucleotide Polymorphism) markers from the nuclear genomes. These data were used for phylogenetic analysis of Baracoffea with Coffea species from Madagascar and Africa. Our new data support the monophyletic origin of Baracoffea within the Coffea of Madagascar, but also reveal a divergence with a sister clade of four species: C. augagneurii, C. ratsimamangae, C. pervilleana and C. Mcphersonii (also called C. vohemarensis), belonging to the Subterminal botanical series and living in dry or humid forests of northern Madagascar. Based on a bioclimatic analysis, our work suggests that Baracoffea may have diverged from a group of Malagasy Coffea from northern Madagascar and adapted to the specific dry climate and low rainfall of western Madagascar. The genomic data generated in the course of this work will contribute to the understanding of the adaptation mechanisms of these particularly singular species

Partial sequencing reveals the transposable element composition of Coffea genomes and provides evidence for distinct evolutionary stories

International audienceThe Coffea genus, 124 described species, has a natural distribution spreading from inter-tropical Africa, to Western Indian Ocean Islands, India, Asia and up to Australasia. Two cultivated species, C. arabica and C. canephora, are intensively studied while, the breeding potential and the genome composition of all the wild species remained poorly uncharacterized. Here, we report the characterization and comparison of the highly repeated transposable elements content of 11 Coffea species representatives of the natural biogeographic distribution. A total of 994 Mb from 454 reads were produced with a genome coverage ranging between 3.2 and 15.7 %. The analyses showed that highly repeated transposable elements, mainly LTR retrotransposons (LTR-RT), represent between 32 and 53 % of Coffea genomes depending on their biogeographic location and genome size. Species from West and Central Africa (Eucoffea) contained the highest LTR-RT content but with no strong variation relative to their genome size

Linear regression analyses between genome size and environmental data.

Linear regression analyses between genome size and environmental data.</p

Baracoffea genetic diversity assessed using 9,665 variant SNPs.

A. Genetic distance of Coffea species from Madagascar. B. PCA analysis. C. Geographic position of species used in the analysis.</p