RESUMEN
"Simple" 1-way interchromosomal insertions involving an interstitial 1q segment are rare, and therefore, their characterization at the base pair level remains understudied. Here, we describe the genomic characterization of a previously unreported de novo interchromosomal insertion (3;1) entailing an about 12-Mb pure gain of 1q21.3q23.3 that causes typical (microcephaly, developmental delay, and facial dysmorphism) and atypical (interauricular communication, small feet with bilateral deep plantar creases, syndactyly of II-IV toes, and mild pachyonychia of all toes) clinical manifestations associated with this region. Based on our analyses, we hypothesize that the duplication of a subset of morbid genes (including LMNA, USF1, VANGL2, LOR, and POGZ) could account for most clinical findings in our patient. Furthermore, the apparent disruption of a promoter region (between CPNE9 and BRPF1) and a topologically associated domain also suggests likely pathogenic reconfiguration/position effects to contribute to the patient's phenotype. In addition to further expanding the clinical spectrum of proximal 1q duplications and evidencing the phenotypical heterogeneity among similar carriers, our genomic findings and observations suggest that randomness - rather than lethality issues - may account for the paucity of "simple" interchromosomal insertions involving the 1q21.3q23.3 region as genomic donor and distal 3p25.3 as receptor. Moreover, the microhomology sequence found at the insertion breakpoint is consistent with a simple nonhomologous end-joining mechanism, in contrast to a chromothripsis-like event, which has previously been seen in other nonrecurrent insertions. Taken together, the data gathered in this study allowed us to inform this family about the low recurrence risk but not to predict the reproductive prognosis for hypothetical carriers. We highlight that genomic-level assessment is a powerful tool that allows the visualization of the full landscape of sporadic chromosomal injuries and can be used to improve genetic counseling.
Asunto(s)
Anomalías Múltiples/genética , Cromosomas Humanos Par 3/genética , Anomalías Congénitas/genética , Genoma Humano , Adulto , Preescolar , Duplicación Cromosómica/genética , Hibridación Genómica Comparativa , Humanos , Lactante , Recién Nacido , Mapas de Interacción de Proteínas , Secuenciación Completa del GenomaRESUMEN
Actin polymerization, actomyosin ring contraction, and stress fiber formation are examples of relevant actions of the RhoA/B/C pathway as GTPases that regulate the cytoskeleton. However, open questions that remain to be addressed are whether this pathway and/or downstream components protect against or facilitate the formation of DNA double-strand breaks, the most lethal form of DNA damage in cells. Genotoxic drugs are radiomimetic and/or chemotherapeutic agents that are currently used for cancer treatments and are associated with specific methodologies; thus, these compounds should represent good tools to answer these questions. In this chapter, we describe two methods, the alkaline comet assay and homologous/nonhomologous recombination assays, to investigate the mechanism by which the Rho pathway modulates the repair of DNA breaks in tumor epithelial cell lines.