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Funder: Barncancerfonden; FundRef: http://dx.doi.org/10.13039/501100006313, Funder: Norwegian Research Council; FundRef: http://dx.doi.org/10.13039/501100005416, Funder: Knut och Alice Wallenbergs Stiftelse; FundRef: http://dx.doi.org/10.13039/501100004063, Funder: Cancerfonden; FundRef: http://dx.doi.org/10.13039/501100002794, Funder: Instrumentariumin Tiedes����ti��; FundRef: http://dx.doi.org/10.13039/501100008413, Funder: Science for Life Laboratory; FundRef: http://dx.doi.org/10.13039/501100009252, Funder: Academy of Finland; FundRef: http://dx.doi.org/10.13039/501100002341, Precision CRISPR gene editing relies on the cellular homology-directed DNA repair (HDR) to introduce custom DNA sequences to target sites. The HDR editing efficiency varies between cell types and genomic sites, and the sources of this variation are incompletely understood. Here, we have studied the effect of 450 DNA repair protein-Cas9 fusions on CRISPR genome editing outcomes. We find the majority of fusions to improve precision genome editing only modestly in a locus- and cell-type specific manner. We identify Cas9-POLD3 fusion that enhances editing by speeding up the initiation of DNA repair. We conclude that while DNA repair protein fusions to Cas9 can improve HDR CRISPR editing, most need to be optimized to the cell type and genomic site, highlighting the diversity of factors contributing to locus-specific genome editing outcomes.

Summary Essential E3 ubiquitin ligase HUWE1 (HECT, UBA, and WWE domain containing 1) regulates key factors, such as p53. Although mutations in HUWE1 cause heterogenous neurodevelopmental X-linked intellectual disabilities (XLIDs), the disease mechanisms common to these syndromes remain unknown. In this work, we identify p53 signaling as the central process altered in HUWE1-promoted XLID syndromes. By focusing on Juberg-Marsidi syndrome (JMS), one of the severest XLIDs, we show that increased p53 signaling results from p53 accumulation caused by HUWE1 p.G4310R destabilization. This further alters cell-cycle progression and proliferation in JMS cells. Modeling of JMS neurodevelopment reveals majorly impaired neural differentiation accompanied by increased p53 signaling. The neural differentiation defects can be successfully rescued by reducing p53 levels and restoring the expression of p53 target genes, in particular CDKN1A/p21. In summary, our findings suggest that increased p53 signaling underlies HUWE1-promoted syndromes and impairs XLID JMS neural differentiation., Graphical abstract, Highlights p53 signaling is hyperactivated in HUWE1-promoted intellectual disabilities Neural differentiation is impaired in Juberg-Marsidi syndrome with HUWE1 p.G4310R Juberg-Marsidi brain organoids have reduced size and aberrant cellular organization Restored p53 signaling rescues neural development in Juberg-Marsidi syndrome, Using transcriptome analysis, Aprigliano et al. identify increased p53 signaling as the most significantly deregulated process in HUWE1-promoted XLIDs. Modeling of a severe HUWE1-promoted XLID, Juberg-Marsidi syndrome, reveals impaired neural differentiation capacity, which is successfully rescued by p53 knockdown and restored expression of p53 target genes, such as CDKN1A/p21.

SUMMARYEssential E3 ubiquitin ligase HUWE1 (HECT, UBA and WWE domain containing 1) regulates key factors, as p53. Mutations in HUWE1 have been associated with neurodevelopmental X-linked intellectual disabilities (XLIDs), however the pathomechanism at the onset of heterogenous XLIDs remains unknown. In this work, we identify p53 signaling as the process hyperactivated in lymphoblastoid cells from patients with HUWE1-promoted XLIDs. The hiPSCs-based modeling of the severe HUWE1-promoted XLID, the Juberg Marsidi syndrome (JMS), reviled majorly impaired neural differentiation, accompanied by increased p53 signaling. The impaired differentiation results in loss of cortical patterning and overall undergrowth of XLID JMS patient-specific cerebral organoids, thus closely recapitulating key symptoms, as microcephaly. Importantly, the neurodevelopmental potential of JMS hiPSCs is successfully rescued by restoring p53 signaling, upon reduction of p53 levels. In summary, our findings indicate that increased p53 signaling leads to impaired neural differentiation and is the common cause of neurodevelopmental HUWE1-promoted XLIDs.

Faciogenital dysplasia 5 (FGD5) amplification drives tumor cell proliferation, and is present in 9.5% of breast cancers. We describe FGD5 expression, assess associations between FGD5 amplification and FGD5 expression, and assess FGD5 expression in relation to proliferation and prognosis. FGD5 immunohistochemistry was done on primary tumors (n=829) and lymph node metastases (n=231) from a cohort of Norwegian patients. We explored associations between FGD5 amplification, FGD5 expression, and proliferation, and analyzed the prognostic value of FGD5 expression by estimating cumulative risks of death and hazard ratios (HRs). We identified nuclear and cytoplasmic expression in 64% and 73% of primary tumors, respectively, and found an association between gene amplification and nuclear expression (p=0.02). The proportion of cases with FGD5 expression was higher in lymph node metastases, compared with primary tumors (p=0.004 for nuclear and p=0.001 for cytoplasmic staining). Neither proliferation nor prognosis was associated with FGD5 expression (age-adjusted HR 1.12 [95% confidence interval = 0.89–1.41] for nuclear expression; and 0.88 [95% CI = 0.70–1.12] for cytoplasmic expression). FGD5 is expressed in a high proportion of breast cancers and lymph node metastases. There was a correlation between FGD5 amplification and nuclear expression, but no association between FGD5 expression and proliferation or prognosis. © 2018. This is the authors' accepted and refereed manuscript to the article. The final authenticated version is available online at: https://doi.org/10.1369/0022155418792032

Parkinson's disease is characterized by the death of dopaminergic neurons, mainly in the substantia nigra, and causes serious locomotor dysfunctions. It is likely that the oxidative damage to cellular biomolecules is among the leading causes of neurodegeneration that occurs in the disease. Selenium is an essential mineral for proper functioning of the brain, and mainly due to its antioxidant activity, it is possible to exert a special role in the prevention and in the nutritional management of Parkinson's disease. Currently, few researchers have investigated the effects of selenium on Parkinson´s disease. However, it is known that very high or very low body levels of selenium can (possibly) contribute to the pathogenesis of Parkinson's disease, because this imbalance results in increased levels of oxidative stress. Therefore, the aim of this work is to review and discuss studies that have addressed these topics and to finally associate the information obtained from them so that these data and associations serve as input to new research. RESUMO A doença de Parkinson é caracterizada pela morte dos neurônios dopaminérgicos, principalmente na substância negra, e causa sérias disfunções locomotoras. É provável que o dano oxidativo às biomoléculas celulares esteja entre as principais causas da neurodegeneração que ocorre nessa doença. O selênio é um mineral essencial para o adequado funcionamento do encéfalo e, principalmente devido a sua atividade antioxidante, é possível que exerça um papel especial na prevenção e no manejo nutricional da doença de Parkinson. Atualmente, poucos pesquisadores têm investigado os efeitos do selênio sobre a doença de Parkinson. Entretanto, sabe-se que níveis corporais muito altos ou muito baixos de selênio podem (possivelmente) contribuir para a patogênese da doença de Parkinson, uma vez que esse desbalanço resulta no aumento dos níveis de estresse oxidativo. Dessa forma, o objetivo deste trabalho é revisar e discutir os estudos que abordaram esses tópicos e então associar as informações obtidas através deles para que esses dados e associações sejam usadas para o estabelecimento de novas pesquisas.

DNA damage and repair activity is increasingly recognized in atherogenesis. By facilitating DNA strand joining by catalyzing phosphodiestere bond formation, DNA ligase is crucial in single/double stranded DNA break repair. We aimed to identify differential DNA ligase activity as well as a