NPHP4 Variants Are Associated With Pleiotropic Heart Malformations

Rationale: Congenital heart malformations are a major cause of morbidity and mortality, especially in young children. Failure to establish normal left-right (L-R) asymmetry often results in cardiovascular malformations and other laterality defects of visceral organs. Objective: To identify genetic mutations causing cardiac laterality defects. Methods and Results: We performed a genome-wide linkage analysis in patients with cardiac laterality defects from a consanguineous family. The patients had combinations of defects that included dextrocardia, transposition of great arteries, double-outlet right ventricle, atrioventricular septal defects, and caval vein abnormalities. Sequencing of positional candidate genes identified mutations in NPHP4. We performed mutation analysis of NPHP4 in 146 unrelated patients with similar cardiac laterality defects. Forty-one percent of these patients also had laterality defects of the abdominal organs. We identified 8 additional missense variants that were absent or very rare in control subjects. To study the role of nphp4 in establishing L-R asymmetry, we used antisense morpholinos to knockdown nphp4 expression in zebrafish. Depletion of nphp4 disrupted L-R patterning as well as cardiac and gut laterality. Cardiac laterality defects were partially rescued by human NPHP4 mRNA, whereas mutant NPHP4 containing genetic variants found in patients failed to rescue. We show that nphp4 is involved in the formation of motile cilia in Kupffer's vesicle, which generate asymmetrical fluid flow necessary for normal L-R asymmetry. Conclusions: NPHP4 mutations are associated with cardiac laterality defects and heterotaxy. In zebrafish, nphp4 is essential for the development and function of Kupffer's vesicle cilia and is required for global L-R patterning

Histone H3.3 beyond cancer: Germline mutations in Histone 3 Family 3A and 3B cause a previously unidentified neurodegenerative disorder in 46 patients

Although somatic mutations in Histone 3.3 (H3.3) are well-studied drivers of oncogenesis, the role of germline mutations remains unreported. We analyze 46 patients bearing de novo germline mutations in histone 3 family 3A (H3F3A) or H3F3B with progressive neurologic dysfunction and congenital anomalies without malignancies. Molecular modeling of all 37 variants demonstrated clear disruptions in interactions with DNA, other histones, and histone chaperone proteins. Patient histone posttranslational modifications (PTMs) analysis revealed notably aberrant local PTM patterns distinct from the somatic lysine mutations that cause global PTM dysregulation. RNA sequencing on patient cells demonstrated up-regulated gene expression related to mitosis and cell division, and cellular assays confirmed an increased proliferative capacity. A zebrafish model showed craniofacial anomalies and a defect in Foxd3-derived glia. These data suggest that the mechanism of germline mutations are distinct from cancer-associated somatic histone mutations but may converge on control of cell proliferation

Building Europe on transnational infrastructures

Eindhoven University of Technology, Netherlands Abstract: The Treaty of Rome signed in 1957 considered the development of a Common Transport Policy (CTP) pivotal to the European integration process. Treaty articles 74-84 targeted the road sector, along with railways and inland waterways, as a core competence of the newly established European Economic Community. The CTP did not get off the ground, however. Instead it languished in obscurity for much of the first forty years of European integration. In 1985 this neglect even led to a ruling of the European Court of Justice, the so-called inactivity verdict. Several histories of the transport policy of the European Union (EU) and its predecessors have lamented the obvious lack of action. To explain the years of impasse they pointed to a strong divergence of interest among the signatories. They argued that 'the Six' (the Benelux countries Belgium, Luxembourg and the Netherlands, as well as France, Germany and Italy) advanced their own strongly divergent national transport policies, and were sceptical of a liberal regime, except for the Dutch, because this would support their highly export-oriented transport sector. This argumentation fits into the analysis by historians of European integration such as Alan Milward, who have argued that the nation states have consistently controlled the integration process, and evidenced that the anticipated expansive and irresistible integration logic, as suggested by neofunctionalist theory and promoters of the European integration process, never happened. Transport is particularly interesting for this debate because, in the processes leading to the treaty, it was seen as one of the sectors that would integrate first. As this did not happen, the transport case seems to validate the conclusion that Europe is first and foremost a Europe of nation states