A founder CEP120 mutation in Jeune asphyxiating thoracic dystrophy expands the role of centriolar proteins in skeletal ciliopathies.

Jeune asphyxiating thoracic dystrophy (JATD) is a skeletal dysplasia characterized by a small thoracic cage and a range of skeletal and extra-skeletal anomalies. JATD is genetically heterogeneous with at least nine genes identified, all encoding ciliary proteins, hence the classification of JATD as a skeletal ciliopathy. Consistent with the observation that the heterogeneous molecular basis of JATD has not been fully determined yet, we have identified two consanguineous Saudi families segregating JATD who share a single identical ancestral homozygous haplotype among the affected members. Whole-exome sequencing revealed a single novel variant within the disease haplotype in CEP120, which encodes a core centriolar protein. Subsequent targeted sequencing of CEP120 in Saudi and European JATD cohorts identified two additional families with the same missense mutation. Combining the four families in linkage analysis confirmed a significant genome-wide linkage signal at the CEP120 locus. This missense change alters a highly conserved amino acid within CEP120 (p.Ala199Pro). In addition, we show marked reduction of cilia and abnormal number of centrioles in fibroblasts from one affected individual. Inhibition of the CEP120 ortholog in zebrafish produced pleiotropic phenotypes characteristic of cilia defects including abnormal body curvature, hydrocephalus, otolith defects and abnormal renal, head and craniofacial development. We also demonstrate that in CEP120 morphants, cilia are shortened in the neural tube and disorganized in the pronephros. These results are consistent with aberrant CEP120 being implicated in the pathogenesis of JATD and expand the role of centriolar proteins in skeletal ciliopathies

The KOUNCIL Consortium: From Genetic Defects to Therapeutic Development for Nephronophthisis

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Mechanical loading inhibits cartilage inflammatory signalling via an HDAC6 and IFT-dependent mechanism regulating primary cilia elongation

Objective: physiological mechanical loading reduces inflammatory signalling in numerous cell types including articular chondrocytes however the mechanism responsible remains unclear. This study investigates the role of chondrocyte primary cilia and associated intraflagellar transport (IFT) in the mechanical regulation of interleukin-1β (IL-1β) signalling.Design: isolated chondrocytes and cartilage explants were subjected to cyclic mechanical loading in the presence and absence of the cytokine IL-1β. Nitric oxide (NO) and prostaglandin E2 (PGE2) release were used to monitor IL-1β signalling whilst Sulphated glycosaminoglycan (sGAG) release provided measurement of cartilage degradation. Measurements were made of HDAC6 activity and tubulin polymerisation and acetylation. Effects on primary cilia were monitored by confocal and super resolution microscopy. Involvement of IFT was analysed using ORPK cells with hypomorphic mutation of IFT88.Results: mechanical loading suppressed NO and PGE2 release and prevented cartilage degradation. Loading activated HDAC6 and disrupted tubulin acetylation and cilia elongation induced by IL-1β. HDAC6 inhibition with tubacin blocked the anti-inflammatory effects of loading and restored tubulin acetylation and cilia elongation. Hypomorphic mutation of IFT88 reduced IL-1β signalling and abolished the anti-inflammatory effects of loading indicating the mechanism is IFT-dependent. Loading reduced the pool of non-polymerised tubulin which was replicated by taxol which also mimicked the anti-inflammatory effects of mechanical loading and prevented cilia elongation.Conclusions: this study reveals that mechanical loading suppresses inflammatory signalling, partially dependent on IFT, by activation of HDAC6 and post transcriptional modulation of tubulin

An FDA-Approved Drug Screen for Compounds Influencing Craniofacial Skeletal Development and Craniosynostosis

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Multivesicular assemblies as real-world testbeds for embryogenic evolutionary systems

Embryogenic evolution emulates in silico cell-like entities to get more powerful methods for complex evolutionary tasks. As simulations have to abstract from the biological model, implicit information hidden in its physics is lost. Here, we propose to use cell-like entities as a real-world in vitro testbed. In analogy to evolutionary robotics, where solutions evolved in simulations may be tested in real-world on macroscale, the proposed vesicular testbed would do the same for the embryogenic evolutionary tasks on mesoscale. As a first step towards a vesicular testbed emulating growth, cell division, and cell differentiation, we present a modified vesicle production method, providing custom-tailored chemical cargo, and present a novel self-assembly procedure to provide vesicle aggregates of programmable composition

Towards tailored communication networks in assemblies of artificial cells

Living Technology is researching novel IT making strong use of programmable chemical systems. These chemical systems shall finally converge to artificial cells resulting in evolvable complex information systems. We focus on procedural manageability and information processing capabilities of such information systems. Here, we present a novel resource-saving formation, processing, and examination procedure to generate and handle single compartments representing preliminary stages of artificial cells. Its potential is exemplified by testing the influence of different glycerophospholipids on the stability of the compartments. We discuss how the procedure could be used both in evolutionary optimization of self-assembling amphiphilic systems and in engineering tailored communication networks enabling life-like information processing in multicompartment aggregates of programmable composition and spatial configuration