Synostosis

Timothy syndrome (TS) is a severe, multisystem disorder characterized by autism, epilepsy, long-QT syndrome and other neuropsychiatric conditions1. TS type 1 (TS1) is caused by a gain-of-function variant in the alternatively spliced and developmentally enriched CACNA1C exon 8A, as opposed to its counterpart exon 8. We previously uncovered several phenotypes in neurons derived from patients with TS1, including delayed channel inactivation, prolonged depolarization-induced calcium rise, impaired interneuron migration, activity-dependent dendrite retraction and an unanticipated persistent expression of exon 8A2-6. We reasoned that switching CACNA1C exon utilization from 8A to 8 would represent a potential therapeutic strategy. Here we developed antisense oligonucleotides (ASOs) to effectively decrease the inclusion of exon 8A in human cells both in vitro and, following transplantation, in vivo. We discovered that the ASO-mediated switch from exon 8A to 8 robustly rescued defects in patient-derived cortical organoids and migration in forebrain assembloids. Leveraging a transplantation platform previously developed7, we found that a single intrathecal ASO administration rescued calcium changes and in vivo dendrite retraction of patient neurons, suggesting that suppression of CACNA1C exon 8A expression is a potential treatment for TS1. Broadly, these experiments illustrate how a multilevel, in vivo and in vitro stem cell model-based approach can identify strategies to reverse disease-relevant neural pathophysiology.

Skull development coincides with the onset of cerebrospinal fluid (CSF) circulation, brain-CSF perfusion, and meningeal lymphangiogenesis, processes essential for brain waste clearance. How these processes are affected by craniofacial disorders such as craniosynostosis are poorly understood. We report that raised intracranial pressure and diminished CSF flow in craniosynostosis mouse models associate with pathological changes to meningeal lymphatic vessels that affect their sprouting, expansion, and long-term maintenance. We also show that craniosynostosis affects CSF circulatory pathways and perfusion into the brain. Further, craniosynostosis exacerbates amyloid pathology and plaque buildup in Twist1+/-:5xFAD transgenic Alzheimer's disease models. Treating craniosynostosis mice with Yoda1, a small molecule agonist for Piezo1, reduces intracranial pressure and improves CSF flow, in addition to restoring meningeal lymphangiogenesis, drainage to the deep cervical lymph nodes, and brain-CSF perfusion. Leveraging these findings, we show that Yoda1 treatments in aged mice with reduced CSF flow and turnover improve lymphatic networks, drainage, and brain-CSF perfusion. Our results suggest that CSF provides mechanical force to facilitate meningeal lymphatic growth and maintenance. Additionally, applying Yoda1 agonist in conditions with raised intracranial pressure and/or diminished CSF flow, as seen in craniosynostosis or with ageing, is a possible therapeutic option to help restore meningeal lymphatic networks and brain-CSF perfusion.

IntroductionGrowing concerns over health and welfare impacts from extreme phenotypes in dogs have created an urgent need for reliable demographic information on the national breed structures of dogs.MethodsThis study included all dogs under primary veterinary care in the UK during 2019 at practices participating in VetCompass. Demographic data on these dogs were analysed to report on the frequency of common breeds and also to report on conformation, bodyweight, sex and neuter associations with these breeds.ResultsThe study included 2,237,105 dogs under UK veterinary care in 2019. Overall, 69.4% (n = 1,551,462) were classified as purebred, 6.7% (149,308) as designer-crossbred and 24.0% (536,335) as nondesigner-crossbred. Across 800 unique breed names, the most frequent breeds at any age were nondesigner-crossbred (n = 536,335, 24.0%), Labrador Retriever (154,222, 6.9%) and Jack Russell Terrier (101,294, 4.5%). Among 229,624 (10.3%) dogs aged under one year, the most frequent breeds were nondesigner-crossbred (n = 45,995, 20.0%), French Bulldog (16,036, 7.0%) and Cockapoo (14,321, 6.2%). Overall, based on breed characteristics, 17.6% (395,739) were classified as brachycephalic, 43.1% (969,403) as mesaticephalic and 8.3% (186,320) as dolichocephalic. Of 1,551,336 dogs that were classifiable based on breed, 52.6% (815,673) were chondrodystrophic. Of 1,462,925 dogs that were classifiable, there were 54.6% (n = 798,426) short haired, 32.6% (476,883) medium haired and 12.8% (186,934) long haired. Of 1,547,653 dogs that were classifiable for ear carriage, 24.5% (n = 379,581) were erect, 28.1% (434,273) were semi-erect, 19.7% (305,475) were v-shaped drop and 27.7% (428,324) were pendulous. Overall, there was a 1.09:1.00 ratio of male (n = 1,163,512; 52.2%) to female dogs (n = 1,067,552; 47.8%).ConclusionsHealth and welfare issues linked to popular breeds with extreme phenotypes suggest that there is much work to do to help owners to make more welfare-friendly decisions when choosing which type of dog to own.

Mutations in the gene encoding cytoskeleton-associated protein 2-like (CKAP2L) have been identified as a causative factor for Filippi syndrome, a rare developmental disorder characterized by facial dysmorphism, syndactyly, and microcephaly. However, the cellular and molecular mechanisms by which CKAP2L contributes to the pathogenesis of this syndrome remain largely unknown. Here, we generated a Ckap2l knockout mouse model to investigate the in vivo and cellular roles of CKAP2L. Interestingly, Ckap2l knockout mice show no overt developmental abnormalities, with the exception of reduced male fertility, evidenced by decreased sperm count, impaired motility, and abnormally elongated flagella. At the cellular level, CKAP2L is a bona fide microtubule-associated protein that localizes to microtubule-based organelles, including the centrosome, mitotic spindle, and ciliary basal body. Depletion of CKAP2L leads to shortened mitotic spindles and cytokinesis failure, resulting in multinucleation. Furthermore, we uncover a conserved function for CKAP2L as a negative regulator of primary cilium length; its loss markedly increases ciliary length in both human and mouse cells. Collectively, these findings position CKAP2L as a multifunctional regulator of microtubule-based organelles and suggest that Filippi syndrome can be classified as a 'centrosomopathy' arising from concurrent defects in cell proliferation and ciliary function.