Formin tails act as a switch, inhibiting or enhancing processive actin elongation.

Formins are large, multidomain proteins that nucleate new actin filaments and accelerate elongation through a processive interaction with the barbed ends of filaments. Their actin assembly activity is generally attributed to their eponymous formin homology (FH) 1 and 2 domains; however, evidence is mounting that regions outside of the FH1FH2 stretch also tune actin assembly. Here, we explore the underlying contributions of the tail domain, which spans the sequence between the FH2 domain and the C terminus of formins. Tails vary in length from ∼0 to >200 residues and contain a number of recognizable motifs. The most common and well-studied motif is the ∼15-residue-long diaphanous autoregulatory domain. This domain mediates all or nothing regulation of actin assembly through an intramolecular interaction with the diaphanous inhibitory domain in the N-terminal half of the protein. Multiple reports demonstrate that the tail can enhance both nucleation and processivity. In this study, we provide a high-resolution view of the alternative splicing encompassing the tail in the formin homology domain (Fhod) family of formins during development. While four distinct tails are predicted, we found significant levels of only two of these. We characterized the biochemical effects of the different tails. Surprisingly, the two highly expressed Fhod-tails inhibit processive elongation and diminish nucleation, while a third supports activity. These findings demonstrate a new mechanism of modulating actin assembly by formins and support a model in which splice variants are specialized to build distinct actin structures during development.

Single-cell RNA-seq supports a developmental hierarchy in human oligodendroglioma.

Although human tumours are shaped by the genetic evolution of cancer cells, evidence also suggests that they display hierarchies related to developmental pathways and epigenetic programs in which cancer stem cells (CSCs) can drive tumour growth and give rise to differentiated progeny. Yet, unbiased evidence for CSCs in solid human malignancies remains elusive. Here we profile 4,347 single cells from six IDH1 or IDH2 mutant human oligodendrogliomas by RNA sequencing (RNA-seq) and reconstruct their developmental programs from genome-wide expression signatures. We infer that most cancer cells are differentiated along two specialized glial programs, whereas a rare subpopulation of cells is undifferentiated and associated with a neural stem cell expression program. Cells with expression signatures for proliferation are highly enriched in this rare subpopulation, consistent with a model in which CSCs are primarily responsible for fuelling the growth of oligodendroglioma in humans. Analysis of copy number variation (CNV) shows that distinct CNV sub-clones within tumours display similar cellular hierarchies, suggesting that the architecture of oligodendroglioma is primarily dictated by developmental programs. Subclonal point mutation analysis supports a similar model, although a full phylogenetic tree would be required to definitively determine the effect of genetic evolution on the inferred hierarchies. Our single-cell analyses provide insight into the cellular architecture of oligodendrogliomas at single-cell resolution and support the cancer stem cell model, with substantial implications for disease management.

Drosophila and human FHOD family formin proteins nucleate actin filaments.

Formins are a conserved group of proteins that nucleate and processively elongate actin filaments. Among them, the formin homology domain-containing protein (FHOD) family of formins contributes to contractility of striated muscle and cell motility in several contexts. However, the mechanisms by which they carry out these functions remain poorly understood. Mammalian FHOD proteins were reported not to accelerate actin assembly in vitro; instead, they were proposed to act as barbed end cappers or filament bundlers. Here, we show that purified Drosophila Fhod and human FHOD1 both accelerate actin assembly by nucleation. The nucleation activity of FHOD1 is restricted to cytoplasmic actin, whereas Drosophila Fhod potently nucleates both cytoplasmic and sarcomeric actin isoforms. Drosophila Fhod binds tightly to barbed ends, where it slows elongation in the absence of profilin and allows, but does not accelerate, elongation in the presence of profilin. Fhod antagonizes capping protein but dissociates from barbed ends relatively quickly. Finally, we determined that Fhod binds the sides of and bundles actin filaments. This work establishes that Fhod shares the capacity of other formins to nucleate and bundle actin filaments but is notably less effective at processively elongating barbed ends than most well studied formins.

Analysis of early angiographic outcome using unique large diameter coils in comparison with standard coils in the embolization of cerebral aneurysms: a retrospective review.

BACKGROUND: The initial experience with the large diameter Penumbra Coil 400 (PC400) system has been positive regarding safety, efficacy, improved packing density and cost effectiveness, but follow-up data are limited. METHODS: This is a single-center retrospective review of 76 aneurysms treated with PC400 coils compared with 301 aneurysms treated with a variety of different bare platinum and bioactively coated coils. Atypical and giant aneurysms were excluded as well as those that had undergone previous treatment. Occlusion classification was determined immediately after the procedure and at short-term follow-up. RESULTS: Compared with controls, in the PC400 group fewer coils were used (3.53 vs 5.44, p<0.05), procedure time was decreased (48 vs 64 min, p<0.05) and packing density was increased (31.7% vs 24.8%, p<0.05). There were more grade III aneurysms (71.1% vs 38.2%, p<0.05) and fewer grade I aneurysms (13.2% vs 30.2%, p<0.05) in the PC400 group than in the control group immediately after the procedure. At first follow-up, however, more aneurysms in the PC400 group improved (51.3% vs 28.7%, p<0.05) in angiographic grade leading to similar rates of acceptable outcome (grades I or II) at first follow-up (PC400 79.5% vs control 77.2%). The adverse event rate was similar in the two groups. CONCLUSIONS: Large diameter PC400 coils can be used to safely treat cerebral aneurysms with fewer coils, decreased procedure time and increased packing density compared with standard coils. The early angiographic outcome is similar to that achieved with standard coils.

An update to the Raymond-Roy Occlusion Classification of intracranial aneurysms treated with coil embolization.

BACKGROUND: The Raymond-Roy Occlusion Classification (RROC) is the standard for evaluating coiled aneurysms (Class I: complete obliteration; Class II: residual neck; Class III: residual aneurysm), but not all Class III aneurysms behave the same over time. METHODS: This is a retrospective review of 370 patients with 390 intracranial aneurysms treated with coil embolization. A Modified Raymond-Roy Classification (MRRC), in which Class IIIa designates contrast within the coil interstices and Class IIIb contrast along the aneurysm wall, was applied retrospectively. RESULTS: Class IIIa aneurysms were more likely to improve to Class I or II than Class IIIb aneurysms (83.34% vs 14.89%, p<0.001) and were also more likely than Class II to improve to Class I (52.78% vs 16.90%, p<0.001). Class IIIb aneurysms were more likely to remain incompletely occluded than Class IIIa aneurysms (85.11% vs 16.67%, p<0.001). Class IIIb aneurysms were larger with wider necks while Class IIIa aneurysms had higher packing density. Class IIIb aneurysms had a higher retreatment rate (33.87% vs 6.54%, p<0.001) and a trend toward higher subsequent rupture rate (3.23% vs 0.00%, p=0.068). CONCLUSIONS: We propose the MRRC to further differentiate Class III aneurysms into those likely to progress to complete occlusion and those likely to remain incompletely occluded or to worsen. The MRRC has the potential to expand the definition of adequate coil embolization, possibly decrease procedural risk, and help endovascular neurosurgeons predict which patients need closer angiographic follow-up. These findings need to be validated in a prospective study with independent blinded angiographic grading.

Initial experience with Penumbra Coil 400 versus standard coils in embolization of cerebral aneurysms: a retrospective review.

BACKGROUND AND PURPOSE: The Penumbra Coil 400 System (PC 400) is the newly introduced platinum coil system designed specifically to enhance filling efficiency by increasing coil diameter. Our goal was to study the packing and treatment advantage of the PC 400's unique geometric configuration compared with conventional coils (controls). MATERIALS AND METHODS: 16 aneurysms embolized with the PC 400 in 2011 were compared with 79 equally matched aneurysms embolized with conventional coils from 2004 to 2011. Primary outcomes assessed were acute packing density, embolization time, and the number of coils required for aneurysm occlusion. RESULTS: Aneurysm embolization with the PC 400 achieved a higher packing density (36.8% vs 28.1%; p<0.005) and with fewer coils (an average of 3.9 vs 6.1 coils per aneurysm; p<0.05). In addition, the total procedural time for the subjects treated with the PC 400 was significantly less (45.7 vs 64.1 min; p<0.05). There were no procedural complications associated with the PC 400. CONCLUSIONS: Compared with conventional coils, the PC 400 is more efficient in the embolization of cerebral aneurysms, achieving greater packing density with fewer coils and less time without compromising safety.