CRISPR-Cas9 correction of OPA1 c.1334G>A: p.R445H restores mitochondrial homeostasis in dominant optic atrophy patient-derived iPSCs.

Autosomal dominant optic atrophy (DOA) is the most common inherited optic neuropathy in the United Kingdom. DOA has an insidious onset in early childhood, typically presenting with bilateral, central visual loss caused by the preferential loss of retinal ganglion cells. 60%-70% of genetically confirmed DOA cases are associated with variants in OPA1, a ubiquitously expressed GTPase that regulates mitochondrial homeostasis through coordination of inner membrane fusion, maintenance of cristae structure, and regulation of bioenergetic output. Whether genetic correction of OPA1 pathogenic variants can alleviate disease-associated phenotypes remains unknown. Here, we demonstrate generation of patient-derived OPA1 c.1334G>A: p.R445H mutant induced pluripotent stem cells (iPSCs), followed by correction of OPA1 through CRISPR-Cas9-guided homology-directed repair (HDR) and evaluate the effect of OPA1 correction on mitochondrial homeostasis. CRISPR-Cas9 gene editing demonstrated an efficient method of OPA1 correction, with successful gene correction in 57% of isolated iPSCs. Correction of OPA1 restored mitochondrial homeostasis, re-establishing the mitochondrial network and basal respiration and ATP production levels. In addition, correction of OPA1 re-established the levels of wild-type (WT) mitochondrial DNA (mtDNA) and reduced susceptibility to apoptotic stimuli. These data demonstrate that nuclear gene correction can restore mitochondrial homeostasis and improve mtDNA integrity in DOA patient-derived cells carrying an OPA1 variant

Eupatilin Improves Cilia Defects in Human CEP290 Ciliopathy Models

The photoreceptor outer segment is a highly specialized primary cilium that is essential for phototransduction and vision. Biallelic pathogenic variants in the cilia-associated gene CEP290 cause non-syndromic Leber congenital amaurosis 10 (LCA10) and syndromic diseases, where the retina is also affected. While RNA antisense oligonucleotides and gene editing are potential treatment options for the common deep intronic variant c.2991+1655A>G in CEP290, there is a need for variant-independent approaches that could be applied to a broader spectrum of ciliopathies. Here, we generated several distinct human models of CEP290-related retinal disease and investigated the effects of the flavonoid eupatilin as a potential treatment. Eupatilin improved cilium formation and length in CEP290 LCA10 patient-derived fibroblasts, in gene-edited CEP290 knockout (CEP290 KO) RPE1 cells, and in both CEP290 LCA10 and CEP290 KO iPSCs-derived retinal organoids. Furthermore, eupatilin reduced rhodopsin retention in the outer nuclear layer of CEP290 LCA10 retinal organoids. Eupatilin altered gene transcription in retinal organoids by modulating the expression of rhodopsin and by targeting cilia and synaptic plasticity pathways. This work sheds light on the mechanism of action of eupatilin and supports its potential as a variant-independent approach for CEP290-associated ciliopathies

The 2011 Magnitude 9.0 Tohoku-Oki Earthquake: Mosaicking the Megathrust from Seconds to Centuries

Geophysical observations from the 2011 moment magnitude (M_w) 9.0 Tohoku-Oki, Japan earthquake allow exploration of a rare large event along a subduction megathrust. Models for this event indicate that the distribution of coseismic fault slip exceeded 50 meters in places. Sources of high-frequency seismic waves delineate the edges of the deepest portions of coseismic slip and do not simply correlate with the locations of peak slip. Relative to the M_w 8.8 2010 Maule, Chile earthquake, the Tohoku-Oki earthquake was deficient in high-frequency seismic radiation-a difference that we attribute to its relatively shallow depth. Estimates of total fault slip and surface secular strain accumulation on millennial time scales suggest the need to consider the potential for a future large earthquake just south of this event

Imaging an Underwater Basin and its Resonance Modes using Optical Fiber Distributed Acoustic Sensing

International audienceDistributed acoustic sensing is an ideal tool for ambient noise tomography owing to the dense spatial measurements and the ability to continuously record in harsh environments, such as underwater. We demonstrate the ability to image a complex underwater basin using ambient noise recorded on a fiber deployed offshore Greece. A two-dimensional shear-wave velocity model was derived by analyzing Scholte-wave dispersion. In addition, extremely detailed frequency-dependent resonance and wave propagation characteristics were revealed by computing power spectral densities (PSD) and auto-correlations (AC), respectively. These observations provide crucial information on lateral and vertical wave propagation, and were used to further constrain the velocity model. The analysis reveals significant lateral variations across the short 2.5 km long fiber segment, including basin edge effects and scattered waves. Waveform simulations further support the obtained model. Our results demonstrate the advantages of incorporating PSD and AC observations into ambient noise-based imaging

Monitoring Deep Sea Currents With Seafloor Distributed Acoustic Sensing

Abstract Underwater fiber optic cables commonly traverse a variety of seafloor conditions, which leads to an uneven mechanical coupling between the cable and the ocean bottom. On rough seafloor bathymetry, some cable portions might be suspended and thus susceptible to vortex‐induced vibrations (VIV) driven by deep ocean currents. Here, we examine the potential of distributed acoustic sensing (DAS) to monitor deep‐sea currents along suspended sections of underwater telecom fiber optic cables undergoing VIV. Oscillations of a seafloor fiber optic cable located in southern France are recorded by DAS along cable sections presumably hanging. Their characteristic frequencies are lower than 1 Hz, at different ocean depths, and have an amplitude‐dependency consistent with the driving mechanism being VIV. Based on a theoretical proportionality between current speed and VIV frequencies, we derive ocean current speed time series at 2,390 m depth from the vortex shedding frequencies recorded by DAS. The DAS‐derived current speed time series is in agreement with recordings by a current meter located 3.75 km away from the hanging cable section (similar dominant period, high correlation after time shift). The DAS‐derived current speed time series displays features, such as characteristic periods and spectral decay, associated with the generation of internal gravity waves and weak oceanic turbulence in the Mediterranean Sea. The results demonstrate the potential of DAS along hanging segments of fiber optic cables to monitor a wide range of oceanography processes, at depths barely studied with current instrumentation

Quantifying microseismic noise generation from coastal reflection of gravity waves recorded by seafloor DAS

International audienceSecondary microseisms are the most energetic noise in continuous seismometer recordings. They are generated by interactions between ocean waves, including between gravity waves incident on and reflected from the coast. Coastal reflections of ocean waves leading to coastal microseismic sources are hard to estimate in various global numerical wave models, and independent quantification of these coastal sources through direct measurements can greatly improve these models. Here, we exploit a 41 km long submarine optical fiber cable located offshore Toulon, France, using Distributed Acoustic Sensing (DAS). We record both the amplitude and frequency of seafloor strains induced by ocean surface gravity waves, as well as secondary microseisms caused by the interaction of gravity waves incident and reflected from the coast. By leveraging the spatially distributed nature of DAS measurements, additional fundamental information is recovered such as the velocity and azimuth of the waves. We find that on average 30 per cent of the gravity waves are reflected at the coast generating local sources of secondary microseisms that manifest as Scholte waves. These local sources represent the most energetic contribution to the seismic noise recorded along the optical fiber and by an onshore broadband station located near the DAS interrogator. Furthermore, we estimate a coastal reflection coefficient of ocean surface gravity waves R2 of about 0.07, which provides improved constraints for seismic noise generation models. In addition, we show that new local sources of microseisms can be generated when gravity waves characteristics (azimuth and frequency content) change and lead to some delays between the optical fiber (OF) cable and buoy recordings. These analyses pave the way for a wide use of DAS data to monitor ocean-solid earth interactions as they provide a wealth of information on the reflection of gravity waves, coastal microseismic sources, and new constraints for numerical models of microseismic noise

Assessing the relationship between lung density and function with oxygen-enhanced magnetic resonance imaging in a mouse model of emphysema

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Purpose: A magnetic resonance imaging method is presented that allows for the simultaneous assessment of oxygen delivery, oxygen uptake, and parenchymal density. The technique is applied to a mouse model of porcine pancreatic elastase (PPE) induced lung emphysema in order to investigate how structural changes affect lung function. Method: Nine-week-old female C57BL6 mice were instilled with saline or PPE at days 0 and 7. At day 19, oxygen delivery, oxygen uptake, and lung density were quantified from T1 and proton-density measurements obtained via oxygen-enhanced magnetic resonance imaging (OE-MRI) using an ultrashort echo-time imaging sequence. Subsequently, the lungs were sectioned for histological observation. Blood-gas analyses and pulmonary functional tests via FlexiVent were performed in separate cohorts. Principal Findings: PPE-challenged mice had reduced density when assessed via MRI, consistent with the parenchyma loss observed in the histology sections, and an increased lung compliance was detected via FlexiVent. The oxygenation levels, as assessed via the blood-gas analysis, showed no difference between PPE-challenged animals and control. This finding was mirrored in the global MRI assessments of oxygen delivery and uptake, where the changes in relaxation time indices were matched between the groups. The heterogeneity of the same parameters however, were increased in PPE-challenged animals. When the oxygenation status was investigated in regions of varying density, a reduced oxygen-uptake was found in low-density regions of PPE-challenged mice. In high-density regions the uptake was higher than that of regions of corresponding density in control animals. The oxygen delivery was proportional to the oxygen uptake in both groups. Conclusions: The proposed method allowed for the regional assessment of the relationship between lung density and two aspects of lung function, the oxygen delivery and uptake. When compared to global indices of lung function, an increased sensitivity for detecting heterogeneous lung disorders was found. This indicated that the technique has potential for early detection of lung dysfunction-before global changes occur

Eupatilin Improves Cilia Defects in Human CEP290 Ciliopathy Models

The photoreceptor outer segment is a highly specialized primary cilium that is essential for phototransduction and vision. Biallelic pathogenic variants in the cilia-associated gene CEP290 cause non-syndromic Leber congenital amaurosis 10 (LCA10) and syndromic diseases, where the retina is also affected. While RNA antisense oligonucleotides and gene editing are potential treatment options for the common deep intronic variant c.2991+1655A>G in CEP290, there is a need for variant-independent approaches that could be applied to a broader spectrum of ciliopathies. Here, we generated several distinct human models of CEP290-related retinal disease and investigated the effects of the flavonoid eupatilin as a potential treatment. Eupatilin improved cilium formation and length in CEP290 LCA10 patient-derived fibroblasts, in gene-edited CEP290 knockout (CEP290 KO) RPE1 cells, and in both CEP290 LCA10 and CEP290 KO iPSCs-derived retinal organoids. Furthermore, eupatilin reduced rhodopsin retention in the outer nuclear layer of CEP290 LCA10 retinal organoids. Eupatilin altered gene transcription in retinal organoids by modulating the expression of rhodopsin and by targeting cilia and synaptic plasticity pathways. This work sheds light on the mechanism of action of eupatilin and supports its potential as a variant-independent approach for CEP290-associated ciliopathies