Death after High-Dose rAAV9 Gene Therapy in a Patient with Duchenne's Muscular Dystrophy.

We treated a 27-year-old patient with Duchenne's muscular dystrophy (DMD) with recombinant adeno-associated virus (rAAV) serotype 9 containing dSaCas9 (i.e., "dead" Staphylococcus aureus Cas9, in which the Cas9 nuclease activity has been inactivated) fused to VP64; this transgene was designed to up-regulate cortical dystrophin as a custom CRISPR-transactivator therapy. The dose of rAAV used was 1×1014 vector genomes per kilogram of body weight. Mild cardiac dysfunction and pericardial effusion developed, followed by acute respiratory distress syndrome (ARDS) and cardiac arrest 6 days after transgene treatment; the patient died 2 days later. A postmortem examination showed severe diffuse alveolar damage. Expression of transgene in the liver was minimal, and there was no evidence of AAV serotype 9 antibodies or effector T-cell reactivity in the organs. These findings indicate that an innate immune reaction caused ARDS in a patient with advanced DMD treated with high-dose rAAV gene therapy. (Funded by Cure Rare Disease.).

Pocket CLARITY enables distortion-mitigated cardiac microstructural tissue characterization of large-scale specimens.

Molecular phenotyping by imaging of intact tissues has been used to reveal 3D molecular and structural coherence in tissue samples using tissue clearing techniques. However, clearing and imaging of cardiac tissue remains challenging for large-scale (>100 mm3) specimens due to sample distortion. Thus, directly assessing tissue microstructural geometric properties confounded by distortion such as cardiac helicity has been limited. To combat sample distortion, we developed a passive CLARITY technique (Pocket CLARITY) that utilizes a permeable cotton mesh pocket to encapsulate the sample to clear large-scale cardiac swine samples with minimal tissue deformation and protein loss. Combined with light sheet auto-fluorescent and scattering microscopy, Pocket CLARITY enabled the characterization of myocardial microstructural helicity of cardiac tissue from control, heart failure, and myocardial infarction in swine. Pocket CLARITY revealed with high fidelity that transmural microstructural helicity of the heart is significantly depressed in cardiovascular disease (CVD), thereby revealing new insights at the tissue level associated with impaired cardiac function.

Accelerated in Vivo Cardiac Diffusion-Tensor MRI Using Residual Deep Learning-based Denoising in Participants with Obesity.

PURPOSE: To develop and assess a residual deep learning algorithm to accelerate in vivo cardiac diffusion-tensor MRI (DT-MRI) by reducing the number of averages while preserving image quality and DT-MRI parameters. MATERIALS AND METHODS: In this prospective study, a denoising convolutional neural network (DnCNN) for DT-MRI was developed; a total of 26 participants, including 20 without obesity (body mass index [BMI] < 30 kg/m2; mean age, 28 years ± 3 [standard deviation]; 11 women) and six with obesity (BMI ≥ 30 kg/m2; mean age, 48 years ± 11; five women), were recruited from June 19, 2019, to July 29, 2020. DT-MRI data were constructed at four averages (4Av), two averages (2Av), and one average (1Av) without and with the application of the DnCNN (4AvDnCNN, 2AvDnCNN, 1AvDnCNN). All data were compared against the reference DT-MRI data constructed at eight averages (8Av). Image quality, characterized by using the signal-to-noise ratio (SNR) and structural similarity index (SSIM), and the DT-MRI parameters of mean diffusivity (MD), fractional anisotropy (FA), and helix angle transmurality (HAT) were quantified. RESULTS: No differences were found in image quality or DT-MRI parameters between the accelerated 4AvDnCNN DT-MRI and the reference 8Av DT-MRI data for the SNR (29.1 ± 2.7 vs 30.5 ± 2.9), SSIM (0.97 ± 0.01), MD (1.3 µm2/msec ± 0.1 vs 1.31 µm2/msec ± 0.11), FA (0.32 ± 0.05 vs 0.30 ± 0.04), or HAT (1.10°/% ± 0.13 vs 1.11°/% ± 0.09). The relationship of a higher MD and lower FA and HAT in individuals with obesity compared with individuals without obesity in reference 8Av DT-MRI measurements was retained in 4AvDnCNN and 2AvDnCNN DT-MRI measurements but was not retained in 4Av or 2Av DT-MRI measurements. CONCLUSION: Cardiac DT-MRI can be performed at an at least twofold-accelerated rate by using DnCNN to preserve image quality and DT-MRI parameter quantification.Keywords: Adults, Cardiac, Obesity, Technology Assessment, MR-Diffusion Tensor Imaging, Heart, Tissue CharacterizationSupplemental material is available for this article.© RSNA, 2021.