Antisense oligonucleotide (ASO)-based drugs for splicing modulation were recently approved for various genetic diseases with unmet need. Here we aimed to develop an ASO-based splicing modulation therapy for Cystic Fibrosis (CF) patients carrying the 3849+10 kb C-to-T splicing mutation in the CFTR gene. We have screened, in FRT cells expressing the 3849+10 kb C-to-T splicing mutation, ~30 2'-O-Methyl-modified phosphorothioate ASOs, targeted to prevent the recognition and inclusion of a cryptic exon generated due to the mutation. The effect of highly potent ASO candidates on the splicing pattern, protein maturation and CFTR function was further analyzed in well differentiated primary human nasal and bronchial epithelial cells, derived from patients carrying at least one 3849+10 kb C-to-T allele. A highly potent lead ASO, efficiently delivered by free uptake, was able to significantly increase the level of correctly spliced mRNA and completely restore the CFTR function to wild type levels in cells from a homozygote patient. This ASO led to CFTR function with an average of 43% of wild type levels in cells from various heterozygote patients. Optimized efficiency of the lead ASO was further obtained with 2'-Methoxy Ethyl modification (2'MOE). The highly efficient splicing modulation and functional correction, achieved by free uptake of the selected lead ASO in various patients, demonstrate the ASO therapeutic potential benefit for CF patients carrying splicing mutations and is aimed to serve as the basis for our current clinical development.

To assess the frequency, severity, and determinants of residual respiratory events during continuous positive airway therapy (CPAP) for obstructive sleep apnea (OSA) as determined by device output. Subjects were consecutive OSA patients at an American Academy of Sleep Medicine accredited multidisciplinary sleep center. Inclusion criteria included CPAP use for a minimum of 3 months, and a minimum nightly use of 4 hours. Compliance metrics and waveform data from 217 subjects were analyzed retrospectively. Events were scored manually when there was a clear reduction of amplitude (≥ 30%) or flow-limitation with 2–3 larger recovery breaths. Automatically detected versus manually scored events were subjected to statistical analyses included Bland-Altman plots, correlation coefficients, and logistic regression exploring predictors of residual events. The mean patient age was 54.7 ± 14.2 years; 63% were males. All patients had a primary diagnosis of obstructive sleep apnea, 26% defined as complex sleep apnea. Residual flow measurement based apnea-hypopnea index (AHIFLOW) > 5, 10, and 15/h was seen in 32.3%, 9.7%, and 1.8% vs. 60.8%, 23%, and 7.8% of subjects based on automated vs. manual scoring of waveform data. Automatically detected versus manually scored average AHIFLOW was 4.4 ± 3.8 vs. 7.3 ± 5.1 per hour. In a logistic regression analysis, the only predictors for a manual AHIFLOW > 5/h were the absolute central apnea index (CAI), (odds ratio [OR]: 1.5, p: 0.01, CI: 1.1–2.0), or using a CAI threshold of 5/h of sleep (OR: 5.0, p: < 0.001, CI: 2.2–13.8). For AHIFLOW > 10/h, the OR was 1.14, p: 0.03 (CI: 1.1–1.3) per every CAI unit of 1/hour. Residual respiratory events are common during CPAP treatment, may be missed by automated device detection and predicted by a high central apnea index on the baseline diagnostic study. Direct visualization of flow data is generally available and improves detection. Reiter J, Zleik B, Bazalakova M, Mehta P, Thomas RJ. Residual events during use of CPAP: prevalence, predictors, and detection accuracy. J Clin Sleep Med 2016;12(8):1153–1158.