Functional Rescue of Dystrophin Deficiency in Mice Caused by Frameshift Mutations Using <i>Campylobacter jejuni</i> Cas9

Koo, Taeyoung,Lu-Nguyen, Ngoc B.,Malerba, Alberto,Kim, Eunji,Kim, Daesik,Cappellari, Ornella,Cho, Hee-Yeon,Dickson, George,Popplewell, Linda,Kim, Jin-Soo American Society of GeneCell Therapy 2018 Molecular therapy Vol.26 No.6

<▼1><P>Duchenne muscular dystrophy (DMD) is a fatal, X-linked muscle-wasting disease caused by mutations in the <I>DMD</I> gene. In 51% of DMD cases, a reading frame is disrupted because of deletion of several exons. Here, we show that CjCas9 derived from <I>Campylobacter jejuni</I> can be used as a gene-editing tool to correct an out-of-frame <I>Dmd</I> exon in <I>Dmd</I> knockout mice. Herein, we used Cas9 derived from <I>S. pyogenes</I> to generate <I>Dmd</I> knockout mice with a frameshift mutation in <I>Dmd</I> gene. Then, we expressed CjCas9, its single-guide RNA, and the EGFP gene in the <I>tibialis anterior</I> muscle of the <I>Dmd</I> knockout mice using an all-in-one adeno-associated virus (AAV) vector. CjCas9 cleaved the target site in the <I>Dmd</I> gene efficiently <I>in vivo</I> and induced small insertions or deletions at the target site. This treatment resulted in conversion of the disrupted <I>Dmd</I> reading frame from out of frame to in frame, leading to the expression of dystrophin in the sarcolemma. Importantly, muscle strength was enhanced in the CjCas9-treated muscles, without off-target mutations, indicating high efficiency and specificity of CjCas9. This work suggests that <I>in vivo DMD</I> frame correction, mediated by CjCas9, has great potential for the treatment of DMD and other neuromuscular diseases.</P></▼1><▼2><P>Koo et al. demonstrate that CjCas9 derived from <I>Campylobacter jejuni</I> can be used as a gene-editing tool to correct an out-of-frame <I>Dmd</I> exon in <I>Dmd</I> knockout mice. This study provides the therapeutic utility of CjCas9 for the treatment of Duchenne muscular dystrophy and other neuromuscular diseases.</P></▼2>

Sofosbuvir and Ledipasvir is Associated with High Sustained Virologic Response and Improvement of Health- Related Quality-of-Life in East-Asians with Hepatitis C

( Zobair Younossi ),( Maria Stepanova ),( Linda Henry ),( Kwanghyub Han ),( Sang Ahn ),( Youngsuk Lim ),( Wanloung Chuang ),( Jia Horng Kao ),( Nguyen Kinh ),( Ching Lung Lai ),( Man Fung Yuen ),( Hen 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: We aim to assess HRQL in east Asian (EA) HCV patients treated with different anti-HCV sofosbuvir (SOF)-based regimens. Methods: 1070 EA HCV subjects were enrolled in two phase 3 clinical trials [China: 55.6%, S. Korea: 20.7%, Taiwan: 16.1%, Vietnam: 4.7%, and Hong Kong: 2.9%]. Patients received IFN+SOF+RBV for 12 weeks (n=155, GT 1 and 6) or SOF+RBV for 12-24 weeks (n=531, GT 1, 2, 3 and 6) or IFN-free RBV-free LDV/SOF (n=384, GT1 only). The SVR-12 rates were 95.5%, 96.0%, and 99.2%, respectively (P=0.008). EA HCV patients completed Short Form-36 before, during, and after treatment and HRQL scores compared between regimens. Results: Baseline HRQL scores were similar between treatment groups. After 2 weeks of treatment, HRQL scores for the IFN+RBV- containing regimen became significantly lower as compared to the IFN-free regimens (average decline up to -11.2 points, P<0.0001). By the end of treatment, IFN-treated group experienced significant declines in most HRQL scores (up to -13.3 points, P<0.02 for 7/8 HRQL scales). Patients on SOF+RBV had milder HRQL impairments (up to -5.4 points, P<0.05 for 5/8 scales). However, patients receiving LDV/SOF had improvement in their HRQL scores (up to +4.3 points by the end of treatment, P<0.001 for 3/8 scales). Achieving SVR-12 with IFN+RBV+ SOF and SOF+RBV was associated with improvement in General Health (GH) and Vitality (VT) (up to +2.9 points, P<0.05), while SVR-12 with LDV/SOF was associated with improvement in Physical Functioning, GH, VT, Mental Health, and Role Emotional (up to +5.9 points, P<0.03). In multivariate analysis, receiving IFN was independently associated with HRQL impairment during treatment (β: -10.4 to -17.3 points, P<0.0001). Conclusions: Treatment of EA HCV patients with IFN or RBV containing regimens is associated with HRQL impairment, while treatment with LDV/SOF is associated with improvement of HRQL during treatment. SVR-12 was associated with greater improvements in HRQL.

In response to: Steatotic liver disease-know your enemies

Michael H. Le,Linda Henry,Mindie H. Nguyen 대한간학회 2024 Clinical and Molecular Hepatology(대한간학회지) Vol.30 No.2

Forecasted 2040 global prevalence of nonalcoholic fatty liver disease using hierarchical bayesian approach

Michael H. Le,Yee Hui Yeo,Biyao Zou,Scott Barnet,Linda Henry,Ramsey Cheung,Mindie H. Nguyen 대한간학회 2022 Clinical and Molecular Hepatology(대한간학회지) Vol.28 No.4

Background/Aims: Due to increases in obesity and type 2 diabetes, the prevalence of nonalcoholic fatty liver disease (NAFLD) has also been increasing. Current forecast models may not include non-obese NAFLD. Here, we used the Bayesian approach to forecast the prevalence of NAFLD through the year 2040. Methods: Prevalence data from 245 articles involving 2,699,627 persons were used with a hierarchical Bayesian approach to forecast the prevalence of NAFLD through 2040. Subgroup analyses were performed for age, gender, presence of metabolic syndrome, region, and smoking status. Sensitivity analysis was conducted for clinical setting and study quality. Results: The forecasted 2040 prevalence was 55.7%, a three-fold increase since 1990 and a 43.2% increase from the 2020 prevalence of 38.9%. The estimated average yearly increase since 2020 was 2.16%. For those aged <50 years and ≥50 years, the 2040 prevalence were not significantly different (56.7% vs. 61.5%, P=0.52). There was a significant difference in 2040 prevalence by sex (males: 60% vs. 50%) but the trend was steeper for females (annual percentage change: 2.5% vs. 1.5%, P=0.025). There was no difference in trends overtime by region (P=0.48). The increase rate was significantly higher in those without metabolic syndrome (3.8% vs. 0.84%, P=0.003) and smokers (1.4% vs. 1.1%, P=0.011). There was no difference by clinical/community setting (P=0.491) or study quality (P=0.85). Conclusions: By 2040, over half the adult population is forecasted to have NAFLD. The largest increases are expected to occur in women, smokers, and those without metabolic syndrome. Intensified efforts are needed to raise awareness of NAFLD and to determine long-term solutions addressing the driving factors of the disease.

High Prevalence of Slow Transit Constipation in Patients With Gastroparesis

Thomas A Zikos,Afrin N Kamal,Leila Neshatian,George Triadafilopoulos,John O Clarke,Monica Nandwani,Linda A Nguyen 대한소화기 기능성질환∙운동학회 2019 Journal of Neurogastroenterology and Motility (JNM Vol.25 No.2

Background/Aims Current evidence suggests the presence of motility or functional abnormalities in one area of the gastrointestinal tract increases the likelihood of abnormalities in others. However, the relationship of gastroparesis to chronic constipation (slow transit constipation and dyssynergic defecation) has been incompletely evaluated. Methods We retrospectively reviewed the records of all patients with chronic dyspeptic symptoms and constipation who underwent both a solid gastric emptying scintigraphy and a high-resolution anorectal manometry at our institution since January 2012. When available, X-ray defecography and radiopaque marker colonic transit studies were also reviewed. Based on the gastric emptying results, patients were classified as gastroparesis or dyspepsia with normal gastric emptying (control group). Differences in anorectal and colonic findings were then compared between groups. Results Two hundred and six patients met the inclusion criteria. Patients with gastroparesis had higher prevalence of slow transit constipation by radiopaque marker study compared to those with normal emptying (64.7% vs 28.1%, P = 0.013). Additionally, patients with gastroparesis had higher rates of rectocele (88.9% vs 60.0%, P = 0.008) and intussusception (44.4% vs 12.0%, P = 0.001) compared to patients with normal emptying. There was no difference in the rate of dyssynergic defecation between those with gastroparesis vs normal emptying (41.1% vs 42.1%, P = 0.880), and no differences in anorectal manometry findings. Conclusions Patients with gastroparesis had a higher rate of slow transit constipation, but equal rates of dyssynergic defecation compared to patients with normal gastric emptying. These findings argue for investigation of possible delayed colonic transit in patients with gastroparesis and vice versa.