Systematic gene therapy derived from an investigative study of AAV2/8 vector gene therapy for Fabry disease.

BACKGROUND: Fabry disease (FD) is a progressive multisystemic disease characterized by a lysosomal enzyme deficiency. A lack of α-galactosidase A (α-Gal A) activity results in the progressive systemic accumulation of its substrates, including globotriaosylceramide (Gb3) and globotriaosylsphingosine (Lyso-Gb3), which results in renal, cardiac, and/or cerebrovascular disease and early death. Enzyme replacement therapy (ERT) is the current standard of care for FD; however, it has important limitations, including a low half-life, limited distribution, and requirement of lifelong biweekly infusions of recombinant enzymes. METHODS: Herein, we evaluated a gene therapy approach using an episomal adeno-associated viral 2/8 (AAV2/8) vector that encodes the human GLA cDNA driven by a liver-specific expression cassette in a mouse model of FD that lacks α-Gal A activity and progressively accumulates Gb3 and Lyso-Gb3 in plasma and tissues. RESULTS: A pharmacology and toxicology study showed that administration of AAV2/8-hGLA vectors (AAV2/8-hGLA) in FD mice without immunosuppression resulted in significantly increased plasma and tissue α-Gal A activity and substantially normalized Gb3 and Lyso-Gb3 content. CONCLUSIONS: Moreover, the plasma enzymatic activity of α-Gal A continued to be stably expressed for up to 38 weeks and sometimes even longer, indicating that AAV2/8-hGLA is effective in treating FD mice, and that α-Gal A is continuously and highly expressed in the liver, secreted into plasma, and absorbed by various tissues. These findings provide a basis for the clinical development of AAV2/8-hGLA.

Development of Lanzyme as the Potential Enzyme Replacement Therapy Drug for Fabry Disease.

Fabry disease (FD) is a progressive multisystemic disease characterized by lysosomal enzyme deficiency. Enzyme replacement therapy (ERT) is one of the most significant advancements and breakthroughs in treating FD. However, limited resources and the high cost of ERT might prevent patients from receiving prompt and effective therapy, thereby resulting in severe complications. Future progress in ERT can uncover promising treatment options. In this study, we developed and validated a recombinant enzyme (Lanzyme) based on a CHO-S cell system to provide a new potential option for FD therapy. Our results indicated that Lanzyme was heavily glycosylated, and its highest activity was similar to a commercial enzyme (Fabrazyme®). Our pharmacokinetic assessment revealed that the half-life of Lanzyme was up to 11 min, which is nearly twice that of the commercial enzyme. In vivo experiments revealed that Lanzyme treatment sharply decreased the accumulation levels of Gb3 and lyso-Gb3 in various tissues of FD model mice, with superior or comparable therapeutic effects to Fabrazyme®. Based on these data, Lanzyme may represent a new and promising treatment approach for FD. Building this enzyme production system for ERT can offer additional choice, potentially with enhanced efficacy, for the benefit of patients with FD.

One-step construction of a food-grade expression system based on the URA3 gene in Kluyveromyces lactis.

Proteins from food-grade expression systems can be used in food products and medical applications. Herein, we describe a one-step method of constructing an expression vector in Kluyveromyces lactis by combining a URA3-deficient strain and a plasmid vector with no drug-resistant selection. Adjacent DNA elements of the vector were assembled in a targeted manner through a reaction with a special recombinase to form a plasmid vector using a one-step reaction. The unnecessary fragments containing the pUC origin and the ampicillin resistance gene were removed, and the vector was isolated and purified before transformation. A single transformation of the vector can produce a URA3-deficient strain. PCR assay, sequencing, and western blot analysis all indicated that the method of vector construction and target protein expression (mCherry and human serum albumin) were successful. This method may potentially be applied to any species containing the URA3 gene; this system has the potential to become a safe and powerful tool for promoting protein expression in food-safe species.

Cardiometabolic Risk Factors and Preclinical Target Organ Damage Among Adults in Ghana: Findings From a National Study.

Background Although sub-Saharan Africa has a high prevalence of cardiovascular diseases (CVDs), there remains a lack of systematic and comprehensive assessment of risk factors and early CVD outcomes in adults in sub-Saharan Africa. Methods and Results Using a stratified multistage random sampling method, we recruited 1106 men and women, aged >18 years, from the general population in Ghana to participate in a national health survey from 2016 to 2017. In Ghanaian adults, the age-standardized prevalence of known CVD risk factors was 15.1% (95% CI, 12.9%-17.3%) for obesity, 6.8% (95% CI, 5.1%-8.5%) for diabetes mellitus, 26.1% (95% CI, 22.9%-29.4%) for hypertension, and 9.3% (95% CI, 7.1%-11.5%) for hyperuricemia. In addition, 10.1% (95% CI, 7.0%-13.2%) of adults had peripheral artery disease, 8.3% (95% CI, 6.7%-10.0%) had carotid thickening, 4.1% (95% CI, 2.9%-5.2%) had left ventricular hypertrophy, and 2.5% (95% CI, 1.5%-3.4%) had chronic kidney disease. Three CVD risk factors appeared to play prominent roles in the development of target organ damage, including obesity for peripheral artery disease (odds ratio [OR], 2.22; 95% CI, 1.35-3.63), hypertension for carotid thickening (OR, 1.92; 95% CI, 1.22-3.08), and left ventricular hypertrophy (OR, 5.28; 95% CI, 2.55-12.11) and hyperuricemia for chronic kidney disease (OR, 5.49; 95% CI, 2.84-10.65). Conclusions This comprehensive health survey characterized the baseline conditions of a national cohort of adults while confirming the prevalence of CVD risk factors, and early CVD outcomes have reached epidemic proportions in Ghana. The distinct patterns of risk factors in the development of target organ damage present important challenges and opportunities for interventions to improve cardiometabolic health among adults in Ghana.

[Prevalence of acute coronary heart disease among farmers in Panyu, Guangzhou: a 20-year population-based study].

OBJECTIVE: To monitor the incidence change of acute coronary heart events in the all-ages farmers in Panyu District, Guangzhou City during 1991-2001 and 2010-2011. METHODS: The surveillance on the same defined population as that from the PRC-USA cooperative epidemiologic project on the cardiovascular and pulmonary diseases 30-year ago was carried out in Panyu, Guangzhou in 1991-2001 and 2010-2011. The crude incidence of acute coronary events and the age-standardized incidence rate were calculated by the year, gender and age, and standardized with the world standard population age distribution. Incidences at the two different periods were compared. The annual average changing rate of the incidence was obtained by the regression analysis methods. RESULTS: In the 11 consecutive years of 1991-2001, a upward trend on the incidence of acute coronary events among the farmers in women in Panyu District was found (P < 0.05). The incidence of the acute coronary events in the year of 2010-2011 was significantly higher than that in the year of 1991 to 2001 [34.06 per 100 000 (age-adjusted rate as 28.50 per 100 000) versus 16.14 per 100 000 (age-adjusted rate as 16.57 per 100 000), P < 0.05]. The age-adjusted rate increased by 83.04%. Peak incidence of acute coronary events in males was noticed in 75-79 age group, and in 80-84 age group in females. Comparing to the period of 1991-2001, the largest incidence increases appeared in the age groups of 35-39 and 75-79 years in males, while in the age groups of 50-54 and 65-69 years in females. Up to 47.37% (36/76) events occurred on the age group older than 75 years, raised by 40.44% comparing to that in 1991-2001 (33.73%, 56/166). CONCLUSIONS: The incidence of acute coronary events among farmers in Panyu District is at middle or low level of China but there is an increasing trend in acute coronary incidence from 1991 to 2011. Our results suggest that the prevention and treatment on acute coronary syndrome should be strengthened, and especially on the age group with the largest increase of disease incidence.

[Matrix metalloproteinase-3 genotype distribution and its relation with blood pressure profiles in Guangzhou rural population].

OBJECTIVE: To analyze the distribution of matrix metalloproteinase-3 (MMP-3) genotype and its association with the blood pressure profiles in Guangzhou rural population. METHODS: This study was conducted among 680 rural residents aged 40-70 years (including 314 men and 366 women) from 3 villages in an rural area of Guangzhou. The blood pressures of the subjects were measured and blood samples were collected for genotype analysis using polymerase chain reaction and direct sequencing of the MMP-3 gene promoter region to detect the 5 adenines (5A)/6 adenines (6A) polymorphisms. RESULTS: The frequencies of MMP-3 genotypes 6A/6A, 5A/5A, and 5A/6A were 82.6%, 1.8% and 15.6% among these residents, respectively. The distribution of MMP-3 genotypes and allele frequencies showed no significant gender- or age-related variations. The men with different genotypes (6A/6A vs 5A/6A+5A/5A) did not show significant differences in blood pressure levels, whereas the women with 5A/6A+5A/5A genotypes had higher systolic and diastolic blood pressures than those with a 6A/6A genotype. The allele 5A was highly frequent in the hypertensive residents as compared with the normotensive residents. CONCLUSIONS: The 6A homozygote is the predominant genotype of MMP-3 in Guangzhou rural population, which has a significantly lower proportion of 5A homozygote than the Western populations. The 5A allele is associated with a high risk of hypertension especially in women and may affect both systolic and diastolic blood pressures.

Adverse trends of cardiovascular risk factors among low risk populations (1983-1994)--a cohort study of workers and farmers in Guangzhou, China.

BACKGROUND: The levels and trends of cardiovascular risk factors vary greatly throughout China. We examine 10-year trends of cardiovascular risk factors (1983-1994) and the factors related to these trends among low-risk cohorts of workers and farmers in Guangzhou, China. METHODS: This is a cohort study of 3,131 workers and 3,493 farmers aged 25-64 years at baseline with 10 years of follow-up. We performed a longitudinal analysis to account for the aging of the cohorts and the repeated measures of the same individual. RESULTS: At baseline the prevalence of overweight (including obese) ranged from 1.0% to 11.8%, hypertension ranged from 3.8% to 10.5%, and mean serum total cholesterol (TC) ranged from 155.4 mg/dl to 187.2 mg/dl. Although prevalence of smoking declined, blood pressure levels and body mass index (BMI) increased significantly, and lipid profiles changed unfavorably during the 10-year follow-ups. The prevalence of hypertension increased from 5.0 percentage points (female farmers) to 12.3 percentage points (male farmers). Mean TC increased significantly (e.g., +22.8 mg/dl and +17.0 mg/dl in male and female farmers, respectively). In the longitudinal data analyses, increase in BMI was associated with increase in blood pressure levels and TC. Significant adverse trends of risk factors persisted after adjustment for aging, education, BMI, smoking, and alcohol intake. CONCLUSION: Urgent action is needed to prevent and reverse the unhealthy trends occurring among these low risk Chinese workers and farmers.

[Epidemiological survey of rheumatic heart disease in schoolchildren in Guangdong and Xinjiang].

OBJECTIVE: To investigate the prevalence of rheumatic heart disease (RHD) among schoolchildren in Guangdong Province and Xinjiang Uygur Autonomous Region. METHODS: Using a cluster sampling method, an epidemiological survey of RHD was conducted in 16 682 primary and high school students by auscultation in Guangdong Province and Xinjiang Uygur Autonomous Region from 2005 to 2006. Review of the clinical records, RHD survey in adults, and examination of the positivity rate of group A beta-hemolytic Streptococcus (GAS) by throat swab cultures in the students aged between 9 and 12 years in the sampled schools were also carried out. RESULTS: No RHD patient was found in the sampled population. In Xinjiang, the prevalence of RHD was 12.9/1000 among adults, higher than that (2.2/1000) in Guangdong Province. The GAS-positive rate in the schoolchildren in Xinjiang ranged from 9.8% to 12.6%, higher than that in Guangdong (2.3%-3.9%). CONCLUSION: The GAS-positive rate among children and incidence of RHD in adults are higher in Xinjiang than in Guangdong. The prevalence of RHD among the schoolchildren shows a reduction compared with that in 1994.

A school-based program for control of group a streptococcal upper respiratory tract infections: a controlled trial in Southern China.

A prospective, school-based study included daily monitoring for incidence of symptomatic streptococcal-associated pharyngitis and monthly determinations of group A streptococcal prevalence. A treatment group received penicillin/erythromycin therapy at school for positive throat cultures; the control group sought medical care from their regular provider. Prevalence and incidence of group A streptococcal pharyngitis were significantly lower among the treatment group than in the controls.