Effect of Temperature and Precipitation on Acute Appendicitis Incidence in Seoul: A Time Series Regression Analysis.

PURPOSE: This study aimed to investigate the relationship between meteorological factors, specifically temperature and precipitation, and the incidence of appendicitis in Seoul, South Korea. METHODS: Using data from the National Health Insurance Service spanning 2010-2020, the study analyzed 165,077 appendicitis cases in Seoul. Time series regression modeling with distributed-lag non-linear models was employed. RESULTS: Regarding acute appendicitis and daily average temperature, the incidence rate ratio (IRR) showed an increasing trend from approximately - 10 °C to 10 °C. At temperatures above 10 °C, the increase was more gradual. The IRR approached a value close to 1 at temperatures below - 10 °C and above 30 °C. Both total and complicated appendicitis exhibited similar trends. Increased precipitation was negatively associated with the incidence of total acute appendicitis around the 50 mm/day range, but not with complicated appendicitis. CONCLUSIONS: The findings suggest that environmental factors, especially temperature, may play a role in the occurrence of appendicitis. This research underscores the potential health implications of global climate change and the need for further studies to understand the broader impacts of environmental changes on various diseases.

COVID-19 Outbreak and Its Association with Healthcare Workers' Emotional Stress: a Cross-Sectional Study.

BACKGROUND: The coronavirus disease 2019 (COVID-19) outbreak has exposed healthcare workers to extreme physical workloads and psychological challenges. Thus, we aimed to assess the immediate correlates of emotional stress and to identify which specific jobs, departments, and exposure types are risk factors for emotional stress in healthcare workers. METHODS: In this cross-sectional study conducted from April 2 to 10, 2020, university hospital workers were administered self-reported questionnaires that covered general characteristics and included the Patient Health Questionnaire, Generalized Anxiety Disorder scale, and a visual analog scale. At-risk groups for depression and anxiety were identified, and the odds ratios for depression and anxiety were analyzed after adjusting for age, gender, education, marital status, and duration of employment. RESULTS: The data of 1,003 participants were analyzed. Of these, 14.2% worked in wards for confirmed COVID-19 cases and 15.2% had had direct contact with these patients. Treating patients with COVID-19 was associated with depression and anxiety, while dealing with COVID-19 test samples was associated with depression. Exposure to random or unspecified patients was also associated with depression. Lastly, social rejection and other negative experiences were associated with depression and anxiety. CONCLUSION: The COVID-19 outbreak is correlated with healthcare workers' emotional stress, and specific types of jobs and duties involving close contact with these patients can be risk factors. Interestingly, even low-exposure groups reported significant depression and anxiety as a result of social stigma and uncertainty. Adequate and timely management measures for emotional stress are required for vulnerable and at-risk groups.

Adverse pregnancy outcomes in healthcare workers: a Korean nationwide population-based study.

PURPOSE: To assess female healthcare workers' pregnancy complications and outcomes including abortion, fetal screening abnormalities, intrauterine growth retardation (IUGR), and preterm labor using nationwide population data and compare these results with those of the general population in Korea. METHODS: Korean National Health Insurance (NHI) claim data was used. We choose 3 different reference groups for comparison: (1) dependents of employees insured by NHI, i.e. non-working women, (2) all insured employees, i.e. all working women, and (3) workers in the education division. To compare the groups, logistic regression was used for multivariate analysis after adjusting for age and income level. RESULTS: Overall, healthcare workers showed a higher adjusted OR (odds ratio) in almost all obstetrical consequences. Miscarriage, threatened abortion, preterm labor, fetal screening abnormalities, and IUGR showed a higher adjusted OR in the working group than in the non-working group. We also observed similar results in a comparison of both the working groups. Regarding workers in the education division, ORs for obstetrical outcomes were also high, except for preterm labor. CONCLUSIONS: Health care workers have a higher risk of adverse pregnancy outcomes such as miscarriage, IUGR, and fetal screening abnormalities.

Targeted inversion and reversion of the blood coagulation factor 8 gene in human iPS cells using TALENs.

Hemophilia A, one of the most common genetic bleeding disorders, is caused by various mutations in the blood coagulation factor VIII (F8) gene. Among the genotypes that result in hemophilia A, two different types of chromosomal inversions that involve a portion of the F8 gene are most frequent, accounting for almost half of all severe hemophilia A cases. In this study, we used a transcription activator-like effector nuclease (TALEN) pair to invert a 140-kbp chromosomal segment that spans the portion of the F8 gene in human induced pluripotent stem cells (iPSCs) to create a hemophilia A model cell line. In addition, we reverted the inverted segment back to its normal orientation in the hemophilia model iPSCs using the same TALEN pair. Importantly, we detected the F8 mRNA in cells derived from the reverted iPSCs lines, but not in those derived from the clones with the inverted segment. Thus, we showed that TALENs can be used both for creating disease models associated with chromosomal rearrangements in iPSCs and for correcting genetic defects caused by chromosomal inversions. This strategy provides an iPSC-based novel therapeutic option for the treatment of hemophilia A and other genetic diseases caused by chromosomal inversions.

Genome-editing technologies for gene correction of hemophilia.

Hemophilia is caused by various mutations in blood coagulation factor genes, including factor VIII (FVIII) and factor IX (FIX), that encode key proteins in the blood clotting pathway. Although the addition of therapeutic genes or infusion of clotting factors may be used to remedy hemophilia's symptoms, no permanent cure for the disease exists. Moreover, patients often develop neutralizing antibodies or experience adverse effects that limit the therapy's benefits. However, targeted gene therapy involving the precise correction of these mutated genes at the genome level using programmable nucleases is a promising strategy. These nucleases can induce double-strand breaks (DSBs) on genomes, and repairs of such induced DSBs by the two cellular repair systems enable a targeted gene correction. Going beyond cultured cell systems, we are now entering the age of direct gene correction in vivo using various delivery tools. Here, we describe the current status of in vivo and ex vivo genome-editing technology related to potential hemophilia gene correction and the prominent issues surrounding its application in patients with monogenic diseases.

Incidence and severity of medication-related osteonecrosis of the jaw in patients with osteoporosis using data from a Korean nationwide sample cohort in 2002 to 2019: a retrospective study.

BACKGROUND: Medication-related osteonecrosis of the jaw (MRONJ) is a significant concern, particularly among patients taking bisphosphonates (BPs), denosumab, and selective estrogen receptor modulators (SERMs) for osteoporosis. Despite the known risks, large-scale cohort studies examining the incidence and severity of MRONJ are lacking. We aimed to ascertain the incidence and risk of MRONJ among these patients, whom we stratified by age groups, medication types, and duration of use. METHODS: We utilized data from the National Health Insurance Service's sample cohort database, focusing on patients aged 40 years and above diagnosed with osteoporosis. The patients were divided into three groups: those prescribed BPs only, those prescribed SERMs only, and those prescribed both. RESULTS: The overall incidence rate of MRONJ was 0.17%. A significantly higher incidence rate was observed among those taking osteoporosis medications, particularly among females with a relative risk of 4.99 (95% confidence interval, 3.21-7.74). The SERM group also had an incidence rate comparable to that of the BP group. Severity was assessed based on the invasiveness of the treatment methods, with 71.3% undergoing invasive treatment in the medication group. CONCLUSION: This study provides valuable insights into the incidence and severity of MRONJ among a large cohort of patients with osteoporosis. It underscores the need for comprehensive guidance on MRONJ risks across different medication groups and sets the stage for future research focusing on specific populations and treatment outcomes.

Preclinical and dose-ranging assessment of hESC-derived dopaminergic progenitors for a clinical trial on Parkinson's disease.

Human embryonic stem cell (hESC)-derived midbrain dopaminergic (mDA) cell transplantation is a promising therapeutic strategy for Parkinson's disease (PD). Here, we present the derivation of high-purity mDA progenitors from clinical-grade hESCs on a large scale under rigorous good manufacturing practice (GMP) conditions. We also assessed the toxicity, biodistribution, and tumorigenicity of these cells in immunodeficient rats in good laboratory practice (GLP)-compliant facilities. Various doses of mDA progenitors were transplanted into hemi-parkinsonian rats, and a significant dose-dependent behavioral improvement was observed with a minimal effective dose range of 5,000-10,000 mDA progenitor cells. These results provided insights into determining a low cell dosage (3.15 million cells) for human clinical trials. Based on these results, approval for a phase 1/2a clinical trial for PD cell therapy was obtained from the Ministry of Food and Drug Safety in Korea, and a clinical trial for treating patients with PD has commenced.

The Impact of Aircraft Noise on the Cognitive Function of Elementary School Students in Korea.

Background: This study evaluated the effects of chronic exposure to aircraft noise on the cognitive functions of Korean elementary school students attending an elementary school around a military airfield and clarified the relationship between noise exposure and cognitive functions. Methods: Five schools with average weight equivalent continuous perceived noise levels (WECPNL) of ≥75 dB were selected from four regions in Korea. Each of these schools was matched with a non-exposed school. The Korean Intelligence Test Primary (KIT-P) was used to measure the scores of four subcategories and the intelligence quotient (IQ). The noise exposure groups were divided into high-exposure (WECPNL ≥ 80 dB) and medium-exposure (75 ≤ WECPNL < 80) groups. The period of exposure during the school year was collected. A linear mixed model, with matched pairs of schools, was used for statistical analysis. Results: In the multivariable linear mixed model adjusted by possible confounders, the reasoning score was significantly lower in the high-exposure group of students than in the no-exposure group. Other scores and IQ were lower in the noise exposure groups, although these were not statistically significant. The duration of exposure did not show a significant relationship with cognitive functions. Conclusion: Chronic exposure to noise from military airfields may influence the cognitive functions, thereby reducing the learning performance of Korean children.

Increase of blood mercury level with shark meat consumption: A repeated-measures study before and after Chuseok, Korean holiday.

Sharks are known to contain high levels of mercury in their meat. However, few studies have directly assessed the changes in mercury concentration in the human body according to shark meat intake. One hundred and ninety-seven participants that traditionally consume shark meat during the Chuseok holiday were recruited from two areas of Gyeongsangbuk-do, South Korea to examine their blood mercury level before and after the holiday season. Blood mercury levels were measured before and after the holiday season. Characteristics such as the consumption of shark meat, intake amount, and the effect on mercury concentration were assessed during the survey. Univariable and multivariable analysis (Linear Mixed Model) were done for assessing the association between shark meat consumption of holiday season and blood mercury level. Among the total participants, 83 consumed shark meat during holiday. In the univariable analysis, a significant increase in blood mercury levels before and after Chuseok was observed only for the group that consumed shark meat during holiday. The multivariable analysis (adjusted for identified confounders that affect both exposure and outcome considering repeated measurements) showed that consuming shark meat was significantly associated with increased blood mercury levels by 3.56 µg/L (95% confidence interval [CI], 2.64-4.67 µg/L). In the model considering the amount consumed as two group, the level of increase was 2.61 µg/L (95% CI, 1.63-3.58 µg/L) for those consuming <100 g, and 6.20 µg/L (95% CI, 4.77-7.62 µg/L) for those consuming ≥100 g compared to group without consuming shark meat. Considering amount consumed as continuous value, 0.02 µg/L (95% CI, 0.01-0.02 µg/L) of blood mercury increase was significantly associated with consuming 1 g. Consumption of shark meat significantly elevated blood mercury levels, exceeding commonly suggested reference concentrations in less than 2 weeks. These findings suggest the need for public health warnings and regulations regarding shark meat consumption.

Risk assessment of heavy metals in tuna from Japanese restaurants in the Republic of Korea.

Background: Studies on the risk of mercury (Hg) in Korean fishery products focus primarily on total Hg levels as opposed to methylmercury (MeHg) levels. None of the few studies on MeHg in tuna investigated tuna from Japanese restaurants. Few have evaluated lead (Pb), cadmium (Cd) and arsenic (As) in tuna. Thus, this study aimed to conduct a risk assessment by evaluating heavy metal concentrations in tuna from Japanese restaurants. Methods: Thirty-one tuna samples were collected from Japanese restaurants in the Republic of Korea. They were classified according to region and species. The concentration of heavy metals in the samples was analyzed using the Ministry of Food and Drug Safety Food Code method. The rate of exceedance of maximum residue levels (MRLs) and the risk compared to the provisional tolerable weekly intake (PTWI) set by the Joint Food and Agriculture Organization/World Health Organization Expert Committee on Food Additives (%PTWI) were evaluated for risk assessment. Results: The mean of MeHg, Pb, Cd and As concentrations were 0.56 ± 1.47 mg/kg, 33.95 ± 3.74 µg/kg, 14.25 ± 2.19 µg/kg and 1.46 ± 1.89 mg/kg, respectively. No sample exceeded the MRLs of Pb and Cd, but 9.7% of the samples exceeded the MRL of MeHg. The %PTWIs of MeHg, Pb, Cd and As were 4.2037, 0.0162, 0.0244 and 1.1627, respectively. The %PTWI of MeHg by age group and sex was highest among men aged 19-29 years (10.6494), followed by men aged 30-49 years (7.2458) and women aged 19-29 years (4.8307). Conclusions: We found that 3 out of 31 samples exceeded the MRL of MeHg. The %PTWI of MeHg showed significant differences based on age and sex, and the value was likely to exceed a safe level depending on individuals' eating behaviors. Therefore, improved risk management for MeHg is required.

Association of heavy metal complex exposure and neurobehavioral function of children.

Background: Exposure to heavy metals is a public health concern worldwide. Previous studies on the association between heavy metal exposure and neurobehavioral functions in children have focused on single exposures and clinical manifestations. However, the present study evaluated the effects of heavy metal complex exposure on subclinical neurobehavioral function using a Korean Computerized Neurobehavior Test (KCNT). Methods: Urinary mercury, lead, cadmium analyses as well as symbol digit substitution (SDS) and choice reaction time (CRT) tests of the KCNT were conducted in children aged between 10 and 12 years. Reaction time and urinary heavy metal levels were analyzed using partial correlation, linear regression, Bayesian kernel machine regression (BKMR), the weighted quantile sum (WQS) regression and quantile G-computation analysis. Results: Participants of 203 SDS tests and 198 CRT tests were analyzed, excluding poor cooperation and inappropriate urine sample. Partial correlation analysis revealed no association between neurobehavioral function and exposure to individual heavy metals. The result of multiple linear regression shows significant positive association between urinary lead, mercury, and CRT. BMKR, WQS regression and quantile G-computation analysis showed a statistically significant positive association between complex urinary heavy metal concentrations, especially lead and mercury, and reaction time. Conclusions: Assuming complex exposures, urinary heavy metal concentrations showed a statistically significant positive association with CRT. These results suggest that heavy metal complex exposure during childhood should be evaluated and managed strictly.

An outbreak of neurologic symptoms among patients exposed to an unknown stench in a high school near an industrial complex: an epidemiological investigation.

OBJECTIVES: Seven students at a high school in Korea visited the emergency room with non-specific neurological symptoms after a stench was noticed during a school entrance ceremony. In relation to this incident, 105 patients visited medical institutions over 5 days. A team of environmental and epidemiological experts was assembled to investigate the incident. METHODS: Our team of experts participated in the investigation 1 month post-incident. Previously, only air samples had been analyzed. We received results of air samples analyzed by other investigators, medical records of some students, and data from police interviews of patients. Additional investigation and interviews were conducted, and the events were reconstructed in spatial and temporal order. RESULTS: A cluster of patients was observed on the south side and parts of the north side of the upper floor. A stench like that reported during the incident had been noticed for about 2 years near the school. Students consistently described a similar stench occurring frequently in the vicinity of the school. According to student statements, the odor mainly resembled something burning. The carboxyhemoglobin levels of some students were observed to be >1.5%. CONCLUSIONS: In the investigation, 2 suspected sources were identified: a science room storing chemicals downstairs from the auditorium and various industrial facilities near the school. Combining the scattered evidence, we considered a toxic puff of gas, perhaps from brief incineration or leakage in a specific area, to be the likely cause of the incident. We describe our approach and the limitations encountered during the investigation.

Preventive Measures Against COVID-19 in Small- and Mid-sized Enterprises from an Early Stage of the Epidemic in Daegu and Gyeongsangbuk-do.

Background: In the early stage of the coronavirus disease 2019 (COVID-19) epidemic, small- and mid-sized enterprises (SMEs) may be an important transmission consideration. The study aimed to identify the pattern of COVID-19 prevention measures during the outbreaks in Daegu and Gyeongsangbuk-do at the early stage of COVID-19. Moreover, we investigated whether SME size and past experiences affected the preventive measures implemented in the region. Methods: A survey detailing the general characteristics and implementation of 12 preventive activities was conducted in 122 SMEs in Daegu and Gyeongsangbuk-do. The survey was analyzed by size and operation period. Results: The study subjects consisted of 53 (43.4%) workplaces with 1-5 employees, 50 (40.9%) workplaces with 6-30 employees, and 19 (15.6%) workplaces with 31-49 employees. The lowest three items among those surveyed were 'symptomatic workers to stay home for 3-4 days' (17.2%), 'work remotely' (18.9%), and 'video meetings' (20.5%). There were significant differences in the rate of several preventive measures implemented. The larger sized SMEs, the higher the number of implementations (p < 0.01). The operation period had no significant relationship with the implementation of preventive measures. The same pattern was observed in multiple generalized linear regression with covariate adjustment. Conclusion: Preventive measures among SMEs with fewer than 50 employees were identified. Even within SMEs, a gap in preventive measures according to size was confirmed. To prevent the spread of infection and protect workers' right to health, different support for different sized SMEs is necessary.

Association between exposure to particulate matter and school absences in Korean asthmatic adolescents.

Background: Because particulate matter (PM) and asthma are closely related, the prevalence of school absence among adolescents with asthma can be affected by the concentration of PM. We aimed to investigate the relationship between school absences due to asthma and the total number of days that the PM concentration exceeded the standard. Methods: We used the data from the 16th Korea Youth Risk Behavior Survey and the PM levels of 17 metropolitan cities and provinces gathered from the AirKorea. Information on the characteristics of asthmatic adolescents and the prevalence of school absence was obtained using a questionnaire, while the PM levels based on the total number of days with poor and very poor PM grades were collected from the AirKorea website. Both χ2 test and logistic regression analysis were performed using the weights presented in the original dataset. Results: In the case of particulate matter of 10 microns in diameter or smaller (PM10), the odds ratio (OR) after adjusting for confounders (sex, school year, body mass index, smoking history, diagnosis of allergic rhinitis, diagnosis of atopic dermatitis and city size) was 1.07 (95% confidence interval [CI]: 1.01-1.13) for absents due to asthma when the total days of poor and very poor grades of PM10 (81 µg/m3 or higher) increased by 1 day. In the analysis of particulate matter of 2.5 microns in diameter or smaller (PM2.5), the OR after adjusting for confounders was 1.01 (95% CI: 1.00-1.03) for absents due to asthma when the total number of days with poor and very poor PM2.5 grades (36 µg/m3 or higher) increased by 1 day. Conclusions: A significant association was observed between the total number of days of poor and very poor PM10 and PM2.5 grades and school absence due to asthma; PM can cause asthma exacerbation and affect the academic life.

How Does the Involuntary Choice of Self-Employment Affect Subjective Well-Being in Small-Sized Business Workers? A Cross-Sectional Study from the Fifth Korean Working Conditions Survey.

In South Korea, self-employed workers comprise 24.6% of the working population-among which 99.7% were found to operate with less than 50 employees. However, few studies have investigated the effects of an involuntary choice of self-employment. In this study, based on the fifth Korean Working Conditions Survey, the factors affecting subjective well-being and mental health in small business owners with less than 50 employees among service/sales workers, who account for more than half of the self-employed population in Korea, were analyzed by the World Health Organization-Five Well-Being Index, using the Student's t-test, ANOVA and logistic regression analysis. Results showed that the well-being level for those who opted for self-employment involuntarily was lower than those who chose it voluntarily. Then, participants were separated into two groups according to sex; the degree corresponding to the poor well-being score group was compared, and that of the group who chose self-employment because they could not find any other work was significantly higher than that of the group who chose it voluntarily, in both men and women, and this was similar even after correcting for covariance. As the number of people being forced to choose self-employment is expected to further increase after the outbreak of COVID-19, future studies should be conducted to improve subjective well-being of such workers.

Risk factors influencing COVID-19 mortality rate in OECD countries: A cross-sectional study.

SUMMARY: The coronavirus disease (COVID-19) has spread globally; however, the COVID-19 mortality rate varies largely across countries. The purpose of this study was to investigate the factors affecting mortality and increase in mortality rate by time trends in 30 member countries of the Organization for Economic Co-operation and Development (OECD). These countries have different national health and medical characteristics in terms of health care use, health equipment, health resource, health risk, and health status at different time points. The results revealed that the lower 25% of countries had an increase in the mortality rate of 27.21% which was higher than the upper 25% of countries' increase in the mortality rate of 20.51%. Therefore, the affected countries should strengthen their medical infrastructure to prepare for such large-scale outbreaks in the future. It is imperative to reduce the health inequality between population groups and achieve health equity, regardless of the income gap, rather than vaccination of specific countries. This will require the management of non-communicable diseases, a solid health insurance system, a stable supply of medical supplies, and strengthening the competency of health care workers.

Transplantation of PSA-NCAM-Positive Neural Precursors from Human Embryonic Stem Cells Promotes Functional Recovery in an Animal Model of Spinal Cord Injury.

BACKGROUND: Spinal cord injury (SCI) results in permanent impairment of motor and sensory functions at and below the lesion site. There is no therapeutic option to the functional recovery of SCI involving diverse injury responses of different cell types in the lesion that limit endogenous nerve regeneration. In this regard, cell replacement therapy utilizing stem cells or their derivatives has become a highly promising approach to promote locomotor recovery. For this reason, the demand for a safe and efficient multipotent cell source that can differentiate into various neural cells is increasing. In this study, we evaluated the efficacy and safety of human polysialylated-neural cell adhesion molecule (PSA-NCAM)-positive neural precursor cells (hNPCsPSA-NCAM+) as a treatment for SCI. METHODS: One hundred thousand hNPCsPSA-NCAM+ isolated from human embryonic stem cell-derived NPCs were transplanted into the lesion site by microinjection 7 days after contusive SCI at the thoracic level. We examined the histological characteristics of the graft and behavioral improvement in the SCI rats 10 weeks after transplantation. RESULTS: Locomotor activity improvement was estimated by the Basso-Beattie-Bresnahan locomotor rating scale. Behavioral tests revealed that the transplantation of the hNPCsPSA-NCAM+ into the injured spinal cords of rats significantly improved locomotor function. Histological examination showed that hNPCsPSA-NCAM+ had differentiated into neural cells and successfully integrated into the host tissue with no evidence of tumor formation. We investigated cytokine expressions, which led to the early therapeutic effect of hNPCsPSA-NCAM+, and found that some undifferentiated NPCs still expressed midkine, a well-known neurotrophic factor involved in neural development and inflammatory responses, 10 weeks after transplantation. CONCLUSION: Our results demonstrate that hNPCsPSA-NCAM+ serve as a safe and efficient cell source which has the potential to improve impaired motor function following SCI.

Physical Activity and Quality of Life in Adult Men and Women with Distorted Perception of Weight Status: Nationwide Surveys (KNHANES 2016-2018).

This study analyzed the distorted perception of weight in adults aged 20 years or older and investigated the difference in the amount of physical activity and health-related quality of life (HRQOL). This study examined 21,326 adults regarding their body mass index (BMI), subjective body recognition (SBR), physical activity (according to the Global Physical Activity Questionnaire; GPAQ), and HRQOL (EuroQol-5 Dimension; EQ-5D) from the 7th Korea National Health and Nutrition Examination Survey (2016−2018). Independent t-test, analysis of variance (ANOVA), chi-square test, and multiple regression analysis were conducted. The group with the same BMI and SBR significantly showed a long time of activity vigorous-intensity (F = 21.25, p = 0.003) and moderate-intensity time (F = 17.24, p < 0.001). In the 'BMI = SBR' group, the sub-group with normal BMI and normal SBR showed the highest vigorous-intensity (mean ± SD = 7.20 ± 26.05, F = 37.86, p < 0.001) and moderate-intensity (mean ± SD = 13.89 ± 30.18, F = 43.27, p < 0.001) activity times. The sub-group with normal BMI and normal SBR had the highest percentage of responding as normal in the five EQ-5D sub-items. For the score of the EQ-5D Index, the highest score was shown in the group that felt subjectively more obese than the actual BMI (F = 56.83, p < 0.001). In the 'BMI = SBR' group, these factors related to physical activity (vigorous-intensity, moderated-intensity, and walking) are factors influencing health-related quality of life in this regression model (F = 396.57, p < 0.001, R2 = 0.165). Various health promotion programs and policy recommendations to reduce the distorted perception of weight are required.

Therapeutic correction of hemophilia A using 2D endothelial cells and multicellular 3D organoids derived from CRISPR/Cas9-engineered patient iPSCs.

The bleeding disorder hemophilia A (HA) is caused by a single-gene (F8) defect and its clinical symptom can be substantially improved by a small increase in the plasma coagulation factor VIII (FVIII) level. In this study, we used F8-defective human induced pluripotent stem cells from an HA patient (F8d-HA hiPSCs) and F8-corrected (F8c) HA hiPSCs produced by CRISPR/Cas9 genome engineering of F8d-HA hiPSCs. We obtained a highly enriched population of CD157+ cells from CRISPR/Cas9-edited F8c-HA hiPSCs. These cells exhibited multiple cellular and functional phenotypes of endothelial cells (ECs) with significant levels of FVIII activity, which was not observed in F8d-HA hiPSC-ECs. After transplantation, the engineered F8c-HA hiPSC-ECs dramatically changed bleeding episodes in HA animals and restored plasma FVIII activity. Notably, grafting a high dose of ECs substantially reduced the bleeding time during multiple consecutive bleeding challenges in HA mice, demonstrating a robust hemostatic effect (90% survival). Furthermore, the engrafted ECs survived more than 3 months in HA mice and reversed bleeding phenotypes against lethal wounding challenges. We also produced F8c-HA hiPSC-derived 3D liver organoids by assembling three different cell types in microwell devices and confirmed its therapeutic effect in HA animals. Our data demonstrate that the combination of genome-engineering and iPSC technologies represents a novel modality that allows autologous cell-mediated gene therapy for treating HA.

EW-7203, a novel small molecule inhibitor of transforming growth factor-ß (TGF-ß) type I receptor/activin receptor-like kinase-5, blocks TGF-ß1-mediated epithelial-to-mesenchymal transition in mammary epithelial cells.

Recently, small molecule inhibitors of transforming growth factorß (TGF-ß) type I receptor kinase / activin receptor-like kinase-5 (ALK5) have been developed to target TGF-ß signalling as a therapeutic strategy for combating cancer. In the present study, the authors examined a novel small molecule inhibitor of ALK5, 3-((5- ([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(6-methylpyridin-2-yl)thiazol-2-ylamino)methyl)benzonitrile (EW-7203) in breast cancer cells to determine if it has potential for cancer treatment. The inhibitory effects of EW-7203 on TGF-ß-induced Smad signalling and epithelial- to-mesenchymal transition (EMT) were investigated in mammary epithelial cells using luciferase reporter assays, immunoblotting, confocal microscopy and wound healing assays. In addition, the suppressive effects of EW-7203 on mammary cancer metastasis to the lung were examined using a Balb / c xenograft model system. The novel ALK5 inhibitor, EW-7203, inhibited the TGF-ß1-stimulated transcriptional activation of p3TP-Lux and pCA-GA12- Luc. In addition, EW-7203 decreased phosphorylated Smad2 levels and the nuclear translocation of Smad2 was increased by TGF-ß1. In addition, EW-7203 inhibited TGF-ß1-induced EMT and wound healing of NMuMG cells. Furthermore, in xenografted Balb / c mice, EW-7203 inhibited metastasis to the lung from breast tumors. The novel ALK5 inhibitor, EW-7203, efficiently inhibited TGF-ß1-induced Smad signalling, EMT and breast tumor metastasis to the lung in vivo, demonstrating that EW-7203 has therapeutic potential for breast cancer metastasis to the lung.