Long-term culture of SH-SY5Y neuroblastoma cells in the absence of neurotrophins : a novel model of neuronal ageing

LS is sponsored by a Wellcome Trust ISSF studentship and YC is a recipient of a China Scholarship Council award.Background Studying human ageing is of increasing importance due to the worldwide ageing population. However, it faces the challenge of lengthy experiments to produce an ageing phenotype. Often, to recreate the hallmarks of ageing requires complex empirical conditions that can confound data interpretation. Indeed, many studies use whole organisms with relatively short life spans, which may have little, or limited, relevance to human ageing. There has been extensive use of cell lines to study ageing in human somatic cells, but the modelling of human neuronal ageing is somewhat more complex in vitro. New Method We cultured the well-characterised SH-SY5Y human neural cell line to produce high purity cultures of cells differentiated to express a neuronal phenotype, and designed a protocol to maintain these cells in culture until they accumulated biomarkers of cellular ageing. Results Our data validate a novel and simple technique for the efficient differentiation and long-term maintenance of SH-SY5Y cells, expressing markers of neuronal differentiation and demonstrating electrical activity in culture. Over time in vitro, these cells progressively accumulate markers of ageing such as enhanced production of reactive oxygen species and accumulation of oxidative damage. Comparison to Existing Methods In comparison to existing techniques to model neuronal ageing our method is cost effective, requiring no specialist equipment or growth factors. Conclusions We demonstrate that SH-SY5Y cells, grown under these culture conditions, represent a simple model of neuronal ageing that is amenable to cell biological, biochemical and electrophysiological investigation.Publisher PDFPeer reviewe

"Including us, talking to us and creating a safe environment" - Youth Patient & Public Involvement and the Walking In ScHools (WISH) Study: Lessons learned

© 2023 The authors. This is an open access article available under a Creative Commons licence. The published version can be accessed at the following link on the publisher’s website: https://doi.org/10.1111/hex.13885Background: Young people have the right to be informed and consulted about decisions affecting their lives. Patient and public involvement (PPI) ensures that research is carried out ‘with’ or ‘by’ young people rather than ‘to’, ‘about’ or ‘for’ them. The aim of this paper is to outline how youth PPI can be embedded within a physical activity intervention, reflect on the impact of PPI and provide recommendations for future PPI in a similar context. Methods: A Youth Advisory Group (YAG) was set up within the Walking In ScHools (WISH) Study to involve adolescent girls in the delivery, implementation and dissemination of a physical activity intervention targeted at adolescents. Schools invited pupils aged 12–14 years and 15–18 years to YAG meetings (n3, from 2019 to 2023). Participative methods were used to inform recruitment strategies and data collection methods for the WISH Study. Results: Across the three YAG meetings, n51 pupils from n8 schools were involved. Pupils enjoyed the YAG meetings, felt that their feedback was valued and considered the meetings a good way to get young people involved in research. The YAG advised on specific issues and although measuring impact was not the primary aim of the YAG meetings, over the course of the study there were many examples of the impact of PPI. Recruitment targets for the WISH Study were exceeded, the attrition rate was low and pupils were engaged in data collection. Conclusion: Youth PPI is a developing field and there are few physical activity studies that report the PPI work undertaken. Within the WISH Study, three YAG meetings were held successfully, and the views of adolescent girls were central to the development of the study. Considering the specific issues that the YAG advised on (study recruitment, attrition and data collection), there was evidence of a positive impact of PPI. Patient or Public Contribution: Pupils from post-primary schools interested/participating in the WISH Study were invited to attend YAG meetings. YAG meetings were set up to consult adolescent girls on the delivery, implementation and dissemination of the WISH intervention.The WISH Study was funded from INTERREG VA funding of €8.84m (including a 15% contribution from the Department of Health in NI and Republic of Ireland), which had been awarded to the HSC Research & Development Division of the Public Health Agency Northern Ireland and to the Health Research Board in Ireland for the Cross-border Healthcare Intervention Trials in Ireland Network project.Published versio

Key findings from The Walking In ScHools (WISH) Study, a peer-led walking intervention for adolescent girls

Project Description: The effects of walking interventions on adult health are known, however the potential of walking to promote physical activity (PA) in adolescents is less known. This study evaluated the effectiveness of a novel, school-based walking intervention at increasing PA levels of adolescent girls. Female pupils aged 12-14 years, were recruited from eighteen (mixed or single-sex) schools across the Border Region of Ireland/Northern Ireland. Schools were randomised to control (usual physical activity; n9) or intervention (n9). In intervention schools, female pupils (15-18 years) were trained as walk leaders and led the younger pupils in 10-15min walks before school, at break and lunchtime. Walks were in school grounds and pupils were encouraged to join as many walks as possible. Excluding holidays, the intervention was delivered for a full school year (18-21 weeks). Accelerometers measured PA and the primary outcome was total PA (cpm). Impact: In total, 589 pupils were recruited (intervention: n286; control n303). Baseline moderate-vigorous PA (MVPA) for the intervention group was (median [IQR]) 36.1 (23.0) mins per day and 35.3 (19.8) mins per day in the control group. At baseline, only 15% (n37) of intervention and 10% (n29) of control participants met PA guidelines (60 mins MVPA per day). Post-intervention total PA (cpm) did not statistically differ between groups when adjusted for age, body mass index z-scores and baseline PA (mean difference, -33.5, 95% CI= -21.2 to 88.1; p=0.213). Learning: ‘Scaling-up’ PA interventions is challenging. Despite a promising feasibility study, the results of this fully powered trial indicate that in this context, the walking programme did not increase PA. Since the COVID-19 pandemic, school environments have changed and although pupils enjoyed the programme, attendance at walks was low. There is a need to better understand the implementation of interventions such as WISH within schools

S06-1 Putting young people at the heart of physical activity research design: The Walking In ScHools (WISH) Study

BACKGROUND: Young people have the right to be informed and consulted about decisions affecting their lives. Youth Patient and Public Involvement (PPI) should be encouraged to ensure research is carried out ‘with' or ‘by' young people rather than ‘to', ‘about' or ‘for' them. PPI can ensure research is relevant, results are accessible and recruitment rates are improved. Young people have had limited involvement in the design, implementation and dissemination of public health research and there have been calls for a greater focus on youth PPI in research. METHODS: Following the WISH feasibility study that consulted young people pre and post-intervention, a Youth Advisory Group (YAG) was set up within the main trial. The WISH study is a clustered randomised controlled trial in which a peer-led, school-based, brisk walking intervention is compared to usual physical activity in adolescent females. The YAG was introduced to inform intervention delivery and provide researchers with an understanding of what would encourage/discourage participation. Schools were asked to invite pupils aged 12-14 years (participants) and 15-18 years (walk leaders). Participative methods were used to develop and review study documentation. The YAG completed a short questionnaire and recruitment rates were monitored. RESULTS: Fourteen pupils from 3 schools attended the 2019 YAG meeting. The YAG agreed the meeting was a good way of getting young people involved in research (93%) and attendees enjoyed the meeting (100%). As a result, changes were made to study documentation, incentives were purchased and recruitment materials developed. Participant recruitment was higher in schools who participated in the YAG (54%) compared to those who did not (47%). In 2021 the second YAG occurred and 1 teacher, 12 participants and 10 walk leaders from 2 schools provided feedback on the trials COVID-19 contingency plan. The girls felt their feedback was valued (100%) and it was important young people had the chance to contribute to research studies (100%). CONCLUSIONS: The views of young people have been central to the development of the WISH Study and although youth PPI is not without challenges, there are many benefits for researchers, the study and the young people involved

S06-5 The Walking In ScHools (WISH) study: Development and evaluation of a peer-led school-based walking intervention in adolescent girls from pilot to fully-powered trial

BACKGROUND: Walking interventions, delivered within the school setting, have the potential to increase physical activity (PA) in adolescents. Previous research has shown that walking is an acceptable form of PA for adolescent girls, and that walking interventions may be effective at increasing PA in this group. Findings from the Walking In ScHools (WISH) pilot study (n199 female participants) found the intervention was effective in increasing light intensity PA in adolescent girls, but further research is needed to examine the effects of walking on overall PA and the role of peer leaders in delivering school-based interventions. The present study aims to build upon this pilot work and evaluate the effectiveness of a novel, low-cost, peer-led school-based walking intervention, delivered across the school year, at increasing accelerometer-measured PA levels of adolescent girls. METHODS: The WISH study is a school-based cluster randomised controlled trial targeting adolescent girls (aged 12-14 years) within the post-primary school setting. Data will be collected at four timepoints, baseline, mid-intervention, post-intervention, and 13 months post-baseline. Following baseline data collection, schools were randomly allocated to intervention (n = 9) or control (n = 9). In intervention schools, older pupils (aged 15-18 years) were trained as walk leaders and led the younger girls in 10-15min walks before school, at break, and during lunch, across the school year (20-22 weeks). The primary outcome measure is accelerometer-measured total PA (post-intervention) and secondary outcomes include anthropometry measures, and wellbeing. RESULTS: Some 590 participants (mean(SD) age 12.6(0.64)years) were recruited from 18 schools across Northern Ireland (n = 9) and the Border region of the Republic of Ireland (n9). Within the intervention schools, 149 walk leaders were trained. At baseline (n = 535), mean(SD) time spent in moderate to vigorous PA (MVPA) was 39.2(17.07)mins/day and 66 (12%) girls achieved PA guidelines of 60 minutes MVPA per day. Data collection and analysis is ongoing. CONCLUSIONS: This research has outlined the development of a novel, peer-led walking intervention and demonstrated its effectiveness at increasing light intensity PA in adolescent girls. The ongoing fully powered trial will build upon this pilot work and further evaluate the effects of the WISH study on increasing PA in adolescent girls

Exploring the Bone Proteome to Help Explain Altered Bone Remodeling and Preservation of Bone Architecture and Strength in Hibernating Marmots

Periods of physical inactivity increase bone resorption and cause bone loss and increased fracture risk. However, hibernating bears, marmots, and woodchucks maintain bone structure and strength, despite being physically inactive for prolonged periods annually. We tested the hypothesis that bone turnover rates would decrease and bone structural and mechanical properties would be preserved in hibernating marmots (Marmota flaviventris). Femurs and tibias were collected from marmots during hibernation and in the summer following hibernation. Bone remodeling was significantly altered in cortical and trabecular bone during hibernation with suppressed formation and no change in resorption, unlike the increased bone resorption that occurs during disuse in humans and other animals. Trabecular bone architecture and cortical bone geometrical and mechanical properties were not different between hibernating and active marmots, but bone marrow adiposity was significantly greater in hibernators. Of the 506 proteins identified in marmot bone, 40 were significantly different in abundance between active and hibernating marmots. Monoaglycerol lipase, which plays an important role in fatty acid metabolism and the endocannabinoid system, was 98-fold higher in hibernating marmots compared with summer marmots and may play a role in regulating the changes in bone and fat metabolism that occur during hibernation