Project description:Definitive parameters or mechanisms underlying the severity of COVID-19 in elderly people remain confused. Thus, this study seeks to elucidate the mechanism behind the increased vulnerability of elderly individuals to severe COVID-19. We employed an aged mouse model with a mouse-adapted SARS-CoV-2 strain to mimic the severe symptoms observed in elderly patients with COVID-19. Comprehensive analyses of the whole lung were performed using transcriptome and proteome sequencing, comparing data from aged and young mice. For transcriptome analysis, bulk RNA sequencing was conducted using an Illumina sequencing platform.
Project description:Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in a global health crisis, particularly affecting the elderly, who are more susceptible to severe outcomes. However, definitive parameters or mechanisms underlying the severity of COVID-19 in elderly people remain confused. Thus, this study seeks to elucidate the mechanism behind the increased vulnerability of elderly individuals to severe COVID-19. We employed an aged mouse model with a mouse-adapted SARS-CoV-2 strain to mimic the severe symptoms observed in elderly patients with COVID-19. Comprehensive analyses of the whole lung were performed using transcriptome and proteome sequencing, comparing data from aged and young mice. For transcriptome analysis, bulk RNA sequencing was conducted using an Illumina sequencing platform. Proteomic analysis was performed using mass spectrometry following protein extraction, digestion, and peptide labeling. We analyzed the transcriptome and proteome profiles of young and aged mice and discovered that aged mice exhibited elevated baseline levels of inflammation and tissue damage repair. After SARS-CoV-2 infection, aged mice showed increased antiviral and inflammatory responses; however, these responses were weaker than those in young mice, with significant complement and coagulation cascade responses. In summary, our study demonstrates that the increased vulnerability of the elderly to severe COVID-19 can be attributed to an attenuated antiviral response and the overactivation of complement and coagulation cascades.
Project description:One inevitable consequence of the effect of age on our bodies is the graduated deterioration of physical function and exercise capacity, driven, in part by the adverse effect of age on muscle tissue. Our primary purpose was to determine the relationship between patterns of gene expression in skeletal muscle and loss of physical function. We hypothesized that some genes that change expression with age would correlate with functional decline, or conversely with preservation of function. Male C57Bl/6 mice were randomly selected from large cohorts [RNA isolated from: adults (6-7 months old, n=9), older (24-25 months old, n=9), and elderly (28+ months of age, n=9)} that were tested for physical ability using a comprehensive functional assessment battery [CFAB, a composite scoring system: comprised of the rotarod (overall motor function), grip strength (fore-limb strength), inverted cling (4-limb strength/endurance), voluntary wheel running (activity rate/volitional exercise), and treadmill tests (endurance)]. We extracted RNA from the tibialis anterior muscles, ran RNAseq to examine the transcriptome using an Illumina NextSeq 550, comparing adults (n=7) to older (n=7) and elderly mice (n=9). Age resulted in gene expression differences of 1.5 log2 fold change or greater (p<0.01) in 46 genes in the older mice and in 252 genes in the elderly (both compared to adults). Current ongoing work is examining the physiological relevance of these genes to age-related loss of physical function. We are in the process of using linear regression to determine which of the genes with age-related changes in expression are associated (R>0.5 and p<0.05) with functional status as measured by CFAB.
Project description:In the scientific landscape, there is a growing interest in defining the role of several biomolecules and humoral indicators of the aging process and in the modifications of these biomarkers induced by physical activity and exercise. The main aim of the present narrative review is to collect the available evidence on the biohumoral indicators that could be modified by physical activity (PA) in the elderly. Online databases including Pubmed, Web of science (Medline), and Scopus were searched for relevant articles published in the last five years in English. Keywords and combination of these used for the search were the following: "biological", "indicators", "markers", "physical", "activity", and "elderly". Thirty-four papers were analyzed for inclusion. Twenty-nine studies were included and divided into four categories: cardiovascular (CV) biomarkers, metabolic biomarkers, inflammatory markers-oxidative stress molecules, and other markers. There are many distinct biomarkers influenced by PA in the elderly, with promising results concerning the metabolic and CV indexes, as a growing number of studies demonstrate the role of PA on improving parameters related to heart function and CV risk like atherogenic lipid profile. Furthermore, it is also a verified hypothesis that PA is able to modify the inflammatory status of the subject by decreasing the levels of pro-inflammatory cytokines, including interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-?). PA seems also to be able to have a direct effect on the immune system. There is a strong evidence of a positive effect of PA on the health of elderly people that could be evidenced and "quantified" by the modifications of the levels of several biohumoral indicators.
Project description:Aging is an important risk factor for disease and is accompanied by the decline of many physiological characteristics, including physical performance. One of the most powerful tests of physical ability in elderly humans is the short physical performance battery (SPPB) (Guralnik et al., 1994), an assessment of the maximum exercise capacity of lower extremities over a short period of time. A similar metric of maximum velocity (MV) in a short period (30 s) is proposed as an equally informative measurement in C. elegans (Hahm et al., 2015). MV declines with age, correlates well with longevity, accurately reports movement ability, and importantly, is predictive of future longevity (Hahm et al., 2015). Here, we characterized the differences in gene expression between high MV group and low MV group of the same age in C. elegans. The genes related to nucleosomes and involved in regulatory processes, including transcriptional regulation (chromatin assembly, regulation of RNA metabolic process and regulation of transcription), and neuronal signaling (neurogenesis, axon guidance, and regulation of neurotransmitter levels) were up-regulated in low MV group compared to high MV group. In contrast, genes belonged to metabolic processes and mitochondria were down-regulated in low MV group compared to high MV group.
Project description:Illumina Infinium EPIC HumanMethylation BeadChip data from saliva DNA samples from a healthy elderly cohort with individuals in the age range 70-95 in Southwest Sweden. The cohort was stratified into study groups based on participants´answers to a questionnaire of different lifestyle factors including vitamin supplementations, smoking and drinking habits, physical activity (per year), sun exposure and eating habits. Vitamin D intake was evaluated from the vitamin D supplementation (alone or in a multivitamin complex), dietary vitamin D intake (fish and seafood frequency) and vitamin D synthesis in the skin (sunlight exposure and use of sunscreen). Differential methylation analysis was performed for all the study groups and the combination of different factors with vitamin D supplementation. Gender, age, smoking and alcohol (SD and frequency) were used as covariates in the analyses. Only the study groups referred to the conclusions of the study are shown.
Project description:Prime editing is a versatile genome-editing technique that shows great promise for the generation and repair of patient mutations. However, some genomic sites are difficult to edit and optimal design of prime-editing tools remains elusive. Here we present a fluorescent prime editing and enrichment reporter (fluoPEER), which can be tailored to any genomic target site. This system rapidly and faithfully ranks the efficiency of prime edit guide RNAs (pegRNAs) combined with any prime editor variant. We apply fluoPEER to instruct correction of pathogenic variants in patient cells and find that plasmid-editing enriches for genomic editing up to 3-fold compared to conventional enrichment strategies. DNA repair and cell cycle-related genes are enriched in the transcriptome of edited cells. Stalling cells in the G1/S boundary increases prime editing efficiency up to 30%. Together, our results show that fluoPEER can be employed for rapid and efficient correction of patient cells, selection of gene-edited cells, and elucidation of cellular mechanisms needed for successful prime editing.