Paolo Garagnani

Mandibuloacral dysplasia: A premature ageing disease with aspects of physiological ageing

Mandibuloacral dysplasia (MAD) is a rare genetic condition characterized by bone abnormalities including localized osteolysis and generalized osteoporosis, skin pigmentation, lipodystrophic signs and mildly accelerated ageing. The molecular defects associated with MAD are mutations in LMNA or ZMPSTE24 (FACE1) gene, causing type A or type B MAD, respectively. Downstream of LMNA or ZMPSTE24 mutations, the lamin A precursor, prelamin A, is accumulated in cells and affects chromatin dynamics and stress response. A new form of mandibuloacral dysplasia has been recently associated with mutations in POLD1 gene, encoding DNA polymerase delta, a major player in DNA replication. Of note, involvement of prelamin A in chromatin dynamics and recruitment of DNA repair factors has been also determined under physiological conditions, at the border between stress response and cellular senescence. Here, we review current knowledge on MAD clinical and pathogenetic aspects and highlight aspects typical of physiological ageing.

The Continuum of Aging and Age-Related Diseases: Common Mechanisms but Different Rates

Geroscience, the new interdisciplinary field that aims to understand the relationship between aging and chronic age-related diseases (ARDs) and geriatric syndromes (GSs), is based on epidemiological evidence and experimental data that aging is the major risk factor for such pathologies and assumes that aging and ARDs/GSs share a common set of basic biological mechanisms. A consequence is that the primary target of medicine is to combat aging instead of any single ARD/GSs one by one, as favored by the fragmentation into hundreds of specialties and sub-specialties. If the same molecular and cellular mechanisms underpin both aging and ARDs/GSs, a major question emerges: which is the difference, if any, between aging and ARDs/GSs? The hypothesis that ARDs and GSs such as frailty can be conceptualized as accelerated aging will be discussed by analyzing in particular frailty, sarcopenia, chronic obstructive pulmonary disease, cancer, neurodegenerative diseases such as Alzheimer and Parkinson as well as Down syndrome as an example of progeroid syndrome. According to this integrated view, aging and ARDs/GSs become part of a continuum where precise boundaries do not exist and the two extremes are represented by centenarians, who largely avoided or postponed most ARDs/GSs and are characterized by decelerated aging, and patients who suffered one or more severe ARDs in their 60s, 70s, and 80s and show signs of accelerated aging, respectively. In between these two extremes, there is a continuum of intermediate trajectories representing a sort of gray area. Thus, clinically different, classical ARDs/GSs are, indeed, the result of peculiar combinations of alterations regarding the same, limited set of basic mechanisms shared with the aging process. Whether an individual will follow a trajectory of accelerated or decelerated aging will depend on his/her genetic background interacting lifelong with environmental and lifestyle factors. If ARDs and GSs are manifestations of accelerated aging, it is urgent to identify markers capable of distinguishing between biological and chronological age to identify subjects at higher risk of developing ARDs and GSs. To this aim, we propose the use of DNA methylation, N-glycans profiling, and gut microbiota composition to complement the available disease-specific markers.

Aberrant methylation patterns in colorectal cancer: a meta-analysis

Colorectal cancer is among the leading causes of cancer death worldwide. Despite numerous molecular characterizations of the phenomenon, the exact dynamics of its onset and progression remain elusive. Colorectal cancer onset has been characterized by changes in DNA methylation profiles, that, owing to the stability of their patterns, are promising candidates to shed light on the molecular events laying at the base of this phenomenon. To exploit this stability and reinforce it, we conducted a meta-analysis on publicly available DNA methylation datasets generated on: normal colorectal, adenoma (ADE) and adenocarcinoma (CRC) samples using the Illumina 450k array, in the systems medicine frame, searching for tumor gene episignatures, to produce a carefully selected list of potential drivers, markers and targets of the disease. The analysis proceeds from a differential meta-analysis of the methylation profiles using an analytical pipeline recently developed by our group [1], through network reconstruction, topological and functional analyses, to finally highlight relevant epigenomic features. Our results show that genes already highlighted for their genetic or transcriptional alteration in colorectal cancer are also differentially methylated, reinforcing -regardless of the level of cellular control- their role in the complex of alterations involved in tumorigenesis. These findings were finally validated in an independent cohort from The Cancer Genome Atlas (TCGA).

Assessing the combined effect of extremely low-frequency magnetic field exposure and oxidative stress on LINE-1 promoter methylation in human neural cells

Extremely low frequency magnetic fields (ELF-MF) have been classified as “possibly carcinogenic”, but their genotoxic effects are still unclear. Recent findings indicate that epigenetic mechanisms contribute to the genome dysfunction and it is well known that they are affected by environmental factors. To our knowledge, to date the question of whether exposure to ELF-MF can influence epigenetic modifications has been poorly addressed. In this paper, we investigated whether exposure to ELF-MF alone and in combination with oxidative stress (OS) can affect DNA methylation, which is one of the most often studied epigenetic modification. To this end, we analyzed the DNA methylation levels of the 5′untranslated region (5′UTR) of long interspersed nuclear element-1s (LINE-1 or L1), which are commonly used to evaluate the global genome methylation level. Human neural cells (BE(2)C) were exposed for 24 and 48xa0h to extremely low frequency pulsed magnetic field (PMF; 50xa0Hz, 1xa0mT) in combination with OS. The methylation levels of CpGs located in L1 5′UTR region were measured by MassARRAY EpiTYPER. The results indicate that exposures to the single agents PMF and OS induced weak decreases and increases of DNA methylation levels at different CpGs. However, the combined exposure to PMF and OS lead to significant decrease of DNA methylation levels at different CpG sites. Most of the changes were transient, suggesting that cells can restore homeostatic DNA methylation patterns. The results are discussed and future research directions outlined.

Genes associated with Type 2 Diabetes and vascular complications

Type 2 Diabetes (T2D) is a chronic disease associated with a number of micro- and macrovascular complications that increase the morbidity and mortality of patients. The risk of diabetic complications has a strong genetic component. To this end, we sought to evaluate the association of 40 single nucleotide polymorphisms (SNPs) in 21 candidate genes with T2D and its vascular complications in 503 T2D patients and 580 healthy controls. The genes were chosen because previously reported to be associated with T2D complications and/or with the aging process. We replicated the association of T2D risk with IGF2BP rs4402960 and detected novel associations with TERT rs2735940 and rs2736098. The addition of these SNPs to a model including traditional risk factors slightly improved risk prediction. After stratification of patients according to the presence/absence of vascular complications, we found significant associations of variants in the CAT, FTO, and UCP1 genes with diabetic retinopathy and nephropathy. Additionally, a variant in the ADIPOQ gene was found associated with macrovascular complications. Notably, these genes are involved in some way in mitochondrial biology and reactive oxygen species regulation. Hence, our findings strongly suggest a potential link between mitochondrial oxidative homeostasis and individual predisposition to diabetic vascular complications.

A bio-cultural approach to the study of food choice: The contribution of taste genetics, population and culture

The study of food choice, one of the most complex human traits, requires an integrated approach that takes into account environmental, socio-cultural and biological diversity. We recruited 183 volunteers from four geo-linguistic groups and highly diversified in terms of both genetic background and food habits from whom we collected genotypes and phenotypes tightly linked to taste perception. We confirmed previous genetic associations, in particular with stevioside perception, and noted significant differences in food consumption: in particular, broccoli, mustard and beer consumption scores were significantly higher (Adjusted Pxa0=xa00.02, Adjusted Pxa0<xa00.0001 and Adjusted Pxa0=xa00.01, respectively) in North Europeans, when compared to the other groups. Licorice and Parmesan cheese showed lower consumption and liking scores in the Sri Lankan group (Adjusted Pxa0=xa00.001 and Adjusted Pxa0<xa00.001, respectively). We also highlighted how rs860170 (TAS2R16) strongly differentiated populations and was associated to salicin bitterness perception. Identifying genetic variants on chemosensory receptors that vary across populations and show associations with taste perception and food habits represents a step towards a better comprehension of this complex trait, aimed at improving the individual health status. This is the first study that concurrently explores the contribution of genetics, population diversity and cultural aspects in taste perception and food consumption.

Three Reportedly Unrelated Families With Liddle Syndrome Inherited From a Common Ancestor

Liddle syndrome is considered a rare Mendelian hypertension. We have previously described 3 reportedly unrelated families, native of an Italian area around the Strait of Messina, carrying the same mutation (&bgr;P617L) of the epithelial sodium channel. The aims of our study were (1) to evaluate whether a close genomic relationship exists between the 3 families through the analysis of mitochondrial DNA and Y chromosome; and (2) to quantify the genomic relatedness between the patients with Liddle syndrome belonging to the 3 families and assess the hypothesis of a mutation shared through identity by descent. HVRI (the hypervariable region I) of the mitochondrial DNA genome and the Y chromosome short tandem repeats profiles were analyzed in individuals of the 3 families. Genotyping 542 585 genome-wide single nucleotide polymorphisms was performed in all the patients with Liddle syndrome of the 3 families and some of their relatives. A panel of 780 healthy Italian adult samples typed for the same set of markers was used as controls. espite different lineages between the 3 families based on the analysis of mitochondrial DNA and Y chromosome, the 3 probands and their 6 affected relatives share the same ≈5 Mbp long haplotype which encompasses the mutant allele. Using an approach based on coalescent theory, we estimate that the 3 families inherited the mutant allele from a common ancestor ≈13 generations ago and that such an ancestor may have left ≈20 carriers alive today. The prevalence of Liddle syndrome in the region of origin of the 3 families may be much higher than that estimated worldwide.

Physical Activity and Nutrition INfluences In ageing (PANINI):: consortium mission statement

decade in poor health. Consequently, there is an urgent need to understand how lifestyle factors can influence age-related changes from gene to society level and how they may be integrated into a net effect of healthy ageing. It is also crucial to develop and validate interventions and health policies to ensure that more of our older adults have a healthy and active later life. This is an urgent and cross-cutting research priority in Europe, and to achieve this, it is vital to increase research capacity in this area to push forward the frontiers of scientific understanding. The Horizon 2020 funded Marie Curie Sklodowska Innovative Training Network—PANINI is addressing this capacity issue by focusing on research and training in two major interacting lifestyle factors with impact at multiple levels, namely, physical activity and nutrition. The aims of PANINI are, therefore, to:

Epigenetic DNA methylation changes in episodic and chronic migraine

According to International Headache Classification (ICHD-3), migraine is classified in episodic migraine (EM) and chronic migraine (CM) based on the frequency of attacks. Medication overuse headache (MOH) is a secondary chronic headache disorder defined as a frequent headache (15 days per month or more) induced by the overuse of analgesics. Currently, the best treatment options include a detoxification programme based on the abrupt withdrawal of overused painkillers and eventually a preventive treatment. Despite these therapeutic efforts, the relapse rate of MOH is high, and no valid biomarkers are available to detect among patients who suffer from EM and CM those at greatest risk of develop MOH. According to available data, certain groups of people are more vulnerable to develop chronic pain conditions and poor adaptation to stress. Based on the scientific knowledge, a behavioural model of headache was proposed considering the strong association between pain, homeostatic imbalance, and affective behaviour. The pain and stress vulnerability is determined by genetic and non-genetic but heritable factors under the effect of environmental exposures as well as stochastic events during development. Differences between individuals’ DNA sequences can predispose toward maladaptive behaviours in many disorders comorbid with MOH, conferring a risk for chronic pain and by functioning as a type of molecular memory. Several evidences confirmed that pain vulnerability and attitude to chronic pain sensitivity are heritable via genetic but also epigenetic pathways though changes in DNA expression. A main epigenetic mechanism is DNA methylation, the covalent addition of a methyl group to the fifth carbon of cytosine residues, which is typically associated with gene silencing. Epigenetic mechanisms would have the potential to link early life events, neuro-inflammation, and brain plasticity in the aetiology of migraine chronification [1]. It has been hypothesised that frequent headache attacks may lower the threshold for subsequent headache attacks through epigenetic mechanisms. The suggested hypothesis is that neuronal activity can cause epigenetic changes altering synaptic plasticity, and the frequent migraine attacks in a feed-forward loop may promote stable epigenetic changes which altering synaptic plasticity supporting migraine chronification. Similarly, psychological acute and chronic stress and female sex hormones, which have been implicated in migraine genesis, are known to exert their physiological effects partly through epigenetic mechanisms [1]. Finally, drug addiction can be viewed as maladaptive neural plasticity that occurs in vulnerable individuals in response to repeated exposure to a drug of abuse. Evidences suggested that this vulnerability is partly determined by nongenetic factors which include environmental exposures as well as stochastic events during development which act through epigenetic mechanisms. However, there are no studies in MOH. The reduction of nucleic acid sequencing costs and the availability of cost-effective microarray solutions for the analysis of DNA methylation has favoured the implementation of epigenomic studies. DNA methylation, miRNA, and histone modifications have proven to be a potential source of powerful and robust biomarkers. Taken together, both the new genetic and epigenetic omic approaches have the potential to provide new molecular insight in the aetiology of migraine chronification, patient stratification, and therapy [2]. Recently, the first genome-wide study of DNA methylation in headache chronification was published. Several potentially implicated loci and processes were identified, but in the combined meta-analysis, the strongest associated CpG sites were related to SH2D5 and NPTX2, two brain-expressed genes * Rossana Terlizzi rossana.terlizzi2@unibo.it

Molecular Aging of Human Liver: An Epigenetic/Transcriptomic Signature

The feasibility of liver transplantation from old healthy donors suggests that this organ is able to preserve its functionality during aging. To explore the biological basis of this phenomenon, we characterized the epigenetic profile of liver biopsies collected from 45 healthy liver donors ranging from 13 to 90 years old using the Infinium HumanMethylation450 BeadChip. The analysis indicates that a large remodeling in DNA methylation patterns occurs, with 8,823 age-associated differentially methylated CpG probes. Notably, these age-associated changes tended to level off after the age of 60, as confirmed by Horvaths clock. Using stringent selection criteria, we further identified a DNA methylation signature of aging liver including 75 genomic regions. We demonstrated that this signature is specific for liver compared to other tissues and that it is able to detect biological age-acceleration effects associated with obesity. Finally, we combined DNA methylation measurements with available expression data. Although the intersection between the two omic characterizations was low, both approaches suggested a previously unappreciated role of epithelial-mesenchymal transition and Wnt-signaling pathways in the aging of human liver.