The contribution of the non-coding genome to disease and its therapeutic potential have been largely unexplored. Recently, several epigenetic drugs developed for cancer treatment have been described to mediate therapeutic effects through the reactivation of the expression of transposable elements in cancer cells. This event activates innate immunity-related pathways and promotes the generation of neoantigens in tumor cells, improving the efficacy of immunotherapeutic treatments. This review focuses on the regulation of transposable elements by epigenetic inhibitors and its implications for immuno-oncology. Traditionally, studies addressing the etiology and treatment of human diseases have focused on the role of coding genes. However, coding genes represent only 2% of the human genome . EffortsBurnet and Thomas suggested back in 1957 that the immune system has the capacity to detect and destroy abnormal cells, however it has taken several decades to develop the first successful immunotherapeutic approaches to treat cancer . A key eIn addition, cancer cells can express modulatory signals that reinforce or prevent the immune reaction against the tumor. “Hot” tumors are typically enriched in interferon (IFN) related pathways, secrete pro-inflammatory cytokines and immuno-attractant chemokines, and are infiltrated by immune cells. However, these immune responses are not sustained for a long time as they may result in autoimmunity. Activated T cells eventually upregulate surface inhibitory receptors such as the programmed cell death protein 1 (PD-1) and the cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) . These rImmunotherapy has achieved very good responses in a percentage of cancer patients; however, many patients have not experienced clinical benefits. Novel targets or combination with other therapies might improve these responses. Despite the traditional assumption that inhibitors of epigenetic repressors affect cancer cell growth through the reactivation of tumor suppressor genes, successful combinations of epigenetic drugs and immune checkpoint inhibitors in animal models and early clinical trials suggest that these drugs have immunomodulatory properties. Unexpectedly, the effects of these epigenetic inhibitors appear to be mediated, at least in part, through the regulation of non-coding elements of the genome.More than 50% of the human genome is made of transposable elements (TEs) which are highly repetitive DNA sequences  Figure A. DespitAn abundant type of transposons are retrotransposons, which use an RNA intermediate to copy and paste themselves into genomic locations . RetrotrRetrotransposons of the non-long terminal repeat (non-LTR) category comprise long and short interspersed nuclear elements  . The mosSINEs are short elements that probably evolved from RNA genes, such as 7SL and tRNA . The mosAbout 8% of the genome is made of LTR retrotransposons commonly represented by human endogenous retroviruses (HERVs) . HERV seThe survival of potentially dangerous retroviruses in the human genome along evolution can be partially explained by their ability to reinvent and participate in cellular functions, a phenomenon called co-option . The LTRIn addition, to combat the potentially harmful effects of TEs, the genome has evolved epigenetic silencing mechanisms to suppress their activity. These mechanisms include small silencing RNAs, histone and DNA modifications, as well as sequence-specific repressors such as KRAB zinc-finger proteins.Although many repetitive elements are silenced by means of mutations and/or epigenetic silencing, it is now clear that some of these elements are expressed in cells under normal circumstances. The transcription of retroelements frequently gives rise to the formation of long double-stranded RNA (dsRNA) . EvidencdsRNAs produced as a result of the transcription of retroelements can accumulate in the cytoplasm and activate antiviral signaling pathways in a process known as “viral mimicry” ,22. ThesCells have developed mechanisms of self-tolerance to dsRNAs consisting of their editing by the adenosine deaminases ADARs that convert adenosine to inosine (A-to-I) . The conOverall, the importance of the fine tuning of dsRNA production, editing, and detection becomes evident in diseases caused by mutations in components involved in its regulation. In humans, MDA5 gain of function mutations cause a broad spectrum of autoinflammatory diseases, including the Aicardi-Goutières syndrome . Not surDifferent mechanisms of epigenetic silencing prevent the transcription of genomic locations. The wrapping of DNA around the histone octamers to form nucleosomes offers itself a barrier for the recruitment and processivity of RNA polymerases . Highly DNA methylation plays a key role in gene silencing, X chromosome inactivation, genome stability, and imprinting. Although most of the interest has been focused in the role of DNA methylation in silencing the expression of tumor suppressor genes in cancer, DNA methylation is likely to play an important role in repeat element regulation since most of the genomic CpG sites subjected to DNA methylation are located at these elements . The DNMTs inhibitors (DNMTi) azacytidine (AZA) and decitabine (DAC) are the most successful and have the longest history of epigenetic drugs used in cancer treatment to date . These cIn addition to showing antiproliferative effects in cancer cells, DNMTi also have been described to improve the effects of immune checkpoint inhibitors in mouse models. Combinations of AZA or the HDACs inhibitor (HDACi) entinostat and anti-PD-1/anti-CTLA-4 antibodies improves the treatment outcome in mouse models of colorectal and breast cancer . In clinMore mechanistic studies revealed that cancer cell lines treated with low doses of AZA induce the expression of genes involved in innate immunity, including antigen presentation, apoptosis, interferon signaling, and anti-viral defense ,35. DeepFine mapping of transcriptional responses to DNMTi in the lung cancer cell line NCI-H1299 revealed that treatment with DAC had no significant direct effects on gene promoter driven transcription. Instead it induced the transcription of thousands of non-annotated transcription start sites (TSSs) that overlapped with TEs . The mosIn addition to the effects of DNMTi in stimulating innate immunity pathways these inhibitors might also enhance the effects of immune checkpoints inhibitors through the induction of expression of tumor-specific antigens ,35. ReceA major drawback of epigenetic therapies that has hampered their progress in the clinic is the inability to select which cancer patients are more likely to respond to therapy. Finding novel markers to predict sensitivity to DNMTi might help to find clinical applications for these drugs in solid tumors. Several lines of evidence suggest a correlation between the expression of TEs, enrichment in innate immunity-related gene signatures and sensitivity to DNMTi treatment. Chiappinelli et al. observed that ovarian serous cancers with high HERV expression were enriched in innate immunity signatures . AdditioHistone lysine specific demethylase LSD1 (KDM1A) was the first discovered histone demethylase . LSD1 waLSD1 has been described to be involved in several types of cancer. High LSD1 expression correlates with malignancy in a range of solid tumors ,44,45. IHaving LSD1 multiple targets and acting both as a coactivator and corepressor has complicated the understanding of how LSD1i block proliferation of cancer cells. Inhibition of transcriptional programs governed by oncogenic transcription factors and induction of differentiation pathways have been reported in solid cancer cell lines and mouse models of hematologic cancers ,52. MoreRecent data suggests that LSD1i might potentiate anti-tumor immune effects through the reactivation of HERVs. Treatment of cancer cell lines with the LSD1i GSK-LSD1 resulted in the induction of interferon-related genes and expression of a subset of HERVs in several cancer cell lines . While iAnalysis of gene expression data in cutaneous melanoma, TNBC, and pHHG patients revealed inverse correlations between LSD1 expression and CD8+ T cell infiltration, prognosis, levels of cytotoxic T cell attracting chemokines expression, PD-L1 levels, and innate immunity signatures ,54,55. TFrom the mechanistic point of view there are still many unresolved questions about the effects of LSD1i in the activation of TEs. A fine dissection of the sites of cryptic transcription induced by these inhibitors is granted to fully understand their effects. This will allow to determine whether DNMTi and LSD1i regulate the same or different types of elements and whether inhibition of both will further stimulate anti-tumor immunity. In addition, further work is needed to explore the effects of LSD1 inhibition in immune cells.EZH2 is the catalytic subunit of the PcG repressor complex 2 (PRC2) that trimethylates lysine 27 on histone 3 (H3K27me3), a mark involved in gene repression. Several lines of evidence have implicated EZH2 in the development and progression of a variety of cancers. EZH2 overexpression has been shown to correlate with aggressiveness and advanced disease in several cancer types ,57,58. AIn addition to the described antiproliferative effects, several studies have shown that the treatment with EZH2i improves the responses to diverse immunotherapy approaches in mouse models of ovarian, bladder cancer, and melanoma ,71,72 anRecent evidence suggests that EZH2 plays a role in silencing TEs in cancer cells. Lower levels of EZH2 and higher levels of innate immunity-related genes expression have been reported in chemotherapy-resistant small-cell lung cancer (SCLC) cell lines compared to parental sensitive cell lines . ReducedHistone acetylation plays an important role in gene expression. Acetylated histones are recognized by epigenetic readers that participate in the recruitment of Pol II to gene promoters. Levels of acetylation at particular genomic locations are the result of an equilibrium between histone acetyl transferases (HATs) and histone deacetylates (HDACs). There are 18 human HDACs grouped into four classes based on their primary homology to yeast HDACs. Among these, class I and II HDACs play a major role in the lysine deacetylation of N-terminal histone tails. Class I comprises HDAC1, 2, 3, and 8. Class II is further divided into two subclasses: IIa  and IIb (HDAC6 and 10). Four inhibitors , which target several HDACs among the different classes, have been FDA approved for the treatment of several hematological malignancies . Other HEvidence obtained in clinical trials suggest that treatment with HDACi improves the efficacy of immunotherapeutic approaches . EffectsG9A is a mammalian histone methyltransferase that catalyzes the methylation histone H3 at lysine 9 (H3K9), participates in gene repression, and is required for the establishment of the silencing of newly integrated proviruses in mESC . G9A is Despite being less explored than the previously discussed inhibitors, combination of G9Ai and DNMTi caused synergistic antitumor effects that correlated with the upregulation of HERVs and viral defense genes in ovarian cancer cell lines with high levels of G9A expression . HERVs uIn addition to G9A, other H3K9 methyltransferases have been involved in the repression of HERVs. For example, the methyltransferase SETDB1 is known to play a role in the silencing of retroelements in mESCs . Knock oRepetitive elements have been considered “junk” DNA for a long time and the study of their regulation and expression largely disregarded. In the last decade, the contribution of transposable elements to the etiology of diseases and their potential to treat cancer is becoming clear. As a field that is emerging, still many critical questions remain unanswered.The evolutionary complexity of retrotransposons suggests that alterations in the epigenetic silencing of these elements might have multiple consequences. In addition to a source of non-coding transcripts and dsRNAs, co-opted HERVS have been described to drive the expression of both oncogenes and tumor suppressors ,88. In aAnother question is to which extent the reactivation of different subfamilies of TEs in response to epigenetic drugs is tumor or cell specific. Activation of the same LTR12C elements by DNMTi and HDACi treatment but not by other epigenetic inhibitors has been seen in different cancer cell lines . HoweverFine dissection of the mechanisms involved in the response to epigenetic drugs is needed to select potential responsive patients as well as successful combinatorial therapies in clinical trials. The expression of TEs in tumors is associated with immune infiltration and increased antigenicity . Reactiv


Increased and changed deposition of extracellular matrix proteins is a key feature of airway wall remodeling in obstructive pulmonary diseases, including asthma and chronic obstructive pulmonary disease. Studies have highlighted that the deposition of various basement membrane proteins in the lung tissue is altered and that these changes reflect tissue compartment specificity. Inflammatory responses in both diseases may result in the deregulation of production and degradation of these proteins. In addition to their role in tissue development and integrity, emerging evidence indicates that basement membrane proteins also actively modulate cellular processes in obstructive airway diseases, contributing to disease development, progression and maintenance. In this review, we summarize the changes in basement membrane composition in airway remodeling in obstructive airway diseases and explore their potential application as innovative targets for treatment development. Chronic respiratory diseases are the third leading cause of death, with approximately 550 million people affected worldwide and 4 million deaths per year Asthma is characterized by variable respiratory symptoms, such as wheeze, shortness of breath, chest tightness and cough, and variable expiratory airflow limitation. In addition, it is usually associated with airway inflammation Both asthma and COPD are associated with cough, dyspnea, mucous hypersecretion, inflammation, airway wall remodeling, and airway hyperresponsiveness. Both inflammation and airway remodeling may lead to airway hyperresponsiveness, defined as an exaggerated obstructive response to various nonspecific stimuli Airway remodeling, defined by structural changes in the airway architecture, is a characteristic feature of both asthma and COPD. Airway remodeling includes increased airway smooth muscle (ASM) mass, increased vascularity, epithelial shedding, goblet cell hyperplasia and increased and changed deposition of extracellular matrix (ECM) proteins, including basement membrane (BM) proteins The BM in the lung is a thin, dense sheet of ECM separating the airway epithelium, endothelial cells or airway and vascular smooth muscle cells from the underlying connective tissue The lamina lucida is the most superficial layer of the BM underlying the epithelium and the endothelium and is comprised mainly of laminins. Binding of laminins to their receptors on the cell membrane initiates a polymerization process resulting in the formation of the primary network of laminin fibers which is essential for BM formation.Laminins are a family of cruciform-like glycoproteins composed of five α, three β and three γ chains forming 16 different laminin heterotrimers Collagen IV binds to the primary network of laminins and forms a secondary network, the lamina densa. The collagen IV is the major protein found in basement membranes and forms a network that is assembled from three distinct heterotrimers that comprise the six collagen IV chains (α1-6); the ubiquitously expressed and the most abundant α1α1α2 isoform and the tissue specific α3α4α5 and α5α5α6 isoforms Nidogens and proteoglycans stabilize the laminin and collagen IV networks Heperan sulphate proteoglycans further stabilize the BMs Physiological turnover of BM proteins may result in the formation of matrikines  The BM has several key functions, including providing 1) a structural scaffold to support cells, 2) a template for tissue repair, 3) a reservoir for growth factors, 4) a selectively permeable barrier for molecules to cross; forming a physical barrier for cells and proteins; and finally forming an adhesive link between the epithelium/endothelium and the interstitial matrix Different BM proteins have specific functions in lung development as well, including directing epithelial branching, smooth muscle differentiation and alveolarization ECM changes are key in remodeling in both asthma and COPD In the airways of asthmatics, increased expression of several laminin chains, including the α2, α3, α5, β1, β2 and γ1 chain in the epithelial BM has been observed  [13], [3In both the large and small airways of patients with COPD, no changes in collagen IV were reported 2-agonist salbutamol Changes in BM proteins in patients with obstructive pulmonary diseases have been shown to correlate with disease characteristics. In patients with asthma, increased expression of laminin in the ASM correlated with a reduced reversibility of airway obstruction in response to the βAlthough no clear changes in collagen IV deposition were observed in biopsy studies, increased turnover of collagen IV, as indicated by measurement of soluble neo-fragments released from collagen IV, has been shown during exacerbations in COPD Maintenance of the ECM composition in the lung is a tightly controlled process. Production, secretion and incorporation of proteins in the ECM is balanced by degradation by proteases, such as matrix metalloproteinases (MMPs), cysteine proteases and serine proteases. An additional layer of regulation is provided by the endogenous inhibitors of these enzymes; the tissue inhibitors of metalloproteinases (TIMPs), serpins and cystatins. In the airways, turnover of some ECM proteins is estimated to occur at a rate of more than 10% per day Laminins are produced and secreted by a variety of structural and inflammatory cells Laminins are degraded by serine proteases and MMPs Turnover may not only result in changed expression patterns of BM proteins in the tissue, but may also expose matricryptic sites, biologically active sites within the ECM molecules protein sequence that are normally not exposed in the mature deposited ECM. In addition, degradation of ECM proteins results in the formation of matrikines. Both matricryptic sites and matrikines may activate (patho)physiological processes which are independent from the parent molecule and contribute to inflammation and remodeling. For review of matrikines in lung health and disease the reader is referred to Collectively, these observations indicate that expression and degradation of BM proteins is tightly balanced by various mechanisms, but maybe be deregulated by external factors, such as inflammation. The precise mechanisms underlying the observed BM changes in asthma and COPD remain to be elucidated.In addition to their physiological role in the lung, BM proteins have also been shown to drive pathophysiological processes in asthma and COPD . The rolAirway inflammation is key in both asthma and COPD pathogenesis and is considered to contribute to variable airway hyperresponsiveness, induction of airway remodeling and to the development, progression and maintenance of disease. Inflammation in asthma is heterogeneous. Thus far, focus has been mainly on type 2 inflammation involving eosinophils, mast cells, T helper 2 (Th2) lymphocytes and elevated immunoglobulin E levels. The mechanisms involved in non-type 2 inflammation are less well characterized Most evidence about the role of laminins in inflammation comes from studies in the vasculature. Neutrophils, basophils, eosinophils and mast cells have been found to be able to interact with laminins In vivo, extravasation of leukocytes occurs mainly at sites with high laminin α4 expression and little or no laminin α5 expression Increased turn-over of collagen IV may also affect inflammatory responses. Degradation of ECM proteins may result in the formation of matrikines. Tumstatin, a matrikine derived from collagen IV α3, inhibits eosinophilic inflammation in mouse and sheep models of asthma associated with reduced airway hyperresponsiveness, potentially via reduction in blood vessel area and vascular endothelial growth factor expression Epithelial changes including epithelial denudation, epithelial desquamation and goblet cell hyperplasia have been shown to be characteristic features of airway remodeling in asthma and COPD Haemophilus influenzae and Moraxella catarrhalis, and Aspergillus species Epithelial denudation and fragmentation of the BM leaves exposed BM and ECM proteins, with potentially neo-epitopes that are usually hidden within the BM/ECM structural network. The exposure of structural BM proteins may increase the chance of bacterial infections caused by micro-organisms, such as In vitro studies on the effects of specific laminin isoforms on ASM cells have shown that laminin α1 chain inhibits ASM phenotype switching and maintains a contractile phenotype, whereas laminin α2 chains promotes a hypercontractile phenotype In vivo, knock out of laminin α2 protected against allergen-induced airway hyperresponsiveness, which was associated with reduced ASM accumulation and normalization of ASM apoptosis Increased ASM mass is a characteristic feature of airway remodeling in asthma and COPD 2-agonists on collagen IV expression or for collagen IV on β2-agonist signaling Collagen IV deposition by asthmatic ASM cells is reduced compared to non-asthmatic cells Changes in the airway wall in asthma and COPD are associated with vascular changes and angiogenesis In vitro, treatment of ASM cells with tumstatin inhibited the formation of a pro-angiogenic ECM Both ECM proteins and ECM fragments have been shown to play a crucial role in regulating angiogenesis under physiological and pathophysiological conditions Despite its apparent thin presence providing a junction between two zones in many tissue structures the BM still holds many secrets that may be key for understanding the pathogenesis of obstructive lung diseases. Through looking to other fields or disease focuses we can see there are exciting avenues of investigation to pursue for obstructive airway diseases.Recent studies have illustrated that the BM is a highly dynamic environment, with a relatively stable scaffold of collagen IV and laminins supporting a highly motile array of players including nidogen, members of the fibulin family, agrin, spondin, and peroxidasin Epithelial cell interactions with the BM are important for establishing cellular polarization Garnering further insight about the role of the BM niche that influences all the cells that come into contact with it  in obstructive lung disease pathogenesis will open new possibilities for developing novel therapeutic strategies.JKB was supported by a Rosalind Franklin Fellowship co-funded by the University of Groningen and the European Union.Bart G.J. Dekkers: Conceptualization, Investigation, Writing – original draft, Visualization, Supervision. Shehab I. Saad: Conceptualization, Investigation, Writing – original draft. Leah J. van Spelde: Conceptualization, Investigation, Writing – original draft. Janette K. Burgess: Conceptualization, Writing – review & editing, Supervision.


It is today widely accepted that a healthy diet can be one of the fundamental approaches to prevent the risk of cancer. To this aim, nutrigenomics studies are indeed providing a precious source of information, favoring the search for compounds that could affect gene expression in a favorable way. Here we present a computational study to select candidate compounds that could play a role in cancer prevention and care. Starting from analyses of gene expression, we identified 7 genes that have opposite expression trends in apoptotic treatments when compared with 8 different cancer types. In addition, based on structure similarity with 6 compounds that affect the expression patterns of these genes in a favorable way against 8 cancer types, we selected 23 natural compounds as suitable candidates for further tests as possible novel drugs or for the design of functional food for cancer treatment and prevention.It is today widely accepted that a healthy diet is very useful to prevent the risk for cancer or its deleterious effects. Nutrigenomics studies are therefore taking place with the aim to test the effects of nutrients at molecular level and contribute to the search for anti-cancer treatments. These efforts are expanding the precious source of information necessary for the selection of natural compounds useful for the design of novel drugs or functional foods. Here we present a computational study to select new candidate compounds that could play a role in cancer prevention and care. Starting from a dataset of genes that are co-expressed in programmed cell death experiments, we investigated on nutrigenomics treatments inducing apoptosis, and searched for compounds that determine the same expression pattern. Subsequently, we selected cancer types where the genes showed an opposite expression pattern and we confirmed that the apoptotic/nutrigenomics expression trend had a significant positive survival in cancer-affected patients. Furthermore, we considered the functional interactors of the genes as defined by public protein-protein interaction data, and inferred on their involvement in cancers and/or in programmed cell death. We identified 7 genes and, from available nutrigenomics experiments, 6 compounds effective on their expression. These 6 compounds were exploited to identify, by ligand-based virtual screening, additional molecules with similar structure. We checked for ADME criteria and selected 23 natural compounds representing suitable candidates for further testing their efficacy in apoptosis induction. Due to their presence in natural resources, novel drugs and/or the design of functional foods are conceivable from the presented results. In 2021, in the United States, 1,898,160 new cancer cases and 608,570 cancer deaths are expected . In US, Cancer hallmarks include proliferative signaling, elusion of programmed cell death, evasion to immune destruction in favor of replicative immortality, angiogenesis, metastasis, and reprogramming of energy metabolism . The genThe International Cancer Genome Consortium (ICGC) is a global initiative to build a comprehensive catalog of mutational abnormalities encompassing 90 different cancer projects . Eighty-The Cancer Genome Atlas (TCGA) project provides large-scale multi-dimensional analyses of multiple chromosomal aberrations, nucleotide substitutions and epigenetic modifications that drive malignant transformations . In 2014Interestingly, in 2020 , an inteToday, it is widely accepted that cancer risk avoidance as well as health and care, may reside on a proper lifestyle, in which the diet has a fundamental role, in terms of weight control, physical activity and food quality . A healtThe increasing attention towards a proper exploitation of natural compounds also determined the increasing amount of nutrigenomics experiments and associated resources. Among these, the Monarch Initiative  and the http://www.foodb.ca, accessed on 13 May 2021), contains information on more than 28,000 chemicals found in more than 1000 unprocessed food products.FooDB  (accessed on 13 May 2021), we identified foods that contain 4 out of the 23 compounds that resulted potentially useful for the prevention of non-Hodgkin lymphoma and skin melanoma.Based on the appropriate exploitation of currently available bioinformatics resources, we recently identified 734 genes that resulted co-expressed with genes involved in programmed cell death but not reported as implicated in this process by gene ontology (GO) or Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways . We furtWe here present a deeper investigation on the 734 genes from . These gWe investigated the expression of these genes in 231 manually curated results from nutrigenomics human cell-based treatments (treatments with nutrients or bioactive food compounds) analyzed by the Affymetrix microarray, and available from the NutriGenomeDB . Only thp-value < 0.05) when the expression pattern was similar to the one shown in apoptotic-induced treatments, and opposite to cancer.The 149 genes were further investigated for their expression in cancer using GEPIA2 . GEPIA2 Protein-protein interaction analysis on the 7 genes was performed using STRING  that colTo identify additional bioactive natural compounds that could induce similar effects to those caused by the compounds already exploited in the nutrigenomics treatments, we used SwissSimilarity , that alhttp://www.foodb.ca, accessed on 13 May 2021), which contains information on more than 28,000 chemicals found in more than 1000 unprocessed food products. The data in FooDB are collected from textbooks, scientific journals, online food composition or nutrient databases, flavor and aroma databases .Possible unprocessed foods possibly containing one of the bioactive compounds were investigated in FooDB  that, in turn, affect the survival in patients with uterine corpus endometrial carcinoma (CD47), brain lower-grade glioma  and pancreatic adenocarcinoma , respectively  that, in turn, affect the survival plots in patients with uterine corpus endometrial carcinoma (CD47), brain lower-grade glioma (CENPB), kidney renal clear cell carcinoma (PAQR4), kidney renal papillary cell carcinoma (PAQR4), liver hepatocellular carcinoma (PAQR4) and pancreatic adenocarcinoma (SLC44A1)  . Japonicarcinoma . Eusynstarcinoma .The five reference natural compounds (with the exception of the rosemary extract that does not have a defined molecular organization) were used in SwissSimilarity  to identCD47, CENPB, ERGIC1, PPRC1, SLC44A1), that resulted to enhance the survival in uterine corpus endometrial carcinoma (CD47), brain lower-grade glioma  and pancreatic adenocarcinoma . Eleven compounds are similar to Bruceine D, which appears to induce the down-regulation of POMGNT1, and is associated with improved survival in lymphoid neoplasm diffuse large B-cell lymphoma and skin cutaneous melanoma. One compound is similar to Withaferin A, which induces the down-regulation of 4 different genes , associated to an improved survival in uterine corpus endometrial carcinoma (CD47), brain lower-grade glioma (CENPB), kidney renal clear cell carcinoma (PAQR4), kidney renal papillary cell carcinoma (PAQR4), liver hepatocellular carcinoma (PAQR4) and pancreatic adenocarcinoma (SLC44A1). Six compounds are similar to Japonicone A which induces the down-regulation of PPRC1, which enhances the survival in pancreatic adenocarcinoma . Our analysis confirmed the presence in 4 foods for only 4 compounds, all similar in structure with Bruceine D  in 2 cancer types  and highlighted 5 novel marker candidates in 6 cancer types  and in Agaricus bisporus (common mushroom). Noticeably, Oyster mushroom is already known to have an anti-tumoral activity against lymphoma and melanoma [Cynara cardunculus (cardoon) and Cynara scolymus (globe artichoke). Although they are known for their beneficial effects for human health [Among these 4 compounds, O-Acetylcyclocalopin A, Marasmal and Cyclocalopin C1 are predicted to be compounds present in melanoma ,146, whemelanoma . Finallyn health , here, fIn this study, we employed a computational strategy to appropriately exploit bioinformatics data resources and tools to investigate on novel gene candidates involved in apoptosis, and affected in their expression by compounds identified by nutrigenomics experiments. This effort resulted in 7 novel genes that have been shown to be down-regulated in apoptosis and in nutrigenomics treatments inducing programmed cell death and up-regulated with an associated unfavorable outcome in 8 cancer types. Among these 7 genes, 5 can be considered novel prognostic candidate markers for the cancer types here considered as well as suitable targets of dedicated molecular investigations for their roles and their interaction with proteins involved in peculiar cell functionalities. On the other hand, a couple of them finds here an independent confirmation, since they are already considered in cancer treatments. In addition, we selected 6 compounds from nutrigenomics treatments and 23 bioactive compounds similar to 5 of them that responded to ADME  prerequisites for being suitable candidates for translational applications.Finally, a preliminary research for potential functional foods resulted in 4 unprocessed foods containing 4 among the 23 bioactive compounds, all of them similar to Bruceine D, potentially useful against non-Hodgkin lymphoma and skin melanoma.A validation of the candidate marker genes here detected, of the effective bioactivity of the selected compounds in treating at least those cancers here highlighted to be associated to the unfavorable up-regulation of these genes, are mandatory to translate these results into practice. Nevertheless, our results highlight also the relevance of public data resources such as the ones here exploited, since they can strongly impact this field of research. The precious opportunity to move from gene expression data to candidate compounds for cancer treatment and prevention, favored by computer-aided selections of potential useful molecules, paves a faster way to novel drugs and functional food design.


Vasopressin is a ubiquitous molecule playing an important role in a wide range of physiological processes thereby implicated in the pathomechanism of many disorders. Its effect is well characterized through V2 receptors, which regulates the water resorption in kidney, while its vasoconstrictory effect through V1a receptor also received a lot of attention in the maintenance of blood pressure during shock. However, the most striking is its central effect both through the V1b receptors in stress-axis regulation as well as through V1a receptors regulating many aspects of our behavior . Vasopressin has been implicated in the development of depression, due to its connection with chronic stress, as well as schizophrenia because of its involvement in social interactions and memory processes. Epigenetic changes may also play a role in the development of these disorders. The possible mechanism includes DNA methylation, histone modification and/or micro RNAs, and these possible regulations will be in the focus of our present review. Arginine-vasopressin (AVP) is a nonapeptide that is synthesized mainly in the supraoptic (SON), paraventricular (PVN) and suprachiasmatic nucleus of the hypothalamus. In addition, AVP is produced in several other brain areas and organs e.g., medial amygdala , bed nucThe best-known role of AVP, as a hormone, is the regulation of salt-water homeostasis , but this special molecule also serves as a neurohormone within the brain involved in more complex mechanisms . SimilarAVP exerts its effect through different, V1a, V1b and V2 receptors , which hTaking into consideration the high prevalence of stress and its contribution to practically all kind of diseases, we will later focus on the stress-regulatory aspect of AVP.AVP is highly implicated in the regulation of the HPA axis  mostly vAVP-induced stress hormone elevation is less sensitive to negative feedback than a CRH-induced one; therefore, it was suggested that this pathway may play a critical role during chronic stress . This thEarly studies focused only on V1b receptor, and a highly specific antagonist was developed (SSR149415) . This coReceptors aside, one might target stress-induced epigenetic changes in the AVP gene. Based upon Engel’s biopsychosocial model  and its Epigenetic mechanisms underlie the molecular background of neural plasticity and neural network formation, a process that requires stable modulation of gene expression ,36. ExpoIn addition to oxytocin, a well-known regulator of labor, lactation as well as social bounding-pregnancy, labor and motherhood induce dynamic changes in the secretion of AVP in mothers and their offspring ,39,40. IPeriodic infant-mother separation during early postnatal life is one of the most commonly used procedures for inducing ELS in rodents. A daily 3 h separation of the pups from their mothers during the first two weeks of life is already enough to induce changes in the hormone homeostasis ,46. For All in all, these sparse studies indicate that ELS may program the AVP genes through epigenetic mechanism (presumably changes in DNA methylation), which may lead to changes in the neuroendocrine systems leading to alteration in stress adaptation and behavioral dysfunction.Three major mechanisms exist for epigenetic reprogramming. (1) The DNA methylation occurs on cytosine nucleic acids that are followed by guanine in the DNA sequence  and is rather specifically targeting single genes, mostly resulting in reduced expression. (2) By contrast, changes in chromatin structure may occur via covalent modification of histone and may increase (via histone acetylation) or decrease (via histone methylation) the access of large portions of DNA to the natural subcellular machinery that governs transcription and translation. Unlike histone modification, DNA methylation is a stable signature of the epigenomic status and is therefore most suitable for explaining long-term processes . TherefoAs introduced above, ELS can cause epigenetic changes both in animal models and humans.Maternal deprivation in mice induced DNA hypomethylation at CpG rich regulatory island (CGI3) region of the AVP gene . As a reParental separation-induced DNA methylation change has also been found in prairie voles . HoweverFurther investigation also showed plasticity of DNA methylation in the brain regulated by testosterone concentration . In maleAll in all, a special signal (sex hormone) can regulate gene expression pattern in the already developed adult brain most probably through the maintenance of DNA methylation patterns. Direct evidence manipulating the methylation pattern is still missing. Nevertheless, this is a very interesting phenomenon, as it supports the idea that gender differences—also in the AVPergic system —may be uMetamphetamine—a psychoactive drug with high addictive potential—can also influence AVP expression by epigenetic changes: it increased TET1- (Ten-eleven translocation methylcytosine dioxygenase 1: an enzyme that might regulate DNA demethylation) and TET3-dependent DNA hydroxymethylation on AVP and CRH genes in rat nucleus accumbens, which is a part of the reward system  receptor gene measured in the blood cells . Right aA few behavioral studies can be found in the literature in connection with AVP and chromatin/histone modification.Our research group demonstrated the possible role of histone modification in the development of schizophrenia-like behavior in genetically AVP deficient Brattleboro rat  . In AVP Microtus ochrogaster) [As mentioned above, AVP plays a crucial role in social behavior. It is one of the most important neuropeptides in the regulation of partner preference, biparental care of pups and selective aggression in socially monogamous prairie vole (ogaster) . In thisogaster) . I.c.v. These preclinical studies suggest a link between histone modifications and phenotypic alteration through changes in AVP or AVP receptor gene expression. However, further studies using histone modifying agent (a hot topic in cancer research ) are stiMicroRNAs (miRNAs) are short (~22 nt long), non-coding RNA molecules that are responsible for posttranscriptional modification of genes, also known as RNA interference. They are anti-sense and complimentary to the RNA product of target genes and reduce the number of proteins without affecting the number of RNA molecules ,69,70. TNumerous miRNA molecules are expressed in the kidney, which regulate water excretion and re-uptake. It is not surprising that these miRNAs might operate in accordance with the AVPergic system, as the major role of AVP as a hormone is to promote translocation of aquaporin 2 (AQP2) to the cell membrane in the collecting duct, thereby increasing water resorption. After in vitro ddAVP  treatment of the rat inner medullary collecting duct cells, a study using microarray assay found 19 potential miRNA molecules, which responded to the V2 receptor agonist administration with a minimum of 1.3-fold change . Most ofFurther studies focused on the effect of miRNA-132 on AVP system . In a st2+ influx. The effect was confirmed in vivo as well, as rats treated with the extract had an increased mRNA expression of CaSR mRNA and miRNA-137 measured by RT-PCR  activation and Ca RT-PCR  mediated via AVP (ddAVP) and V2 receptor activation. Apparently, this effect was cell selective as the number of ECVs from juxtaglomerular origins did not change in the medium, while it decreased from proximal tubule and cortical collecting duct cell origins. Although the proximal tubule cells were of human origin (HK2 cell lines), and the other two were of mouse origin, the ECV uptake was still induced upon ddAVP incubation. Similar results were obtained when the experiments were repeated in vivo: mice systematically injected with fluorescently labelled ECVs and the V2 receptor antagonist, tolvaptan, had an increase in ECV numbers in the urine. Moreover, the authors confirmed clinical human relevance with a case study of an adolescent patient with diabetes insipidus. After intranasal ddAVP treatment, the ECV amount of glomerular and proximal tubule cell origins increased in the urine samples of this patient. This indicates an evolutionary conservative mechanism, which can be studied both by in vitro and in vivo models . ECVs caPeromyscus californicus) dams on diet containing bisphenol A  before, during and after pregnancy gave birth to pups who had increased expression of AVP mRNA in the hypothalamus and hippocampus (among many other brain areas)  B. An assIn relation to miRNAs, research focused on the classical role of AVP, namely salt-water regulation and blood pressure control through vasoconstriction. However, as an indirect evidence of miRNA-AVP connection during stress, it was shown that ELS—combined with morphine administration—induced an upregulation of AVP mRNA in the hippocampus together with downregulation of several miRNA expression .An increasing body of evidence confirms epigenetic contribution to changes in AVP or AVP receptor mRNA level, not only during the early perinatal period , but also in adulthood. DNA methylation is more targeted on a single gene; therefore, it is better characterized in relation to AVP. However, some hint for bidirectional interaction with histone acetylation was also described. To date, miRNAs are implicated in the hormonal, peripheral role of AVP, and less is known about their interaction regarding behavioral alteration. We have to admit, however, that most of the studies remain at the descriptive level, and confirmation of direct interaction by pharmacological modification of the epigenetic changes is still pending.


As HF and diabetes are recognised inflammatory conditions, with reports of enhanced activation of the NLRP3 inflammasome, we will also consider evidence linking microtubule organisation, inflammation and the association to mitochondrial motility. Diabetes is a global pandemic but with limited treatment options for diabetic cardiomyopathy, therefore we also discuss potential therapeutic approaches to target the mitochondrial-microtubule-inflammatory axis.A pathophysiological consequence of both type 1 and 2 diabetes is remodelling of the myocardium leading to the loss of left ventricular pump function and ultimately heart failure (HF). Abnormal cardiac bioenergetics associated with mitochondrial dysfunction occurs in the early stages of HF. Key factors influencing mitochondrial function are the shape, size and organisation of mitochondria within cardiomyocytes, with reports identifying small, fragmented mitochondria in the myocardium of diabetic patients. Cardiac mitochondria are now known to be dynamic organelles (with various functions beyond energy production); however, the mechanisms that underpin their dynamism are complex and links to motility are yet to be fully understood, particularly within the context of HF. This review will consider how the outer mitochondrial membrane protein Miro1 (Rhot1) mediates mitochondrial movement along microtubules  Diabetic Mellitus (DM) remains a global epidemic, with an estimated 463 million cases worldwide in 2019, and is associated with marked morbidity and mortality rates . RecentlMitochondrial dysfunction is a hallmark of HF  and moreDCM is a chronic low grade inflammatory condition, with inflammation associated with the pathogenesis of HF and linked to the development of mitochondrial dysfunction [as reviewed recently ]. KaludeThis review will summarise evidence for the emerging role of dysregulated mitochondrial movement in HF/DCM, the intersection with aberrant mitochondrial dynamics and NLRP3 activity focusing on the involvement of Miro1; as well as considering the putative mechanisms involved.via an equilibrium of fission and fusion  which upon activation (via phosphorylation) recruits cytosolic Parkin , maintenance and degradation (mitophagy). Over time mitochondria can accumulate damage as a result of multiple mutations in nuclear-encoded mitochondrial genes or oxidative damage c Parkin , 21. Parc Parkin .via the formation of tubular protrusions known as nanotunnels, visualised by live-cell confocal imaging and electron microscopy  are essential for regulating mitochondrial size and shape, important for mitochondrial “quality control,” the movement of mitochondria within the cell is also a crucial factor with distribution tightly linked to cellular energy requirements. An elegant study from Bers and colleagues not only captured, using live imaging, fusion and fission events within cardiomyocytes but also tracked mitochondrial movement . Interes2+ dependent manner  for tissue regeneration in models of anthracycline-induced cardiomyopathy identify the intrinsically high Miro1 content of iPSC-MSCs as essential for facilitating mitochondrial relocation and improved cardiac bioenergetics  ; interac2+ binding to Miro1, via two EF-hands within the primary sequence, elicits a conformational change, mediating the association and dissociation of the Miro1-complex assembly from the microtubules, MTs, . This process underpins mitochondrial moment  via MTs to the perinuclear region where they undergo mitophagy . Ab. Ab2+ bipatients  and assopatients . Mitochox status . Organisergetics , 53. Whiergetics , how impitophagy .The presence of MTs in cardiomyocytes has been known for many years . MTs are2+ transients, leading to arrhythmogenesis , are the sites for NLRP3 assembly and HDAC6 knockout (and loss of ubiquitin-binding) leads to impaired inflammasome assembly and activation (Since the acetylation of α-tubulin/NLRP3 is under the control of HDAC6 it is perhaps not surprising that beneficial effects of using HDAC6 inhibitors in preventing IL-1β generation have been demonstrated . Althougtivation . Clearly2+ homeostasis, which was suggested to lead to the dissociation of Miro1 from mitochondria and subsequent impaired mitochondrial movement, stalled mitophagy and accumulation of damaged ROS producing mitochondria that in turn triggered activation of NLRP3.An indirect link between Miro1 and NLRP3 has also been identified in a rat pancreatic cells using high-fat and high glucose stressors to mimic T2DM conditions . SpecifiHere we have highlighted evidence for the physio-pathological role of Miro1-mediated movement of mitochondria along MTs and while the majority of data is from the study of neurons evidence is emerging to support a similar role for Miro1 in the heart . TherefoAdditionally, we have highlighted a potential link between mitochondrial motility and inflammation and the involvement of deacetylation and HDAC6 . AlthougAK and SK wrote and planned the manuscript. ZA and HD contributed to the writing process. SK generated the figures in consultation with AK. All authors contributed and approved the submitted article.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Using Consensus-Based Standards for the Selection of Health Measurement Instruments guidelines, what is the best patient-reported outcome measure (PROM) to evaluate outpatient postpartum recovery?This systematic review identified 15 PROMs evaluating outpatient postpartum recovery in 46 studies, involving 19 165 patients, with most psychometric measurement properties of included PROMs graded very-low-level or low-level evidence. The best-performing PROMs were the Maternal Concerns Questionnaire, the Postpartum Quality of Life tool, and the World Health Organization Quality of Life-BREF; however, these all have significant limitations.There is a need to robustly develop and validate a PROM that can comprehensively evaluate outpatient postpartum recovery. Outpatient postpartum recovery is an underexplored area of obstetrics. There is currently no consensus regarding which patient-reported outcome measure (PROM) clinicians and researchers should use to evaluate postpartum recovery.To evaluate PROMs of outpatient postpartum recovery using Consensus-Based Standards for the Selection of Health Measurement Instruments (COSMIN) guidelines.An initial literature search performed in July 2019 identified postpartum recovery PROMs and validation studies. A secondary search in July 2020 identified additional validation studies. Both searches were performed using 4 databases , with no date limiters. Studies with PROMs evaluating more than 3 proposed outpatient postpartum recovery domains were considered. Studies were included if they assessed any psychometric measurement property of the included PROMs in the outpatient postpartum setting. The PROMs were assessed for the following 8 psychometric measurement properties, as defined by COSMIN: content validity, structural validity, internal consistency, cross-cultural validity and measurement invariance, reliability, measurement error, hypothesis testing, and responsiveness. Psychometric measurement properties were evaluated in each included study using the COSMIN criteria by assessing (1) the quality of the methods ; (2) overall rating of results ; (3) level of evidence assessed using the Grading of Recommendations, Assessment, Development and Evaluations assessment tool; and (4) level of recommendation, which included class A , class B (not class A or class C), or class C (not recommended).In total, 15 PROMs (7 obstetric specific and 8 non–obstetric specific) were identified, evaluating outpatient postpartum recovery in 46 studies involving 19 165 women. The majority of psychometric measurement properties of the included PROMs were graded as having very-low-level or low-level evidence. The best-performing PROMs that received class A recommendations were the Maternal Concerns Questionnaire, the Postpartum Quality of Life tool, and the World Health Organization Quality of Life–BREF. The remainder of the evaluated PROMs had insufficient evidence to make recommendations regarding their use (and received class B recommendations).This review found that the best-performing PROMs currently available to evaluate outpatient postpartum recovery were the Maternal Concerns Questionnaire, the Postpartum Quality of Life tool, and the World Health Organization Quality of Life–BREF; however, these tools all had significant limitations. This study highlights the need to focus future efforts on robustly developing and validating a new PROM that may comprehensively evaluate outpatient postpartum recovery. This systematic review uses Consensus-Based Standards for the Selection of Health Measurement Instruments guidelines to identify the best available multidomain patient-reported outcome measure tools that assess maternal postpartum recovery beyond delivery hospitalization. Postpartum recovery is a multidimensional process, which is associated with women’s long-term physical, psychosocial, and emotional health and infants’ physical and social development.3 Morbidity in the outpatient setting is common following childbirth and frequently persists beyond 6 months post partum.6 Despite this, for most women, the 6-week postpartum appointment remains the only contact with the health care system following childbirth. Although outpatient postpartum recovery domains (aspects or dimensions constituting postpartum recovery) have been proposed ,7 limited guidance is available to clinicians regarding how to best assess postpartum recovery.An estimated 353 000 mothers give birth each day globally.8 Although PROMs have been developed and validated to assess inpatient postpartum recovery,13 to our knowledge, there is currently no consensus regarding which PROM optimally measures the quality of recovery for outpatient postpartum women.15 Systematic review and psychometric analysis of available PROMs may help inform clinicians and researchers regarding whether a suitable measure exists that is capable of adequately assessing outpatient postpartum recovery. Such a multidomain PROM may be useful for screening and identifying women who are recovering poorly and may benefit from targeted interventions and further physician input. Developing a standardized, multidimensional PROM will address an unmet need, which may help reduce morbidity and improve quality of life in the outpatient postpartum period.Patient-reported outcome measures (PROMs), structured questionnaires enabling patients to report their health status, are now widely regarded as the gold standard means of assessing recovery following surgery.16 to determine the best PROM for use after hospital discharge.We performed a systematic review of validated PROMs used to assess more than 3 domains of outpatient postpartum recovery, using Consensus-Based Standards for the Selection of Health Measurement Instruments (COSMIN) methods,CRD42020180533) and follows the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) reporting guidelines and Consensus-Based Standards for the Selection of Health Measurement Instruments (COSMIN) guidelines for performing systematic reviews of PROMs.16 Recovery for purposes of this review was classified into 13 domains adapted from previously proposed postpartum recovery domains as already stated. A comprehensive list of symptoms for each recovery domain is provided in eTable 1 in the This review was registered with PROSPERO : structural validity ; internal consistency (interrelatedness among PROM items); cross-cultural validity  or measurement invariance (whether respondents from different groups respond similarly to a particular item); reliability (ability of a PROM to distinguish between patients); measurement error ; criterion validity (how PROM scores compare to a gold standard measure); hypothesis testing ; and responsiveness (ability to detect change over time between 2 postpartum time points).To be considered an appropriate measure of outpatient postpartum recovery, the PROM required question items covering more than 3 of the aforementioned 13 postpartum recovery domains . We sought validation studies for outpatient postpartum recovery PROMs. We included studies that assessed 1 or more psychometric measurement properties of the identified PROMs used in the outpatient postpartum setting. Articles were screened to determine whether they included any psychometric assessment of the PROMs. Eight psychometric measurement properties were sought and assessed . We also excluded studies not published in English and studies evaluating recovery in the inpatient or antepartum settings (≤5 days post partum). We excluded editorials, letters, theses, and abstracts from scientific meetings and questionnaires that were developed to measure patient satisfaction. Finally, we excluded PROMs that consisted of free-text answers rather than predetermined item responses, such as with Likert scales, because these answers could not be easily compared between patient cohorts and because of the limited interpretability of such PROMs.17 A standardized data collection tool was used by 3 authors  to extract data from the included studies.Shortlisted articles were entered into the Rayyan reviewing system online.As recommended by the COSMIN checklist, assessment of each PROM was achieved by (1) assessment of each included study for risk of bias associated with each of the 8 psychometric measurement properties, with the quality of individual studies  evaluated based on methods used to assess the psychometric measurement property (or properties) of the PROM in each study; (2) psychometric measurement property assessment of the quality of the PROM using the COSMIN checklist, with the PROM graded as sufficient, insufficient, inconsistent, or indeterminate based on results reported in individual studies ; (3) summarizing pooled results from 1 and 2 for each PROM and each of the 8 psychometric measurement properties, to assess the overall PROM quality as sufficient, insufficient, inconsistent, or indeterminate and grading of quality of evidence as high, moderate, low, or very low based on all available studies; and (4) producing summary of findings tables that were then used to make recommendations regarding the selection of the most appropriate PROM to evaluate outpatient postpartum recovery.Adequate content validity was determined if a PROM evaluated at least 66% of postpartum recovery domains (≥9 of 13 domains) as assessed by study authors. Internal consistency was evaluated using total scores wherever possible. If individual domain scores were assessed, then PROMs for this outcome were graded as sufficient if Cronbach α for most domains was at least 0.7. Similarly, for hypotheses tested in multiple studies, a PROM was graded as “sufficient” if most results were in accordance with the tested hypotheses.Grading of the level of evidence was based on assessment of risk of bias, inconsistency, imprecision, and indirectness. Studies with response rates of 70% or higher were not downgraded for selection bias as decided a priori by study authors. For studies that failed to report missing data, the numbers approached and the response rates were downgraded accordingly owing to potential for selection bias.Recommendations were based on each PROM being classified as A, B, or C as follows: (A) PROMs with evidence for sufficient content validity (any level) and at least low-quality evidence for sufficient internal consistency; (B) PROMs categorized not in A or C; and (C) PROMs with high-quality evidence for an insufficient measurement property. PROMs categorized as A were recommended for use, and results obtained with these PROMs may be trusted. PROMs categorized as B had the potential to be recommended for use but required further research to assess their quality. PROMs categorized as C were not recommended for use.63 There were 1277 postpartum studies registered with ClincalTrials.gov, none of which were validation studies for any of the identified PROMs. Three PROMs from the previous scoping review were excluded . Searching of reference lists led to identification of 1 additional PROM . The secondary search specific to the 15 identified PROMs plus a review of the reference lists of included studies identified 9 additional validation studies. The reasons for study exclusion are provided in eTable 2 in the A flow diagram summarizing the 46 included studies and PROMs evaluated in this review is provided in the 64 and 8 were developed in nonobstetric populations. All included studies performed some psychometric measurement property assessment. In total, 19 165 women were evaluated in the 46 included validation studies; 4657 women were evaluated using the 7 obstetric-specific PROMs in 20 studies, and 14 508 women were evaluated using the 8 non–obstetric-specific PROMs in 26 studies.Of the 15 PROMs used to evaluate postpartum recovery, 7 were developed for use in the obstetric population,The SF-36 and the Inventory of Functional Status After Childbirth had the greatest number of validation studies among the non–obstetric-specific PROMs (9 studies) and obstetric-specific PROMs (7 studies). Studies validating the 12-item Short Form Health Survey in the obstetric setting had the highest patient numbers (n = 7496) among all PROMs evaluated. Most of the studies were performed in primarily English-speaking countries and evaluated English versions of PROMs (27 of 46 studies). Twenty-one of the 46 studies evaluated translated versions.The risk of bias for the assessment of the methods for each psychometric measurement property in each of the included studies is summarized in eTable 4 in the The content validity of each of the 15 included PROMs is summarized in The risk of bias, the results, and the overall rating of the results from individual studies are given in eTable 4 in the Structural validity was adequately evaluated in 3 of the included studies. Only the PQOL and Rural Postpartum Quality of Life (RPQOL) PROMs performed well in this psychometric measurement property .Internal consistency was the most frequently evaluated and assessed psychometric measurement property. Studies reported internal consistency either for total scores or for individual domains. Sufficient internal consistency was shown for all obstetric-specific PROMs and for the SF-36, the WHOQOL-BREF, and the Nottingham Health Profile among the non–obstetric-specific PROMs.Although several studies used and evaluated psychometric measurement properties of translated versions of PROMs, none of these studies assessed cross-cultural validity using multiple group factor analysis or differential item functioning for group factors, in line with COSMIN recommendations. Similarly, none of the included studies reported measurement invariance using this method.Sufficient reliability, shown by intraclass correlation coefficients of 0.7 or higher, was reported only in obstetric-specific PROMs with the Inventory of Functional Status After Childbirth, the Barkin Index of Maternal Functioning, and the PQOL. The WHOQOL-BREF showed inadequate reliability with a moderate level of evidence.Measurement error was not assessed in any of the included validation studies using COSMIN recommended methods. None of the included studies for any of the PROMs defined the minimal clinically important difference in score .Hypothesis testing was performed for all but 2 PROMs . The majority of PROMs evaluated for this psychometric measurement property compared scores between different modes of delivery at various time points in the postpartum period, ranging from 1 week to 12 years post partum. The other groups compared included women requiring intensive care admission, maternal morbidity (such as preeclampsia), postnatal depression, and urinary incontinence. The Inventory of Functional Status After Childbirth, the Barkin Index of Maternal Functioning, the Maternal Postpartum QOL tool, the PQOL, the SF-36, the European quality of life 5-dimension questionnaire, the WHOQOL-BREF (moderate level of evidence), and the World Health Organization Disability Assessment Schedule 2.0 were the only PROMs that reported “sufficient” results associated with the majority of hypotheses evaluated.Responsiveness of PROMs was deemed sufficient with the Inventory of Functional Status After Childbirth and SF-36, which were graded with a low level of evidence. This psychometric measurement property was inconsistently evaluated among the included studies. Several studies reported scores at different postpartum times without formal statistical comparison between groups, thus allowing for only qualitative reporting and evaluation. Those studies were scored as inadequate for methods. None of the PROMs showed sufficient consistency in terms of responsiveness or ability to detect change over time. This is likely attributable to a large range in postpartum time points evaluated between studies and different PROMs. It was a requirement for all domains  to demonstrate change between time points to be considered to have sufficient responsiveness.46 Response rates were not reported in 14 of the included studies. Five studies reported response rates but did not include information regarding numbers of women approached to participate in the study. Twenty-one studies reported numbers approached and response rate without reporting missing data from surveys completed , and although it achieved a class A level of recommendation, this was based on data from a single validation study involving 221 patients in Australia.The MCQ is based on data from 5 non–peer-reviewed theses with the number of questions ranging from 46 to 51 and either very low or low levels of evidence. This PROM is based on input from 19 mothers and 6 nurses. The MCQ fails to evaluate sleep; it has not been used beyond 2 weeks post partum; it has limited data regarding its cross-cultural validity, reliability, measurement error, hypothesis testing, and responsiveness; and it has very low levels of evidence supporting findings of its psychometric measurement properties. The time taken to complete the MCQ is 15 to 20 minutes.The main finding of this systematic review is a lack of adequately validated PROMs for evaluation of postpartum recovery. The best available PROMs for outpatient postpartum recovery are the MCQ, PQOL, and WHOQOL-BREF; however, these PROMs all have significant limitations. None of the 15 PROMs assessed in our review evaluated all 13 outpatient postpartum recovery domains. Therefore, to our knowledge, there are currently no PROMs for outpatient postpartum recovery assessment that can be recommended for widespread use. These findings highlight the need to robustly develop and validate a new PROM that comprehensively evaluates outpatient postpartum recovery.66 The effects of poor postpartum recovery may be associated with impaired long-term women’s health and infant development.4 There is, therefore, a clinical need to assess recovery in the postpartum period. The use of PROMs is considered to be the gold standard to measure postoperative recovery, and scores on PROMs have been used to guide value-based hospital reimbursements in certain health care systems following elective surgery.67 In 2018, the American College of Obstetricians and Gynecologists recommended routine follow-up for all mothers to be performed within 3 weeks of delivery followed by a comprehensive postpartum visit no later than 12 weeks after birth.68 A PROM that performs well in measures of validity, reliability, and responsiveness would be invaluable for obstetricians and may help screen patients prior to these consultations to facilitate the direction of the discussion.Approximately 33% of maternal deaths occur between 1 week and 1 year following childbirth, and impaired postpartum recovery may lead to delays in returning to work.7 The heterogeneity evident among the PROMs used in postpartum recovery studies justifies the need to perform systematic reviews that evaluate the psychometric measurement properties of existing PROMs and help guide clinicians and researchers to decide which PROM to use. Our recommendations may direct future research efforts by encouraging the use of the most appropriate PROMs with adequate content validity and internal consistency. The use of optimal PROMs may maximize efficiency of research efforts by optimizing the quality of data that can be pooled from future studies and by facilitating interpretation of findings.A previous review identified 201 PROMs that have been used to evaluate postpartum recovery.69 This process may help determine which domains should be evaluated and whether specific domains should be weighted according to the level of perceived importance to key stakeholders.This systematic review provides recommendations regarding which PROMs to consider using when assessing outpatient postpartum recovery. Future studies are needed to identify the best PROMs for use by clinicians and researchers to assess individual postpartum recovery domains, such as depression, pain, and sleep. Although this study identified the MCQ, PQOL, and WHOQOL-BREF as the best available PROMs to evaluate postpartum recovery, the 12 PROMs that received class B recommendations require further psychometric measurement property evaluation. However, given the described limitations of the PROMs that received class A recommendations in this analysis, future research efforts should focus on developing and validating a comprehensive PROM of outpatient postpartum recovery. This may be achieved, for example, by using published methods endorsed by the Patient-Reported Outcomes Measurement Information System initiative (PROMIS).Because no gold standard measure of postpartum recovery currently exists, we were unable to assess the criterion validity of included PROMs. We did not exclude studies based on the amount of time after delivery that PROMs were completed, and we acknowledge that this may have influenced the rating for hypothesis testing because the postpartum time point is likely to determine the ability of a PROM to detect differences between groups. We also acknowledge that internal consistency was reported in various ways among the included studies. Some studies reported statistics related to individual domains, whereas other studies reported Cronbach α related to the total PROM score. For studies reporting individual domains, we considered the overall rating to be sufficient if most of the domains had a Cronbach α of 0.7 or higher. We also deemed hypothesis testing to be sufficient if the majority of results presented were in accordance with the hypotheses tested. We believe that, by adopting this pragmatic approach, we consistently assessed the PROMs. Finally, scoring and ranking the quality of the methods, the interpretation of results, and the grading of evidence is a subjective process. However, all included articles were reviewed by 3 authors, and we believe that discussion among these authors resolved discrepancies and reduced variability in interpretation, thereby minimizing the chance of misclassification.This analysis provides a detailed assessment of the psychometric measurement properties of the PROMs that have been used to assess outpatient postpartum recovery. The best-performing PROMs were the MCQ, PQOL, and WHOQOL-BREF; however, all of these tools have significant shortcomings. Therefore, this study highlights the need to robustly develop and validate a new PROM that comprehensively evaluates outpatient postpartum recovery.


Merkel cell polyomavirus (MCPyV) infection causes near-ubiquitous, asymptomatic infection in the skin, but occasionally leads to an aggressive skin cancer called Merkel cell carcinoma (MCC). Epidemiological evidence suggests that poorly controlled MCPyV infection may be a precursor to MCPyV-associated MCC. Clearer understanding of host responses that normally control MCPyV infection could inform prophylactic measures in at-risk groups. Similarly, the presence of MCPyV in most MCCs could imbue them with vulnerabilities that-if better characterized-could yield targeted intervention solutions for metastatic MCC cases. In this review, we discuss recent developments in elucidating the interplay between host cells and MCPyV within the context of viral infection and MCC oncogenesis. We also propose a model in which insufficient restriction of MCPyV infection in aging and chronically UV-damaged skin causes unbridled viral replication that licenses MCC tumorigenesis. Merkel cell polyomavirus (MCPyV) infection can be detected on the skin of most healthy adults  associates with LT in replication centers where it amplifies MCPyV replication by recruiting replication factor C (RFC)   binding sites similar to other polyomaviruses, but this binding activity is not required for transformation of rat fibroblasts  cluster domains   cells, highly expressed miR-M1 transcripts down-regulated the expression of MCPyV T antigens and MCPyV replication  digest collagen fibers and activate chemokines  is dramatically repressed in MCC cell lines and tumor cells  the epidemiology of MCC and MCPyV infections, (2) existing research on fibroblast biology and wounding responses in human skin, and (3) in vivo as they are ex vivo and in vitro . In non-malignant human cells, HPVs, like MCPyV, normally replicate and maintain their genomes as episomes  Grants R01CA187718, R21AR074073, R21AI149761, the NCI Cancer Center Support Grant (NCI P30 CA016520), and the Penn CFAR pilot award (P30 AI 045008).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


In the evolving landscape of highly multiplexed imaging techniques that can be applied to study complex cellular microenvironments, this review characterizes the use of imaging mass cytometry (IMC) to study the human kidney. We provide technical details for antibody validation, cell segmentation, and data analysis specifically tailored to human kidney samples, and elaborate on phenotyping of kidney cell types and novel insights that IMC can provide regarding pathophysiological processes in the injured or diseased kidney. This review will provide the reader with the necessary background to understand both the power and the limitations of IMC and thus support better perception of how IMC analysis can improve our understanding of human disease pathogenesis and can be integrated with other technologies such as single cell sequencing and proteomics to provide spatial context to cellular data. The predominant methods for cell identification during the pathological analysis of formalin-fixed paraffin-embedded (FFPE) samples from human renal core biopsy tissues include cell morphology and immunohistochemistry or immunofluorescence (IF). The limited amount of tissue obtained from renal biopsy confines the type of analysis performed, preventing extensive analysis at the molecular and cellular level. Thus, most analyses are descriptive, with few efforts to provide quantitative information about the tubular, stromal, and nonresident cell populations in disease states . Single-2 per pixel  is a powerful analytical platform in which a high-resolution laser is combined with a mass cytometer that permits mass spectrometry-based, spatially preserved high-dimensional analysis of intact FFPE and frozen tissues at a resolution of 1 μmer pixel . In IMC, imaging .2 resolution provides the ability to identify basic subcellular localization of antigens (nucleus vs. cytoplasm), while the large number of antibodies that can be simultaneously analyzed supports both cell identification and cell activation state determination. The technique can therefore be employed to identify cellular markers that provide fundamental tissue architectural layout, as well as secondary cellular responses such as protein modifications, signaling pathway activation, cell injury states, and cell proliferation. IMC allows immune cell markers to be investigated in both healthy and diseased tissues with their distribution pattern and proximity to tissue resident cells spatially mapped  a means to investigate the cellular heterogeneity of the tumor microenvironment and understanding of cancer progression and resistance to current therapies ; (ii) a Currently there are three publications showing IMC analysis of human kidney tissue  and one in vitro cell expression, morphologic features identified by a pathologist, and cell co-localization of more than one cell-specific marker. Kidney-MAPPS was then employed to successfully generate a two-dimensional quantitative atlas of 22 distinct cell populations that were identified in the normal human kidney. In addition to well-characterized parts of the tubule, interstitium, and glomerulus, the analysis revealed several cell populations that had not yet been described. A population of megalin (low), aquaporin-1 (AQP1)+, aquaporin-2 (AQP2)−, cytokeratin-7 (CK7)+ tubular cells in the medulla were assigned to represent the thin descending limb of the loop of Henle. A cluster of megalin+, AQP1+, vimentin+ cells was suggestive of an injured, fibrotic, or regenerative cell type, while CK7+, AQP2+ cells were felt to represent principal cells. Moreover, the analysis identified cells in connecting tubular segments with morphologically larger diameter than the distal convoluted tubule (DCT) as calbindin+, CK7+, AQP2+ transition cells between the DCT and collecting duct. The Kidney-MAPPS protocol accurately identified, quantified, and localized ~92% of all cells in the human kidney, and quality control measures for validity and reproducibility of unsupervised analysis of data showed no significant differences in the manual vs. Kidney-MAPPS assigned phenotypes, out of over 10,000 cells scored  to quantitatively characterize the cellular makeup of the reference human kidney . The ants scored . Furthers scored .+ proximal tubular cells exclusively in deceased donor tissue that was surrounded by macrophage-rich infiltrates and co-expressed markers of injury and proliferation, kidney injury molecule 1 (KIM-1) and Ki67, respectively  following implantation as compared to living donor kidneys that were low risk for subsequent DGF . The resectively . This isectively . Moreoveectively . These r+ macrophages, CD11c+ dendritic cells, and CD19+ B cells in two of the COVID-19 patient kidney samples  to VK and LC. Additional support came from the German Research Foundation to MW (WE6653/1-1).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Wolfram syndrome (WFS) is a rare autosomal recessive neurodegenerative disease whose diagnosis requires diabetes mellitus and optic atrophy (OA). WFS includes a wide spectrum of other possible complications such as diabetes insipidus, sensorineural deafness, urinary tract problems, neurological and psychiatric disorders. Most WFS patients show type 1 syndrome (WFS1) caused by mutations in the WFS1 gene, encoding Wolframin protein, while few patients are affected by WFS type 2 (WFS2) due to a pathogenetic variants in the CISD2 gene encoding an endoplasmic reticulum intermembrane small protein. WFS2 is considered a phenotypic and genotypic variant of WFS, from which differs only for the increased risk of bleeding and presence of peptic ulcers. OA and diabetes are considered cardinal features of WFS. We hereby report the ophthalmologic evaluation in a patient, previously described, with WFS2 after 8 years of follow-up. A 20-year-old white woman was referred to our retinal center for the first time in 2012 following a diagnosis of a novel intragenic exon 2 CISD2 homozygous deletion, for the suspicion of an associated bilateral OA. Fundus examination, spectral-domain optical coherence tomography, visual field, visual evoked potentials were performed and confirmed the presence of an optic neuropathy that remained stable over 8 years follow up. A slowly progressive optic neuropathy, rather than OA can characterize patients with WFS2 and CISD2 intragenic deletion. Wolfram syndrome (WFS) is a rare autosomal recessive neurodegenerative disease characterized by a wide phenotypic spectrum including non-autoimmune diabetes mellitus (DM) , and optOur group reported the first Italian Caucasian girl with WFS2 , carryinLater on, some authors reported other two Italian siblings with a novel homozygous CISD2 mutation mapping within the donor splice site of intron 1. Authors hypothesized that the alteration presumably would have caused a skipping of exon 1 with subsequent disrupting of the mRNA splicing by eliminating exon 2 . Among tWe hereby report the evolution of optic neuropathy over 8 years of follow-up of our patient with WFS2 and CISD2 intragenic deletion.The previous clinical history of our patient, who currently is a young woman of 28 years, has been described in the previous report . She camActually, her diabetes is moderately-controlled  with a very low total daily dose of insulin requirement (<0.5 IU/kg).The annual audiological evaluation showed a stable mild bilateral sensorineural hearing loss on high frequencies, without the need for hearing aids throughout the follow-up period.She is still on pump inhibitor therapy and no longer presented peptic ulcers at endoscopic controls.From July 2014, medical history was characterized by the presence of recurrent febrile urinary tract infections. Renal ultrasonography was performed showing grade IV vesicoureteral reflux (VUR), for which prophylactic antibiotic therapy was started, and bilateral pyelectasis with thickened distended bladder. Voiding cystourethrography and urodynamic test confirmed severe VUR and showed a low-capacity corrugated-walled bladder with sphincteric dyssynergia and pathological post-voiding residue. Diagnosis of neurogenic bladder was made, and the attempt of oxybutynin therapy failed due to drug side effects (blurred and flashing vision). Intermittent bladder catheterization was started. Patient underwent annual endoscopic examinations for peptic ulcers, which did not recur, and had no further bleeding episodes. Apart from the neurogenic bladder, our patient did not develop any other neurological disorders. Currently, she has a successful career as a lawyer.In March 2020, she performed the last brain MRI that showed a slight increase in the antero-posterior diameter of both eyes with mild prominence of the CSF sheaths of both optic nerves, mild lower worm hypoplasia and multiple T2/FLAIR hyperintense areas in the cerebral bi-hemispheric white matter.From 2012 to date, she has been regularly followed up at the Referral Center for Inherited Retinal Dystrophies of the Eye Clinic, University of Campania “Luigi Vanvitelli.” Periodic ophthalmological examinations were performed, including best corrected visual acuity (BCVA), measurements with the Snellen visual chart, slit lamp anterior segment examination, measurements of intraocular pressure, fundus examination, Octopus visual fields assessment (Octopus 900 Haag Streit), electroretinography (ERG) and visual evoked potentials (VEP) . Spectral-domain optical coherence tomography  was performed to evaluate retinal nerve fiber layer (RNFL) thickness and monitoring disease progression.At the last ophthalmic examination in September 2020, patient's BCVA was 20/40 in right eye (RE) and 20/50 in left eye (LE), the same visual acuity presented at the first visit in 2012. Anterior segment slit lamp examination and intraocular pressure (<20 mmHg) were normal bilaterally at each control. Pupils were isochoric, normoreactive to light and no relative afferent pupillary defect was detected. Patient presented no signs of nystagmus, strabismus, and dyschromatopsia on color test evaluation. Fundus examination was stable over time showing bilateral optic disk temporal pallor with normal macula in both eyes and no signs of diabetic retinopathy . MacularTo the best of our knowledge, this is the longest follow-up in the literature in a patient with WFS2 and CISD2 intragenic deletion.In this report we show that WFS2 can be characterized by an optic neuropathy rather than an OA, with a very slow progression over the time.To date, few cases of WFS2 with mutations in CISD2 have been reported in addition to our case: 3 consanguineous families of Jordanian descent , two ItaOur patient was followed up yearly and the clinical picture including all macular and optic nerve examinations along with the BCVA remained stable over time. In our patient the mutation detected probably led to encode a protein that doesn't cause OA but optic neuropathy.Our findings further confirm that the involvement of the optic nerve in patients with WFS2 does not always manifest with OA. Additionally, the optic neuropathy progression over the years can be very slow allowing for a stable visual acuity.Ophthalmological evaluations in WFS2 patients with CISD2 intragenic deletion should be performed longitudinally. All retinal examinations may be helpful in distinguish the patients presenting with OA from those presenting with optic neuropathy, considering the differences in term of prognosis. Indeed, WFS symptoms have a negative impact on individuals' daily function and quality of life. Apart from the finding of a neurogenic bladder, the clinical picture of our patient has been stable over the years, especially as regards the optic neuropathy which allowed her to maintain a good visual acuity and not go blind. She also managed to graduate, due to the stability of the clinical picture over time. The long follow-up of our patient allows us to hypothesize that not all WFS2 cases evolve to blindness. Further studies with comprehensive evaluation of the visual function of patients with WFS2 are needed to clarify the results of our study.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. The patients/participants provided their written informed consent to participate in this study.VD and EM were responsible for the initial plan and study design. VD, EM, FR, and FD collected and extracted data. VD and EM interpreted data and drafted the manuscript. VD, EM, FR, FD, RG, FT, CI, AF, and FS did a critical revision of the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


SGK1 mRNA and protein were found to be elevated in the spinal cord and brain in various pathological pain models. Blocking SGK1 significantly attenuated pain-like responses and the development of pathological pain. These studies provide strong evidence that SGK1 plays a role in the development of various types of pathological pain and that targeting SGK1 may be a novel therapeutic strategy for pain management. In this review article, we provide evidence from animal models for the potential role of SGK1 in the regulation of pathological pain caused by inflammation, nerve injury, psychiatric disorders, and chronic opioid exposure.Currently, the management of acute and chronic pain in clinical practice remains unsatisfactory due to the existence of limited effective treatments, and novel therapeutic strategies for pathological pain are urgently needed. In the past few decades, the role of serum and glucocorticoid-inducible kinase 1 (SGK1) in the development of pain and diurnal rhythms has been implicated in numerous studies. The expression levels of  A large body of evidence implies that presurgical psychological vulnerability, such as anxiety, is a powerful risk factor for the development of postsurgical hyperalgesia (Kehlet et al., As mentioned above, chronic pain is also a strong risk factor for the development of negative emotions such as anxiety, depression, and stress (de Heer et al., SGK1 mRNA was increased in the spinal cord of non-fear-conditioned rats following an intraplantar formalin injection. In contrast, the formalin-induced increase in SGK1 mRNA expression, which tended to strongly decrease, was not observed in fear-conditioned rats. Although it failed to reach statistical significance, the suppression of pain-like response might be associated with the failure of formalin-induced upregulation of SGK1 mRNA expression in fear conditions (Olango et al., Fear-conditioned analgesia is a phenomenon in which an animal is exposed to conditioned aversive stimuli, resulting in the profound suppression of pain responses. Olango et al. found that the expression level of SGK1 mRNA was observed in the striatum of mice after acute morphine stimulation, and a selective glucocorticoid receptor antagonist attenuated the morphine-evoked upregulation of SGK1, suggesting that the alteration of SGK1 induced by morphine depends on glucocorticoid receptor signaling (Slezak et al., SGK1 mRNA is upregulated in whole brain lysates following chronic oxycodone administration, a mu opioid receptor agonist (Hassan et al.,  SGK1 mRNA in wild-type mice but not in mu-opioid receptor knockout mice. However, a single injection of morphine did not alter the expression of SGK1 mRNA. These findings indicated that SGK1 was upregulated after chronic treatment with morphine and subsequent mu-opioid receptor activation (Befort et al., Opioids have powerful analgesic effects and are widely used in clinics for the treatment of acute and cancer pain. However, chronic drug exposure unavoidably leads to many side effects, such as addiction and opioid tolerance, thereby limiting opioids’ application in chronic pain management. Chronic opioid-induced cellular alterations remain unclear. Alterations of SGK1 levels in the spinal cord and brain have been implicated after opioid administration. For example, an increase in The role of SGK1 in opioid tolerance has been further defined in a recent study. In this study, Li et al.  showed tIn this review article, we summarize the role of SGK1 in pathological pain. These studies have provided evidence that SGK1 plays a critical role in the development of inflammatory pain, neuropathic pain, emotional dysfunctions related to pain, and opioid tolerance. Treatment with the SGK1 inhibitor GSK-650394 or small interfering RNA could relieve nociceptive hypersensitivity in various pathological pain models, suggesting that SGK1 may represent a potential therapeutic target for pain management. However, there are currently no clinically approved drugs known to inhibit the activity of SGK1, and few clinical trials have studied SGK1 inhibitors in pain therapies. Although the clinical value of analgesia therapy targeting SGK1 has not been clearly confirmed, preclinical studies of SGK1will provide a basis for clinical trials and drug development. Therefore, more selective and clinically relevant drugs targeting SGK1 require further exploration in future studies and clinical trials.BL and NL wrote and edited this review. ZH contributed in production of figures. XZ and GD drafted the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The major challenge of DNA replication is to provide daughter cells with intact and fully duplicated genetic material. However, various endogenous or environmental factors can slow down or stall DNA replication forks; these replication problems are known to fuel genomic instability and associated pathology, including cancer progression. Whereas the mechanisms emphasizing the source and the cellular responses of replicative problems have attracted much consideration over the past decade, the propagation through mitosis of genome modification and its heritability in daughter cells when the stress is not strong enough to provoke a checkpoint response in G2/M was much less documented. Some recent studies addressing whether low replication stress could impact the DNA replication program of the next generation of cells made the remarkable discovery that DNA damage can indeed be transmitted to daughter cells and can be processed in the subsequent S-phase, and that the replication timing program at a subset of chromosomal domains can also be impacted in the next generation of cells. Such a progression of replication problems into mitosis and daughter cells may appear counter-intuitive, but it could offer considerable advantages by alerting the next generation of cells of potentially risky loci and offering the possibility of an adaptive mechanism to anticipate a reiteration of problems, notably for cancer cells in the context of resistance to therapy. Accurate replication of the human genome in S-phase and faithful segregation of sister chromatids in mitosis are fundamental for the maintenance of chromosome stability from one generation of cells to the next . Cells tSpecific chromosomal domains have a higher propensity for DFSs and include intrinsically hard-to-replicate repetitive sequences which form secondary non-B DNA structures , as well as late S-phase replicating regions or loci with a paucity of active and dormant origins . A greatRS is a major cause of pathologies, including cancer, premature ageing, and other disorders associated with genomic instability ,8. WhilePOL H gene mutations. This late DNA replication MiDAS was then confirmed to take place at broken CFS loci by combining EdU detection with fluorescent in situ hybridization (FISH) in cells exposed to low doses of aphidicolin treatment [Cells that have not fully replicated their genomic DNA are found to enter mitosis when an ultimate mechanism to complete DNA replication at these under-replicated loci, called the mitotic DNA synthesis or MiDAS, takes place . In factreatment . The stireatment . MiDAS hreatment ,15.The switch from normal DNA replication to MiDAS seems to be due to the action of the ubiquitin ligase TRAIP that provokes replisome disassembly in response to uncompleted DNA replication and allows the accessibility of nucleases and repair factors . When ceCollectively, these observations established that, while the S-phase was conventionally believed to end before the G2-phase of the cell cycle, it can extend almost to the stage of cell division, at least at some difficult-to-replicate chromosomal domains. A successful replication completion by MiDAS could limit the formation of ultrafine anaphase bridges (UFBs) which impede the proper partition and affect the integrity of chromosomes and the transmission of DNA damage to the next generation of cells (see next paragraphs).When unresolved by MiDAS, late replication intermediates at CFSs following replication stress can lead to physical connections, referred to as ultra-fine DNA bridges (UFBs), between sister chromatids before anaphase that contain a partially denatured, single-stranded or stretched DNA conformation. This affects proper chromosome segregation and results in sister chromatid nondisjunction, a potential source of genome instability . UFBs caWhen not processed during mitosis, under-replicated DNA regions or DNA breaks at CFSs generated by stressed mother cells can be inherited by unstressed daughter cells where they become sequestered and shielded by NBs that contain 53BP1, a large adaptor protein that interacts with the DNA scaffold made of histones and histone-binding proteins involved in DNA double-strand break (DSB) repair ,21. This is not surprising since to repair DNA breakage during the G1 phase of the cell division cycle, the HR pathway must be actively suppressed to avoid genomic changes. Such HR inhibition can be achieved by 53BP1 in order to block the molecular actors that create single-strand DNA at the break sites, in preparation for recombination .These findings reveal the concept that a response to RS may also occur in otherwise unstressed G1 daughter cells. As for MiDAS, such an inherited response was documented in different stress contexts. For example, a significant increase in the frequency of 53BP1 NBs was observed in cells lacking the MUS81 nuclease or BRCA2, and their frequency was further elevated when the two proteins were concomitantly depleted . A signiIt has recently been explored how cells deal with inherited under-replicated DNA sequestered in 53BP1-NBs . Live ceThis mechanism, which highlights how 53BP1-NBs restore the loci at risk and limit genome instability after passage through mitosis, also suggests the idea that full genomic DNA duplication may take more than a single cell division cycle.At the exit of mitosis, the transition from highly compact chromatin to a less compact interphase chromatin overlaps with the loading of replication origin licensing factors, in particular the ORC complex, which are essential for executing proper DNA replication. ORC serves as a scaffold for the subsequent association of CDC6 and CDT1, which together coordinate the loading of the MCM2-7 complex in order to form the prereplication complex (pre-RC) required for replication fork formation and activity . RecentlReplication factories contain clusters of several origins located in physical proximity to each other that fire nearly simultaneously. This organization involves the formation of chromatin loops corresponding to the inter-origin DNA regions, whose size correlates with the length of the replicons, facilitating access of the initiator proteins required to activate origins. We recently reported that RS in mother cells can induce a spatial re-organization of activated origins in G1 daughter cells . Indeed,The changes in MCM loading and origin selection in daughter cells may affect the timing of activation of replication origins of a subset of domains in the subsequent S-phase. To test this hypothesis, mother cells were released from aphidicolin and the replication timing (RT) of the next generation of cells was analysed by using human whole genome microarrays ,28. StriWhereas the mechanisms highlighting the source and the cellular responses of RS have attracted much attention over the last 15 years, the propagation through mitosis of genome modification, and its heritability in daughter cells when the stress is not strong enough to provoke a checkpoint response in G2/M, was much less documented. These recent findings (summarized in Transmission of replication problems may be an essential step to mark such problematic loci as potentially risky and may thus open up the possibility of a “memory” adaptive response among daughter cells to ensure an efficient replication of these hazardous DNA sequences in the next S-phase. During the clonal evolution of cancer cells, which are often defective in checkpoint and show a high degree of oncogene-induced endogenous replicative stress, such a process may be exacerbated and may contribute to the survival of a cellular clone, especially when exposed to the multiple therapeutic treatments used in clinics that target DNA replication.