Aging is a primary risk factor for several diseases, including neurodegenerative disorders and cancer, with DNA damage accumulation being one of the key drivers. Frailty is a severe form of aging that affects many older adults and is characterized by dysfunction across multiple body systems, leading to increased vulnerability and late-life disability. The pathogenic mechanisms differentiating frailty from healthy aging, however, remain unclear. Here, we compared the DNA damage response (DDR) in skin fibroblasts and circulating free DNA (cfDNA) levels in the blood of frail individuals with those of healthy young and elderly controls. To infer a molecular fingerprint of frailty, we also sequenced the methylated cfDNA to identify potential tissue-specific dysfunction. Frail subjects exhibited significantly higher baseline levels of DNA damage, as evidenced by increased γH2AX and 53BP1 foci in fibroblasts under basal conditions. However, no significant differences in DDR capacity were observed post-irradiation between frail and age-matched controls. Elevated cfDNA levels in frail individuals correlated with higher TNF-α levels, suggesting a link between cfDNA, inflammation, and frailty. cfDNA methylation analysis revealed an abundance of hypomethylated fragments and identified a distinctive fingerprint of differentially methylated regions that discriminated frail subjects from age-matched controls. Moreover, deconvolution analysis identified the small intestine as a potential cfDNA tissue of origin, implicating a role of this anatomical site in frailty pathobiology. Our findings provide a molecular basis for understanding frailty and its links to aging-related diseases and may open avenues for biomarker development and targeted interventions.

Field cancerization describes molecular alterations occurring in histologically normal tissues surrounding tumors that may contribute to cancer initiation and progression. In prostate cancer (PCa), the molecular characteristics of peritumoral tissue (PTT) remain incompletely understood. Because microRNAs (miRNAs) play key roles in gene regulation, tumor progression, and microenvironmental remodeling, we investigated miRNA expression patterns and regulatory networks across benign tissue (BT), PTT, and tumor tissue (TT). Small RNA sequencing was performed on matched formalin-fixed paraffin-embedded samples from 40 patients with PCa. Differential expression analysis was conducted using DESeq2, adjusting for age and Gleason grade, while functional enrichment analysis and weighted gene co-expression network analysis (WGCNA) were used to identify dysregulated pathways and conserved miRNA modules. PTT exhibited a molecular profile intermediate between BT and TT, consistent with a field cancerization effect. Compared with BT, 102 miRNAs were differentially expressed in TT and 57 in PTT, with 39 miRNAs (68% of the PTT-associated miRNAs) overlapping the tumor signature. Shared dysregulated pathways included PI3K-Akt, p53, and HIF-1 signaling; whereas, PTT showed additional enrichment in pathways related to epigenetic regulation (Polycomb Repressive Complex) and cellular stress responses (mitophagy, protein processing in ER) exclusively through up-regulated miRNAs; no pathways were uniquely enriched from down-regulated miRNAs in PTT. WGCNA identified conserved miRNA modules enriched for members of the let-7, miR-200, miR-103/107, and miR-106a~363 families, which have established roles in epithelial-mesenchymal transition, tumor progression, and microenvironmental remodeling. Collectively, these findings demonstrate that histologically benign peritumoral tissues harbor tumor-associated miRNA programs and regulatory networks that closely resemble those observed in prostate tumors, providing molecular evidence of field cancerization in PCa and identifying potential miRNA-mediated mechanisms relevant to disease progression and biomarker development.

Background/Aim:Breast cancer is the most common cancer in women worldwide, and early detection remains a significant challenge. Recent studies have identified increased expression of Mammaglobin A (Q13296, Gene: SCGB2A2) mRNA in breast cancer, suggesting its potential as a disease marker, although its function is not fully understood. To elucidate Mammaglobin's role, this study sought to identify co-expressed miRNAs and analyze the biological pathways they regulate. Materials and Methods:Using TCGAbiolinks and Firebrowse, miRNA and gene expression data were collected from 86 patients, including tumor and normal tissue samples from the Cancer Genome Atlas (TCGA) Breast Cancer cohort. Transcriptomic data were analyzed with DESeq2, and a Spearman correlation was calculated for significant p-values, which were further explored using enrichment tools and target gene databases. Results:DESeq2 was used to identify differential expression of miRNAs between normal and tumor breast tissues. Out of 782 miRNAs differentially expressed in breast cancer, hsa-mir-184 and hsa-mir-190b showed a significant positive correlation with SCGB2A expression. These markers were also upregulated in breast cancer tissues compared to normal tissues. Bioinformatics analysis revealed that hsa-mir-184 and hsa-mir-190b play important roles in cancer and cellular proliferation. These miRNAs target a wide range of genes, including sorting nexin 9 (SNX9) and annexin 6 (ANXA6), which are involved in membrane stability, vesicular trafficking, and cell mobility, and they contribute to cancer metastasis. Conclusion:The positive correlation among the expression of hsa-miR-184, hsa-miR-190b, and SCGB2A2 suggests that they may participate in shared biological pathways. These pathways govern critical cellular processes, such as membrane trafficking and cell signaling, which are frequently disrupted in cancer. Consequently, these findings enable a better understanding of the role of Mammaglobin in breast cancer signaling.

Background: MicroRNA (miRNA) dysregulation plays a major role in glioblastoma (GBM) biology, but clinical implications are still incompletely defined. We conducted an exploratory miRNA profiling study of GBM in a Colombian cohort, associating molecular information with clinical characteristics. Methods: We retrospectively analyzed all adult patients treated surgically between 2015 and 2023 in two different institutions. Tumor samples were sequenced for small RNA. Associations with age, Ki67, and clinical variables were analyzed by differential miRNA expression analyses. Pathway enrichment using curated miRNA annotation databases was performed. Survival outcomes were evaluated using univariate Cox models. Results: 29 GBMs were included (23 histology grade 4; 6 molecularly reclassified grade 2–3). Increasing age correlated with downregulation of hsa-miR-17-5p and hsa-miR-92a-3p, while tumors with Ki67 ≥ 20% showed upregulation of hsa-miR-17-5p and hsa-miR-20a-5p, implicating the oncogenic miR-17-92 cluster in age- and proliferation-associated regulation. In the full cohort, only hsa-miR-31-5p had a significant correlation with overall survival. Pathway analyses revealed enrichment of miRNAs annotated to signaling pathways implicated in GBM biology. Conclusions: We identified age- and proliferation-associated miRNA patterns centered on the miR-17-92 cluster. The opposing regulation of this cluster with age (downregulation) versus high proliferative activity (upregulation), coupled with its lack of survival association, indicates that these patterns reflect underlying tumor biological heterogeneity rather than serving as prognostic biomarkers. These findings should be considered hypothesis-generating. Larger studies integrating molecular and surgical variables are necessary to establish the clinical utility of miRNA profiling in GBM

Background/Objectives: The oral-gut-lung axis represents a dynamic system where exosomes carrying mRNAs and non-coding RNAs might help to regulate microbiota and human cell crosstalk to establish transcriptional regulatory networks controlling cellular biological processes and signaling pathways. Methods: We conducted a comprehensive transcriptomic analysis to characterize the molecular cargo of extracellular exosomes in the context of gut and lung cancer. Results: By analyzing gut and lung exosomes cargo with our previous transcriptomic studies from tumoral and inflammatory tissues, we found that exosomes can transport key RNAs that codify specific receptors that facilitate pathogenic interaction with microorganisms and RNAs that are part of interacting gene and transcriptional regulatory networks that control the function of differentially expresses genes, all involved in biological processes like cell cycle, plasticity and growth regulation, invasion, metastasis, microenvironmental remodeling, epigenetic, and microbial and immunological modulation, during the unlocking of phenotypic plasticity for the acquisition of the hallmarks of cancer in the oral-gut-lung axis. Conclusions: Exosomal RNA regulation of transcriptional networks represents a pivotal axis in the interplay between inflammation and cancer, offering opportunities for innovative diagnostic and therapeutic approaches.

Angiogenesis plays a crucial role in solid tumor growth. Ischemia and inflammation induce various angiogenic mediators, and patient metabolic conditions importantly influence this process. Macrophages closely interact with the vascular system and regulate angiogenesis through pro/anti-angiogenic factors. Traditionally, pro-angiogenic activity has been attributed to M2-like macrophages. We question this, as recent evidence suggests that also M1-like macrophages can be pro-angiogenic. Therefore, the aim is to identify the pro/anti-angiogenic gene expression profiles of human polarized macrophages unbiasedly. We also examine the effect of hyperglycemia on angiogenic gene expression, reflecting its role in diabetes and other metabolic conditions. Bioinformatic analysis was performed on the angiogenesis-related gene expression profiles of CD14+ monocyte-derived M1(IFN-γ)- and M2(IL-4)-polarized macrophages. The top differentially expressed genes were selected for validation. Macrophages were generated in vitro and polarized to M1(IFN-γ) and M2(IL-4/IL-6) cells under standard/hyperglycemic conditions. After immunophenotypic confirmation, selected gene expression was quantified using qPCR. IL-4 and IL-6 induce distinct M2-like phenotypes with mixed pro/anti-angiogenic gene expression. Remarkably, IFN-γ stimulation also increases several pro-angiogenic genes. Hyperglycemia affects the angiogenic expression profile in both M1- and M2-like macrophages, although distinctive identities remain intact. The pro-angiogenic phenotype is not limited to M2-polarized macrophages. Both M1- and M2-like macrophages express complex pro/anti-angiogenic gene profiles, which are only mildly influenced by hyperglycemia.

Inflammation can positively and negatively affect tumorigenesis based on the duration, scope, and sequence of related events through the regulation of signaling pathways. A transcriptomic analysis of five pulmonary arterial hypertension, twelve Crohn’s disease, and twelve ulcerative colitis high throughput sequencing datasets using R language specialized libraries and gene enrichment analyses identified a regulatory network in each inflammatory disease. IRF9 and LINC01089 in pulmonary arterial hypertension are related to the regulation of signaling pathways like MAPK, NOTCH, human papillomavirus, and hepatitis c infection. ZNF91 and TP53TG1 in Crohn’s disease are related to the regulation of PPAR, MAPK, and metabolic signaling pathways. ZNF91, VDR, DLEU1, SATB2-AS1, and TP53TG1 in ulcerative colitis are related to the regulation of PPAR, AMPK, and metabolic signaling pathways. The activation of the transcriptomic network and signaling pathways might be related to the interaction of the characteristic microbiota of the inflammatory disease, with the lung and gut cell receptors present in membrane rafts and complexes. The transcriptomic analysis highlights the impact of several coding and non-coding RNAs, suggesting their relationship with the unlocking of cell phenotypic plasticity for the acquisition of the hallmarks of cancer during lung and gut cell adaptation to inflammatory phenotypes.

Background/Objectives: Combined chromosome 7 gain and chromosome 10 loss (+7/−10) is the most frequent cytogenetic alteration and a defining diagnostic criterion for isocitrate dehydrogenase wild-type (IDHwt) glioblastoma. Despite the association with poor prognosis, its clinical and therapeutic significance remains unclear. We aim to systematically review its clinical significance, focusing on prevalence, prognostic value, and potential association with therapeutic resistance in adult patients. Methods: PubMed, Embase, CENTRAL, Scopus, EBSCOhost, and Web of Science were searched from inception to April 2025, using controlled vocabulary and free-text terms. Eligible studies included adult glioblastoma with molecular confirmation of combined chromosome 7 gain and chromosome 10 loss and reported survival or treatment response. Quality was assessed qualitatively, and findings were synthesized descriptively. Results: Of 3249 records, 5 observational studies (523 patients) were included. The signature was present in 60% to 70% of glioblastoma cases and frequently co-occurred with epidermal growth factor receptor amplification and telomerase reverse transcriptase promoter mutations. This alteration was consistently associated with shorter survival (mean, 8–70 weeks) compared with tumors lacking the alteration (19–170 weeks). In one study, the signature was more common in radioresistant tumors (9/20 vs. 1/10). Molecular evidence suggests that this alteration arises early in tumorigenesis. Conclusions: The +7/−10 cytogenetic alteration, common in glioblastoma, is frequently associated with aggressive clinical behavior. While exploratory data suggest a possible association with radiotherapy response, current evidence is insufficient to establish a predictive or therapeutic role. Its principal clinical value lies in diagnosis, molecular classification, and risk stratification. Incorporating cytogenetic testing for this alteration into routine glioblastoma workup may improve risk stratification and guide individualized management.

Background and Objective: The discovery of novel molecular biomarkers via next-generation sequencing technologies has revolutionized how glioblastomas (GBMs) are classified nowadays. This has resulted in more precise diagnostic, prognostic, and therapeutic approaches to address this malignancy. The present work examines the applications of single-cell RNA sequencing (scRNA-seq) in GBM, focusing on its potential to address tumor complexity and therapeutic resistance and improve patient outcomes. Methods: A scoping review of original studies published between 2009 and 2024 was conducted using the PUBMED and EMBASE databases. Studies in English or Spanish related to single-cell analysis and GBM were included. Key Findings: The database search yielded 453 publications. Themes related to scRNA-seq applied for the diagnosis, prognosis, treatment, and understanding of the cancer biology of GBM were used as criteria for article selection. Of the 24 studies that were included in the review, 11 focused on the tumor microenvironment and cell subpopulations in GBM samples, 5 investigated the use of sequencing to elucidate the GBM cancer biology, 3 examined disease prognosis using sequencing models, 3 applied translational research through scRNA-seq, and 2 addressed treatment-related problems in GBM elucidated by scRNA-seq. Conclusions: This scoping review explored the various clinical applications of scRNA-seq technologies in approaching GBM. The findings highlight the utility of this technology in unraveling the complex cellular and immune landscapes of GBM, paving the way for improved diagnosis and personalized treatments. This cutting-edge approach might strengthen treatment strategies against tumor progression and recurrence, setting the stage for multi-targeted interventions that could significantly improve outcomes for patients with aggressive, treatment-resistant GBMs.

Introduction:Gliomas are believed to arise from neuroglial stem cells and are histologically classified based on morphological similarities to normal neuroglial cells. This study aims to describe the clinical, histopathological, and demographic features of glioma patients treated in two reference centers in Colombia. Methods:This descriptive cross-sectional study included all patients with a histologically confirmed glioma treated at two institutions in Bogotá, Colombia, between January 2015 and December 2023. 272 patients with diffuse gliomas were included, and data were collected on sociodemographic characteristics, clinical presentation, histopathologic diagnosis, immunohistochemical markers, extent of resection, functionality, complications, and survival. Results:Amongst all cases, 36.00% were glioblastomas, 14.70% oligodendrogliomas, and 12.10% astrocytomas. 49.10% of patients were females, average age was 48.8 ± 21.0 years. While in the frontal lobe, most glioblastomas (38.95%) and oligodendrogliomas (47.50%) were found, astrocytomas were more frequent in the insula (27.27%). The average follow-up was 11.8 ± 16.0 months. Near-total resection was achieved in 40.10% of patients, followed by subtotal resection (37.00%), gross-total resection (11.45%), and intentional biopsy (11.45%). 31.25% of patients had new-onset motor deficits, and only 3% persisted after the 3-month follow-up. Overall survival was higher in females (males: 28.57% vs. females: 55.00%) (p = 0.0013). The 2-year overall survival for glioblastoma was 21%, 5-year for glioma, NOS 38%, for astrocytoma 15%, and 8-year for oligodendroglioma 5% (p < 0.0001). Conclusions:We present the largest study to date of diffuse glioma in Colombia's population. Clinical findings and overall survival trends are similar to those reported worldwide, however, further molecular analysis is needed for adequate diagnosis and classification.

Aging is the main risk factor for Parkinson's disease (PD), yet our understanding of how age-related mechanisms contribute to PD pathophysiology remains limited. We conducted a longitudinal analysis of blood samples from the Parkinson's Progression Markers Initiative cohort to investigate DNA damage in PD. Patients with PD exhibited disrupted DNA repair pathways and biased suppression of longer transcripts, indicating age-related, transcription-stalling DNA damage. Notably, at the intake visit, this DNA damage signature was detected only in patients with more severe progression of motor symptoms over 3 years, suggesting its potential as a predictor of disease severity. We validated this signature in independent PD cohorts and confirmed increased DNA damage in peripheral blood cells and dopamine neurons of the substantia nigra pars compacta in postmortem PD brains. Our study sheds light on an aging-related mechanism in PD pathogenesis and identifies potential markers of disease progression, providing a diagnostic platform to prognosticate disease progression.

Parkinson's Disease (PD) is an age-related neurodegenerative disorder that has been associated with increased DNA damage. To test if PD is associated with increased somatic mutations, we analyzed RNA-seq data in whole blood from 5 visits of the Parkinson's Progression Markers Initiative for clonally amplified somatic variants. Comprehensive analysis of RNA-sequencing data revealed a total of 5,927 somatic variants (2.4 variants per sample on average). Mutation frequencies were significantly elevated in PD subjects as compared to age-matched controls at the time of the last visit. This was confirmed by RNA analysis of substantia nigra. By contrast, the fraction of carriers with clonal hematopoiesis, was significantly reduced in old PD patients as compared to old healthy controls. These results indicate that while the overall mutation rate is higher in PD, specific clonally amplified mutations are protective against PD, as has been found for Alzheimer's Disease.

Glioblastoma (GBM) is the most common malignant central nervous system tumor. The emerging field of epigenetics stands out as particularly promising. Notably, the discovery of micro RNAs (miRNAs) has paved the way for advancements in diagnosing, treating, and prognosticating patients with brain tumors. We aim to provide an overview of the emergence of miRNAs in GBM and their potential role in the multifaceted management of this disease. We discuss the current state of the art regarding miRNAs and GBM. We performed a narrative review using the MEDLINE/PUBMED database to retrieve peer-reviewed articles related to the use of miRNA approaches for the treatment of GBMs. MiRNAs are intrinsic non-coding RNA molecules that regulate gene expression mainly through post-transcriptional mechanisms. The deregulation of some of these molecules is related to the pathogenesis of GBM. The inclusion of molecular characterization for the diagnosis of brain tumors and the advent of less-invasive diagnostic methods such as liquid biopsies, highlights the potential of these molecules as biomarkers for guiding the management of brain tumors such as GBM. Importantly, there is a need for more studies to better examine the application of these novel molecules. The constantly changing characterization and approach to the diagnosis and management of brain tumors broaden the possibilities for the molecular inclusion of novel epigenetic molecules, such as miRNAs, for a better understanding of this disease.

Climate change is currently one of humanity’s greatest threats. To help scholars understand the psychology of climate change, we conducted an online quasi-experimental survey on 59,508 participants from 63 countries (collected between July 2022 and July 2023). In a between-subjects design, we tested 11 interventions designed to promote climate change mitigation across four outcomes: climate change belief, support for climate policies, willingness to share information on social media, and performance on an effortful pro-environmental behavioural task. Participants also reported their demographic information (e.g., age, gender) and several other independent variables (e.g., political orientation, perceptions about the scientific consensus). In the no-intervention control group, we also measured important additional variables, such as environmentalist identity and trust in climate science. We report the collaboration procedure, study design, raw and cleaned data, all survey materials, relevant analysis scripts, and data visualisations. This dataset can be used to further the understanding of psychological, demographic, and national-level factors related to individual-level climate action and how these differ across countries.

The FMR1 5' regulation gene region harbors a CGG trinucleotide repeat expansion (CGG-TRE) that causes Fragile X syndrome (FXS) when it expands to more than 200 repetitions. Ricaurte is a small village in southwestern Colombia, with an FXS prevalence of 1 in 38 men and 1 in 100 women (~100 times higher than the worldwide reported prevalence), defining Ricaurte as the largest FXS cluster in the world. In the present study, using next-generation sequencing of whole exome capture, we genotype 55 individuals from Ricaurte (49 with either full mutation or with premutation), four individuals from neighboring villages (with either the full mutation or with the premutation), and one unaffected woman, native of Ricaurte, who did not belong to any of the affected families. With advanced clustering and haplotype reconstruction, we modeled a common haplotype of 33 SNPs spanning 83,567,899 bp and harboring the FMR1 gene. This reconstructed haplotype was found in all the men from Ricaurte who carried the expansion, demonstrating that the genetic conglomerate of FXS in this population is due to a founder effect. The definition of this founder effect and its population outlining will allow a better prediction, follow-up, precise and personalized characterization of epidemiological parameters, better knowledge of the disease's natural history, and confident improvement of the clinical attention, life quality, and health interventions for this community.

Aging is the main risk factor for Parkinson’s disease (PD), yet our understanding of how age-related mechanisms contribute to PD pathophysiology remains limited. We conducted a longitudinal analysis of the Parkinson’s Progression Markers Initiative cohort to investigate the involvement of DNA damage in PD. Our findings revealed that PD patients exhibit disrupted DNA repair pathways and biased suppression of longer transcripts, indicating the presence of age-related, transcription-stalling DNA damage. Notably, this DNA damage signature was only detected in patients with more severe motor symptom progression over a three-year period, suggesting its potential as a predictor of disease severity. We further validated this signature in independent PD cohorts and confirmed increased signs of DNA damage in dopamine neurons of the substantia nigra pars compacta through histopathological analysis of PD brains. Our study sheds light on an aging-related mechanism in PD pathogenesis and identifies markers of disease progression providing a readily applicable diagnostic platform to prognosticate disease progression. One Sentence Summary Parkinson’s disease patients display a DNA damage signature in blood that is predictive of disease progression. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement This study was supported by the Michael J Fox Foundation (Pier Giorgio Mastroberardino and Jan Vijg). Daisy Sproviero was supported by Fondazione Veronesi. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The Parkinson's Progression Markers Initiative (PPMI) is an observational, international, multicenter study and the dataset is available after request on the website described below. This study was conducted in accordance with the Declaration of Helsinki and Good Clinical Practice guidelines after approval of local ethics committees of the participating sites as described in <https://www.michaeljfox.org/news/ppmi-rna-sequencing-project>. Written informed consent was obtained from all participants before undergoing any study evaluations. Formalin fixed paraffin embedded fully anonymous human midbrain tissue sections derived from PD patients and age-matched healthy controls were kindly provided by the Queen Square Brain Bank for Neurological Disorders, London, UK according to Material transfer agreement number 10-2019. The Ethics Committee made up by Prof. Marco Pedrazzi, Prof.ssa Francesca Caloni, Prof.ssa Monica Ferraroni, Prof. Carlo Flamigni, Prof. Fabrizio Gardoni, Prof.ssa Daniela Milani, Prof. Emanuele Montanari, Prof. Gianfranco Mormino, Prof.ssa Francesca Poggi, Prof. Giuseppe Testa, Prof. Giovanni Ziccardi (all affiliated to Universita degli Studi di Milano Statale, Milan, Italy) examined the research project, Neurodegenerative proteinopathies: mechanical stress and nuclear deformation, presented by Dr. Domenico Delia, co-author on the manuscript, who also performed the experiments. The Ethics Committee during the session on 23rd September 2019, considered the received proposal valid, adequately documented, as well as in line with ethical standards. All the informations are described in document Allegato 4 (Protocol n-39-19). I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present study are available upon reasonable request to the authors <https://ida.loni.usc.edu/pages/access/geneticData.jsp#441>

Effectively reducing climate change requires marked, global behavior change. However, it is unclear which strategies are most likely to motivate people to change their climate beliefs and behaviors. Here, we tested 11 expert-crowdsourced interventions on four climate mitigation outcomes: beliefs, policy support, information sharing intention, and an effortful tree-planting behavioral task. Across 59,440 participants from 63 countries, the interventions' effectiveness was small, largely limited to nonclimate skeptics, and differed across outcomes: Beliefs were strengthened mostly by decreasing psychological distance (by 2.3%), policy support by writing a letter to a future-generation member (2.6%), information sharing by negative emotion induction (12.1%), and no intervention increased the more effortful behavior-several interventions even reduced tree planting. Last, the effects of each intervention differed depending on people's initial climate beliefs. These findings suggest that the impact of behavioral climate interventions varies across audiences and target behaviors.

Purpose Angiogenesis is a vital process involved in the formation of new blood vessels from existing ones. Macrophages play a crucial role in initiating endothelial activation and inflammation, and are involved in the pathological angiogenesis. Traditionally, macrophages have been classified, with the pro-angiogenic activity attributed to the M2 phenotype. However, recent evidence challenges the notion that only M2 macrophages possess pro-angiogenic properties. This study aims to investigate the pro- and anti-angiogenic properties of human polarized macrophages in normo- and hyper-glycemic conditions, in order to gain a better insight into the angiogenic capacity of M1- and M2-like macrophages in diabetes.Methods A comprehensive bioinformatic analysis of pro- and anti-angiogenic gene expression profiles related in M1-vs. M2-polarized macrophages was performed based on a large previously published dataset. The most contributing differentially expressed genes in angiogenesis were selected for further validation. Macrophages were generated and polarized by culturing CD14+ monocytes and their stimulation with any of IFN-γ, IL-4, or IL-6 cytokines. Polarized macrophages were immunophenotyped using flow cytometry, and their expression of the selected genes were measured using qPCR. Finally, the proangiogenic capacity of the cells was assessed in an in vitro 3-D endothelial tubule formation assay, containing GFP-expressing human retinal endothelial cells, pericytes, and pro-angiogenic growth factors.Results IL-4 and IL-6 induce distinct M2-like phenotypes in macrophages with mixed pro- and anti-angiogenic gene expressions. Hyperglycemia has a mild negative effect on the expression of M2-associated markers, however it does not significantly affect the angiogenic properties of macrophages.Conclusion Our data support the concept of a spectrum model for macrophage polarization, indicating that the angiogenic status of polarized macrophages is not limited to the M2-phenotype, but is rather mediated by microenvironmental cues, and can result in diverse phenotypic characteristics. The effect of hyperglycemia on the angiogenic capacity of macrophages requires more comprehensive investigation.### Competing Interest StatementThe authors have declared no competing interest.

PURPOSE Prostate cancer is a significant global health issue. The tumor microenvironment significantly influences cancer progression, treatment response, and metastasis. There's a lack of studies directly comparing adjacent tissue behavior to both tumor and normal tissue. An analytical strategy was developed to identify genes altered in the adjacent tissue to the tumor without the need for comparison with tumor or normal tissue. METHODS Data sets containing information on both adjacent tumor and normal tissue were searched. Also, transcriptomic data from the prostate cancer project in TCGA for adjacent tumor samples were searched. Differentially expressed genes (DEGs) were identified from the dataset enabling direct comparison between adjacent tissue and normal tissue. The DEGs were utilized as probes to identify other genes of interest within the TCGA database. We hypothesized that genes showing correlated expression with the DEGs would also exhibit altered expression in adjacent tissue within TCGA.A co-expression network was constructed using all genes displaying a correlation coefficient greater than 0.6 with at least one of the DEGs. Finally, a network analysis was conducted. RESULTS GSE6919 was the only dataset that allowed direct comparison of adjacent tissue to prostate cancer with normal tissue. Six DEGs were identified TRIB1, C18orf54, ARF1, NFKBIZ, and LINC00662, and MAFB. The transcriptomes of the 54 adjacent tissues to prostate cancer from TCGA were interrogated, revealing that the expression profiles of 3316 genes had a Pearson correlation coefficient greater than 0.6 with at least one of the 6 DEGs. With these genes, a co-expression network was constructed, retaining 3254 genes. Two networks were identified, one with 2234 genes and another with 395 genes. The first network was enriched with genes related to protein synthesis and vesicle transport from the endoplasmic reticulum to the Golgi apparatus. The second network was enriched with genes from the IL17 signaling pathway and the TNF signaling pathway. CONCLUSION The proposed method proved effective in identifying genes potentially altered in adjacent tissue to the tumor, even in the absence of normal tissue for comparison. Some pathways identified have been previously associated with cancer progression, metastasis, and treatment response. Further studies are warranted to determine whether the behavior of these and novel discovered pathways in adjacent tissue has clinical implications for prostate cancer.