Non-immune Cells Research Articles

Single cell transcriptome profiling reveals pathogenesis of bullous pemphigoid

Bullous pemphigoid (BP) triggers profound functional changes in both immune and non-immune cells in the skin and circulation, though the underlying mechanisms remain unclear. In this study, we conduct single-cell transcriptome analysis of lesional and non-lesional skin, as well as blood samples from BP patients. In lesional skin, non-immune cells upregulate pathways related to metabolism, wound healing, immune activation, and cell migration. LAMP3+DCs from cDC2 show stronger pro-inflammatory signatures than those from cDC1, and VEGFA+ mast cells, crucial for BP progression, are predominantly in lesional skin. As BP patients transition from active to remission stages, blood B cell function shifts from differentiation and memory formation to increased type 1 interferon signaling and reduced IL-4 response. Blood CX3CR1+ ZNF683+ and LAG3+ exhausted T cells exhibit the highest TCR expansion among clones shared with skin CD8+T cells, suggesting their role in fueling skin CD8+T cell clonal expansion. Clinical BP severity correlates positively with blood NK cell IFN-γ production and negatively with amphiregulin (AREG) production. NK cell-derived AREG mitigates IFN-γ-induced keratinocyte apoptosis, suggesting a crucial balance between AREG and IFN-γ in BP progression. These findings highlight functional shifts in BP pathology and suggest potential therapeutic targets.

Improving the Wound Healing Process: Pivotal role of Mesenchymal stromal/stem Cells and Immune Cells.

Wound healing, a physiological process, involves several different types of cells, from immune cells to non-immune cells, including mesenchymal stromal/stem cells (MSC), and their interactions. Immune cells including macrophages, neutrophils, dendritic cells (DC), innate lymphoid cells (ILC), natural killer (NK) cells, and B and T lymphocytes participate in wound healing by secreting various mediators and interacting with other cells. MSCs, as self-renewing, fast proliferating, and multipotent stromal/stem cells, are found in a wide variety of tissues and critically involved in different phases of wound healing by secreting various molecules that help to improve tissue healing and regeneration. In this review, first, we described the four main phases of wound healing, second, we reviewed the function of MSCs, MSC secretome and immune cells in improving the progress of wound repair (mainly focusing on skin wound healing), third, we explained the immune cells/MSCs interactions in the process of wound healing and regeneration, and finally, we introduce clinical applications of MSCs to improve the process of wound healing.

Multiomics Analysis Reveals Molecular Changes during Early Progression of Precancerous Lesions to Lung Adenocarcinoma in Never-Smokers.

Lung cancer is the most common cause of cancer mortality globally, and the prevalence of lung adenocarcinoma, the most common lung cancer subtype, has increased sharply in East Asia. Early diagnosis leads to better survival rates, but this requires an improved understanding of the molecular changes during early tumorigenesis, particularly in nonsmokers. In this study, we performed whole-exome sequencing and RNA sequencing of samples from 94 East Asian patients with precancerous lesions [25 with atypical adenomatous hyperplasia (AAH); 69 with adenocarcinoma in situ (AIS)] and 73 patients with early invasive lesions [minimally invasive adenocarcinoma (MIA)]. Cellular analysis revealed that the activities of endothelial and stromal cells could be used to categorize tumors into molecular subtypes within pathologically defined types of lesions. The subtypes were linked with the radiologically defined type of lesions and corresponded to immune cell infiltration throughout the early progression of lung adenocarcinoma. Spatial transcriptomic analysis revealed the distribution of epithelial cells, endothelial cells, fibroblasts, and plasma cells within MIA samples. Characterization of the molecular lesion subtypes identified positively selected mutational patterns and suggested that angiogenesis in the late-stage AIS type potentially contributes to tissue invasion of the MIA type. This study offers a resource that may help improve early diagnosis and patient prognosis, and the findings suggest possible approaches for early disease interception. Significance: Integrative analysis of multiomics data revealed coordination between immune and nonimmune cells during early progression of precancerous lesions to lung adenocarcinomas and shed light on the molecular characteristics of clinically defined subtypes.

TNIP1 Impacts Prognosis by Modulating the Immune Microenvironment in BRCA.

Breast invasive carcinoma (BRCA) affects women worldwide, and despite advancements in diagnosis, prevention, and treatment, outcomes remain suboptimal. TNIP1, a novel target involved in multiple immune signaling pathways, influences tumor development and survival. However, the connection between BRCA and TNIP1 remains unclear. Analysis of data from the TCGA, GEO, Sangerbox, and Ualcan databases revealed that TNIP1 is underexpressed in BRCA tissues. This finding was corroborated by RT-PCR and immunohistochemistry. Furthermore, data from the TCGA and GEPIA2 databases, along with Sangerbox, identified TNIP1 as a marker of poor prognosis in BRCA patients. TNIP1 expression shows significant positive correlations with the BRCA Tumor Microenvironment (TME) StromalScore (R = 0.22), ImmuneScore (R = 0.25), and ESTIMATEScore (R = 0.27). Various algorithms have demonstrated a strong association between TNIP1 expression and BRCA tumor-infiltrating immune cells (TIICs). Further analysis using EPIC, TIMER, MCPCounter, QUANTISEQ, xCell, and other computational tools revealed that elevated TNIP1 expression is significantly associated with increased immune cell scores. TNIP1 expression in BRCA tumor tissues also shows a strong correlation with immune checkpoint markers. Data from the HAP database indicate that TNIP1 expression is predominantly involved in the normal skin microenvironment. Subsequent analysis using the TISCH platform with the BRCA single-cell dataset demonstrated that TNIP1 exhibits higher expression levels in immune cells compared to non-immune cells in BRCA patients. This expression is significantly positively correlated with inflammation (R = 0.25) and differentiation (R = 0.28) within the TME, while showing negative correlations with BRCA stemness (R = -0.34) and invasion (R = -0.22). Consequently, TNIP1 is proposed as a potential prognostic marker and therapeutic target for BRCA.

Immunometabolic landscaping in Crohn’s disease creeping fat – Does fat matter?

Abstract Background and aims The study proposed herein aims to explore the complex immunometabolic landscape of Creeping Fat (CF) in Crohn's disease (CD) ileitis, an area that remains largely unexplored. Our hypothesis posits the existence of distinct sub-entities within mesenteric adipose tissue, characterized by unique metabolic states in immune cells, non-immune cells, and adipocytes. The primary objective is to elucidate the immunometabolic environment within ileal CF, leveraging an advanced Imaging Mass Cytometry (IMC)-based methodology. Methods Employing a 37-marker IMC panel, we will conduct comprehensive spatial and metabolic profiling of adipose and intestinal tissues from a cohort of 50 patients, including those with CD and ulcerative colitis (UC), and compare these profiles with samples from bariatric surgery patients. This approach enables single-cell level analysis with spatial resolution, uncovering the metabolic states and signatures of various cell types. Previous work in our group has already demonstrated the utility of this technique in revealing significant variations in cell metabolism across different inflammatory states and cell types in CD. Our methodology integrates the study of immune cell clusters, metabolic states, and subtypes with distinct metabolic signatures. By comparing metabolic signatures in adipose tissues with those in inflamed ileal mucosal tissue, we aim to establish an understanding of metabolic processes within CF. Anticipated impact Anticipated impact includes the identification of unique immunometabolic signatures in CF, providing new insights into the pathomechanisms of CD. This could lead to novel therapeutic targets focusing on the metabolism of specific cell types within CF. The study holds the potential to reshape our understanding of CD, paving the way for more targeted and effective treatments for this debilitating disease. Given our established expertise, robust methodology, and comprehensive cohort, this research is poised to offer significant contributions to the field of immunometabolism in IBD, enhancing our understanding of CF and its role in CD.

The Role of Innate Immunity in Healthy Aging Through Antimicrobial Peptides.

In a super-aging society, the increase in the elderly population is closely tied to a rise in infectious diseases due to factors such as weakened immune systems and decreased vaccine efficacy in older adults. Various opportunistic pathogens commonly encountered in everyday life can cause infections and diseases when an individual's immune defence is weakened due to aging. These factors underscore the importance of preventive measures against pathogenic infections and the aging of immune systems in the elderly. The immune response acts as the defence mechanism against foreign substances, including pathogens and abnormal cells. Specifically, the innate immune response is the body's first line of defence, offering a rapid and nonspecific response to pathogens. Advances in the study of innate immunity's regulatory functions in both immune and non-immune cells have broadened our understanding of innate immune responses' impact on health. This includes a focus on immune effectors like antimicrobial peptides (AMPs) and their potential implications for health and longevity. This review summarises the common principles and evolutionary adaptations of innate immunity via AMPs, in mammals and invertebrates. Especially, this review discusses the conserved mechanisms regulating AMP production and the role of AMPs in modulating aging and diseases from invertebrate to human. Therefore, it highlights the potential role of innate immunity in addressing aging through AMPs.

Parenchymal and Inflammatory Responses to Ozone Exposure in the Aging Healthy and Surfactant Protein C Mutant Lung.

Ozone (O3) is a ubiquitous pollutant known to produce acute, transient inflammation through oxidative injury and inflammation. These effects are exacerbated in susceptible populations, such as the elderly and those exhibiting genetic mutations in central nodes of pulmonary function. To comprehend the impact of these predisposing factors, the present study examines structural, mechanical, and immunological responses to single acute O3 exposure (0.8 ppm, 3h) in young (8-14 week old), middle-aged (44-52 week old), and old (>80 week old) mice. Furthermore, this work compares the impact of a clinically relevant mutation in the gene encoding for the alveolar epithelial type 2 specific surfactant protein C. Aging was associated with reduced lung resistance and increases in respiratory elastic properties, the latter of which was exacerbated in SP-C mutant mice. Ozone exposure produced focal injury localized at the terminal bronchiole-to-alveolar junctions and enlarged alveoli in aged SP-C mutant lungs. Flow cytometric analysis revealed increases in mononuclear myeloid abundance in aged SP-C mutant lungs, paired with a contraction in CD8+ expressing cells. Expansion of tertiary lymphoid tissues was also noted in aged groups, more evident in the mutant mice. Spatial transcriptomics of CD68+ macrophages and CD45- non-immune parenchymal cells highlighted age-dependent shifts in inflammatory and extracellular matrix organization signaling, and enrichment in senescence and chromatin remodeling pathways. These results illustrate the structural and immunological impact of O3 in the aging wild type and mutant lung and emphasize the significance of modeling environmental exposure in at-risk populations.

Roles for Exosomes in the Pathogenesis, Drug Delivery and Therapy of Psoriasis.

Psoriasis is a chronic, recurrent and inflammatory skin disease. Although conventional immunosuppressants can ameliorate psoriatic symptoms, it tends to relapse over time. Previous studies have shown that exosomes from both immune and non-immune cells participate in psoriatic immunopathology. The biologically active cargoes in exosomes accelerate psoriasis progression by altering gene profiles and signaling pathways of neighboring cells. On the other hand, exosomes can be utilized as drug delivery platforms for psoriasis treatment. Especially, engineered exosomes may serve as drug delivery systems for effective delivery of proteins, nucleic acids or other drugs due to their low immunogenicity, good stability and ability to fuse with target cells. Therefore, investigation into the mechanisms underlying intercellular communications mediated by exosomes in skin lesions likely helps design drugs for therapy of psoriasis. In this review, we have summarized recent advances in the biogenesis of exosomes and their potential roles in the pathogenesis and treatment of psoriasis and further discussed their challenges and future directions in psoriasis treatment. In particular, this review highlights the immunoregulatory function of exosomes derived from immune or non-immune cells and exosome-based therapeutic applications in psoriasis, including their drug delivery systems. Thus, this review may help accelerate applications of exosomes for drug delivery and treatment of psoriasis.

Association between TLR-2 and TLR-9 gene polymorphisms (rs5743708 and rs5743836) and susceptibility to psoriatic arthritis in Egyptian patients

Psoriasis (PsO) is a chronic immune-mediated disease of the skin. Psoriatic arthritis (PsA) is a prevalent chronic inflammatory disease that is associated with joint destruction and disability. The presence of PsO is the single greatest risk factor for the development of PsA. Toll-like receptors (TLRs) are trans-membrane proteins coded by the toll genes family. They are expressed in different cell types including immune and non-immune cells. Polymorphisms in TLR genes that lead to changes in these receptors or interfere with the transcription rates of their messenger ribonucleic acid (mRNA), may be involved in the chronic inflammatory immune response observed in PsA. This study involved 50 patients with PsA, 50 patients with cutaneous PsO and 50 age and sex matched normal subjects as controls. We aimed to assess TLR-2 (rs5743708) and TLR-9 (rs5743836) gene polymorphisms as potential risk factors for PsA in Egyptian patients with cutaneous PsO. Genotyping and allele frequencies were performed using Real Time polymerase chain reaction (qRT-PCR). Toll-like receptor-2 (TLR-2) rs5743708 and TLR-9 rs5743836 polymorphisms were associated with increased risk of PsO as an autoimmune disease, however they were not related to increase susceptibility to PsA in this cohort study of Egyptian patients.

Key Interleukins in Inflammatory Bowel Disease-A Review of Recent Studies.

Inflammatory bowel disease (IBD) is an immune disorder of the gastrointestinal tract with a complex aetiopathogenesis, whose development is influenced by many factors. The prevalence of IBD is increasing worldwide, in both industrialized and developing countries, making IBD a global health problem that seriously affects quality of life. In 2019, there were approximately 4.9 million cases of IBD worldwide. Such a large number of patients entails significant healthcare costs. In the treatment of patients with IBD, the current therapeutic target is mucosal healing, as intestinal inflammation often persists despite resolution of abdominal symptoms. Treatment strategies include amino salicylates, corticosteroids, immunosuppressants, and biologic therapies that focus on reducing intestinal mucosal inflammation, inducing and prolonging disease remission, and treating complications. The American College of Gastroenterology (ACG) guidelines also indicate that nutritional therapies may be considered in addition to other therapies. However, current therapeutic approaches are not fully effective and are associated with various limitations, such as drug resistance, variable efficacy, and side effects. As the chronic inflammation that accompanies IBD is characterized by infiltration of a variety of immune cells and increased expression of a number of pro-inflammatory cytokines, including IL-6, TNF-α, IL-12, IL-23 and IFN-γ, new therapeutic approaches are mainly targeting immune pathways. Interleukins are one of the molecular targets in IBD therapy. Interleukins and related cytokines serve as a means of communication for innate and adaptive immune cells, as well as nonimmune cells and tissues. These cytokines play an important role in the pathogenesis and course of IBD, making them promising targets for current and future therapies. In our work, we review scientific studies published between January 2022 and November 2024 describing the most important interleukins involved in the pathogenesis of IBD. Some of the papers present new data on the precise role that individual interleukins play in IBD. New clinical data have also been provided, particularly on blocking interleukin 23 and interleukin 1beta. In addition, several new approaches to the use of different interleukins in the treatment of IBD have been described in recent years.

Glucose-dependent insulinotropic polypeptide receptor signaling alleviates gut inflammation in mice.

Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 (GLP-1) are gut-derived peptide hormones that potentiate glucose-dependent insulin secretion. The clinical development of GIP receptor (GIPR)-GLP-1 receptor (GLP-1R) multi-agonists exemplified by tirzepatide and emerging GIPR antagonist-GLP-1R agonist therapeutics such as maritide is increasing interest in the extra-pancreatic actions of incretin therapies. Both GLP-1 and GIP modulate inflammation, with GLP-1 also acting locally to alleviate gut inflammation in part through anti-inflammatory actions on GLP-1R+ intestinal intraepithelial lymphocytes. In contrast, whether GIP modulates gut inflammation is not known. Here, using gain and loss of function studies, we show that GIP alleviates 5-fluorouracil (5FU)-induced gut inflammation, whereas genetic deletion of Gipr exacerbates the proinflammatory response to 5FU in the murine small bowel (SB). Bone marrow (BM) transplant studies demonstrated that BM-derived Gipr-expressing cells suppress 5FU-induced gut inflammation in the context of global Gipr deficiency. Within the gut, Gipr was localized to non-immune cells, specifically stromal CD146+ cells. Hence, the extra-pancreatic actions of GIPR signaling extend to the attenuation of gut inflammation, findings with potential translational relevance for clinical strategies modulating GIPR action in people with type 2 diabetes or obesity.

Molecular Mechanisms of Rett Syndrome: Emphasizing the Roles of Monoamine, Immunity, and Mitochondrial Dysfunction

Rett syndrome (RTT), which predominantly affects females, arises in most cases from mutations in the Methyl-CpG-binding Protein-2 (MECP2) gene. When MeCP2 is impaired, it disrupts the regulation of numerous genes, causing the production of dysfunctional proteins associated with various multi-systemic issues in RTT. In this review, we explore the current insights into molecular signaling related to monoamines, immune response, and mitochondrial function, and their implications for the pathophysiology of RTT. Research has shown that monoamines—such as dopamine, norepinephrine, epinephrine, serotonin, and histamine—exhibit alterations in RTT, contributing to a range of neurological symptoms. Furthermore, the immune system in RTT individuals demonstrates dysfunction through the abnormal activity of microglia, macrophages, lymphocytes, and non-immune cells, leading to the atypical release of inflammatory mediators and disruptions in the NF-κB signaling pathway. Moreover, mitochondria, essential for energy production and calcium storage, also show dysfunction in this condition. The delicate balance of producing and scavenging reactive oxygen species—termed redox balance—is disrupted in RTT. Targeting these molecular pathways presents a promising avenue for developing effective therapies.

Amoeboid cells undergo durotaxis with soft end polarized NMIIA.

Cell migration towards stiff substrates has been coined as durotaxis and implicated in development, wound healing, and cancer, where complex interplays between immune and non-immune cells are present. Compared to the emerging mechanisms underlying the strongly adhesive mesenchymal durotaxis, little is known about whether immune cells - migrating in amoeboid mode - could follow mechanical cues. Here, we develop an imaging-based confined migration device with a stiffness gradient. By tracking live cell trajectory and analyzing the directionality of T cells and neutrophils, we observe that amoeboid cells can durotax. We further delineate the underlying mechanism to involve non-muscle myosin IIA (NMIIA) polarization towards the soft-matrix-side but may not require differential actin flow up- or down-stiffness gradient. Using the protista Dictyostelium, we demonstrate the evolutionary conservation of amoeboid durotaxis. Finally, these experimental phenomena are theoretically captured by an active gel model capable of mechanosensing. Collectively, these results may shed new lights on immune surveillance and recently identified confined migration of cancer cells, within the mechanically inhomogeneous tumor microenvironment or the inflamed fibrotic tissues.

Activation of STAT6 in Intestinal Epithelial Cells Predisposes to Gut Inflammation.

Ulcerative colitis (UC) is an inflammatory bowel disease (IBD) often associated with a Type 2 immune response. Although previous reports hint at a role for signal transducer and activator of transcription (STAT) 6 signaling in non-immune cells, the contribution of STAT6-activation particularly in intestinal epithelial cells (IECs) is still unknown. Dextran sodium sulfate (DSS)-induced colitis is a model for UC in mice that we applied here on animals with expression of a constitutively active version of STAT6 in IECs (VillinCre_STAT6vt mice). We report increased pathology and mortality due to enhanced and systemic inflammation in these mice. Bulk RNA sequencing of colonic tissue from naïve VillinCre_STAT6vt mice showed differential expression of more than 140 genes compared to control mice. Gene set enrichment analysis revealed STAT6-regulated expression of the unfolded protein response, MTORC- and MYC-signaling, and protein secretion pathways. A comparison of gene expression in the colon of naïve VillinCre_STAT6vt mice and a human single-cell RNA sequencing dataset of a patient cohort with IBD revealed overlapping changes in the epithelial and macrophage compartment compared to corresponding controls. In conclusion, we found that activation of STAT6 in the intestinal epithelium predisposes to exacerbated colitis and gut inflammation.

Amino Acid Metabolism and Autophagy in Atherosclerotic Cardiovascular Disease.

Cardiovascular disease is the most common cause of mortality globally, accounting for approximately one out of three deaths. The main underlying pathology is atherosclerosis, a dyslipidemia-driven, chronic inflammatory disease. The interplay between immune cells and non-immune cells is of great importance in the complex process of atherogenesis. During atheroprogression, intracellular metabolic pathways, such as amino acid metabolism, are master switches of immune cell function. Autophagy, an important stress survival mechanism involved in maintaining (immune) cell homeostasis, is crucial during the development of atherosclerosis and is strongly regulated by the availability of amino acids. In this review, we focus on the interplay between amino acids, especially L-leucine, L-arginine, and L-glutamine, and autophagy during atherosclerosis development and progression, highlighting potential therapeutic perspectives.

Left atrial single-cell transcriptomics reveals amphiregulin as a surrogate marker for atrial fibrillation

Atrial fibrillation (AF) is strongly associated with strokes, heart failure, and increased mortality. This study aims to identify the monocyte–macrophage heterogeneity and interactions of these cells with non-immune cells, and to identify functional biomarkers in patients with AF. Therefore, we assess the single cell landscape of left atria (LA), using a combination of single cell and nucleus RNA-seq. Myeloid cells in LA tissue are categorized into five macrophage clusters, three monocyte clusters, and others. Cell-Chat analysis revealed that monocytes and IL1B+ macrophages send epidermal growth factor (EGF) signals to fibroblasts. Amphiregulin (AREG) is the most upregulated gene in monocytes and IL1B+ macrophages in the AF group, compared with healthy controls from other groups. Serum AREG levels are higher in patients with persistent AF. These data suggested that EGF signaling pathway could be a therapeutic target for AF and serum AREG levels provide an effective biomarker for predicting persistent AF.

Universal Immune Learning Ability in Immune and Non-Immune Cells (A Review)

Universal Immune Learning Ability in Immune and Non-Immune Cells (A Review)

324 Phenotyping of immune and non-immune cells in cutaneous sarcoidosis compared to homeostatic conditions and after mTOR inhibition

324 Phenotyping of immune and non-immune cells in cutaneous sarcoidosis compared to homeostatic conditions and after mTOR inhibition

Cutibacterium acnes strains associated with bone prosthesis infections cannot evade the host immune system.

Cutibacterium acnes is a commensal skin bacterium that is involved in bone prosthesis infections (BPIs) and presents low-grade clinical symptoms. C. acnes has been thought to escape the immune system at bone sites. Our study was carried out on a laboratory strain and two BPI-related clinical strains, one of which surprisingly induced clinical symptoms of inflammation in the patient. We investigated the ability of these C. acnes strains to trigger in vitro human primary neutrophils (PMN) response through inflammatory mediators measurements (antibody arrays, ELISA, RT-qPCR, zymography) and activation status assessment (flow cytometry), and to induce in vivo PMN recruitment from the bloodstream in mice air-pouch model. PMN-mediated inflammation was also studied in an original in vitro model mimetic of an infected bone site that combine titanium alloy, human primary osteoblasts, human primary neutrophils and C. acnes strains. We demonstrated for the first time that both C. acnes planktonic and biofilm cultures, triggered an effective immune response by neutrophils in vitro and their recruitment in vivo. This host response was enhanced when using a strain from a patient with inflammatory signs. In an original infected prosthesis mimetic model, osteoblasts and neutrophils were able to detect C. acnes, but their response to the clinical C. acnes inflammatory strain decreased. This work provides the first evidence showing that the immune cell response to pathogenic C. acnes may be tuned by nonimmune cells at the infected site, such as osteoblasts, which may promote bacterial persistence.

Extracellular Vesicles as Novel Immunomodulatory Agents: Potential for Autoimmune and Inflammatory Diseases

Extracellular vesicles (EVs) are nanoscale membrane-bound particles released by various cell types, including immune and non-immune cells, and play crucial roles in intercellular communication. Emerging evidence suggests that EVs have significant immunomodulatory properties, influencing various aspects of immune responses and contributing to the pathogenesis of autoimmune and inflammatory diseases. This review explores the biogenesis, composition, and functional roles of EVs, focusing on their potential as novel immunomodulatory agents. We discuss how EVs can modulate immune cell activation, differentiation, and function, highlighting their roles in promoting immune tolerance and regulating inflammatory processes. Furthermore, we explore the therapeutic applications of EVs in managing autoimmune and inflammatory diseases, including their potential as biomarkers, drug delivery vehicles, and therapeutic agents. We also address the challenges and limitations associated with EV research, including isolation techniques, standardization, and the need for a better understanding of their mechanisms of action. This review emphasizes the potential of EVs as innovative therapeutic strategies for treating autoimmune and inflammatory conditions, paving the way for future research to harness their immunomodulatory capabilities. Keywords: Extracellular vesicles, Immunomodulation, Autoimmune diseases, Inflammatory diseases, Intercellular communication