Clinical science最新文献

Coronary microvascular dysfunction (CMD) is prevalent in diabetes. Ten-Eleven Translocation-2 (TET2) as the major demethylase in endothelial cells (ECs) is decreased in diabetic CMD, and the role warrants further exploration. In this study, a multi-modality imaging, consist of transthoracic Doppler echocardiography and artery spin labeling cardiac magnetic resonance, is assessed for coronary microvascular function. The expression of TET2 is down-regulated in the heart of diabetic CMD mice. ECs TET2 conditional knockout increases the severity of CMD in diabetic mice. Mechanistically, TET2 deficiency declined the expression of CMPK2, a mtDNA synthetase. Additionally, the mitochondrial electron transport chain complexes encoded by mtDNA are downregulated, which contributes to the excessive production of reactive oxygen species. This, in turn, exacerbated mitochondrial dysfunction, manifesting as mitochondrial membrane potential depolarization, aberrant opening of the mitochondrial permeability transition pore, and structural abnormalities in mitochondria. Therapeutic research demonstrates that Vitamin C improves ECs mitochondrial function in diabetic CMD through the TET2-CMPK2 pathway, revealing its potential clinical therapeutic value. In conclusion, we show that loss of endothelial TET2 impairs endothelial mitochondrial function and exacerbated diabetic CMD by regulating the expression of mitochondria enzyme CMPK2.

Rheumatoid arthritis (RA) is a chronic autoimmune disease primarily characterized by persistent synovial inflammation, hyperplasia, and joint destruction. Despite advancements in current treatment options, approximately 80% of RA patients fail to achieve optimal therapeutic outcomes, and the precise pathogenesis of RA remains unclear. Therefore, identifying new therapeutic targets is crucial for improving the management of RA. This study aims to explore the role of IL-33 in the synovial microenvironment of RA, particularly its key function in promoting the differentiation of pro-inflammatory macrophages, providing new experimental evidence to address the persistent synovial inflammation in some treatment-resistant RA patients. We measured the levels of IL-33 in serum and synovial fluid from matched RA patients and found that, in both the active disease group and the remission group, the synovial fluid IL-33 levels were significantly higher than those in serum. Furthermore, synovial fluid IL-33 levels were more strongly correlated with disease activity (Disease Activity Score 28, Erythrocyte Sedimentation and key autoimmune markers (anti-CCP antibodies, rheumatoid factor, immunoglobulins IgG, immunoglobulins IgM, Anti-keratin antibody). These results suggest that IL-33 plays a central role in the immune dysregulation of the synovial microenvironment in RA patients. Further multiplex immunohistochemical analysis revealed a significant increase in IL-33, ST2, and CD86-positive macrophages in the synovium of patients with active RA, with the proportion of CD86+ macrophages closely correlated with disease activity and the IL-33 concentration in the synovial fluid. Single-cell RNA sequencing data further indicated that IL-33 plays an important role in the development of pro-inflammatory macrophages in the synovium during the active phase of RA. KEGG enrichment analysis and cell experiments demonstrated that the IL-33/ST2 signaling pathway promotes the differentiation of monocytes into a pro-inflammatory macrophage phenotype through the MAPK/NF-κB pathway. The pro-inflammatory macrophages secrete IL-33 and TNF, further exacerbating synovial inflammation, creating a vicious cycle that leads to disease persistence. These findings provide new evidence for the progression and treatment of synovial disease in RA, highlighting the critical role of IL-33 in the synovial microenvironment, particularly in assessing disease activity and treatment evaluation in RA patients.

Intrarenal renin angiotensin system (iRAS) activation has been implicated in tubulopathy in diabetic kidney disease (DKD), its underlying mechanisms remain unclear. Type 1 diabetic Akita mice and Akita mice with angiotensinogen (Agt) deletion in renal tubules (Akita AgtRT KO) and their respective controls were studied at 42 weeks. Akita mice exhibit increased AGT expression, oxidative stress, tubular cell size and luminal dilation in proximal tubules (PTs), while reduced in Akita AgtRT KO mice. Elevated senescence-associated β-galactosidase (SA-β-gal) activity and senescence-associated secretory phenotype (SASP) along with increased senescence marker p16 expression in distal tubules (DTs) were also observed in Akita mice, all normalized in Akita AgtRT KO mice. Human CKD/DKD datasets confirmed AGT positively correlated with CDKN2A/p16 expression. Akita mice also showed elevated NADPH oxidase 4 (NOX4) expression and mitochondrial cristae disruption in PTs, accompanied by significant oxidative DNA damage, renal inflammation, fibrosis and apoptosis cf. Akita AgtRT KO mice. In vitro, high glucose and angiotensin II (Ang II) triggered NOX4-mediated mitochondrial oxidative stress and dysfunction in proximal tubular (HK-2) cells. In addition, Ang II induced p16-dependent senescence in distal tubular (MDCK) cells. Conditioned medium from senescent MDCK cells triggered epithelial-to-mesenchymal transition in HK-2 cells, which was reversed by losartan or N-acetylcysteine. These findings suggest that iRAS promotes tubulopathy in DKD through NOX4-induced mitochondrial oxidative stress and dysfunction in PTs and oxidative DNA damage-induced senescence in DTs, providing new therapeutic targets.

This study aimed to investigate the therapeutic efficacy of tolerogenic dendritic cells (tolDCs) in IgA nephropathy (IgAN) mice. Male C57BL/6 mice were used to construct an IgAN model via oral mucosal immunization, while tolDCs were generated by exposing bone marrow-derived dendritic cells (BMDCs) to IL-10 and characterized for their immunosuppressive phenotype and function; wild-type (WT) and IgAN mice then received either PBS or tolDCs treatments, followed by comprehensive assessments of renal functional parameters, histopathological changes, and inflammatory cytokines profiles. The IgAN mouse model was successfully established by week 8 post-immunization, exhibiting characteristic renal pathology including glomerular mesangial IgA deposition, accompanied by mesangial cells hyperplasia and mesangial matrix expansion. IL-10-induced tolDCs exhibited a stable immunosuppressive phenotype: Compared with BMDCs, tolDCs exhibited a 3.5-fold upregulation in PD-L1 surface expression and a 7.7-fold increase in IL-10 protein level, while IL-12 protein expression remained unchanged. Functionally, tolDCs demonstrated targeted migration to the kidneys and promoted regulatory T cells (Tregs) differentiation in IgAN mice. Therapeutic administration of tolDCs significantly attenuated disease progression: Compared to the IgAN-PBS group, the IgAN-tolDCs group showed 59.3% and 55.4% reductions in glomerular and tubulointerstitial IgA deposition, respectively, accompanied by 5.5-fold and 2.9-fold increases in Tregs infiltration. At the inflammatory cytokines level, the mRNA and protein expression of renal IL-10 was up-regulated, while renal IL-12 and TGF-β was down-regulated in the IgAN-tolDCs group. Renal function parameters remained stable in the IgAN-tolDCs group, confirming the safety of tolDCs immunotherapy. In conclusion, IL-10-induced tolDCs exhibit a stable immunosuppressive phenotype and can migrate to the kidneys of IgAN mice, and their therapeutic administration alleviates renal IgA deposition, promotes renal Treg accumulation, and suppresses pro-inflammatory cytokine expression, collectively attenuating renal immunoinflammatory injury, supporting tolDCs as a promising targeted therapy for IgAN.

Chimeric antigen receptor (CAR) T cell therapy has emerged as a groundbreaking advancement in cancer immunotherapy, demonstrating remarkable success in treating hematologic malignancies. However, its application in solid tumors remains challenging. The complex manufacturing process and severe treatmentrelated toxicities further limit its broader clinical application. To address these challenges, researchers are investigating alternative CAR-engineered immune cells, including CAR-NK cells, CAR-γδ T cells, and CARmacrophages (CAR-M), which offer distinct advantages over conventional CAR-T therapy. Notably, CAR-NK and CAR-γδ T cells exhibit HLA-independent cytotoxicity, making them promising 'off-the-shelf' therapeutic options. Meanwhile, CAR-M not only phagocytose tumor cells and present antigens but also remodel the immunosuppressive tumor microenvironment. Despite their potential, these innovative therapies still face several challenges in clinical application. This review systematically summarizes recent advances in CAR-T cells, CAR-NK cells, CAR-γδ T cells, and CAR-M for cancer treatment, providing a comprehensive analysis of their respective strengths, limitations, and future optimization strategies to support the clinical translation of next-generation CAR-based immunotherapies.

Pulmonary iron levels are increased in COPD, possibly due to increased red blood cell leakage from the microvasculature. Neutrophils cause endothelial cell damage which may cause vascular dysfunction and iron dysregulation in COPD. We investigate the relationships between neutrophilic inflammation, iron metabolism and vascular dysfunction in COPD. Using gene and protein analysis, associations between neutrophilic inflammation, iron dysregulation and vascular dysfunction were investigated in two COPD bronchoscopy cohorts: EvA (n=51) and Manchester (n=33). Patients were sub-grouped based on bronchoalveolar lavage (BAL) neutrophil percentage (neutrophilhigh≥3% and neutrophillow<3%). Heme was measured in BAL by LC-MS. BAL cell gene expression of neutrophilic inflammation markers such as C-X-C Motif Chemokine Ligand 8 (CXCL8) and Interleukin 6 Receptor (IL6R) were significantly increased in neutrophilhigh compared to neutrophillow patients in both cohorts; fold change (FC) differences 1.06 - 17. We found increased markers of iron and iron trafficking including Lactoferrin (LTF), Lipocalin-2 (LCN2) and Myoglobin (MB) in neutrophilhigh patients in both cohorts. BAL cell gene expression and BAL fluid protein levels of the vascular dysfunction marker, Vascular Endothelial Growth Factor (VEGF), were significantly higher in neutrophilhigh compared to neutrophillow patients. Fibrinogen and heme were significantly increased in neutrophilhigh BAL fluid. In vitro experiments revealed that blood neutrophils had significantly increased expression of LTF and VEGFA following LPS-stimulation and heme induces endothelial dysfunction. COPD patients with distal lung neutrophilic inflammation have dysregulated iron metabolism which may be a consequence of increased vascular leakage into the airways.

Soluble (pro)renin receptor (sPRR), the product of site-1 protease (S1P)-mediated cleavage of PRR, has emerged as an important player in the physiological and pathophysiological processes in the kidney. However, a potential role of S1P-derived sPRR in acute ischemia-reperfusion injury (IRI) still needs to be explored. Hence, the current study comprehensively examined the involvement of S1P-derived sPRR in the pathogenesis of renal IRI in mice. The mouse model of IRI was generated by inducing 30 min of bilateral ischemia, followed by reperfusion. Various parameters of renal injury were assessed at 24 hours after acute kidney injury (AKI). The production of sPRR was blocked by pharmacological inhibition of S1P using PF-429242 or mutagenesis of the cleavage site of PRR (PRRR279V/L282V). The severity of AKI was estimated by plasma creatinine, blood urea nitrogen (BUN), urine microalbumin/creatinine ratio, and tubular injury. Administration of PF-429242 significantly improved IRI-induced renal dysfunction, albuminuria, and tubular injury, accompanied by suppressed macrophage infiltration and M1 polarization. In parallel, IRI elevated plasma sPRR and urinary renin levels, which were both blunted by PF-429242 treatment. These findings were all recapitulated in PRRR279V/L282V mice. Together, these results suggest that S1P-derived sPRR plays a key role in the pathogenesis of renal IRI through macrophage M1 polarization.

Sex differences critically influence the renal response to ischemic injury, yet the mechanisms underlying differing recovery between males and females remain incompletely understood. Using a unilateral ischemia-reperfusion model with contralateral nephrectomy (uIRIx), we performed a longitudinal analysis of the transition from acute kidney injury (AKI) to chronic kidney disease (CKD) in male and female mice following unilateral ischemia-reperfusion injury with contralateral nephrectomy (uIRIx) over 98 days. Male mice developed sustained renal dysfunction, characterized by persistent proteinuria, a marked reduction in glomerular filtration rate, and progressive increases in urinary albumin/creatinine ratio (uACR), consistent with an ongoing functional decline. Histologically, males displayed extensive tubular dilation, interstitial fibrosis, and elevated kidney injury molecule-1 (KIM-1) expression, together with persistent macrophage and T-cell infiltration indicative of unresolved inflammation. In contrast, females exhibited partial functional recovery with improved GFR, reduced proteinuria, and attenuated structural damage, including less fibrosis and tubular injury across all timepoints. Morphometric analysis revealed smaller glomerular cross-sectional areas in males at day 14, suggesting early maladaptive remodelling, whereas females demonstrated adaptive hypertrophy that may preserve filtration capacity. Assessment of peritubular capillaries (CD31) indicated more effective microvascular preservation in females, consistent with estrogen-mediated endothelial protection. Collectively, these findings demonstrate that females are protected from the maladaptive progression of ischemic AKI to CKD, highlighting longitudinal sex-specific dynamics in renal repair and chronic disease development.

Epilepsy is a debilitating neurological disorder characterized by recurrent seizures, affecting millions of patients worldwide. Retrospective studies in Temporal lobe epilepsy (TLE) patients have shown a high incidence of an initial precipitating event (IPE) in early childhood, followed by a silent period where epileptogenesis occurs to end up in chronic epilepsy. Epileptogenesis, the process through which a normal brain undergoes structural and functional changes leading to epilepsy, is not completely understood . We hypothesized that epigenetics may be involved in epileptogenesis, specifically affecting astrocytes through pathological remodeling. To study this process, we used three approaches: The lithium-pilocarpine model of TLE in rats, primary astroglial cultures exposed to epileptogenic DAMP named HMGB1, and brain tissue samples resected from TLE patients with drug-resistant epilepsy. We found that the IPE achieved by lithium-pilocarpine treatment (127/30 mg/kg IP) induced the DNA methylation of astrocytes at 7-, 21-, and 35 days post-IPE, indicating persistent epigenetic alterations in astrocytes during the epileptogenic period. In addition, we observed the downregulation of homeostatic astroglial genes, including AQP4, glutamine synthase (GS), and Kir4.1, along with increased expression of proinflammatory genes (C3, MAFG) and DNA methyltransferases (DNMT). These alterations were mimicked in primary astrocyte cultures exposed to the epileptogenic HMGB1 (500 ng/ml; 18 hours) which resulted in the hypermethylation of homeostatic astroglial genes and repression of homeostatic genes. HMGB1-induced repression of astroglial homeostatic genes was prevented by the treatment with DNMT inhibitor decitabine. Interestingly, astrocytes from TLE patients brains showed reactive astrogliosis, increased DNA methylation, and downregulation of homeostatic genes Kir4.1 and GS. Taken together, these findings show that astrocytes are pathologically altered during the epileptogenic period by epigenetic modifications, combining the proinflammatory gain of function with the loss of homeostatic profile. This may contribute to the long-term alterations underlying epileptogenesis.

Impaired glucose tolerance (IGT) and insulin resistance (IR), including prediabetes and diabetes, increase risk of developing age-related disorders, such as cardiovascular disorders, kidney disorders, and Alzheimer's disease. We analyzed mitochondrial bioenergetics of platelets collected from 208 adults, 55 years and older, with IGT and IR and without normoglycemic (NG). Platelets from IGT participants exhibited unique mitochondrial bioenergetic profiles exemplified by higher mitochondrial respiration compared with NG. IGT platelets exhibited higher glucose-dependent maximal respiration (Max) and spare respiratory capacities (SRCs) and higher fatty acid oxidation (FAO)-dependent maximal coupled (MaxOXPHOS) and uncoupled (maximal electron transport system) respiration compared with NG. Correlating mitochondrial bioenergetics from all 208 participants with measures of glucose tolerance (oral glucose tolerance test values measured 120 min after glucose administration, and oral glucose tolerance test area under the curve), and historical glucose measures [hemoglobin A1 (HbA1c)] revealed significant positive associations. Most associations were unaltered with age, sex, and body mass index adjustments. Examining NG and IGT participants separately, we found platelet respiration and HbA1c exhibited positive association in NG participants. Significant positive associations emerged between platelet SRC, FAO, FAO+CI (oxygen flux due to FAO + complex I activities), and HbA1c. No significant associations were observed in the IGT group. Given the utilization of blood-based mitochondrial bioenergetic profiling strategies in clinical research, this work provides new insights into the clinical features of IR that can affect platelet mitochondrial bioenergetics.