Tubulointerstitial fibrosis is a critical and irreversible process of chronic kidney disease. Dedifferentiated proximal tubular cells (PTCs) after injury are important for tubulointerstitial fibrosis. Hepatocyte nuclear factor 4 alpha (HNF4A) is the main regulatory factor for PTC differentiation. However, its role in PTC dedifferentiation and kidney fibrosis remains unclear. To investigate the role of HNF4A in kidney fibrosis, bioinformatics analysis and in vivo models were used to evaluate its expression in kidney tissues. The mechanisms through which the HNF4A P2 isoform inhibits kidney fibrosis were examined by using both in vivo and in vitro models. In this study, we revealed that the sustained downregulation of HNF4A expression was a key characteristic of abnormally repaired PTCs after injury and was associated with cell dedifferentiation. It was confirmed that the HNF4A P2 isoform, rather than the P1 isoform, inhibited TGF-β1-induced PTC dedifferentiation. The activation of fibroblasts, which was induced by dedifferentiated PTCs through paracrine signalling, was also inhibited. In vivo experiments confirmed that HNF4A P2 was more effective than HNF4A P1 was in alleviating kidney fibrosis. Mechanistically, on one hand, HNF4A P2 antagonized the TGF-β1-induced dedifferentiation of PTCs by inhibiting the JAG1/NOTCH pathway. On the other hand, the distinct structure of HNF4A P2 from that of P1 made it unaffected by TGF-β1-activated SRC, allowing HNF4A P2 to perform transcriptional regulatory functions. These findings suggest that targeting the HNF4A P2 isoform could serve as a novel therapeutic strategy to alleviate kidney fibrosis.

Framework nucleic acids effectively meet the demands for precise size control and accurate targeting in the design of drug delivery systems, while developing a controllable drug delivery system with low immunogenicity and high efficiency for delivering nucleic acid drugs to the tumor immune microenvironment (TIME) remains significant challenge. Inspired by ancient Chinese mortise and tenon joint structures, this study develops an intelligent self-assembling DNA tetrahedron (TDN@siCSF-1R), which consists of a gapped DNA tetrahedron (TDN) and a therapeutic siRNA against Colony-Stimulating Factor-1 Receptor (siCSF-1R) that non-covalently bind with TDN via its gap, aiming to target tumor-associated macrophages (TAMs) and inhibit the CSF-1R pathway. Additionally, a CD206 mRNA-responsive sequence is introduced into the gapped TDN, triggering the site-specific release of siCSF-1R in M2-like TAMs, thereby achieving the precise targeting of CSF-1R in M2-like TAMs and reducing off-target effect. The mortise-and-tenon-like TDN@siCSF-1R synchronously combines the self-assembly flexibility and structural stability, significantly inhibiting 4T1 tumor growth, lung metastasis, and tumor recurrence after resection in vivo. Furthermore, it repolarizes M2-like TAMs and activates infiltrating T cells in TIME, thereby reshaping the immunosuppressive microenvironment, and offering a promising strategy for the clinical application of cancer immunotherapy.

Immune checkpoint therapy for prostate cancer (PCa), a classic 'immune-cold' tumor characterized by an immunosuppressive tumor microenvironment, failed previously in clinical trials, but the underlying causes remain elusive. Here we find that YY1+, immunosuppressive macrophages aggregate in the hypoxic areas of PCa. Mechanistically, hypoxia promotes the phase separation of YY1 in the nucleus, where YY1 binds to NUSAP1 and promotes the SUMOylation, phase separation and stabilization of HIF-1α. Either myeloid-specific conditional knockout of YY1 or a treatment with tenapanor for decreasing the YY1-NUSAP1-HIF-1α interaction impairs subcutaneous PCa tumor formation in mouse prostate tumor models. Lastly, a first-generation tetrahedral DNA nanostructure based on the proteolysis targeting chimera technique, termed YY1-DcTAC, allows targeting and degrading YY1 in tumor-associated macrophages for inducing antitumor effects and CD8+ T cell tumor infiltration in mouse tumor models. In summary, our findings underscore the pivotal role of YY1 in the hypoxia/HIF-1α pathway in tumor-associated macrophages and support the targeting of YY1 for treating PCa.

It has been claimed that microRNA 503-5p (miR-503-5p) is the key to the future diagnosis and treatment of cardiac hemangioma (CH), but the relationship between the two has not been fully validated. In this study, we analyzed the effect of miR-503-5p targeting type IA bone morphogenetic protein receptor (BMPR1A) on CH to inform future diagnosis and treatment of CH. First, miR-503-5p and BMPR1A abnormal expression sequences (vectors) were transfected into human hemangioma-derived endothelial cells (HemECs) and human umbilical vein endothelial cells (HUVECs) to observe alterations in cell biological behavior, adhesion, and epithelial mesenchymal transition (EMT). We found enhanced proliferative, invasive and migrating abilities of HemECs and HUVECs after silencing miR-503-5p or increasing BMPR1A, accompanied by reduced apoptosis, elevated intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1), and accelerated EMT; after increasing miR-503-5p or silencing BMPR1A, the cells exhibited reduced apoptosis, elevated ICAM-1 and VCAM-1, and accelerated EMT (P<0.05). Subsequently, a dual-luciferase reporter assay was performed to analyze the targeting relationship between miR-503-5p and BMPR1A. The results showed that miR-503-5p inhibited BMPR1A-wild type (WT) fluorescence activity (P<0.05). Through the rescue experiment, it was observed that the biological behavior of the cells with simultaneous elevation or simultaneous silencing of miR-503-5p and BMPR1A was not different from that of cells transfected with BMPR1A empty vector (P>0.05), indicating that the effect of BMPR1A on cells was reversed by miR-503-5p. Finally, in the analysis of clinical records, we found that CH cases exhibited lower miR-503-5p and higher BMPR1A levels than healthy controls (P<0.05). The expression of the two genes was negatively correlated (P<0.05). These results suggest that miR-503-5p participates in CH growth by targeted sponging of BMPR1A.

Immune checkpoint therapy for prostate cancer (PCa) has failed in clinical trials; however, the precise underlying mechanisms involved remain elusive. PCa, a classic "immune-cold" tumor, is characterized by an immunosuppressive tumor microenvironment. Within this milieu, macrophages, the predominant immune cell population, have a propensity to infiltrate the hypoxic zones of tumors. In a previous study, we showed that Yin Yang 1 (YY1) is highly expressed in macrophages in PCa tissues. Here, through multiplexed imaging mass cytometry (IMC) of a PCa tissue microarray, we further demonstrate that YY1+ macrophages aggregate in hypoxic areas of tumors and that hypoxia promotes the phase separation of YY1 in the nucleus by increasing YY1 tyrosine phosphorylation in macrophages. Furthermore, YY1 binds to NUSAP1 and promotes the SUMOylation of HIF-1α, which promotes phase separation and stabilization of the HIF-1α protein. We also demonstrated that either treatment with a small molecule inhibitor (tenapanor) to decrease the YY1-NUSAP1-HIF-1α interaction or myeloid-specific YY1 gene knockout impairs subcutaneous PCa tumor formation. Furthermore, we present a first-generation tetrahedral DNA nanostructure (TDN) based on the proteolysis targeting chimera (PROTAC) technique, named YY1-DcTAC, which targets and degrades YY1 in tumor-associated macrophages. In a PCa mouse model, YY1-DcTAC exhibited prolonged drug efficacy, robust macrophage-specific responsiveness, potent antitumor effects, and increased CD8+ T cell tumor infiltration. In summary, our findings underscore the pivotal role of YY1 within the hypoxia/HIF-1α pathway in tumor-associated macrophages and affirm the therapeutic potential of targeting YY1 for treating PCa. ### Competing Interest Statement The authors have declared no competing interest.

PURPOSE:Cisplatin-containing combination chemotherapy has been the standard of care since the late 1980s, but the response rate is <50%. Studies have shown that the efficiency of chemotherapy differs among molecular subtypes of bladder cancer. In this study, we aimed to correlate FOXA1, a marker for differentiation of the basal and luminal subtypes, with tumor immune cell infiltration and the effect of chemotherapy in bladder cancer. MATERIALS AND METHODS:Eighty-three patients with bladder cancer treated with chemotherapy were reviewed. Clinicopathological variables for each case were recorded. FOXA1, M2 tumor-associated macrophage (TAM), dendritic cell (DC), and cytotoxic T lymphocyte (CTL) were examined by immunohistochemistry. The relationship between FOXA1, immune cell infiltration, and clinical response to chemotherapy was assessed. RESULTS:The overall objective response rate was 34%. The objective response rate for tumors with lower FOXA1 expression was 58% and for tumors with higher FOXA1 expression was 12%. Tumors with infiltrated M2 TAM proportion <3% had a higher objective response rate compared with infiltrated M2 TAM proportion >3% tumors (46% vs. 21%, p = 0.02). Tumors with infiltrated CTL proportion >5% had a higher objective response rate compared with infiltrated CTL proportion <5% tumors (50% vs. 17%, p = 0.002). DCs showed no significant differences. We found that the objective response rate for tumors with lower FOXA1 expression, proportion <3% M2 TAM infiltration, and proportion >5% CTL infiltration is 82%. Lower FOXA1 expression was associated with low M2 TAM infiltration and high CTL infiltration. CONCLUSIONS:Thus, we showed that in patients with bladder cancer who received chemotherapy, the higher clinical response rate is associated with low FOXA1 expression, low M2 TAM infiltration, and high CTL infiltration.

Figure S5 shows human splenic and skin expressions of CCR8

Figure S4 shows donor immune cell reconstitution in the blood post Ccr8-/- donor bone marrow transplant

Figure S6 shows lower FC receptor transcription in normal skin versus tumor tissues

Binding, blocking and Fc engaging characteristics of anti-mouse CCR8 antibodies

Figure S8 shows binding and CD16 cross-linking characteristics of anti-human CCR8 antibodies

Figure S7 shows 4T1 mouse tumor reductions resulting from anti-mCCR8 or anti-mCCR8/anti-mPD1 treatments

BackgroundErectile dysfunction is a condition with a high incidence among adult men. Lycopene has been shown to lower blood glucose and reduce weight in diabetic patients because of its antioxidant and anti-inflammatory properties. However, the association between lycopene and the incidence of erectile dysfunction is unclear. ObjectiveThe aim of this study was to examine the dietary lycopene intake and its association with erectile dysfunction risk in the US population. Materials and methodsWe investigated the lycopene intake of adult participants with complete information on clinical variables from the National Health and Nutrition Examination Survey between 2001 and 2004. Dose-response curve analysis was applied to explore the association between lycopene intake and erectile dysfunction. Logistic regression models were used to adjust for confounders. Different ethnicities, body mass index level, hypertension status, diabetes status, and smoking status were analyzed as subgroups. Propensity score matching was employed to eliminate the effects of potential confounders to confirm the reliability of the results. ResultsA total of 3265 participants with lycopene consumption data were included in our study, including 931 individuals with erectile dysfunction and 2334 without erectile dysfunction during National Health and Nutrition Examination Survey 2001-2004. We found more consumption of lycopene in the non-erectile dysfunction group than in the erectile dysfunction group. Dose-response curve analysis revealed a significant negative association between lycopene intake and erectile dysfunction prevalence. After adjusting for age, race, cigarette smoking, body mass index, annual family income, education, physical activity, hypertension, diabetes, depression, and testosterone level, we found that increased lycopene intake reduced the odds ratio of erectile dysfunction. Low lycopene intake was positively related to erectile dysfunction in almost all subgroups, especially in Mexican American, non-Hispanic white, body mass index <25, hypertension positive, diabetes mellitus negative, and smoke negative. Furthermore, the results were confirmed in the 1:1 matched group. ConclusionOur national data suggest that lower dietary lycopene intake is positively associated with an increased risk of erectile dysfunction in US men.

Supplemental materials and methods

Figure S2 shows single cell RNAseq analysis of CCR8+ and CCR8- subpopulations of tumor CD4+FOXP3+ T cells

Figure S1 shows that FOXP3+ cells lacking CCR8 are composed of effector T cells and less activated Tregs

BackgroundTumor-associated macrophages are mainly polarized into the M2 phenotype, remodeling the tumor microenvironment and promoting tumor progression by secreting various cytokines.MethodsTissue microarray consisting of prostate cancer (PCa), normal prostate, and lymph node metastatic samples from patients with PCa were stained with Yin Yang 1 (YY1) and CD163. Transgenic mice overexpressing YY1 were constructed to observe PCa tumorigenesis. Furthermore, in vivo and in vitro experiments, including CRISPR-Cas9 knock-out, RNA sequencing, chromatin immunoprecipitation (ChIP) sequencing, and liquid-liquid phase separation (LLPS) assays, were performed to investigate the role and mechanism of YY1 in M2 macrophages and PCa tumor microenvironment.ResultsYY1 was highly expressed in M2 macrophages in PCa and was associated with poorer clinical outcomes. The proportion of tumor-infiltrated M2 macrophages increased in transgenic mice overexpressing YY1. In contrast, the proliferation and activity of anti-tumoral T lymphocytes were suppressed. Treatment targeting YY1 on M2 macrophages using an M2-targeting peptide-modified liposome carrier suppressed PCa cell lung metastasis and generated synergistic anti-tumoral effects with PD-1 blockade. IL-4/STAT6 pathway regulated YY1, and YY1 increased the macrophage-induced PCa progression by upregulating IL-6. Furthermore, by conducting H3K27ac-ChIP-seq in M2 macrophages and THP-1, we found that thousands of enhancers were gained during M2 macrophage polarization, and these M2-specific enhancers were enriched in YY1 ChIP-seq signals. In addition, an M2-specific IL-6 enhancer upregulated IL-6 expression through long-range chromatin interaction with IL-6 promoter in M2 macrophages. During M2 macrophage polarization, YY1 formed an LLPS, in which p300, p65, and CEBPB acted as transcriptional cofactors.ConclusionsPhase separation of the YY1 complex in M2 macrophages upregulated IL-6 by promoting IL-6 enhancer-promoter interactions, thereby increasing PCa progression.

Castration-resistant prostate cancer (CRPC) is the most common malignant tumor of the male urinary system. Nanodrug delivery systems (NDDS) have been widely applied in drug delivery for tumor therapy; however, nanotherapeutics encounter various biological barriers that prevent successful accumulation of drugs, specifically at diseased sites. Therefore, there is an urgent need to develop a CRPC-targeting nanocomposite with fine biocompatibility for penetrating various biological barriers, delivering sufficient drugs to the targeting site and improving therapeutic efficiency. In this work, CRPC cell membranes were firstly adapted as biomimetic vectors for the encapsulating PEG−PLGA polymer containing the chemotherapy drug docetaxel (DTX). The CRPC membrane-camouflaged nanoparticles can easily escape early recognition by the immune system, penetrate the extracellular barrier, and evade clearance by the circulatory system. In addition to the characteristics of traditional nanoparticles, the CRPC cell membrane contains an arsenal of highly specific homotypic moieties that can be used to recognize the same cancer cell types and increase the targeted drug delivery of DTX. In vivo fluorescence and radionuclide dual-model imaging were fulfilled by decorating the biomimetic nanosystem with near-infrared dye and isotope, which validated the homotypic targeting property offered by the CRPC cell membrane coating. Importantly, remarkably improved therapeutic efficacy was achieved in a mice model bearing CRPC tumors. This homologous cell membrane enabled an efficient drug delivery strategy and enlightened a new pathway for the clinical application of tumor chemotherapy drugs in the future.

Acetylcholinesterase (AChE) plays an important role in the process of chemotherapy drug-induced cell apoptosis and is also an apoptosis marker to diagnose disease and assay the effect of the drug. Thus, we designed and synthesized a novel near-infrared fluorescent probe (NFL-SF) to detect AChE activity, which contained near infrared (NIR) fluorophore NFL-OH and 2-thienylformyl chloride as the recognition unit. The probe has higher specificity and sensitivity to AChE and excellent biocompatibility. With the increasing concentration of AChE, the probe displayed an enhanced fluorescence signal at 745 nm for 30 min, with the detection limit of 0.2 mU/mL. In addition, NFL-SF not only detected exogenous AChE activity, but also effectively monitored the endogenous AChE activity of chemotherapeutic drug-induced cell apoptosis in living cells. When the AChE activity was inhibited with donepezil, the fluorescence intensity of the NIR channel was significantly reduced. Notably, the probe was successfully applied to in vivo imaging of nude mice. We believe that this probe could provide a strategy for rapidly and visually monitoring AChE activity.

前列腺癌是男性较为常见的恶性肿瘤,其发病率及病死率呈逐年上升趋势.文献报道,骨骼是前列腺癌远处转移的最常见靶器官,但有关骨髓转移的报道少见,有关其治疗鲜有报道[1].我院收治了1例以血小板显著减少为首发症状的前列腺癌伴骨髓转移患者,现报道如下.