ABSTRACT Background Although epidermal growth factor receptor tyrosine kinase inhibitors (EGFR TKIs) are effective in treating NSCLC with EGFR mutations, the development of resistance limits their long‐term efficacy. Autophagy has been implicated as a potential mechanism behind this acquired resistance. This study aims to investigate the role of Stanniocalcin 2 (STC2) in mediating autophagy and its contribution to EGFR TKI resistance. Methods STC2 expression was manipulated to examine its effects on autophagy and EGFR TKI resistance both in vitro and in vivo. Autophagic activity was assessed by measuring LC3B‐II expression levels, the number of LC3 puncta, and the accumulation of autophagic vacuoles. Pharmacological inhibitors and small interfering RNA (siRNA) were used to assess whether the regulatory relationship between STC2 and Beclin 1 was involved in STC2‐mediated autophagy and EGFR TKI resistance. Results Here, we found that STC2 was associated with increased autophagic activity, as evidenced by elevated LC3B‐II levels, enhanced LC3 puncta formation, and increased accumulation of autophagic vacuoles. Conversely, STC2 knockdown resulted in reduced autophagic activity. We identified that STC2 is associated with ERK1/2 activation and Beclin 1–related autophagic activity. Importantly, inhibiting autophagy reversed the resistance to EGFR TKIs induced by STC2, as demonstrated in vitro and in vivo xenograft mouse models. Conclusion These findings suggest that STC2 appears to promote cytoprotective autophagy, and targeting the STC2‐associated ERK/Beclin 1 signaling axis may offer a promising strategy to overcome resistance to EGFR TKIs.

Background:Diagnosing primary cutaneous T-cell lymphoma (PCTCL) relies on the comprehensive evaluation of clinical presentation, histopathology, immune profile, and molecular testing. Regarding molecular testing, the BIOMED-2 assay is most commonly used for detecting monoclonality of T-cell receptor (TCR) genes. However, its sensitivity is not ideal for PCTCLs, and a supplementary test is required. We recently found that the ratios of non-recombined TCR sequences J2-2P and J2-3 of the TCRβ gene were higher in T-cell lymphoma (TCL) samples than in non-TCL cases. Here, we aim to evaluate the diagnostic value of this novel biomarker for PCTCL via digital polymerase chain reaction (dPCR) as a supplementary method to the BIOMED-2 clonality assay. Methods:Formalin-fixed paraffin-embedded tissue samples of clinicopathologically proven PCTCL cases with a negative BIOMED-2 clonality result were retrospectively retrieved, and the clinicopathological characteristics were collected. After RNA extraction and quality checking, dPCR was performed on PCTCL specimens and control samples to quantify the J2-2P and J2-3 sequences. Results:We collected 35 skin biopsy samples from patients with early-stage PCTCL and 11 inflammatory dermatoses as controls. Ratios of the J2-2P to J2-3 concentration in controls ranged from 0.03 to 0.55% (mean ± standard deviation: 0.21 ± 0.17%). In contrast, 25 of the 35 (71.42%) PCTCL specimens exhibited J2-2P/J2-3 ratios exceeding the defined upper limit of the control group. Conclusion:Our dPCR assay on the non-recombined TCRβ sequences revealed that a majority of the PCTCL cases with a negative BIOMED-2 result had higher ratios of the J2-2P to J2-3 concentration than inflammatory dermatoses. Therefore, this approach may supplement the BIOMED-2 clonality assay for diagnosing PCTCL.

PurposeVascular restenosis following angioplasty remains a major clinical challenge driven by excessive vascular smooth muscle cell (VSMC) migration, proliferation, and fibrosis. Thrombin, a potent signaling molecule beyond its coagulative function, promotes neointimal hyperplasia. Dapagliflozin, a sodium-glucose cotransporter 2 (SGLT2) inhibitor, has been shown to exert cardiovascular protection independent of its glucose-lowering effects, yet its role in thrombin-mediated fibrotic signaling remains unclear.MethodsCultured A10 vascular smooth muscle cells were used to evaluate thrombin- stimulated A Disintegrin And Metalloprotease 17 (ADAM17), ERK1/2, c-Jun, and connective tissue growth factor (CTGF) by immunoblotting and migration analyses. Enzymatic activity (ADAM17 and thrombin) assays were tested by cell-free system kit. Pharmacological inhibition of ADAM17 by TAPI-1 and extracellular signal-regulated kinase (ERK) by U0126 were employed to delineate pathway hierarchy. An in vivo rat carotid artery balloon injury model was used to assess the effects of dapagliflozin on neointimal formation.ResultsThrombin stimulation markedly increased ADAM17 phosphorylation, ERK1/2 and c-Jun activation, and CTGF expression in A10 cells. Dapagliflozin significantly suppressed these responses in a concentration-dependent manner without affecting thrombin enzymatic activity. Inhibition of ADAM17 recapitulated the effects of dapagliflozin, confirming ADAM17 as a key upstream mediator of thrombin-induced ERK/c-Jun/CTGF signaling. Functionally, dapagliflozin attenuated VSMC migration in vitro and significantly reduced neointimal hyperplasia in the rat carotid artery balloon injury model.ConclusionDapagliflozin inhibits thrombin-induced ADAM17 activation and downstream ERK1/2-c-Jun-CTGF signaling, thereby limiting VSMC migration and neointimal formation after vascular injury. This SGLT2-independent mechanism provides new insight into the pleiotropic vasculoprotective actions of dapagliflozin and identifies ADAM17 as a potential therapeutic target for preventing vascular restenosis and fibrosis-related cardiovascular disease.

The spatial role of M1 and M2 tumor-associated macrophages (M1/M2 TAMs) in precision medicine remains unclear. EGFR and TP53 are among the most frequently mutated genes in lung adenocarcinoma. We characterized the mutation status and density of M1/M2 TAMs within tumor islets and stroma in 117 lung adenocarcinomas using next-generation sequencing and immunohistochemistry, respectively. Stromal M1 TAMs were positively correlated with disease progression and smoking history. In contrast, islet M1/M2 TAMs were predominantly found in tumors with wild-type TP53 (wtp53) but not associated with EGFR status. The presence of wtp53 was associated with the spatial distribution of M1/M2 TAMs in tumor islets and stroma. Additionally, dominance of islet M1 TAMs and M1-signature were significantly associated with improved survival in patients with wtp53 lung adenocarcinoma, unlike in those with mutant TP53. Conditioned medium from M1 macrophages (M1 CM) induced apoptosis in wtp53 cells through increased p53 accumulation. We found that interferons in M1 CM activate JAK1/TYK2 via IFNARs, leading to enhanced STAT1 expression and Y701 phosphorylation. This activation facilitates p53-STAT1 interactions, reduces the interaction between p53 and MDM2, and subsequently decreases p53 ubiquitination. M1 CM inhibited tumorigenesis, and silencing p53 reduced the anti-tumor efficacy of polyinosinic:polycytidylic acid (poly I:C) in vivo. Furthermore, higher M1-signature was significantly associated with better responses and survival following anti-PD1 treatment in wtp53 melanomas. IFNs/STAT1/p53 signaling was critical for the anti-tumor activity of M1 macrophages. These findings suggest that p53 modulates the spatial balance of M1/M2 TAMs, and the tumoricidal effects of M1 TAMs depend on p53 status. Thus, p53 companion diagnostics could facilitate the development of M1-oriented therapies, which may be particularly beneficial for wtp53 patients when combined with immunotherapy.

The tumor microenvironment in breast cancer is typically immunosuppressive. Combined neoadjuvant chemotherapy and immunotherapy is the standard of care for high-risk, triple-negative breast cancer patients, but it is not routinely applied to other breast cancer subtypes. Somatic mutation of STK11 (serine/threonine kinase 11), a tumor suppressor, occurs in the MSK-IMPACT platform in 1.9

INTRODUCTION:In the field of interventional cardiology, coronary in-stent restenosis (ISR) continues to present a clinical hurdle, even with the progress made in stent design and pharmacological interventions. While drug-eluting stents (DESs) and drug-eluting balloons (DEBs) have markedly decreased the occurrence of ISR when compared to bare-metal stents, the condition persists as a complication in revascularization, contributing to increased patient morbidity and challenging long-term treatment outcomes. Thus, a deeper understanding of ISR mechanisms and the development of novel therapeutic approaches are crucial for improving patient outcomes. METHODS:In this study, we utilized the A10 cell as an in vitro model and induced common carotid artery balloon dilation injury in Sprague-Dawley rats as an animal model to explore the potential clinical applications of SCH79797, particularly in the treatment of ISR. RESULTS:SCH79797, a protease-activated receptor-1 antagonist, induced apoptosis of smooth muscle cells through various pathways. SCH79797 promoted apoptosis via JNK/c-Jun and p53 upregulation in the cytosol. We also observed an increased Bax/Bcl-2 ratio in mitochondria, p53 translocation to mitochondria, and changes in the mitochondrial membrane potential to mitochondrial membrane permeabilization. Our comparative analysis with vorapaxar revealed the apoptotic effects of SCH79797 to be independent of its PAR-1 antagonist activity. Furthermore, SCH79797 administration significantly reduced common carotid artery restenosis and thrombosis following balloon injury in vivo. DISCUSSION:Our study has been the first to demonstrate SCH79797 to directly induce VSMC apoptosis via the p53-mediated mitochondrial pathway, providing a novel mechanistic insight into ISR treatment. Unlike traditional anti-proliferative agents used in DESs, SCH79797 uniquely combines apoptotic induction with antithrombotic effects, making it a dual-action therapeutic candidate. This research study has laid the groundwork for localized drug-eluting strategies that can leverage SCH79797's properties to prevent ISR more effectively while minimizing systemic side effects. CONCLUSION:Our findings have established SCH79797 as a promising candidate for reducing ISR through apoptosis modulation. By leveraging the p53-mediated mitochondrial apoptotic pathway, SCH79797 may provide a groundbreaking approach to reducing restenosis. These findings could offer significant implications for the future development of targeted drug-eluting strategies by locally delivering SCH79797 in a controlled manner using DES or DEB, presenting SCH79797 as a transformative candidate in interventional cardiology.

AbstractBackgroundIdentifying breast cancer markers with superior sensitivity, cost‐effectiveness, and practicality is imperative. Circulating immune cells and plasma cytokines hold promise as potential breast cancer markers.AimsTo search and validate subtype of circulating immune cells and plasma cytokines as biomarkers for breast cancer patients.Materials & MethodsUsing flow cytometry, we investigated circulating immune cell profiles in patients with breast cancer and healthy controls. To validate clinical observations, an orthotopic breast cancer model was established.ResultsWe analyzed 19 healthy controls and 27 patients (22 testing group and 5 validation group) with breast cancer and revealed distinct populations, including CD3+CD4+ T lymphocytes, cytotoxic T lymphocytes (CTLs; CD3+CD8+), polymorphonuclear myeloid‐derived suppressor cells (PMN‐MDSCs), and monocytic (M)‐myeloid‐derived suppressive cells. Patients with breast cancer exhibited reduced CD3+CD4+ T lymphocyte, CD3+CD8+ CTL, and CD33+CD15− M‐MDSC levels compared with healthy controls. Diminished CD3+CD8+ CTL levels correlated with advanced cancer grade, extensive intraductal components, and positive lymphatic tumor emboli. Treatment effects included decreased T lymphocyte/PMN‐MDSC levels, contrasting with elevated circulating CD3+CD8+ cell levels posttreatment, subsequently declining upon recurrence in the validation group. Elevated plasma chemokine (C–C motif) ligand 2 (CCL2) levels distinguished patients with breast cancer from healthy controls. Our orthotopic model supported decreased circulating CD3+CD8+ CTL levels in cancer‐bearing mice, followed by a postresection increase.DiscussionWe found that circulating CD3+CD8+ CTL levels decreased in patients with breast cancer, increased after treatment, and decreased again upon recurrence.ConclusionCirculating CD3+CD8+ CTL emerged as promising prognostic biomarkers and therapeutic targets in breast cancer management.

Tumor relapse and metastasis are the major causes of mortality associated with urothelial cancer. In the tumor microenvironment, negative regulatory molecules and various immune cell subtypes suppress antitumor immunity. The inflammatory microenvironment, associated with neutrophils and neutrophil extracellular traps (NETs), promotes tumor metastasis. However, no drugs are currently available to specifically inhibit neutrophils and NETs. In this study, we first demonstrated that icaritin (ICT), a Chinese herbal remedy that is a first-line treatment for advanced and incurable hepatocellular carcinoma, reduces NETs caused by suicidal NETosis and prevents neutrophil infiltration in the tumor microenvironment. Mechanistically, ICT binds to and inhibits the expression of PADI2 in neutrophils, thereby suppressing PADI2-mediated histone citrullination. Moreover, ICT inhibits ROS generation, suppresses the MAPK signaling pathway, and inhibits NET-induced tumor metastasis. Simultaneously, ICT inhibits tumoral PADI2-mediated histone citrullination, which consequently suppresses the transcription of neutrophil-recruiting genes such as GM-CSF and IL-6. The downregulation of IL-6 expression, in turn, forms a regulatory feedback loop through the JAK2/STAT3/IL-6 axis. Through a retrospective study of clinical samples, we found a correlation between neutrophils, NETs, UCa prognosis, and immune evasion. Combining ICT with immune checkpoint inhibitors may have synergistic effects. In summary, our study demonstrated that ICT could be a novel inhibitor of NETs and a novel UCa treatment.

Yes-associated protein 1 (YAP1) mediated hippo pathway has attracted several research importance in various types of malignancies. The hampered Hippo-YAP1 axis in bladder cancer (BC) was identified as a major driver of BC progression and oncogenesis. The activity of Hippo pathway is controlled via regulation of phosphorylation cascade of MST1/2-LATS1/2-YAP1, while other modification as ubiquitination of hippo pathway protein also mediated the activity of Hippo pathway through the co-regulation of E3 ligases and deubiquitinases. In this study, we identified USP20 as a Hippo/YAP1 pathway activity related deubiquitinase through combined analysis of siRNA screening and deubiquitinase over-expression. Further analysis identified that USP20 directed regulated the YAP1 expression and target gene of YAP1, connective tissue growth factor (CTGF) as well as cysteine-rich angiogenic inducer 61 (CYR61). Tissue microarray assay confirmed that USP20 was elevated in tumor tissue and correlated with YAP1 expression. Mechanism study identified that USP20 directly interacted with YAP1 protein and promoted the stability of YAP1 protein through hampering the K48-linked poly-ubiquitination. Our findings in this study revealed that USP20 as a novel deubiquitinase in regulating the Hippo-YAP1 pathway in BC.

Background and objectiveCryoablation is a traditional antitumor therapy with good prospects for development. The efficacy of endoscopic management as a kidney-sparing surgery for high-risk upper tract urothelial carcinoma (UTUC) remains controversial. Our aim was to evaluate the impact of endoscopic cryoablation (ECA) versus radical nephroureterectomy (RNU) on survival outcomes, renal function, and complications.MethodsWe retrospectively analyzed data for 116 patients with newly diagnosed high-risk UTUC who underwent either ECA (n = 13) or RNU (n = 103) from March 25, 2019 to December 8, 2021. Propensity score matching (1:4) using the nearest neighbor method was performed before analysis. The primary outcome was overall survival (OS). Secondary outcomes included progression-free survival (PFS), intravesical recurrence-free survival (RFS), the change in renal function, and treatment-emergent adverse events (TEAEs).Key findings and limitationsAt median follow-up of 28.2 mo for the ECA group and 27.6 mo for the RNU group, 2-yr OS (82% vs 84%), PFS (73% vs 71%), and intravesical RFS (81% vs 83%) rates after matching did not significantly differ. A decline in renal function was observed after RNU, but not after ECA. Five (41.7%) patients in the ECA group reported six TEAEs, and 17 patients (35.4%) in the RNU group reported 20 TEAEs.Conclusions and clinical implicationsIn comparison to RNU, ECA for UTUC resulted in noninferior oncological outcomes and superior preservation of renal function.Patient summaryOur study suggests that a treatment called endoscopic cryoablation for high-risk cancer in the upper urinary tract can help in preserving kidney function, with similar survival outcomes to those after more extensive surgery. This option can be considered for selected patients with a strong preference for kidney preservation.

Background: This study aims to investigate the clinical characteristics and the short-term consequences of COVID19 in older people with epilepsy. Methods: From January 28 to March 6,2023,a telephone follow-up survey was conducted to a group of older people with epilepsy after COVID-19 outbreak in Shanghai. Data on infection symptoms,duration,COVID-19 vaccination, and clinical characteristics and management of epilepsy were collected. Results: Eighty-two participants aged >= 50 years with SARS-CoV-2 infection were recruited. Common symptoms included fever (72.0%) and cough (63.4%).Overall, anti-seizure medication(ASM) accessibility was sufficient (95.1%),and medication adherence was good (93.9%).Three participants (3.7%)reported 1-3 seizures after virus infection. No statistically significant differences were observed in terms of clinical characteristics and consequences of COVID-19 in subgroups with participants aged <63 and >= 63years,with and without fever, antipyretic drugs users and non-users, and with >= 4 and <4 seizures/year before COVID-19. Conclusion: Our study found few cases of seizure onset and good ASM adherence after SARS-CoV-2 infection in older people with epilepsy. Further prospective studies are needed to understand the long-term consequences of COVID-19.

BackgroundThe influences of blood lipids and lipid-regulatory medications on the risk of bladder cancer have long been suspected, and previous findings remain controversial. We aimed to assess the causality between blood lipids or lipid-regulatory medications and bladder cancer susceptibility by means of a comprehensive Mendelian Randomization (MR) study.MethodsGenetic proxies from genome-wide association studies (GWAS) of four blood lipid traits and lipid-lowering variants in genes encoding the targets of lipid-regulatory medications were employed. The largest ever GWAS data of blood lipids and bladder cancer involving up to 440,546 and 205,771 individuals of European ancestry were extracted from UK Biobank and FinnGen Project Round 6, respectively. A two-sample bidirectional MR study was performed using the inverse variance weighted as the main method. The heterogeneity, horizontal pleiotropy, MR Steiger, and leave-one-out analyses were also conducted as sensitivity tests.ResultsThere was indicative evidence that genetically predicted low-density lipoprotein cholesterol (LDL-C) affected bladder cancer susceptibility based on 146 single nucleotide polymorphisms (SNPs) with an odds ratio (OR) of 0.776 (95% confidence interval [CI] = 0.625–0.965, p = 0.022). However, this result became non-significant after two SNPs that possibly drove the effect were removed as demonstrated by leave-one-out analysis. The reversed MR analysis suggested that bladder cancer could not affect serum lipid levels. No causal relationship was found between the lipid-lowering effect of lipid-regulatory medications (fibrates, probucol, statins, ezetimibe, proprotein convertase subtilisin/kexin type 9 [PCSK9] inhibitors, and evinacumab) and the risk of bladder cancer. No heterogeneity or pleiotropy was found (all p > 0.05).ConclusionThis MR study revealed for the first time, using the most recent and comprehensive GWAS data to date, that genetically predicted total cholesterol (TC) and the lipid-lowering effect of lipid-regulatory medications had no causal association with bladder cancer susceptibility. We also verified claims from early studies that low-density lipoprotein cholesterol (HDL-C), LDL-C, and triglyceride (TG) are not related to bladder cancer susceptibility either. The current study indicated that lipid metabolism may not be as important in the tumorigenesis of bladder cancer as previously believed.

Background: Resistance to standard chemotherapy is a critical problem for breast cancer patients. The ATP-binding cassette (ABC) superfamily transporters actively pump out drugs and play an important role in chemoresistance. ABCB1 (ABC subfamily B, member 1, also named as multidrug resistance protein 1, MDR1) and suppressive myeloid-derived suppressor cells (MDSCs) potentially involve in chemoresistance of breast cancer. The relationship between ABCB1 and immune genes in breast cancer has not been widely studied.Methods: Microarray and RNA sequencing data were obtained from The Cancer Genome Atlas Breast Invasive Carcinoma in Genomic Data Commons Data Portal and Gene Expression Omnibus database. A patient-derived xenograft (PDX) model of HER2+ breast cancer was established to investigate the association between ABCB1 and immune genes in breast cancer.Results: Expression of ABCB1 increased in doxorubicin-selected MCF-7/ADR cells. High expression of ABCB1 mRNA is correlated with lymph-node metastasis and worse overall survival in patients with breast cancer. ABCB1 is positively correlated with IL6, CSF1, CSF3, and PTGS2. In the HER2+ stage IIA breast cancer PDX model, both doxorubicin and paclitaxel suppressed growth of P2 tumors. IL6, CSF1, CSF3, and PTGS2 expression were suppressed by paclitaxel but not doxorubicin. Intrasplenic MDSCs, including CD11b+Ly6G+ and CD11b+Ly6C+ cells, were more abundant than intratumor MDSCs in PDX-carrying nude mice. Clinically, the patient developed cancer recurrence after adjuvant chemotherapy with doxorubicin-based regimen and was well controlled after paclitaxel-trastuzumab combined therapy.Conclusion: ABCB1 was a poor predictor of HER2+ LN- breast cancer. Regulation of immune genes by ABCB1 contributed to cancer recurrence and treatment effect. The PDX model was suitable for investigation the expression of target genes and expansion of immune cells.

EGFR exon 19 deletion (Del‐19) comprises multiple advanced NSCLC subtypes. EGFR‐tyrosine kinase inhibitor (TKI) efficacy and T790M acquisition in various Del‐19 subtypes is unknown. We prospectively collected tissue samples from patients harboring NSCLC with Del‐19 between 2006 and 2020. We evaluated EGFR‐TKI treatment effectiveness among the different Del‐19 subtypes. We collected 1391 NSCLC samples from 892 patients with Del‐19, and the most common subtype was del E746‐A750 (67.5%). 741 patients had taken first‐ or second‐generation EGFR‐TKIs. There were no significant differences in response rates between patients with different Del‐19 subtypes (P = .630). Patients with indel E746 had the longest median PFS (14.6 months), but those with non‐LRE deletions had the shortest PFS (8.9 months; P = .002). For OS analysis, patients with indel E746 also had the longest OS (34.1 months), but those with non‐LRE deletions had the shortest OS (21.1 months; P = .046). Patients with different Del‐19 subtypes showed no significant differences in the T790M acquisition rates (P = .443). Among the 151 patients with acquired T790M who received third‐generation EGFR‐TKIs, the Del‐19 subtype was not associated with different RR and PFS. In vitro cellular viability and activation of the EGFR pathway analysis were consistent with the clinical findings. In conclusion, compared with del E746‐A750, indel E746 was associated with longer PFS and OS, but the non‐LRE subtype was correlated with shorter survival prognosis. There were no significant differences in the acquired T790M rate and treatment effectiveness of subsequent third‐generation EGFR‐TKIs between various Del‐19 subgroups.

Background: Assessing the prognosis preoperatively in patients with upper tract urothelial carcinoma (UTUC) remains a challenge for urologists. Gross hematuria (GH) and flank pain (FP) are the 2 most common and easily perceived symptoms of UTUC. Therefore, we aimed to investigate the prognostic values of GH and FP in patients with UTUC after undergoing radical nephroureterectomy (RNU). Methods: This article retrospectively analyzed 179 patients with UTUC who underwent RNU and examined the associations between the FP, GH, and long-term survival. After dividing patients into 4 subgroups (presenting as GH without FP, FP without GH, no FP and GH, FP with GH), we focused on the prognostic values of the 4 subgroups using univariate and multivariate analyses. We then proposed a risk stratification model for UTUC based on the independent prognostic factors for cancer-specific survival (CSS) with external validation (146 additional UTUC patients formed the validation cohort). Results: Patients with FP had worse oncological outcomes than those without FP (P < .05). After dividing the 179 patients into 4 subgroups, the “FP without GH” subgroup suffered the worst oncological outcomes (P < .001). The Cox multivariate regression analysis showed that “FP without GH” (P < .001), tumor multifocality (P = .005), and pathological stage (P = .004) were independent prognostic factors for CSS. Good performance of the risk stratification model was achieved in both the training and external validation cohorts. Conclusion: The presence of “flank pain without gross hematuria” was one of the independent risk factors of CSS and OS besides the pathological stage and tumor multifocality. To our knowledge, this is the first study that adding complaint to risk stratification model in UTUC.

BackgroundRecurrence and metastasis of bladder cancer are major factors affecting patient prognosis. Endoscopic cryoablation achieved a better clinical outcome among clinical patients and could be synergistic with ICIs. Thus, this study aimed to evaluate the immunological mechanism of cryoablation for bladder cancer to reveal the therapeutic mechanism. MethodsWe systematically reviewed the clinical prognosis of patients underwent cryoablation at Huashan Hospital in these first-in-human studies (ChiCTR-INR-17013060). Murine models were constructed to explore cryoablation-induced tumour-specific immunity, which was further confirmed by primary bladder tumour organoids and autologous lymphocytes cocultured system. ResultsCryoablation improved progression-free survival and recurrence-free survival respectively. Assessment of murine models after cryoablation confirmed microenvironment remodelling and tumour-specific T cells expansion. Enhanced antitumour effects were found after coculture of organoids with autologous lymphocytes collected from post-cryoablation. We also demonstrated cryoablation-induced tumour elimination required IFNGR expression on tumour cells. In addition, a long-lasting antitumour memory response is achieved by cryoablation and could be enhanced after combination with ICIs. ConclusionsThis study revealed endoscopic cryoablation is an efficient and safe therapy for bladder tumour treatment. The tumour-specific immune responses induced by cryoablation could reduce tumour recurrence and metastasis.

Background:Upper tract urothelial carcinoma (UTUC) is a relatively rare disease with a poor prognosis. A growing body of evidence demonstrates that inflammation and the inflammatory microenvironment play a crucial role in tumorigenesis and tumor progression. Our aim was to evaluate the prognostic value of blood inflammation markers and develop a prediction model that incorporates inflammation markers in order to predict overall survival (OS) of UTUC.Methods:We included 304 localized UTUC patients from two medical institutions who had undergone radical nephroureterectomy (RNU) (167 in the training cohort, 137 in the validation cohort). Univariate and multivariate Cox regression analyses were performed to screen the prognostic factors, and a nomogram and a web-based calculator were generated based on these predictors. The Harrell's concordance index (C-index), the area under the receiver operating characteristic (ROC) curve, the calibration curve, and decision curve analysis (DCA) were used to evaluate the performance of the nomogram.Results:Independent predictors incorporated in the nomogram were pathological stage, surgical margin, albumin-to-globulin ratio (AGR), and hemoglobin-to-red cell distribution width ratio (HRR). The c-index value was 0.726 in the training cohort and 0.761 in the validation cohort. The area under the ROC of the nomogram at 1-, 3- and 5-year in the training and validation sets were 0.765, 0.755, 0.763, and 0.791, 0.833, 0.802, respectively. Both the internal and external validation calibration plots showed a subtle distinction between the predicted and the actual probabilities. And it appears to provide incremental benefits for clinical decision-making in comparison to the American Joint Committee of Cancer (AJCC) staging system.Conclusions:In patients with UTUC after RNU, lower preoperative AGR and HRR were independent predictors of inferior survival. In addition, we created a novel blood inflammation marker-based dynamic nomogram that may be useful for surgeons or oncologists in risk stratification and patient selection for more intensive therapy and closer follow-up.

Metabolism vulnerability of cisplatin resistance in BCa cells remains to be discovered, which we applied integrated multi-omics analysis to elucidate the metabolism related regulation mechanism in bladder cancer (BCa) microenvironment. Integrated multi-omics analysis of metabolomics and proteomics revealed that MAT2A regulated methionine metabolism contributes to cisplatin resistance in BCa cells. We further validated MAT2A and cancer stem cell markers were up-regulated and circARHGAP10 was down-regulated through the regulation of MAT2A protein stability in cisplatin resistant BCa cells. circARHGAP10 formed a complex with MAT2A and TRIM25 to accelerate the degradation of MAT2A through ubiquitin-proteasome pathway. Knockdown of MAT2A through overexpression of circARHGAP10 and restriction of methionine up-take was sufficient to overcome cisplatin resistance in vivo in immuno-deficiency model but not in immuno-competent model. Tumor-infiltrating CD8+ T cells characterized an exhausted phenotype in tumors with low methionine. High expression of SLC7A6 in BCa negatively correlated with expression of CD8. Synergistic inhibition of MAT2A and SLC7A6 could overcome cisplatin resistance in immuno-competent model in vivo. Cisplatin resistant BCa cells rely on methionine for survival and stem cell renewal. circARHGAP10/TRIM25/MAT2A regulation pathway plays an important role in cisplatin resistant BCa cells while circARHGAP10 and SLC7A6 should be evaluated as one of the therapeutic target of cisplatin resistant BCa.

Prostate cancer is the most prevalent type of cancer among men worldwide. The importance of circular RNA (circRNA) in prostate cancer and its connection to malignancy has been steadily recognized. circRNA expression was obtained by circRNA sequencing of prostate cancer. circRNA and its function were further analysed. The results were verified by qRT-PCR, RIP assay, FISH, RNA pulldown, WB, CCK-8, colony formation assay and wound-healing assay. BALB/c Nude mice were used for xenograft hosts. Low expression of circDHRS3 was assessed in prostate cancer. Overexpression of circDHRS3 inhibited prostate cancer growth and migration in vitro. Additionally, miR-421 was shown to be the downstream target of circDHRS3, as shown by fluorescence in situ hybridization and dual-luciferase experiments. The rescue assay results for the PC3 and Du145 cell lines demonstrated that circDHRS3 inhibits prostate cancer cell lines' ability to proliferate and metastasize by modulating MEIS2 expression through the circDHRS3/miR-421/MEIS2 axis. In vivo investigations confirmed that the overexpression of circDHRS3 could inhibit both the lung and bone metastasis of prostate cancer cells. circDHRS3 has the potential to become a biomarker and a targeted therapeutic site for prostate cancer, particularly in the malignant stage. Our study indicates that circDHRS3 inhibits prostate cancer cell proliferation and metastasis through the circDHRS3/miR-421/MEIS2 axis.