Radiation proctopathy is a severe dose-limiting complication of pelvic radiotherapy that lacks effective interventions. Here, we investigate whether iron accumulation contributes to radiation-induced rectal fibrosis and elucidate the underlying mechanism. In a mouse model of radiation proctopathy, we observed significant ferritin-mediated iron accumulation in fibrotic rectal tissues, which was negatively correlated with vitamin D receptor (VDR) downregulation. In vitro, iron overload promoted activation of human and mouse rectal fibroblasts by suppressing VDR expression, which relieved inhibition of YAP and triggered its nuclear translocation, leading to upregulation of pro-fibrotic target genes (Ctgf and Ankrd1). Notably, treatment with the iron chelator deferoxamine (DFO) effectively cleared iron accumulation, restored VDR expression, inhibited YAP activation, and significantly alleviated radiation-induced collagen deposition and fibroblast activation in vivo. Collectively, these findings reveal a critical role of the 'iron accumulation-VDR-YAP' regulatory axis in radiation proctopathy and suggest that iron chelation represents a potential therapeutic strategy warranting further preclinical and clinical investigation.

PURPOSE:Patients with identical stages or similar tumor volumes can vary significantly in their responses to radiation therapy (RT) due to individual characteristics, making personalized RT for non-small cell lung cancer (NSCLC) challenging. This study aimed to develop a deep learning model by integrating pretreatment and midtreatment computed tomography (CT) to predict the treatment response in NSCLC patients. METHODS AND MATERIALS:We retrospectively collected data from 168 NSCLC patients across 3 hospitals. Data from Shanghai General Hospital (SGH, 35 patients) and Shanxi Cancer Hospital (SCH, 93 patients) were used for model training and internal validation, while data from Linfen Central Hospital (LCH, 40 patients) were used for external validation. Deep learning, radiomics, and clinical features were extracted to establish a varying time interval long short-term memory network for response prediction. Furthermore, we derived a model-deduced personalize dose escalation (DE) for patients predicted to have suboptimal gross tumor volume regression. The area under the receiver operating characteristic curve (AUC) and predicted absolute error were used to evaluate the predictive Response Evaluation Criteria in Solid Tumors classification and the proportion of gross tumor volume residual. DE was calculated as the biological equivalent dose using an /α/β ratio of 10 Gy. RESULTS:The model using only pretreatment CT achieved the highest AUC of 0.762 and 0.687 in internal and external validation respectively, whereas the model integrating both pretreatment and midtreatment CT achieved AUC of 0.869 and 0.798, with predicted absolute error of 0.137 and 0.185, respectively. We performed personalized DE for 29 patients. Their original biological equivalent dose was approximately 72 Gy, within the range of 71.6 Gy to 75 Gy. DE ranged from 77.7 to 120 Gy for 29 patients, with 17 patients exceeding 100 Gy and 8 patients reaching the model's preset upper limit of 120 Gy. CONCLUSIONS:Combining pretreatment and midtreatment CT enhances prediction performance for RT response and offers a promising approach for personalized DE in NSCLC.

Gliomas are the most lethal types of adult brain tumors with a devastating prognosis, but many therapies have failed to exert good therapeutic benefits because of the extremely hypoxic and immunosuppressive tumor microenvironment. To address these challenges, we herein present a semiconducting polymer (SP)-based small interfering RNA (siRNA) nanosystem with the loading of oxygen self-supplying perfluorohexane (PFH) and conjugation of siRNA via a singlet oxygen (1O2)-cleavable linker. The nanosystems are further camouflaged with a macrophage membrane to obtain the final RM@SPN-siRNA. RM@SPN-siRNA displays an enhanced enrichment at the orthotopic glioma site due to surface cell membrane camouflaging. PFH provides sufficient oxygen to relieve tumor hypoxia, which boosts the production of 1O2 by the SP working as the radiosensitizer under external X-ray irradiation. The generated 1O2 destroys the 1O2-cleavable linker and disrupts the membrane structure to enable in situ siRNA release at the tumor site and subsequent activatable programmed death ligand-1 (PD-L1) silencing for tumor cells. As a consequence, an immunological effect is triggered to effectively inhibit tumor growths in an orthotopic glioma mouse model. This study offers an X-ray-responsive siRNA nanosystem for precise protein silencing and treatment of deep-seated orthotopic tumors.

While various tactics have been adopted to treat glioma, the clinical therapeutic efficacy of glioma is still poor due to its aggressiveness, blood–brain barrier (BBB), drug resistance and highly immunosuppressive microenvironment. We herein present X-ray-activatable hitchhiking polymer nanodrugs (HPNFcN) that can enable controlled ferroptosis and immunization for rejecting orthotopic glioma. HPNFcN are formed via loading a newly synthesized ferroptosis-inducing ferrocene prodrug and an immunotherapeutic drug (NLG919) into reactive oxygen species (ROS)-responsive semiconducting polymer nanosystems with surface embellishment of a neutrophil targeting ligand. We found that neutrophil-targeted HPNFcN could achieve effective delivery into the orthotopic glioma sites via acrossing BBB. Low-dose X-ray irradiation of HPNFcN resulted in the generation of ROS by semiconducting polymer, which not only activated the ferrocene prodrug to cause ferroptosis, but also wrecked the ROS-responsive nanosystems to on-demand deliver NLG919. Moreover, the ferroptosis effect triggered immunogenic cell death (ICD) to promote immunization, which synergized with the NLG919-based blockade for reinforcing antitumor immune response. Such a precise and effective therapeutic strategy could observably reject orthotopic glioma without significant side effects. This study demonstrates the superiorities of HPNFcN for glioma treatment via neutrophil-mediated effective delivery and X-ray-activatable therapeutic actions of ferroptosis and immunization.

Osteoporosis, characterized by low BMD, is a highly heritable metabolic bone disorder. Although single nucleotide variations (SNVs) have been extensively studied, they explain only a fraction of BMD heritability. Although genomic structural variations (SVs) are large-scale genomic alterations that contribute to genetic diversity in shaping phenotypic variations, the role of SVs in osteoporosis susceptibility remains poorly understood. This study aims to identify and prioritize genes that harbor BMD-related SVs. We performed whole genome sequencing on 4982 subjects from the Louisiana Osteoporosis Study. To obtain high-confidence SVs, the detection of SVs was performed using an ensemble approach. The SVs were tested for association with BMD variation at the hip (HIP), femoral neck (FNK), and lumbar spine (SPN), respectively. Additionally, we conducted co-occurrence analysis using multi-omics approaches to prioritize the identified genes based on their functional importance. Stratification was employed to explore the sex- and ethnicity-specific effects. We identified significant SV-BMD associations: 125 for FNK-BMD, 99 for SPN-BMD, and 83 for HIP-BMD. We observed SVs that were commonly associated with both FNK and HIP BMDs in our combined and stratified analyses. These SVs explain 13.3% to 19.1% of BMD variation. Novel bone-related genes emerged, including LINC02370, ZNF family genes, and ZDHHC family genes. Additionally, FMN2, carrying BMD-related deletions, showed associations with FNK or HIP BMDs, with sex-specific effects. The co-occurrence analysis prioritized an RNA gene LINC00494 and ZNF family genes positively associated with BMDs at different skeletal sites. Two potential causal genes, IBSP and SPP1, for osteoporosis were also identified. Our study uncovers new insights into genetic factors influencing BMD through SV analysis. We highlight BMD-related SVs, revealing a mix of shared and specific genetic influences across skeletal sites and gender or ethnicity. These findings suggest potential roles in osteoporosis pathophysiology, opening avenues for further research and therapeutic targets.

Although many important advances have been made in the treatment of nasopharyngeal carcinoma (NPC) in recent years, local recurrence and distant metastasis remain the main factors affecting NPC prognosis. Biomarkers for predicting the prognosis of NPC need to be urgently identified. Here, we used whole-exon sequencing (WES) to determine whether PICK1 mutations are associated with the prognosis of NPC. Functionally, PICK1 inhibits the proliferation and metastasis of NPC cells both in vivo and in vitro. Mechanistically, PICK1 inhibited the expression of proteins related to the Wnt/β-catenin signaling pathway. PICK1 restrained the nuclear accumulation of β-catenin and accelerated the degradation of β-catenin through the ubiquitin-proteasome pathway. The reduced PICK1 levels were significantly associated with poor patient prognosis. Hence, our study findings reveal the mechanism by which PICK1 inactivates the Wnt/β-catenin signaling pathway, thereby inhibiting the progression of NPC. They support PICK1 as a potential tumor suppressor and prognostic marker for NPC.

Blood pressure (BP)-relevant non-coding SNPs identified by GWAS may influence the expression of distal protein-coding genes. Our previous study detected 195 SNPs (out of 26585) in CTCF-binding sites. CTCF and Cohesin are known to modulate cell-type-specific gene expression from a distance by forming chromatin loops. We hypothesized that both may regulate gene expression in BP-relevant cell types. To test this, we depleted CTCF and Rad21 (Cohesin subunit) in human induced pluripotent stem cells-derived endothelial cells (iEC) and performed genome-wide transcriptomic analysis. We detected 366 and 4184 differentially expressed genes (DEGs) in CTCF and Rad21 depleted iEC (CTCF-/Rad21-KD-iEC) compared to iEC control, respectively. Among them, 111 DEGs are common between the CTCF-KD and Rad21-KD-iEC, though majority showed opposite regulatory effects (Figure 1A). We found 5619 DEGs in the CTCF-KD vs Rad21-KD-iEC (Figure 1B). Pathway analysis detected elevated cell cycle (G2M checkpoint), DNA replication (E2F targets), and repair pathways in Rad21-KD compared to the CTCF-KD-iEC. Rad21 plays an important role in chromosome partitioning and repair, so its depletion may cause dysregulation of these pathways. While the CTCF-KD-iEC showed enrichment of the Epithelial-to-mesenchymal transition (EMT), inflammatory (IFN-gamma, IFN-alpha), and hypoxia pathways (Figure 1C). As CTCF maintains the chromosome boundaries, CTCF-KD may affect chromatin compactness and trigger abnormal gene expression related to these pathways. In conclusion, our study detected different effects of depletion of CTCF and Rad21 on gene expression in a BP-relevant cell type. The findings provide a basis to explore the role of chromatin looping in BP-relevant gene regulation.

Background: Tacrolimus is one of the most commonly used basic immunosuppressants nowadays, but the high variability of tacrolimus blood concentration often leads to kidney transplant recipients frequently experiencing drug concentrations above or below the target concentration, resulting in renal toxicity or rejection of the transplanted kidney. The aim of this study is to explore the correlation of renal function with intra-patient variability (IPV) of tacrolimus blood concentration among recipients of renal transplants at 1-, 3-, 5-, and 10-year post-transplantation. Methods: Recipients of renal transplants who were treated with tacrolimus for immunosuppression at the Shanghai General Hospital between January 2001 and December 2009, and followed up until 2019 were included in this retrospective study. Demographic characteristics and laboratory investigation results at their 1-, 3-, 5-, and 10-year follow-up visits were collected from their hospital medical records. Patients were divided into a low or high IPV group based on the IPV of their tacrolimus concentrations. Results: A total of 167 kidney transplant recipients were included in the study. At the 3-year followup visit, patients in the low IPV group had significantly lower blood urea nitrogen (BUN) (6.3 +/- 1.8 vs. 8.2 +/- 6.2 mu mol/L, P=0.04), serum creatinine (Scr) (88.8 +/- 23.6 vs. 104.8 +/- 39.6 mu mol/L, P=0.009), and blood uric acid (UA) (329.1 +/- 80.2 vs. 375.9 +/- 95.1 mu mol/L, P=0.004), as well as significantly higher estimated glomerular filtration rate (eGFR) values than patients in the high IPV group. Blood UA levels were significantly lower in patients in the low IPV group than the high IPV group at the 10-year follow-up (362.7 +/- 92.6 vs. 398.5 +/- 105.2 mu mol/L, P=0.042). There was no significant difference between the low and high IPV groups with respect to BUN, Scr, UA, or eGFR at the 1- and 5-year follow-up. Conclusions: Recipients of renal transplants with lower IPV in tacrolimus concentration appeared to have better renal function over time. Controlling IPV may contribute to improved renal outcomes post- transplantation.

Background The enriched proteins within in vitro fertilisation (IVF)-generated human embryonic microenvironment could reverse progestin resistance in endometrial cancer (EC). Methods The expression of thymic stromal lymphopoietin (TSLP) in EC was evaluated by immunoblot and IHC analysis. Transcriptome sequencing screened out the downstream pathway regulated by TSLP. The role of TSLP, androgen receptor (AR) and KANK1 in regulating the sensitivity of EC to progestin was verified through a series of in vitro and in vivo experiments. Results TSLP facilitates the formation of a BMP4/BMP7 heterodimer, resulting in activation of Smad5, augmenting AR signalling. AR in turn sensitises EC cells to progestin via KANK1. Downregulation of TSLP, loss of AR and KANK1 in EC patients are associated with tumour malignant progress. Moreover, exogenous TSLP could rescue the anti-tumour effect of progestin on mouse in vivo xenograft tumour. Conclusions Our findings suggest that TSLP enhances the sensitivity of EC to progestin through the BMP4/Smad5/AR/KANK1 axis, and provide a link between embryo development and cancer progress, paving the way for the establishment of novel strategy overcoming progestin resistance using embryo original factors.

BACKGROUND: Hypertension or elevated blood pressure (BP) is a worldwide clinical challenge and the leading primary risk factor for kidney dysfunctions, heart failure, and cerebrovascular disease. The kidney is a central regulator of BP by maintaining sodium-water balance. Multiple genome-wide association studies revealed that BP is a heritable quantitative trait, modulated by several genetic, epigenetic, and environmental factors. The SNPs identified in genome-wide association studies predominantly (>95%) reside within noncoding genomic regions, making it difficult to understand how they regulate BP. Given the central role of the kidney in regulating BP, we hypothesized that chromatin-accessible regions in renal tissue would be enriched for BP-associated single nucleotide polymorphisms. METHODS: We manually dissected 2 important kidney segments that maintain the sodium-water balance: proximal tubules and medullary thick ascending limbs from the human and rat kidneys. To delineate their chromatin and transcriptomic profiles, we performed the assay for transposase-accessible chromatin and RNA sequencing, respectively. RESULTS: The chromatin accessibility maps revealed the shared and unique cis -regulatory elements that modulate the chromatin accessibility in proximal tubule and medullary thick ascending limbs of humans and rats. We developed a visualization tool to compare the cross-species epigenomic maps to identify potential regulatory targets for hypertension pathogenesis. We also identified a significant enrichment of BP-associated single nucleotide polymorphisms (1064 for human proximal tubule and 1172 for human medullary thick ascending limbs) within accessible chromatin regions of both segments, including rs1173771 and rs1421811 at the NPR3 locus and rs1800470 at the TGFb1 locus. CONCLUSIONS: Collectively, this study lays a foundation for interrogating how intergenic single nucleotide polymorphisms may regulate polygenic traits such as BP.

PDF file - 123K, Ezh2 regulates E-cadherin expression.

Essential hypertension, a multifaceted disorder, is a worldwide health problem. A complex network of genetic, epigenetic, physiological, and environmental components regulates blood pressure (BP), and any dysregulation of this network may result in hypertension. Growing evidence suggests a role for epigenetic factors in BP regulation. Any alterations in the expression or functions of these epigenetic regulators may dysregulate various determinants of BP, thereby promoting the development of hypertension. Histone posttranslational modifications are critical epigenetic regulators that have been implicated in hypertension. Several studies have demonstrated a clear association between the increased expression of some histone-modifying enzymes, especially HDACs (histone deacetylases), and hypertension. In addition, treatment with HDAC inhibitors lowers BP in hypertensive animal models, providing an excellent opportunity to design new drugs to treat hypertension. In this review, we discuss the potential contribution of different histone modifications to the regulation of BP.

BACKGROUND:Various kidney diseases such as acute kidney injury, chronic kidney disease, polycystic kidney disease, renal cancer, and kidney stones, are an important part of the global burden, bringing a huge economic burden to people around the world. Ferroptosis is a type of nonapoptotic iron-dependent cell death caused by the excess of iron-dependent lipid peroxides and accompanied by abnormal iron metabolism and oxidative stress. Over the past few decades, several studies have shown that ferroptosis is associated with many types of kidney diseases. Studying the mechanism of ferroptosis and related agonists and inhibitors may provide new ideas and directions for the treatment of various kidney diseases.SUMMARY:In this review, we discuss the differences between ferroptosis and other types of cell death such as apoptosis, necroptosis, pyroptosis, cuprotosis, pathophysiological features of the kidney, and ferroptosis-induced kidney injury. We also provide an overview of the molecular mechanisms involved in ferroptosis and events that lead to ferroptosis. Furthermore, we summarize the possible clinical applications of this mechanism among various kidney diseases.KEY MESSAGE:The current research suggests that future therapeutic efforts to treat kidney ailments would benefit from a focus on ferroptosis.

Aims: Hypoxia is an important feature of nasopharyngeal carcinoma (NPC). "Protein interacting with PRKCA 1" (PICK1) is commonly downregulated in human malignancies and is functionally related to poor prognosis. However, there is a limited understanding of the upstream mechanisms regulating PICK1 currently.Main methods: PICK1 and HIF-1a expression levels were analyzed by Immunohistochemistry (IHC), western blotting, and quantitative real-time PCR assay. Protein stability and ubiquitin assays were used to investigate PICK1 protein degradation. Immunofluorescence and co-immunoprecipitation assays were used to demonstrate the interaction between RBCK1 and PICK1. Gene knockdown by siRNA transfection was used to investigate the role of HIF-1a and RBCK1 in hypoxia-induced PICK1 degradation. Cell Counting Kit-8 (CCK-8), 5-Ethynyl-2 & PRIME;- deoxyuridine (EdU) assays and subcutaneous xenograft nude models were used to explore the roles of RBCK1 and PICK1 in NPC cell proliferation.Key findings: PICK1 expression in NPC tissue was negatively relative to that of HIF-1a. HIF-1a downregulated PICK1 expression by facilitating its ubiquitination by the E3 ligases RANBP2-type and C3HC4-type zinc finger containing 1 (RBCK1), thereby enhancing proteasome-mediated PICK1 degradation. RBCK1 knockdown inhibited NPC cell proliferation, which was ameliorated by double knockdown of RBCK1/PICK1.Significance: These data provide evidence for an NPC cell adaptation mechanism to hypoxia, where HIF-1a regulates RBCK1, which targets PICK1 for degradation to promote cell proliferation.

PDF file - 63K, Multivariate Cox regression analysis for overall survival of patients with bladder cancer.

PURPOSE:This study is purposed to establish a predictive model for acute severe hematologic toxicity (HT) during radiotherapy in patients with cervical or endometrial cancer and investigate whether the integration of clinical features and computed tomography (CT) radiomics features of the pelvic bone marrow (BM) could define a more precise model. METHODS:A total of 207 patients with cervical or endometrial cancer from three cohorts were retrospectively included in this study. Forty-one clinical variables and 2226 pelvic BM radiomic features that were extracted from planning CT scans were included in the model construction. Following feature selection, model training was performed on the clinical and radiomics features via machine learning, respectively. The radiomics score, which was the output of the final radiomics model, was integrated with the variables that were selected by the clinical model to construct a combined model. The performance of the models was evaluated using the area under the receiver operating characteristic curve (AUC). RESULTS:The best-performing prediction model comprised two clinical features (FIGO stage and cycles of postoperative chemotherapy) and radiomics score and achieved an AUC of 0.88 (95% CI, 0.81-0.93) in the training set, 0.80 (95% CI, 0.62-0.92) in the internal-test set and 0.85 (95% CI, 0.71-0.94) in the external-test dataset. CONCLUSION:The proposed model which incorporates radiomics signature and clinical factors outperforms the models based on clinical or radiomics features alone in terms of the AUC. The value of the pelvic BM radiomics in chemoradiotherapy-induced HT is worthy of further investigation.

PDF file - 67K, siRNA Sequence in the study.

The overall therapeutic output is significantly affected by the route of nanomedicines in vivo, including the successive steps of delivery, exogenous excitation, and final metabolism, thus requiring the elaborate composition and structure of nanomedicines. To develop nanomedicines with the full-route advances, we have designed and architected the biomimetic and biodegradable ultrasmall-in-nano particles (UiNP, 80 nm) with the radiation-photo synergy for the first time, by utilizing cancer cell membrane (CCM) nanovehicles to camouflage multiple ultrasmall particles (UP6, 6 nm) composed of Chlorin e6 (Ce6) and hafnium (Hf) clusters via the covalent coordination. The UiNP with the camouflage effect, homotypic adhesion and suitable size can obtain 4-fold blood circulation half-live that of UP6 and 2.5-fold accumulation level that of nanoparticle (NP180, 180 nm), achieving delivery advance. Moreover, the UiNP not only exhibit a dual-modal X-ray therapy with radio-radiodynamic effect via energy transfer but also afford a dual-modal phototherapy with photodynamic-photothermal effect by adjusting the radiative-nonradiative transition of Ce6 within the confined space, thus balancing the killing efficacy and the penetration depth for therapeutic advance. Furthermore, benefiting from the ultrasmall size and intrinsic biodegradability of UP6 and CCM camouflage, the UiNP are less accumulated in reticuloendothelial organs than NP180 and thereby rapid metabolize via urine and feces, exerting the metabolic advance. Therefore, such novel UiNP architectures with full-route advances are much superior to the conventional agents, which can provide some insights for developing new types of ultrasmall-in-nano agents. (C) 2022 Elsevier Ltd. All rights reserved.

Radiation therapy offers limited clinical benefits for patients with pancreatic cancer, partly as a result of the predominantly immunosuppressive microenvironment characteristic of this specific type of cancer. A large number of abnormal blood vessels and high-density fibrous matrices in pancreatic cancer will lead to hypoxia within tumor tissue and hinder immune cell infiltration. We used low-dose X-ray irradiation, also known as low dose radiation therapy (LDRT), to normalize the blood vessels in pancreatic cancer, while simultaneously administering an inhibitor of focal adhesion kinase (FAK) to reduce pancreatic cancer fibrosis. We found that this treatment successfully reduced pancreatic cancer hypoxia, increased immune cell infiltration, and increased sensitivity to radiation therapy for pancreatic cancer.

Radiation-induced rectal injury is a common side effect of radiotherapy. Hypoxia often occurs after radiotherapy. This study aimed to explore the bystander effect of hypoxia on radiation-induced rectal injury. In vivo, apoptosis increased nearby the highly hypoxic area in the rectal tissues in the mouse models of radiation-induced rectal injury, indicating the potential involvement of hypoxia. In vitro, flow cytometry and Western blotting showed that both hypoxia and hypoxic human intestinal epithelial crypt (HIEC) cell supernatant promoted apoptosis in normoxic HIEC cells. The pro-apoptotic effect of extracellular vesicles (EVs) derived from hypoxic HIEC cell to normoxic HIEC cells was then determined. MiR-122-5p was chosen for further studies through a microRNA (miRNA) microarray assay and apoptosis was alleviated in cells receiving miR-122-5p inhibiting hypoxic EVs. Together, our study demonstrated that the miR-122-5p containing-EVs derived from hypoxic HIEC cells promoted apoptosis in normoxic HIEC cells. Hypoxic EV-derived miR-122-5p plays a critical pathologic role in radiation-induced rectal injury and may be a potential therapeutic target.