PDIA2 has a dual function in promoting androgen deprivation therapy induced venous thrombosis events and castrate resistant prostate cancer progression.

Androgen deprivation therapy (ADT) is the first line of treatment for metastatic prostate cancer (PCa) that effectively delays the tumor progression. However, it also increases the risk of venous thrombosis event (VTE) in patients, a leading cause of mortality. How a pro-thrombotic cascade is induced by ADT remains poorly understood. Here, we report that protein disulfide isomerase A2 (PDIA2) is upregulated in PCa cells to promote VTE formation and enhance PCa cells resistant to ADT. Using various in vitro and in vivo models, we demonstrated a dual function of PDIA2 that enhances tumor-mediated pro-coagulation activity via tumor-derived extracellular vehicles (EVs). It also stimulates PCa cell proliferation, colony formation, and xenograft growth androgen-independently. Mechanistically, PDIA2 activates the tissue factor (TF) on EVs through its isomerase activity, which subsequently triggers a pro-thrombotic cascade in the blood. Additionally, TF-containing EVs can activate the Src kinase inside PCa cells to enhance the AR signaling ligand independently. Androgen deprivation does not alter PDIA2 expression in PCa cells but enhances PDIA2 translocation to the cell membrane and EVs via suppressing the clathrin-dependent endocytic process. Co-recruitment of AR and FOXA1 to the PDIA2 promoter is required for PDIA2 transcription under androgen-deprived conditions. Importantly, blocking PDIA2 isomerase activity suppresses the pro-coagulation activity of patient plasma, PCa cell, and xenograft samples as well as castrate-resistant PCa xenograft growth. These results demonstrate that PDIA2 promotes VTE and tumor progression via activating TF from tumor-derived EVs. They rationalize pharmacological inhibition of PDIA2 to suppress ADT-induced VTE and castrate-resistant tumor progression.

Neutrophil Extracellular Traps Delay Diabetic Wound Healing by Inducing Endothelial-to-Mesenchymal Transition via the Hippo pathway.

Diabetic foot ulcers (DFUs) are among the most frequent complications of diabetes with significant morbidity and mortality. Diabetes can trigger neutrophils to undergo histone citrullination by protein arginine deiminase 4 (encoded by Padi4 in mice) and release neutrophil extracellular traps (NETs). The specific mechanism of NETs-mediated wound healing impairment in diabetes remains unknown. In this study, we show neutrophils are more susceptible to NETosis in diabetic wound environments. Via in vitro experiments and in vivo models of wound healing using wide-type and Padi4 -/- mice, we demonstrate NETs can induce the activation of PAK2 via the membrane receptor TLR-9. Then PAK2 phosphorylates the intracellular protein Merlin/NF2 to inhibit the Hippo-YAP pathway. YAP binds to transcription factor SMAD2 and translocates from the cytoplasm into the nucleus to promote endothelial-to-mesenchymal transition (EndMT), which ultimately impedes angiogenesis and delays wound healing. Suppression of the Merlin/YAP/SMAD2 pathway can attenuate NET-induced EndMT. Inhibition of NETosis accelerates wound healing by reducing EndMT and promoting angiogenesis. Cumulatively, these data suggest NETosis delays diabetic wound healing by inducing EndMT via the Hippo-YAP pathway. Increased understanding of the molecular mechanism that regulates NETosis and EndMT will be of considerable value for providing cellular targets amenable to therapeutic intervention for DFUs.

Interleukin 6-regulated macrophage polarization controls atherosclerosis-associated vascular intimal hyperplasia.

Vascular intimal hyperplasia (VIH) is an important stage of atherosclerosis (AS), in which macrophages not only play a critical role in local inflammation, but also transform into foam cells to participate into plaque formation, where they appear to be heterogeneous. Recently, it was shown that CD11c+ macrophages were more associated with active plaque progression. However, the molecular regulation of phenotypic changes of plaque macrophages during VIH has not been clarified and thus addressed in the current study. Since CD11c- cells were M2a-polarized anti-inflammatory macrophages, while CD11c+ cells were M1/M2b-polarized pro-inflammatory macrophages, we used bioinformatics tools to analyze the CD11c+ versus CD11c- plaque macrophages, aiming to detect the differential genes associated with M1/M2 macrophage polarization. We obtained 122 differential genes that were significantly altered in CD11c+ versus CD11c- plaque macrophages, regardless of CD11b expression. Next, hub genes were predicted in these 122 genes, from which we detected 3 candidates, interleukin 6 (Il6), Decorin (Dcn) and Tissue inhibitor matrix metalloproteinase 1 (Timp1). The effects of these 3 genes on CD11c expression as well as on the macrophage polarization were assessed in vitro, showing that only expression of Il6, but not expression of Dcn or Timp1, induced M1/M2b-like polarization in M2a macrophages. Moreover, only suppression of Il6, but not suppression of either of Dcn or Timp1, induced M2a-like polarization in M1/M2b macrophages. Furthermore, pharmaceutical suppression of Il6 attenuated VIH formation and progression of AS in a mouse model that co-applied apolipoprotein E-knockout and high-fat diet. Together, our data suggest that formation of VIH can be controlled through modulating macrophage polarization, as a promising therapeutic approach for prevent AS.

RNA Splicing of the Abi1 Gene by MBNL1 contributes to macrophage-like phenotype modulation of vascular smooth muscle cell during atherogenesis.

BACKGROUND: Vascular smooth muscle cells (VSMC) switch to macrophage-like cells after cholesterol loading, and this change may play an important role in atherogenesis. Muscleblind-like splicing regulator 1 (MBNL1) is a well-known splicing factor that has been implicated in many cellular processes. However, the role of MBNL1 in VSMC macrophage-like transdifferentiation is largely unknown. In this study, we aim to characterize the role of MBNL1-induced gene splicing during atherogenesis. METHODS: The expression of MBNL1 and Abelson interactor 1 (Abi1) splice variants (Abi1-e10 and Abi1-Δe10) was compared between artery tissues from healthy donors and atherosclerosis patients. Regulatory mechanisms of MBNL1-induced Abi1 gene splicing were studied, and the signal pathways mediated by Abi1 splice variants were investigated in VSMC. RESULTS: Loss of MBNL1 was found in the macrophage-like VSMC (VSMC-M) in artery wall from atherosclerosis patients. In vitro and in vivo evidence confirmed that Abi1 is one of the MBNL1 target genes. Loss of MBNL1 significantly induces the Abi1-Δe10 isoform expression. Compared to the known actin organization activities of the Abi1 gene, we discovered a novel action of Abi1-Δe10, whereby Abi1-Δe10 activates Rac1 independent of upstream stimulation and triggers the Rac1-NOX1-ROS pathway, which results in increased expression of transcription factor Kruppel-like factor 4 (KLF4). While Abi1-Δe10 inhibits contractile VSMC biomarkers expression and cell contraction, it stimulates VSMC proliferation, migration and macrophage-like transdifferentiation. CONCLUSION: Loss-of-function of MBNL1 activates VSMC-M transdifferentiation to promote atherogenesis through regulating Abi1 RNA splicing.

Clinical efficacy of one-stage thrombus removal via contralateral femoral and ipsilateral tibial venous access for pharmacomechanical thrombectomy in entire-limb acute deep vein thrombosis.

OBJECTIVE: In the present study, we compared the early results between different approaches for pharmacomechanical thrombectomy (PMT) in the treatment of entire-limb acute deep vein thrombosis (DVT). METHODS: The present retrospective cohort study included patients with entire-limb acute DVT who had undergone PMT from January 2016 to March 2019 at two independent vascular centers. At the first center (Renji Hospital), the vascular surgeons used contralateral femoral venous access or ipsilateral tibial venous access (CFVA/ITVA). All consecutive patients with entire-limb acute DVT had undergone PMT through CFVA/ITVA at the first center. At the second center (Affiliated Hangzhou First People's Hospital), the vascular surgeons had conducted PMT using the traditional approach via ipsilateral popliteal venous access (IPVA). All consecutive patients had undergone PMT through IPVA at the second center. The primary endpoint was the incidence of post-thrombotic syndrome (PTS). The secondary endpoints included thrombus removal grade, venous primary patency rate, and the incidence of moderate-to-severe PTS. RESULTS: A total of 73 patients were enrolled in the present study, including 37 patients with CFVA/ITVA at the first center and 36 patients with IPVA at the second center. No significant difference was detected between the two groups in age, gender, onset time, affected limb, or risk factors. The proportion of patients who had undergone catheter-directed thrombolysis was significantly lower in the CFVA/ITVA group than in the IPVA group (P = .010). Thrombus removal grade III was achieved more often in the CFVA/ITVA group than in the IPVA group (P = .007). The PTS incidence was significantly lower in the CFVA/ITVA group than in the IPVA group (P = .043). The thrombus removal grade and access type were independent factors associated with the development of PTS. Patients with complete thrombus removal (grade III) and CFVA/ITVA had a significantly lower incidence of PTS. CONCLUSIONS: PMT can increase the thrombus clearance rate, reduce the requirement for subsequent catheter-directed thrombolysis, and, potentially, decrease the incidence of PTS using CFVA/ITVA instead of traditional IPVA in the treatment of entire-limb acute DVT.

SREBP1 suppresses the differentiation and epithelial function of hiPSC-derived endothelial cells by inhibiting the microRNA199b-5p pathway.

Human induced pluripotent stem cell (hiPSC)-derived endothelial cell (hiPSC-EC) transplantation is a promising therapy for treating peripheral artery disease (PAD). However, the poor differentiation of hiPSCs limits their clinical application. Therefore, finding key factors that regulate cellular differentiation is crucial for improving the therapeutic efficacy of hiPSC-EC transplantation. Sterol regulatory element binding protein 1 (SREBP1) is a key regulator of lipid metabolism and stem cell differentiation. However, it remains unknown whether SREPBP1 modulates hiPSC differentiation. In this study, we showed that SREBP1 expression was negatively associated with hiPSC differentiation and EC function. The results show that SREBP1 binds to the promoter region of miR199b-5p and suppresses its transcription, resulting in the activation of Notch1 signaling. Blocking SREBP1 increased both hiPSC differentiation and EC angiogenesis. These findings demonstrate a novel role for SREBP1 in hiPSC differentiation and EC angiogenesis.

Neutrophil-Lymphocyte Ratio and Platelet-Lymphocyte Ratio Predict Severity and Prognosis of Lower Limb Arteriosclerosis Obliterans.

BACKGROUND: The neutrophil-lymphocyte ratio (NLR) and platelet-lymphocyte ratio (PLR) are 2 markers of inflammation, which are associated with worse cardiovascular disease outcomes. Here, we aimed to determine the association between these ratios and disease severity and evaluate predictive validity of the NLR and PLR in lower limb arteriosclerosis obliterans (ASO). METHODS: We evaluated 211 patients with a diagnosis of ASO from January 2016 to December 2018 at Shanghai Jiaotong University Renji Hospital. The NLR and PLR were accessed from routinely drawn peripheral venous blood at the ward of vascular surgery during hospitalization. The association between the NLR and PLR with baseline characteristics, disease severity, and one-year outcomes were determined, respectively. RESULTS: Both the NLR and PLR showed significant values on predicting disease severity. A higher NLR (P = 0.001) and PLR (P < 0.001) were associated with lower ankle-brachial index and worse clinical presentation. Both the NLR and PLR are positively correlated with one-year readmission rate (P < 0.001, P = 0.001, respectively). Both the NLR and PLR also positively correlated with the tissue loss rate and one-year mortality (P = 0.007, P = 0.034, respectively). CONCLUSIONS: The NLR and PLR show a positive association with the severity of lower extremity peripheral artery disease, both higher ratios correlate with poor prognosis, especially, the risk of one-year readmission. A higher NLR also correlates with one-year mortality.

Viabahn Stent Graft for the Endovascular Treatment of Occlusive Lesions in the Femoropopliteal Artery: A Retrospective Cohort Study with 4-Year Follow-Up.

BACKGROUND: The polytetrafluoroethylene-covered Viabahn stent may be effective for the endovascular treatment of patients with femoropopliteal artery occlusive lesions. However, the long-term efficacy of Viabahn stent remains unclear. The aim of the study is to evaluate the long-term patency of Viabahn stent grafts in patients with occlusive lesions in the femoropopliteal artery. METHODS: Consecutive patients with occlusive lesions in the femoropopliteal artery who had been treated with Viabahn stent grafts during the period from June 2013 to December 2016 at our center were retrospectively included. Accumulative incidences of primary patency and secondary patency were estimated by Kaplan-Meier survival analysis, and the predictors of primary patency were evaluated by Cox regression analysis. RESULTS: A total of 66 patients underwent successful endovascular treatment and were included in the study. Endovascular treatment with a Viabahn stent was associated with a complication rate of 9.1% and a 30-day mortality rate of 1.5%. Sixty-one patients were followed for a mean duration of 29.5 months. The 1-year, 2-year, 3-year, and 4-year primary patency rates were 81.7%, 74.7%, 67.6%, and 58.9%, respectively. The secondary patency rates were 94.9%, 92.9%, 90.1%, and 90.1%, respectively. The overall major amputation rate was 5.0%. The results of multivariate Cox regression analyses showed that stent location was the only independent predictor of primary patency (P = 0.001). Implantation of a Viabahn stent above the knee, compared with implantation below the knee, was associated with a higher rate of primary patency. CONCLUSIONS: The Viabahn stent graft is associated with a satisfactory rate of long-term patency for the endovascular treatment of occlusive lesions in the femoropopliteal artery, especially for those located above the knee.

Clinical Features and Outcomes of Patients With Acute Mesenteric Ischemia and Concomitant Colon Ischemia: A Retrospective Cohort Study.

BACKGROUND: Early identification of patients with acute mesenteric ischemia (AMI) involving the large bowel may play a decisive role in improving the prognosis of AMI. This study aims to compare the outcomes between patients with isolated AMI and AMI patients with colon involvement (CI) and to identify the predictors of worse outcomes. The different surgical modalities for AMI patients with CI were also evaluated. METHODS: This retrospective cohort study included 199 AMI patients admitted from January 2005 to January 2014. Based on colonoscopy and pathology reports, 39 patients were diagnosed as AMI with CI, and 160 were AMI patients without CI. The clinical outcomes and different surgical modalities were compared. Risk factors of 30-d mortality and short bowel syndrome (SBS) were identified. RESULTS: The 30-d mortality (10% versus 49%, P < 0.01) and SBS incidence (19% versus 49%, P < 0.01) were higher in AMI patients with CI than AMI patients without CI. AMI patients with CI have higher rate of bowel resection (68% versus 95%, P < 0.001) and second-look laparotomy (25% versus 54%, P < 0.001) than patients with AMI alone. For AMI patients with CI, emergent laparotomy was associated with shorter hospital stay (P = 0.04) and less incidence of SBS (74% versus 25%, P < 0.001) than initial endovascular therapy. Patients with ostomy had less repeated bowel resection (11% versus 63%, P = 0.001) and rate of SBS (21% versus 79%, P < 0.001) than patients with primary bowel anastomosis. Serum procalcitonin level and colon ischemia were risk factors of 30-d mortality and SBS for AMI. CONCLUSIONS: AMI patients with CI represent a special cohort of AMI patients with higher risk of poor outcome. Compared to initial endovascular therapy, emergent laparotomy was associated with shorter length of hospital stay and reduced incidence of SBS.

MDCT assessment of resectability in hilar cholangiocarcinoma.

PURPOSE: The purpose of this study is to investigate the value of multidetector computed tomography (MDCT) assessment of resectability in hilar cholangiocarcinoma, and to identify the factors associated with unresectability and accurate evaluation of resectability. METHODS: From January 2007 to June 2015, a total of 77 consecutive patients were included. All patients had preoperative MDCT (with MPR and MinIP) and surgical treatment, and were pathologically proven with hilar cholangiocarcinoma. The MDCT images were reviewed retrospectively by two senior radiologists and one hepatobiliary surgeon. The surgical findings and pathologic results were considered to be the gold standard. The Chi square test was used to identify factors associated with unresectability and accurate evaluation of resectability. RESULTS: The sensitivity, specificity, and overall accuracy of MDCT assessment were 83.3 %, 75.9 %, and 80.5 %, respectively. The main causes of inaccuracy were incorrect evaluation of N2 lymph node metastasis (4/15) and distant metastasis (4/15). Bismuth type IV tumor, main or bilateral hepatic artery involvement, and main or bilateral portal vein involvement were highly associated with unresectability (P < 0.001). Patients without biliary drainage had higher accuracy of MDCT evaluation of resectability compared to those with biliary drainage (P < 0.001). CONCLUSION: MDCT is reliable for preoperative assessment of resectability in hilar cholangiocarcinoma. Bismuth type IV tumor and main or bilateral vascular involvement highly suggest the unresectability of hilar cholangiocarcinoma. Patients without biliary drainage have a more accurate MDCT evaluation of resectability. We suggest MDCT should be performed before biliary drainage to achieve an accurate evaluation of resectability in hilar cholangiocarcinoma.

Successful Parenchyma-Sparing Anatomical Surgery by 3-Dimensional Reconstruction of Hilar Cholangiocarcinoma Combined with Anatomic Variation.

The combination of hilar cholangiocarcinoma and anatomic variation constitutes a rare and complicated condition. Precise understanding of 3-dimensional position of tumor in the intrahepatic structure in such cases is important for operation planning and navigation. We report a case of a 61-year woman presenting with hilar cholangiocarcinoma. Anatomic variation and tumor location were well depicted on preoperative multidetector computed tomography (MDCT) combined with 3-dimensional reconstruction as the right posterior segmental duct drained to left hepatic duct. The common hepatic duct, biliary confluence, right anterior segmental duct, and right anterior branch of portal vein were involved by the tumor (Bismuth IIIa). After carefully operation planning, we successfully performed a radical parenchyma-sparing anatomical surgery of hilar cholangiocarcinoma: Liver segmentectomy (segments 5 and 8) and caudate lobectomy. MDCTcombined with 3-dimensional reconstruction is a reliable non-invasive modality for preoperative evaluation of hilar cholangiocarcinoma.