Intensive Surveillance of Porcine-Rhesus Kidney Xenotransplant Using Different Ultrasound Techniques.

BACKGROUND: Significant progress has been made in kidney xenotransplantation in the past few years, and this field is accelerating towards clinical translation. Therefore, surveillance of the xenograft with appropriate tools is of great importance. Ultrasonography has been widely used in kidney allotransplantation and served as an economical and non-invasive method to monitor the allograft. However, questions remain whether the ultrasonographic criteria established for human kidney allograft could also be applied in xenotransplantation. METHODS: In the current study, we established a porcine-rhesus life sustaining kidney xenotransplantation model. The xenograft underwent intensive surveillance using gray-scale, colorful Doppler ultrasound as well as 2D shear wave elastography. The kidney growth, blood perfusion, and cortical stiffness were measured twice a day. These parameters were compared with the clinical data including urine output, chemistry, and pathological findings. RESULTS: The observation continued for 16 days after transplantation. Decline of urine output and elevated serum creatinine were observed on POD9 and biopsy proven antibody-mediated rejection was seen on the same day. The xenograft underwent substantial growth, with the long axis length increased by 32% and the volume increased by threefold at the end of observation. The resistive index of the xenograft arteries elevated in response to rejection, together with impaired cortical perfusion, while the peak systolic velocity (PSV) was not compromised. The cortical stiffness also increased along with rejection. CONCLUSION: In summary, the ultrasound findings of kidney xenograft shared similarities with those in allograft but possessed some unique features. A modified criteria needs to be established for further application of ultrasound in kidney xenotransplantation.

NPAS2 promotes aerobic glycolysis and tumor growth in prostate cancer through HIF-1A signaling.

BACKGROUND: Prostate cancer (PCa), one of the common malignant tumors, is the second leading cause of cancer-related deaths in men. The circadian rhythm plays a critical role in disease. Circadian disturbances are often found in patients with tumors and enable to promote tumor development and accelerate its progression. Accumulating evidence suggests that the core clock gene NPAS2 (neuronal PAS domain-containing protein 2) has been implicated in tumors initiation and progression. However, there are few studies on the association between NPAS2 and prostate cancer. The purpose of this paper is to investigate the impact of NPAS2 on cell growth and glucose metabolism in prostate cancer. METHODS: Quantitative real-time PCR (qRT-PCR), immunohistochemical (IHC) staining, western blot, GEO (Gene Expression Omnibus) and CCLE (Cancer Cell Line Encyclopedia) databases were used to analyze the expression of NPAS2 in human PCa tissues and various PCa cell lines. Cell proliferation was assessed using MTS, clonogenic assays, apoptotic analyses, and subcutaneous tumor formation experiments in nude mice. Glucose uptake, lactate production, cellular oxygen consumption rate and medium pH were measured to examine the effect of NPAS2 on glucose metabolism. The relation of NPAS2 and glycolytic genes was analyzed based on TCGA (The Cancer Genome Atlas) database. RESULTS: Our data showed that NPAS2 expression in prostate cancer patient tissue was elevated compared with that in normal prostate tissue. NPAS2 knockdown inhibited cell proliferation and promoted cell apoptosis in vitro and suppressed tumor growth in a nude mouse model in vivo. NPAS2 knockdown led to glucose uptake and lactate production diminished, oxygen consumption rate and pH elevated. NPAS2 increased HIF-1A (hypoxia-inducible factor-1A) expression, leading to enhanced glycolytic metabolism. There was a positive correlation with the expression of NPAS2 and glycolytic genes, these genes were upregulated with overexpression of NPAS2 while knockdown of NPAS2 led to a lower level. CONCLUSION: NPAS2 is upregulated in prostate cancer and promotes cell survival by promoting glycolysis and inhibiting oxidative phosphorylation in PCa cells.

Role of Complement System in Kidney Transplantation: Stepping From Animal Models to Clinical Application.

Kidney transplantation is a life-saving strategy for patients with end-stage renal diseases. Despite the advances in surgical techniques and immunosuppressive agents, the long-term graft survival remains a challenge. Growing evidence has shown that the complement system, part of the innate immune response, is involved in kidney transplantation. Novel insights highlighted the role of the locally produced and intracellular complement components in the development of inflammation and the alloreactive response in the kidney allograft. In the current review, we provide the updated understanding of the complement system in kidney transplantation. We will discuss the involvement of the different complement components in kidney ischemia-reperfusion injury, delayed graft function, allograft rejection, and chronic allograft injury. We will also introduce the existing and upcoming attempts to improve allograft outcomes in animal models and in the clinical setting by targeting the complement system.

Kidney Organoids as a Novel Platform to Evaluate Lipopolysaccharide-Induced Oxidative Stress and Apoptosis in Acute Kidney Injury.

Sepsis-associated acute kidney injury (SA-AKI) is a life-threatening syndrome. Lipopolysaccharide (LPS) is a widely used inducer for modeling SA-AKI both in vivo and in vitro. However, due to the innate complexity of the kidney architecture, the mechanisms underlying the pathogenesis of SA-AKI, as well as those involved in LPS-induced kidney injury remain to be clarified. Kidney organoids derived from human pluripotent stem cells (hPSCs) act as a model of multiple types of kidney cells in vitro and eliminate potential confounders in vivo. In the current study, we established LPS-induced kidney injury models both in vivo and in human kidney organoids. Kidney function, pathological changes, and markers of oxidative stress were evaluated with/without the presence of methylprednisolone (MP) treatment both in vivo and in vitro. The extent of LPS-induced oxidative stress and apoptosis in kidney organoids was further investigated in vitro. LPS-induced acute kidney injury in mice, together with pathological changes and increased oxidative stress, as well as enhanced apoptosis in kidney cells were evaluated. These phenomena were ameliorated by MP treatment. Experiments in kidney organoids showed that the LPS-induced apoptotic effects occurred mainly in podocytes and proximal tubular cells. Our experiments demonstrated the efficacy of using kidney organoids as a solid platform to study LPS-induced kidney injury. LPS induced oxidative stress as well as apoptosis in kidney cells independently of changes in perfusion or immune cell infiltration. MP treatment partially alleviated LPS-induced injury by reducing kidney cell oxidative stress and apoptosis.

Snai1-induced partial epithelial-mesenchymal transition orchestrates p53-p21-mediated G2/M arrest in the progression of renal fibrosis via NF-κB-mediated inflammation.

Renal fibrosis is the common feature of all progressive kidney diseases and exerts great burden on public health worldwide. The maladaptive repair mechanism of tubular epithelial cells, an important mediator of renal fibrogenesis, manifests with partial epithelial-mesenchymal transition (EMT) and cell cycle arrest. The aim of this study is to investigate the possible correlation between partial EMT and cell cycle arrest, and elucidate the underlying mechanism. We examined human kidney allograft samples with interstitial fibrosis and three mice renal fibrosis models, unilateral ureter obstruction (UUO), ischemia-reperfusion injury, and Adriamycin nephropathy. The partial EMT process and p53-p21 axis were elevated in both human allograft with interstitial fibrosis, as well as three mice renal fibrosis models, and showed a time-dependent increase as fibrosis progressed in the UUO model. Snai1 controlled the partial EMT process, and led to parallel changes in renal fibrosis, G2/M arrest, and inflammation. p53-p21 axis arrested cell cycle at G2/M, and prompted partial EMT and fibrosis together with inflammation. NF-κB inhibitor Bay11-7082 disrupted the reciprocal loop between Snai1-induced partial EMT and p53-p21-mediated G2/M arrest. We demonstrated the reciprocal loop between partial EMT and G2/M arrest of TECs during renal fibrogenesis and revealed NF-κB-mediated inflammatory response as the underlying mechanism. This study suggests that targeting NF-κB might be a plausible therapeutic strategy to disrupt the reciprocal loop between partial EMT and G2/M arrest, therefore alleviating renal fibrosis.

Advances of Contrast-Enhanced Ultrasonography and Elastography in Kidney Transplantation: From Microscopic to Microcosmic.

Kidney transplantation is the best choice for patients with end-stage renal disease. To date, allograft biopsy remains the gold standard for revealing pathologic changes and predicting long-term outcomes. However, the invasive nature of transplant biopsy greatly limits its application. Ultrasound has been a first-line examination for evaluating kidney allografts for a long time. Advances in ultrasound in recent years, especially the growing number of studies in elastography and contrast-enhanced ultrasonography (CEUS), have shed new light on its application in kidney transplantation. Elastography, including strain elastography and shear wave elastography, is used mainly to assess allograft stiffness and, thus, predict renal fibrosis. CEUS has been used extensively in evaluating blood microperfusion, assessing acute kidney injury and detecting different complications after transplantation. Requiring the use of microbubbles also makes CEUS a novel method of gene transfer and drug delivery, enabling promising targeted diagnosis and therapy. In this review, we summarize the advances of elastography and CEUS in kidney transplantation and evaluate their potential efficiency in becoming a better complement to or even substitute for transplant biopsy in the future.

Cyclic helix B peptide ameliorates renal tubulointerstitial fibrosis induced by unilateral ureter obstruction via inhibiting NLRP3 pathway.

BACKGROUND: Renal fibrosis is the inevitable outcome of all progressive chronic kidney diseases (CKD) and leads to a gradual loss of renal function. We previously reported cyclic helix B peptide (CHBP), a novel synthesized peptide derived from erythropoietin, had shown effective renoprotection. In this study, we investigated the anti-fibrotic and renoprotective effect of CHBP in a murine renal tubulointerstitial fibrosis model induced by unilateral ureter obstruction (UUO). METHODS: Mice were subjected to the UUO model and CHBP was given intraperitoneally. To assess the therapeutic effects of CHBP, pathological injury, deposition of extracellular matrix (ECM) and the progression of epithelial-mesenchymal transition (EMT) were examined in vivo. The anti-fibrotic effects of CHBP was validated in vitro using TCMK-1 cells treated with TGF-ß1. Involvement of the NLRP3 pathway was demonstrated both in vivo and in vitro. RESULTS: CHBP significantly ameliorated renal tubulointerstitial injury and fibrosis in terms of ECM deposition. The EMT process was also alleviated after CHBP treatment. Similar therapeutic effects of CHBP were also observed in vitro in TGF-ß1 treated tubular epithelial cells (TECs). NLRP3/caspase-1/IL-1ß pathway was involved and activated upon injury, both in vivo and in vitro. While the activation of the NLRP3 pathway was found to be in negative correlation with CHBP treatment. CHBP could suppress the activation of NLRP3 and its downstream inflammatory mediators even with addition of extracellular ATP, a direct activator of the NLRP3 inflammasome. CONCLUSIONS: Our results suggest that CHBP could effectively protect the kidney from renal tubulointerstitial fibrosis in the UUO model via counteracting the NLRP3/caspase-1/IL-1ß pathway.

Diagnosis and Treatment of Early Transplant Renal Artery Stenosis: Experience From a Center in Eastern China.

Few studies have focused on the clinical characteristics of transplant renal artery stenosis (TRAS) with early onset. Sixteen cases diagnosed with TRAS in our center from January 2014 to August 2018 were retrospectively analyzed. Sixteen transplant patients without TRAS were selected as controls. The median diagnostic time for TRAS was 47.5 days after transplantation. No significant difference was observed between the TRAS group and the control group. The serum creatinine level (Scr), estimated glomerular filtration rate, systolic/diastolic blood pressure, graft artery peak systolic velocity (PSV), and resistive index of intersegmental artery (RI-ISA) between the 2 groups were (5.55 ± 3.49) and (1.89 ± 0.85) mg/dL; (17.83 ± 14.94) and (49.39 ± 19.96) mL/min; (143.50 ± 9.49)/(86.14 ± 7.38) and (130.38 ± 18.86)/(82.81 ± 12.52) mm Hg; (3.39 ± 1.57) and (1.31 ± 0.51) m/s; and (0.51 ± 0.10) and (0.67 ± 0.13), respectively. All showed statistical significance except the diastolic blood pressure. The Scr, estimated glomerular filtration rate, systolic/diastolic blood pressure, graft artery PSV, and RI-ISA in the TRAS group prior and after treatment were (5.55 ± 3.49) and (3.20 ± 1.50) mg/dL; (17.83 ± 14.94) and (25.60 ± 13.29) mL/min; (143.50 ± 9.49)/(86.14 ± 7.38) and (128.07 ± 16.16)/(75.71 ± 7.56) mm Hg; (3.39 ± 1.57) and (2.00 ± 1.04) m/s; and (0.51 ± 0.10) and (0.61 ± 0.10); all showed statistical significance. Receiver operating characteristic analysis showed an area under curve of 0.8616 for PSV and 0.8535 for RI-ISA in diagnosing TRAS. Patients with TRAS in our center showed a unique characteristic of early onset. The most prominent clinical symptom of TRAS is increasing Scr level instead of refractory hypertension. Screening of color Doppler flow imaging with a graft artery PSV >2.5 m/s and RI-ISA <0.5 could yield a preliminary diagnosis of TRAS. Percutaneous transluminal angioplasty/stenting could effectively improve allograft function as well as color Doppler flow imaging indexes.

Pathogenesis of Chronic Allograft Dysfunction Progress to Renal Fibrosis.

Kidney transplantation is a life-change measurement for the patients of end-stage renal disease (ESRD). However, the renal allograft cannot avoid initial acute kidney injury (AKI) and subsequent chronic allograft dysfunction (CAD), gradually develops fibrosis and eventually loses function. It is imperative to disclose the pathogenesis of AKI and CAD in order to facilitate interventions. We have studied the involvement of immunity, inflammation, and apoptosis in ischemia-reperfusion injury (IRI) and/or immunosuppressant induced AKI models, with associated chronic damage. Our research mainly focused on tubular epithelial cells (TECs) that are passive victims and also active participators in injury and mediate following repair or fibrosis. Targeting not only fibroblasts/myofibroblasts, but also TECs, might be a fundamental strategy to prevent and treat renal fibrosis. We have also evaluated the potential application of siRNA targeting caspase-3 and tissue protective erythropoietin derivatives, HBSP and CHBP, aiming to treat AKI and prevent CAD. Significant improvements have been obtained, but timely diagnosis and precise therapy of AKI and prevention of CAD progressing to ESRD are still very challenging. Modern technologies such as microarray and sequencing analysis have been used to identify biomarkers and potentially facilitate individual cell target treatment for transplant patients.

Prediction of renal allograft chronic rejection using a model based on contrast-enhanced ultrasonography.

OBJECTIVE: To evaluate the application of contrast-enhanced ultrasonography (CEUS) for the diagnosis of renal allograft chronic rejection (CR). METHODS: A total of 104 patients who were suspected to have AR or CR were enrolled in this study (derivation group, n = 66; validation group, n = 38). Before biopsy, all patients received an ultrasound examination. RESULTS: In the CR group, rising time (RT) and time to peak (TTP) of medulla (RTm and TTPm, respectively) were significantly longer compared to those in the AR group. The kidney volume was significantly decreased in the CR group but was increased in the AR group. In the derivation group, age, change in kidney volume, and TTPm were identified as independent predictors by multivariate analysis. Based on the multivariate analysis results and area under receiver operating characteristic (ROC) curves (AUROCs) of individual markers, we constructed a new index as follows: P = -5.424 + 0.074 × age -9.818 × kidney volume change + 0.115 × TTPm; New Index = eP /(1 + eP ). The new index discriminates CR from AR and had better AUROCs than any other parameters. CONCLUSION: In conclusion, the new index provides a new diagnosis model for CR.

Renal tubular epithelial cells: the neglected mediator of tubulointerstitial fibrosis after injury.

Renal fibrosis, especially tubulointerstitial fibrosis, is the inevitable outcome of all progressive chronic kidney diseases (CKDs) and exerts a great health burden worldwide. For a long time, interests in renal fibrosis have been concentrated on fibroblasts and myofibroblasts. However, in recent years, growing numbers of studies have focused on the role of tubular epithelial cells (TECs). TECs, rather than a victim or bystander, are probably a neglected mediator in renal fibrosis, responding to a variety of injuries. The maladaptive repair mechanisms of TECs may be the key point in this process. In this review, we will focus on the role of TECs in tubulointerstitial fibrosis. We will follow the fate of a tubular cell and depict the intracellular changes after injury. We will then discuss how the repair mechanism of tubular cells becomes maladaptive, and we will finally discuss the intercellular crosstalk in the interstitium that ultimately proceeds tubulointerstitial fibrosis.