Validation of the Anti-Inflammatory Effect of Tenebrio Molitor Larva Oil in a Colitis Mouse Model.

Ulcerative colitis is caused by various external factors and is an inflammatory disease that causes decreased intestinal function. Tenebrio molitor larvae contain more than 30 % fat, and the fat component consists of 45 % oleic acid, 20 % linoleic acid and 20 % polyunsaturated fatty acids. In this study, after administering Tenebrio molitor larva oil (TMLO) in a dextran sodium sulphate (DSS)-induced ulcerative colitis mouse model, the pathological findings and inflammatory markers of colitis were analysed to assess whether a colitis mitigation effect was achieved. In the TMLO-administered group, the colon length increased, the spleen weight decreased, and the body weight increased compared with that in the DSS group. In addition, the disease activity index level decreased, the mRNA expression level of inflammatory cytokines in the colon decreased, and the myeloperoxidase activity level significantly decreased. Also, the activity of the NF-κB pathway involved in the regulation of the inflammatory response was lower in the TMLO group than in the DSS group. Taken together, these results suggest that TMLO suppresses occurrence of acute ulcerative colitis in the DSS mouse model. Therefore, TMLO has the potential to be developed as a health food for the prevention and treatment of ulcerative colitis.

Should asymptomatic contralateral inguinal hernia be laparoscopically repaired in the adult population as benefits greatly outweigh risks? A systematic review and meta-analysis.

PURPOSE: When laparoscopically repairing a symptomatic inguinal hernia, surgeons will discover a contralateral asymptomatic hernia in 22% of patients. It is estimated 30% of asymptomatic hernias become symptomatic and require repair. Thus, should they be repaired in a 2-for-1 operation? The main purpose is to examine the evidence and make a recommendation for the need to repair the contralateral asymptomatic inguinal hernia prophylactically in the adult population during unilateral inguinal hernia presentation. METHOD: A systematic literature search was conducted up to 15 February 2021 using PubMed and the Cochrane Library. Management pathway taken, mean operating time, duration of follow-up, pain, duration of hospital stay and perioperative complications were extracted. Risk of bias was assessed using the ROBINS-I tool. RESULTS: Six non-randomised studies (1774 patients) were included; 978 patients had both hernias repaired, 796 patients had only the symptomatic hernia repaired. There was no significant difference in length of hospital stay, return to activities of daily living nor complications. Mean operating time was slightly lower for patients who had unilateral hernia repair (mean difference = - 14.57 min, 95%CI - 25.59, - 3.45). Reported pain scores were lower for patients who only had one hernia repaired (- 0.33 units, 95%CI - 0.48, - 0.18). The overall risk of bias for the six studies were low-to-moderate risk. CONCLUSION: Asymptomatic inguinal hernias can be repaired when found. While there is minimal increase in operation time and pain, no significant difference to total hospital stay. Importantly, this is likely to prevent the need for another operation in almost a third of patients.

[Successful rituximab treatment of recurrent glomerulonephritis associated with antibodies against the glomerular basement membrane]. / Erfolgreiche Rituximabtherapie des Rezidivs einer Glomerulonephritis assoziiert mit Antikörpern gegen die glomeruläre Basalmembran.

This article presents a case of recurrent anti-GBM disease (with antibodies against the glomerular basement membrane [GBM]) in a 17-year-old patient successfully treated with rituximab. Kidney biopsy with detection of linear deposition of immunoglobulin G (IgG) along the basement membrane is the diagnostic gold standard, which should be accompanied by serological testing. However, standard assays for the detection of anti-GBM antibodies have a high rate of false-negative results. In this particular case, an increase in proteinuria despite standard therapy (plasmapheresis, steroids, cyclophosphamide) was the clinical correlate of relapsing disease. The use of rituximab completely resolved the recurrent anti-GBM disease.

Effects of lysophospholipid supplementation to reduced energy, crude protein, and amino acid diets on growth performance, nutrient digestibility, and blood profiles in broiler chickens.

Two experiments investigated the effects of lysophospholipid (LPL) supplementation on low-energy and low-nitrogenous diets for broilers. A total of 300 one-day-old male chicks (Ross 308) was allotted to 5 treatments in a completely randomized design. Each group consisted of 6 replicates with 10 birds each. Experimental diet I included positive control (PC) having 3,025 (starter), 3,150 (grower), and 3,200 kcal/kg (finisher) of ME; negative control (NC) was 150 kcal/kg of ME lower than PC, and LPL-05, LPL-10, and LPL-15 treatments were NC + 0.05%, 0.10%, and 0.15% of LPL supplementation, respectively. Experimental diet II included positive control (PC) having a formulated amount of crude protein including Lys and Met + Cys that met the Ross 308 standards; negative control (NC) was 4% lower CP and AA than PC; other treatments were supplemented with LPL at 0.05% (LPL-05), 0.10% (LPL-10), and 0.15% (LPL-15) into the NC, respectively. Experiment I showed that growth performance linearly increased as the LPL inclusion increased (P < 0.001). Broilers fed LPL-10 and LPL-15 increased digestibility of DM (P < 0.05), crude protein (P < 0.01), and total amino acids (P < 0.01) compared to NC. Serum glucose (P < 0.01) and high-density lipoprotein (P < 0.05) concentrations were greater in groups fed LPL-10 than those fed PC. Furthermore, leg muscle increased in birds fed LPL-10 compared with NC (P < 0.05). Experiment II observed a linear response to LPL supplementation in the whole period, in terms of body weight gain (P = 0.015) and feed conversion ratio (P = 0.027). Feeding of 0.15% LPL had promising effects on digestibility of crude protein and ether extract compared with NC (P < 0.01 and P < 0.05, respectively). Overall, LPL could be considered as a feed additive to reduced energy (-150 kcal/kg) or nitrogenous diets (-5%) in order to improve growth performance and nutrient digestibility without adverse effects on lymphoid organs and hepatic enzyme of broilers.

Effects of lysophospholipid on growth performance, carcass yield, intestinal development, and bone quality in broilers.

A study was conducted to evaluate the effects of supplementing different levels of lysophospholipid (LPL) to normal or reduced energy diets on growth performance, carcass yield, intestinal morphology, and skeletal development in broilers. A total of 960 one-day-old Cobb 500 male birds were allocated using a 2 × 4 factorial arrangement with 2 energy levels (NE: normal and RE: 100 kcal/kg metabolizable energy reduction) and 4 LPL supplement levels (0, 0.025, 0.050, and 0.075%). Three diet phases were fed throughout the trial: starter (days 0 to 7), grower (days 8 to 21), and finisher (days 22 to 42) phases. Body weight (BW), feed intake (FI), and feed conversion ratio were calculated at the end of each phase. At day 7 and 21, duodenum and jejunum samples were collected for intestinal morphology and claudin-3 expression analyses, and tibia were sampled for bone quality analyses. At day 42, 4 birds per replicate were selected to measure carcass yield. The results showed low metabolizable energy diets impaired bird's growth performance, intestine development, and bone quality. The 0.075% LPL supplement in NE improved BW, BW gain, and FI in the finisher and overall period compared with no LPL supplement in NE (P < 0.05). In RE, the 0.025% LPL supplement significantly improved growth performance compared to the other treatments in RE (P < 0.05). The interactions on processing parameters were detected with LPL supplement in NE diets; 0.025, 0.05, and 0.075% LPL supplements significantly increased pectoral major percentages compared to the one without LPL supplement in NE (P < 0.05). The 0.075% LPL supplement increased dressing percentage (cold carcass weight/live BW) compared with the others (P < 0.05). The intestine morphology results showed LPL had positive effects on intestine development mainly during the early age (day 7) and claudin-3 expression at both day 7 and 21. Furthermore, LPL supplement significantly increased the total Ca and P deposition and positively affected the bone structure development. In summary, dietary LPL supplementation promoted growth performance, carcass yield, intestinal development, intestinal health, and bone quality.

Adgrf5 contributes to patterning of the endothelial deep layer in retina.

Neovascularization of the inner retinal space is a major cause of vision loss. In retinal angiomatous proliferation (RAP) syndrome, newly formed vessels originate from the retinal plexus and invade the inner retinal space. However, the molecular pathways preventing subretinal vascularization remain largely unknown. In most murine models of RAP, pathological neovascularization occurs concomitantly with the development of the retinal vasculature. Here, we demonstrate that disturbing the sequence of morphogenetic events that shape the three-layered retinal vascular network leads to subretinal vascularization. Sprouts emerging from the perivenous region after the first postnatal week extended toward the retinal space where they merged into the deep layer. The small GTPase Rac1 was required for the formation of these vascular extensions and the vascular inner plexus is formed coaxially to the overarching veins. The adhesion receptor Adgrf5 was highly expressed in the endothelium of the central nervous system, where it regulates blood-brain barrier formation. The vascular superficial plexus of Adgrf5 mutant mouse retinae exhibited an increased vascular density in the perivenous areas with increased projections toward the inner plexus where they subsequently created hyper-dense endothelial cells (EC) clusters. Disturbing the perivenous pool of EC thus significantly altered the inner plexus formation. These abnormalities culminated in transient vascular protrusions in the inner retinal space. Taken together, these results reveal a previously unobserved vascular morphogenetic defect in Adgrf5 knockout mice, implicating a role for ADGRF5 in the initiation of subretinal vascularization. Our findings also illustrate how vein-derived EC shape the inner retinal layer formation and could control the appearance of angiomatous malformations.

Ultra-deep next-generation sequencing of plasma cell-free DNA in patients with advanced lung cancers: results from the Actionable Genome Consortium.

BACKGROUND: Noninvasive genotyping using plasma cell-free DNA (cfDNA) has the potential to obviate the need for some invasive biopsies in cancer patients while also elucidating disease heterogeneity. We sought to develop an ultra-deep plasma next-generation sequencing (NGS) assay for patients with non-small-cell lung cancers (NSCLC) that could detect targetable oncogenic drivers and resistance mutations in patients where tissue biopsy failed to identify an actionable alteration. PATIENTS AND METHODS: Plasma was prospectively collected from patients with advanced, progressive NSCLC. We carried out ultra-deep NGS using cfDNA extracted from plasma and matched white blood cells using a hybrid capture panel covering 37 lung cancer-related genes sequenced to 50 000× raw target coverage filtering somatic mutations attributable to clonal hematopoiesis. Clinical sensitivity and specificity for plasma detection of known oncogenic drivers were calculated and compared with tissue genotyping results. Orthogonal ddPCR validation was carried out in a subset of cases. RESULTS: In 127 assessable patients, plasma NGS detected driver mutations with variant allele fractions ranging from 0.14% to 52%. Plasma ddPCR for EGFR or KRAS mutations revealed findings nearly identical to those of plasma NGS in 21 of 22 patients, with high concordance of variant allele fraction (r = 0.98). Blinded to tissue genotype, plasma NGS sensitivity for de novo plasma detection of known oncogenic drivers was 75% (68/91). Specificity of plasma NGS in those who were driver-negative by tissue NGS was 100% (19/19). In 17 patients with tumor tissue deemed insufficient for genotyping, plasma NGS identified four KRAS mutations. In 23 EGFR mutant cases with acquired resistance to targeted therapy, plasma NGS detected potential resistance mechanisms, including EGFR T790M and C797S mutations and ERBB2 amplification. CONCLUSIONS: Ultra-deep plasma NGS with clonal hematopoiesis filtering resulted in de novo detection of targetable oncogenic drivers and resistance mechanisms in patients with NSCLC, including when tissue biopsy was inadequate for genotyping.

Successful Treatment of Fibrosing Cholestatic Hepatitis With Daclatasvir and Asunaprevir After Liver Transplantation: A Case Report.

BACKGROUND: Fibrosing cholestatic hepatitis (FCH) is an aggressive form of hepatitis C virus (HCV) recurrence after liver transplantation (LT). Most FCH cases are fatal, occurring as a secondary disease following rapidly progressive liver dysfunction and graft failure. We report a case of early-onset FCH after LT that was successfully treated using daclatasvir and asunaprevir. CASE REPORT: A 59-year-old woman underwent living donor LT for HCV-related liver cirrhosis. However, liver function was not improved after LT and gradually worsened. A liver biopsy was performed at 30 and 47 days after the living donor LT to identify the cause of the liver dysfunction. The first biopsy result showed no specific finding. However, combined treatment with pegylated interferon and ribavirin was started because of a high HCV viral load (> 8.0 log IU/mL). Nevertheless, liver function and HCV viral load deteriorated, and the second biopsy performed on postoperative day 47 revealed FCH. We converted the antiviral agents into daclatasvir and asunaprevir and performed plasmapheresis twice. Since then, the liver dysfunction and HCV viral load gradually improved, and HCV RNA clearance occurred at week 11 after treatment. The patient achieved a sustained virologic response at week 24 after completion of the treatment. CONCLUSION: Daclatasvir combined with asunaprevir can be a useful treatment option in potentially fatal FCH after LT.

Comparison of the efficacy of diegnogest and levonorgestrel-releasing intrauterine system after laparoscopic surgery for endometriosis.

AIM: To compare the efficacy between dienogest and levonorgestrel-releasing intrauterine system (LNG-IUS) after laparoscopic surgery for endometriosis. METHODS: A total of 285 women were diagnosed as endometriosis by laparoscopy between 2011 and 2015. Patients were grouped into no treatment (n = 83), treatment with dienogest (n =130) and treatment with LNG-IUS (n =72) after laparoscopic surgery. The changes of the pain scores were checked at 6, 12 and 24 months after the surgery, and the rates of disease recurrence and treatment discontinuation rate were determined. RESULTS: The participants' mean age was 38.9 years (range 21-54 years). The mean age of LNG-IUS group (43.7 years) was significantly higher than the no treatment and dienogest groups (39.3 vs 33.9 years, respectively). At 6 and 12 months, the median pain scores in treatment (dienogest and LNG-IUS) groups were significantly lower than control group. Both treatment groups had significantly lower recurrence rate than control group (3.8% and 9.7%, respectively, vs 32.5%, P =0.001). No significant difference was found in the recurrence rate between the two treatment groups (P =0.461). Patients in the LNG-IUS group showed lower rate of discontinuation due to complication (27.8%) than those in dienogest group (35.6%, P =0.010). CONCLUSION: LNG-IUS treatment in the patients with endometriosis is effective for postoperative pain control and preventing recurrence, however, the LNG-IUS group is older, it is difficult to compare the efficacy between dienogest and LNG-IUS in present study.

Urgent Living-Donor Liver Transplantation in a Patient With Concurrent Active Tuberculosis: A Case Report.

BACKGROUND: Although active tuberculosis (TB) is considered a contraindication for liver transplantation (LT), this is the only treatment in patients with liver failure and concurrent active TB. We report a case with successful urgent living-donor LT for irreversible liver failure in the presence of active TB. CASE PRESENTATION: A 48-year-old man, with a history of decompensated alcoholic liver cirrhosis, was presented with stupor. At admission, his consciousness had deteriorated to semi-coma, and his renal function also rapidly deteriorated to hepatorenal syndrome. A preoperative computed tomography scan of the chest revealed several small cavitary lesions in both upper lobes, and acid-fast bacillus stain from his sputum was graded 2+. Adenosine deaminase levels from ascites were elevated, suggesting TB peritonitis. A first-line anti-TB drug regimen was started immediately (rifampin, isoniazid, levofloxacin, and amikacin). An urgent living-donor LT was performed 2 days later. After LT, the regimen was changed to second-line anti-TB drugs (amikacin, levofloxacin, cycloserine, and pyridoxine). The sputum acid-fast bacillus stain tested negative on postoperative day 10. His liver function remained well preserved, even after the reversion to first-line anti-TB treatment. The patient recovered without any anti-TB medication-related complications and was discharged. CONCLUSIONS: LT can be prudently performed as a life-saving option, particularly for patients with liver failure and concurrent active TB.

Effect of preoxygenation using non-invasive ventilation before intubation on subsequent organ failures in hypoxaemic patients: a randomised clinical trial.

BACKGROUND: Previous data showed that non-invasive ventilation (NIV) applied for 3 min before tracheal intubation ensured better oxygenation compared with using a non-rebreather bag-valve-mask. We aimed to determine whether preoxygenation using NIV is effective in reducing the incidence of organ dysfunction in hypoxaemic, critically ill patients in intensive care. METHODS: A multicentre, randomised, open-label trial evaluating 100% FiO2 administered with NIV (99 patients) vs with face mask (102 patients) for 3 min before tracheal intubation. The primary endpoint was the maximal value of Sequential Organ Failure Assessment score within 7 days after intubation. RESULTS: The median (inter-quartile range) values of the maximal value of the Sequential Organ Failure Assessment score within 7 days post-intubation were not significantly different between the two randomised groups: nine (6-12) in the NIV group vs 10 (6-12) in the face mask group (P=0.65). In patients treated by NIV prior to the randomisation, there was a significant increase in the occurrence in adverse events in patients randomised to face mask [odds ratio=5.23 (1.61;16.99), P=0.0059]. CONCLUSIONS: This study failed to demonstrate any benefits of using NIV as a preoxygenation method to reduce organ dysfunction compared with usual preoxygenation in hypoxaemic, critically ill patients requiring tracheal intubation for invasive ventilation. NIV should not be discontinued for preoxygenation in the cases of patients treated by NIV before the decision to intubate. CLINICAL TRIAL REGISTRATION: NCT00472160.

Estimation of dispersion curves of water-loaded structures by using approximated acoustic mass.

For coupled structures surrounded by heavy fluids, it is difficult to obtain dispersion curves from an eigenvalue analysis, because the external fluid term in the coupled equation includes transcendental functions for frequency and wavenumber. Thus, in this study, the acoustic mass of the external fluid was approximated as a function of frequency or wavenumber only. The coupled equation can then be used to calculate eigenvalues, and can estimate dispersion curves from an eigenvalue analysis. Because of this assumption, those dispersion curves will contain errors. Accordingly, those errors were evaluated in this study through a comparison with a dispersion curve derived from forced responses. The acoustic mass was also evaluated for a water-loaded plate; this can be formulated theoretically. As a result, the acoustic mass is less sensitive to frequency changes than wavenumber changes, and using the fluid term defined at a low frequency has advantages when estimating the dispersion curve. Finally, the generality of the proposed method was identified through the application for a submerged cylindrical shell.

Efficacy of combining ING4 and TRAIL genes in cancer-targeting gene virotherapy strategy: first evidence in preclinical hepatocellular carcinoma.

Current treatments of hepatocellular carcinoma (HCC) are ineffective and unsatisfactory in many aspects. Cancer-targeting gene virotherapy using oncolytic adenoviruses (OAds) armed with anticancer genes has shown efficacy and safety in clinical trials. Nowadays, both inhibitor of growth 4 (ING4), as a multimodal tumor suppressor gene, and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), as a potent apoptosis-inducing gene, are experiencing a renaissance in cancer gene therapy. Herein we investigated the antitumor activity and safety of mono- and combined therapy with OAds armed with ING4 (Ad-ΔB/ING4) and TRAIL (Ad-ΔB/TRAIL) gene, respectively, on preclinical models of human HCC. OAd-mediated expression of ING4 or TRAIL transgene was confirmed. Ad-ΔB/TRAIL and/or Ad-ΔB/ING4 exhibited potent killing effect on human HCC cells (HuH7 and Hep3B) but not on normal liver cells. Most importantly, systemic therapy with Ad-ΔB/ING4 plus Ad-ΔB/TRAIL elicited more eradicative effect on an orthotopic mouse model of human HCC than their monotherapy, without causing obvious overlapping toxicity. Mechanistically, Ad-ΔB/ING4 and Ad-ΔB/TRAIL were remarkably cooperated to induce antitumor apoptosis and immune response, and to repress tumor angiogenesis. This is the first study showing that concomitant therapy with Ad-ΔB/ING4 and Ad-ΔB/TRAIL may provide a potential strategy for HCC therapy and merits further investigations to realize its possible clinical translation.

Risk factors for anastomotic leakage following ileosigmoid or ileorectal anastomosis.

AIM: Reconstruction with an ileosigmoidal anastomosis (ISA) or ileorectal anastomosis (IRA) is a surgical option after a subtotal colectomy. Anastomotic leakage (AL) is a problematic complication and high rates have been reported, but there is limited understanding of the risk factors involved. The aim of this study was to assess the established and potential predictors of AL following ISA and IRA. METHOD: This was a retrospective cohort study including all patients who had undergone ISA or IRA at three Swedish referral centres for colorectal surgery between January 2007 and March 2015. Data regarding clinical characteristics, treatment and outcome were collected from medical records. Univariate and multivariate logistic regression models were used to determine the association between patient and treatment related factors and the cumulative incidence of AL. RESULTS: In total, 227 patients were included. Overall, AL was detected amongst 30 patients (13.2%). Amongst patients undergoing colectomy with synchronous ISA or IRA (one-stage procedure), AL occurred in 23 out of 120 (19.2%) compared with seven out of 107 (6.5%) after stoma reversal with ISA or IRA (two-stage procedure) (P = 0.004). In addition, the multivariate analyses revealed a statistically significantly lower odds ratio for AL following a two-stage procedure (OR 0.10, 95% CI 0.03-0.41, P = 0.001). CONCLUSIONS: This study confirms high rates of AL following ISA and IRA. In particular, a synchronous procedure with colectomy and ISA/IRA carries a high risk of AL.

B-cell activating factor BAFF reflects patients' immunological risk profile after kidney transplantation.

The B-cell activating factor BAFF plays an important role in the development and maturation of B-lymphocytes, which can contribute to the generation of donor-specific antibodies and thus may influence graft function and graft survival. Inconsistent data on the role of BAFF levels after renal transplantation for the formation of donor-specific antibodies and the contribution for allograft rejection exist. The aim of the current study was to determine to what extent the degree of pre-immunization is reflected by each patient's BAFF levels before transplantation and in the follow-up. Furthermore, the impact of BAFF on allograft rejection frequency as well as severity and resulting allograft function over time was analyzed. Additionally, the impact of viral infections on BAFF levels after transplantation - as a potential confounder - was examined. For this purpose, a group of pre-sensitized patients (PRA>0%, (52±24% on average), n=40) was compared with non-sensitized patients (PRA=0%, n=62) and in a subsequent analysis stratification in accordance to the detected BAFF level was performed. Pre-sensitized patients had significantly higher BAFF levels before transplantation and suffered significantly more often from early steroid-resistant, mainly antibody-mediated rejections. A result which was confirmed also in highly sensitized patients with PRA levels >50%. Additionally, in the follow-up patients with either rising BAFF levels over time or BAFF levels above the median also had significantly more often antibody mediated rejections. Additionally, patients with BAFF levels above detected median even displayed impaired creatinine values as well as an induced eGFR slope up to month 48 after transplantation. The occurrence of viral infections (CMV, BKV) was only an additional influencing factor in the absence of concomitant allograft rejections. Therefore, the B-cell proliferation factor BAFF appears not only to reflect the immunological risk profile of patients in the context of kidney transplantation, it may possibly be further developed as a predictor of patients with an increased risk profile for subsequent allograft rejection and impaired allograft function.

Veno-Arterial-Venous Hybrid Mode of Extracorporeal Membrane Oxygenation for Acute Respiratory Distress Syndrome Combined With Septic Shock in a Liver Transplant Patient: A Case Report.

BACKGROUND: Post-operative respiratory failure is common in liver transplant patients, with a direct effect on graft and recipient outcomes. We present a case of successful treatment for a patient who had acute respiratory distress syndrome combined with septic shock after living-donor liver transplantation with veno-arterial-venous hybrid mode of extracorporeal membrane oxygenation (ECMO) support. METHODS: A 49-year-old male patient underwent successful emergency living-donor liver transplantation for acute-on-chronic liver failure associated with alcoholic liver cirrhosis. The patient was diagnosed with concurrent active pulmonary tuberculosis and tuberculosis peritonitis just before emergency liver transplantation. Intermittent post-operative small-bowel obstruction caused by adhesions from tuberculosis peritonitis was nonresponsive to conservative management. RESULTS: On post-operative day 114, we performed emergency adhesio-lysis because of bowel ischemia signs. However, the patient had progressive deterioration of arterial oxygenation despite conventional ventilatory support and nitric oxide gas inhalation, and septic shock resulted from aspiration pneumonia. Therefore, we decided to administer ECMO. After initiation of ECMO support, hemodynamic status, pneumonic consolidation, and oxygenation status gradually improved, and the patient was weaned from ECMO on the 11th day. Finally, the patient was discharged on post-transplant day 204. CONCLUSIONS: ECMO can be a beneficial rescue option for the management of refractory cardiopulmonary failure in liver transplant recipients. In addition, the hybrid mode of ECMO is a helpful salvage option when conventional modes of ECMO are inadequate.

A clinical drug library screen identifies clobetasol propionate as an NRF2 inhibitor with potential therapeutic efficacy in KEAP1 mutant lung cancer.

The Kelch-like ECH-associated protein 1 (KEAP1)-nuclear factor E2-related factor 2 (NRF2)pathway has a central role in cellular antioxidant defense. NRF2 activation due to KEAP1 or NRF2 mutations occurs frequently in many cancers, suggesting that NRF2 inhibition could be a promising therapeutic strategy. However, no potent NRF2 inhibitors are clinically available to date. To develop potent NRF2 inhibitors for therapeutic purpose, we screened ~4000 clinical compounds and determined clobetasol propionate (CP) as the most potent NRF2 inhibitor. Mechanistically, CP prevented nuclear accumulation and promoted ß-TrCP-dependent degradation of NRF2 in a glucocorticoid receptor- and a glycogen synthase kinase 3 (GSK3)-dependent manner. As a result, CP induced oxidative stress and strongly suppressed the anchorage-independent growth of tumors with KEAP1 mutation, but not with the wild-type KEAP1. Further, CP alone or in combination with rapamycin strongly inhibited the in vitro and in vivo growth of tumors harboring mutations in KEAP1 or both KEAP1 and LKB1 that are frequently observed in lung cancer. Thus, CP could be a repurposed therapeutic agent for cancers with high NRF2 activity. We also proposed that the use CP and rapamycin in combination could be a potential therapeutic strategy for tumors harboring both KEAP1 and LKB1 mutations.

TGFß engages MEK/ERK to differentially regulate benign and malignant pancreas cell function.

While TGFß signals are anti-proliferative in benign and well-differentiated pancreatic cells, TGFß appears to promote the progression of advanced cancers. To better understand dysregulation of the TGFß pathway, we first generated mouse models of neoplastic disease with TGFß receptor deficiencies. These models displayed reduced levels of pERK irrespective of KRAS mutation. Furthermore, exogenous TGFß led to rapid and sustained TGFBR1-dependent ERK phosphorylation in benign pancreatic duct cells. Similar to results that our group has published in colon cancer cells, inhibition of ERK phosphorylation in duct cells mitigated TGFß-induced upregulation of growth suppressive pSMAD2 and p21, prevented downregulation of the pro-growth signal CDK2 and ablated TGFß-induced EMT. These observations suggest that ERK is a key factor in growth suppressive TGFß signals, yet may also contribute to detrimental TGFß signaling such as EMT. In neoplastic PanIN cells, pERK was not necessary for either TGFß-induced pSMAD2 phosphorylation or CDK2 repression, but was required for upregulation of p21 and EMT indicating a partial divergence between TGFß and MEK/ERK in early carcinogenesis. In cancer cells, pERK had no effect on TGFß-induced upregulation of pSMAD2 and p21, suggesting the two pathways have completely diverged with respect to the cell cycle. Furthermore, inhibition of pERK both reduced levels of CDK2 and prevented EMT independent of exogenous TGFß, consistent with most observations identifying pERK as a tumor promoter. Combined, these data suggest that during carcinogenesis pERK initially facilitates and later antagonizes TGFß-mediated cell cycle arrest, yet remains critical for the pathological, EMT-inducing arm of TGFß signaling.