The Proteasomal Deubiquitinating Enzyme PSMD14 Regulates Macroautophagy by Controlling Golgi-to-ER Retrograde Transport

Ubiquitination regulates several biological processes, however the role of specific members of the ubiquitinome on intracellular membrane trafficking is not yet fully understood. Here, we search for ubiquitin-related genes implicated in protein membrane trafficking performing a High-Content siRNA Screening including 1187 genes of the human “ubiquitinome” using amyloid precursor protein (APP) as a reporter. We identified the deubiquitinating enzyme PSMD14, a subunit of the 19S regulatory particle of the proteasome, specific for K63-Ub chains in cells, as a novel regulator of Golgi-to-endoplasmic reticulum (ER) retrograde transport. Silencing or pharmacological inhibition of PSMD14 with Capzimin (CZM) caused a robust increase in APP levels at the Golgi apparatus and the swelling of this organelle. We showed that this phenotype is the result of rapid inhibition of Golgi-to-ER retrograde transport, a pathway implicated in the early steps of the autophagosomal formation. Indeed, we observed that inhibition of PSMD14 with CZM acts as a potent blocker of macroautophagy by a mechanism related to the retention of Atg9A and Rab1A at the Golgi apparatus. As pharmacological inhibition of the proteolytic core of the 20S proteasome did not recapitulate these effects, we concluded that PSMD14, and the K63-Ub chains, act as a crucial regulatory factor for macroautophagy by controlling Golgi-to-ER retrograde transport

Cost Effectiveness of Daclatasvir/Asunaprevir Versus Peginterferon/Ribavirin and Protease Inhibitors for the Treatment of Hepatitis c Genotype 1b Naïve Patients in Chile.

IntroductionDaclatasvir and Asunaprevir (DCV/ASV) have recently been approved for the treatment of chronic hepatitis C virus infection. In association, they are more effective and safer than previous available treatments, but more expensive. It is unclear if paying for the additional costs is an efficient strategy considering limited resources.MethodsA Markov model was built to estimate the expected costs in Chilean pesos (CL$) and converted to US dollars (US$) and benefits in quality adjusted life years (QALYs) in a hypothetic cohort of naive patients receiving DCV/ASV compared to protease inhibitors (PIs) and Peginterferon plus Ribavirin (PR). Efficacy was obtained from a mixed-treatment comparison study and costs were estimated from local sources. Utilities were obtained applying the EQ-5D survey to local patients and then valued with the Chilean tariff. A time horizon of 46 years and a discount rate of 3% for costs and outcomes was considered. The ICERs were estimated for a range of DCV/ASV prices. Deterministic and probabilistic sensitivity analyses were performed.ResultsPIs were extendedly dominated by DCV/ASV. The ICER of DCV/ASV compared to PR was US$16,635/QALY at a total treatment price of US$ 77,419; US$11,581 /QALY at a price of US$ 58,065; US$6,375/QALY at a price of US$ 38,710; and US$1,364 /QALY at a price of US$ 19,355. The probability of cost-effectiveness at a price of US$38,710 was 91.6$ 19,355. Although the results are sensitive to certain parameters, the ICER did not increase above the suggested threshold of 1 GDP per capita.ConclusionsDCV/ASV can be considered cost-effective at any price of the range studied. These results provide decision makers useful information about the value of incorporating these drugs into the public Chilean healthcare system

Expected Hepatitis C complications in patients treated with DCV/ASV versus PR.

<p>HCV: Hepatitis C Virus, PR: Peginterferon plus Ribavirin, PIs: Protease Inhibitors, DCV/ASV: Daclatasvir plus Asunaprevir, F4: cirrhosis, DC: Decompensated cirrhosis, HCC: hepatocellular carcinoma, HT: liver transplant, BCI: Bayesian credibility interval.</p><p>Expected Hepatitis C complications in patients treated with DCV/ASV versus PR.</p

Cost-effectiveness plane for DCV/ASV versus PR: The cost-effectiveness plane represents the base case scenario assuming a DCV/ASV total treatment price of US$ 38,710.

<p>Cost-effectiveness plane for DCV/ASV versus PR: The cost-effectiveness plane represents the base case scenario assuming a DCV/ASV total treatment price of US$ 38,710.</p

Probabilistic sensitivity analysis considering four DCV/ASV prices.

<p>PR: Peginterferon plus Ribavirin, PIs: Protease Inhibitors, DCV/ASV: Daclatasvir plus Asunaprevir, QALY: Quality Adjusted Life Years, ICER: Incremental cost effectiveness ratio, 1 $US = 620 CLP</p><p>^aMontecarlo simulation (5,000 iterations)</p><p>^bConsidering a suggested threshold of 1times GDP per capita</p><p>Probabilistic sensitivity analysis considering four DCV/ASV prices.</p

Acceptability curves for DCV/ASV versus PR for different prices of DCV/ASV: The acceptability curve shows the probability of cost-effectiveness considering four different prices for DCV/ASV (total drug treatment cost): US77,419—US 77,419—US 58,065- US38,710−US 38,710- US 19,355.

<p>Acceptability curves for DCV/ASV versus PR for different prices of DCV/ASV: The acceptability curve shows the probability of cost-effectiveness considering four different prices for DCV/ASV (total drug treatment cost): US$77,419—US$ 58,065- US$38,710- US$ 19,355.</p

Conversion to Mycophenolate Mofetil Monotherapy in Liver Recipients: Calcineurin Inhibitor Levels are Key

The use of calcineurin inhibitors (CNI) after liver transplantation is associated with post-transplant nephrotoxicity. Conversion to myc-ophenolate mofetil (MMF) monotherapy improves renal function, but is related to graft rejection in some recipients. Our aim was to identify variables associated with rejection after conversion to MMF monotherapy. Conversion was attempted in 40 liver transplant recipients. Clinical variables were determined and peripheral mononuclear blood cells were immunophenotyped during a 12-month follow-up. Conversion was classified as successful (SC) if rejection did not occur during the follow-up. MMF conversion was successful with 28 patients (70%) and was associated with higher glomerular filtration rates at the end of study. It also correlated with increased time elapsed since transplantation, low baseline CNI levels (Tacrolimus ≤ 6.5 ng/mL or Cyclosporine ≤ 635 ng/mL) and lower frequency of tacrolimus use. The only clinical variable independently related to SC in multivariate analysis was low baseline CNI levels (p = 0.02, OR: 6.93, 95%, CI: 1.3-29.7). Mean baseline fluorescent intensity of FOXP3+ T cells was significantly higher among recipients with SC. In conclusion, this study suggests that baseline CNI levels can be used to identify recipients with higher probability of SC to MMF monotherapy. Clinicaltrials.gov identification: NCT01321112