Joseph D. Grant Professor and Professor of Microbiology and Immunology

Publications

  • Pegylated Interferon Lambda for Covid-19. Reply. The New England journal of medicine Reis, G., Mills, E. J., Glenn, J. S. 2023; 388 (22): 2108

    View details for DOI 10.1056/NEJMc2303519

    View details for PubMedID 37256990

  • Treatment of chronic hepatitis d with peginterferon lambda - the phase 2 LIMT-1 clinical trial. Hepatology (Baltimore, Md.) Etzion, O., Hamid, S., Lurie, Y., Gane, E. J., Yardeni, D., Duehren, S., Bader, N., Nevo-Shor, A., Channa, S. M., Cotler, S. J., Mawani, M., Parkash, O., Dahari, H., Ingrid, C., Glenn, J. 2023

    Abstract

    Chronic Hepatitis Delta Virus (HDV) infection leads to the most aggressive form of human viral hepatitis for which there is no FDA-approved therapy. PEG IFN-lambda-1a (Lambda) has previously demonstrated a good tolerability profile in HBV and HCV patients compared to PEG IFN-alfa. The goal of the Phase 2 LIMT-1 trial was to evaluate the safety and efficacy of Lambda monotherapy in patients with HDV.Open-label study of Lambda 120 or 180 mcg, administered once weekly by subcutaneous injections for 48 weeks followed by 24 weeks of post-treatment follow-up.33 patients were allocated to Lambda 180 mcg (n=14) or 120 mcg (n=19). Baseline mean values: HDV RNA 4.1 log10 IU/mL (SD±1.4); ALT 106 IU/L (35-364) and bilirubin 0.5 mg/dL (0.2-1.2). Intention to treat rates of virologic response to Lambda 180 mcg and 120 mcg , 24 weeks following treatment cessation were 5 of 14(36%) and 3 of 19 (16%), respectively. Post treatment response rate of 50% was seen in low BL viral load (≤4 log10) on 180 mcg. Common on-treatment AEs included flu-like symptoms and elevated transaminase levels. Eight (24%) cases of hyperbilirubinemia with or without liver enzyme elevation, leading to drug discontinuation were mainly observed in the Pakistani cohort. The clinical course was uneventful, and all responded favorably to dose reduction or discontinuation.Treatment with Lambda in patients with chronic HDV may result in virologic response during and following treatment cessation. Clinical phase 3 development of Lambda for this rare and serious disease is ongoing.

    View details for DOI 10.1097/HEP.0000000000000309

    View details for PubMedID 36800850

  • Early Treatment with Pegylated Interferon Lambda for Covid-19. The New England journal of medicine Reis, G., Moreira Silva, E. A., Medeiros Silva, D. C., Thabane, L., Campos, V. H., Ferreira, T. S., Santos, C. V., Nogueira, A. M., Almeida, A. P., Savassi, L. C., Figueiredo-Neto, A. D., Dias, A. C., Freire Júnior, A. M., Bitarães, C., Milagres, A. C., Callegari, E. D., Simplicio, M. I., Ribeiro, L. B., Oliveira, R., Harari, O., Wilson, L. A., Forrest, J. I., Ruton, H., Sprague, S., McKay, P., Guo, C. M., Limbrick-Oldfield, E. H., Kanters, S., Guyatt, G. H., Rayner, C. R., Kandel, C., Biondi, M. J., Kozak, R., Hansen, B., Zahoor, M. A., Arora, P., Hislop, C., Choong, I., Feld, J. J., Mills, E. J., Glenn, J. S. 2023; 388 (6): 518-528

    Abstract

    The efficacy of a single dose of pegylated interferon lambda in preventing clinical events among outpatients with acute symptomatic coronavirus disease 2019 (Covid-19) is unclear.We conducted a randomized, controlled, adaptive platform trial involving predominantly vaccinated adults with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in Brazil and Canada. Outpatients who presented with an acute clinical condition consistent with Covid-19 within 7 days after the onset of symptoms received either pegylated interferon lambda (single subcutaneous injection, 180 μg) or placebo (single injection or oral). The primary composite outcome was hospitalization (or transfer to a tertiary hospital) or an emergency department visit (observation for >6 hours) due to Covid-19 within 28 days after randomization.A total of 933 patients were assigned to receive pegylated interferon lambda (2 were subsequently excluded owing to protocol deviations) and 1018 were assigned to receive placebo. Overall, 83% of the patients had been vaccinated, and during the trial, multiple SARS-CoV-2 variants had emerged. A total of 25 of 931 patients (2.7%) in the interferon group had a primary-outcome event, as compared with 57 of 1018 (5.6%) in the placebo group, a difference of 51% (relative risk, 0.49; 95% Bayesian credible interval, 0.30 to 0.76; posterior probability of superiority to placebo, >99.9%). Results were generally consistent in analyses of secondary outcomes, including time to hospitalization for Covid-19 (hazard ratio, 0.57; 95% Bayesian credible interval, 0.33 to 0.95) and Covid-19-related hospitalization or death (hazard ratio, 0.59; 95% Bayesian credible interval, 0.35 to 0.97). The effects were consistent across dominant variants and independent of vaccination status. Among patients with a high viral load at baseline, those who received pegylated interferon lambda had lower viral loads by day 7 than those who received placebo. The incidence of adverse events was similar in the two groups.Among predominantly vaccinated outpatients with Covid-19, the incidence of hospitalization or an emergency department visit (observation for >6 hours) was significantly lower among those who received a single dose of pegylated interferon lambda than among those who received placebo. (Funded by FastGrants and others; TOGETHER ClinicalTrials.gov number, NCT04727424.).

    View details for DOI 10.1056/NEJMoa2209760

    View details for PubMedID 36780676

  • Programmable Antivirals and Just-in-Time Vaccines: Biosecurity Implications of Viral RNA Secondary Structure Targeting. Health security Pannu, J., Glenn, J. S. 2022

    View details for DOI 10.1089/hs.2022.0098

    View details for PubMedID 36576394

  • Modeling the Interplay between HDV and HBV in Chronic HDV/HBV Patients MATHEMATICS Mhlanga, A., Zakh, R., Churkin, A., Reinharz, V., Glenn, J. S., Etzion, O., Cotler, S. J., Yurdaydin, C., Barash, D., Dahari, H. 2022; 10 (20)
  • ESTIMATING HEPATITIS DELTA PREVALENCE AMONG HIGH RISK POPULATIONS IN THE UNITED STATES: A SYSTEMATIC REVIEW Wong, R. J., Brosgart, C. L., Wong, S. S., Feld, J. J., Glenn, J., Moraras, K., Hamid, S. S., Ward, J. W., Wedemeyer, H., Yurdaydin, C., Gish, R. G. WILEY. 2022: S254-S255
  • ESTIMATING THE PREVALENCE OF CHRONIC HEPATITIS B AMONG FOREIGN-BORN PERSONS LIVING IN CANADA BY COUNTRY OF ORIGIN Wong, R. J., Brosgart, C. L., Wong, S. S., Feld, J. J., Hirode, G., Glenn, J., Hamid, S. S., Moraras, K., Ward, J. W., Wedemeyer, H., Yurdaydin, C., Gish, R. G. WILEY. 2022: S261-S262
  • FUNCTIONAL CURE WITH LONAFARNIB-BASED THERAPY IN CHRONIC HEPATITIS DELTA Yurdaydin, C., Yurdcu, E., Gumussoy, M., Keskin, O., Gencdal, G., Akyildiz, M., Zeybel, M., Idilman, R., Bozdayi, M., Choong, I., Glenn, J. WILEY. 2022: S297-S298
  • Programmable antivirals to target conserved essential shapes in pandemic viral genomes NATURE MEDICINE Glenn, J. S. 2022

    View details for DOI 10.1038/s41591-022-01910-3

    View details for Web of Science ID 000844603700002

    View details for PubMedID 36008723

    View details for PubMedCentralID PMC9406275

  • Programmable antivirals targeting critical conserved viral RNA secondary structures from influenza A virus and SARS-CoV-2. Nature medicine Hagey, R. J., Elazar, M., Pham, E. A., Tian, S., Ben-Avi, L., Bernardin-Souibgui, C., Yee, M. F., Moreira, F. R., Rabinovitch, M. V., Meganck, R. M., Fram, B., Beck, A., Gibson, S. A., Lam, G., Devera, J., Kladwang, W., Nguyen, K., Xiong, A., Schaffert, S., Avisar, T., Liu, P., Rustagi, A., Fichtenbaum, C. J., Pang, P. S., Khatri, P., Tseng, C., Taubenberger, J. K., Blish, C. A., Hurst, B. L., Sheahan, T. P., Das, R., Glenn, J. S. 2022

    Abstract

    Influenza A virus's (IAV's) frequent genetic changes challenge vaccine strategies and engender resistance to current drugs. We sought to identify conserved and essential RNA secondary structures within IAV's genome that are predicted to have greater constraints on mutation in response to therapeutic targeting. We identified and genetically validated an RNA structure (packaging stem-loop 2 (PSL2)) that mediates in vitro packaging and in vivo disease and is conserved across all known IAV isolates. A PSL2-targeting locked nucleic acid (LNA), administered 3d after, or 14d before, a lethal IAV inoculum provided 100% survival in mice, led to the development of strong immunity to rechallenge with a tenfold lethal inoculum, evaded attempts to select for resistance and retained full potency against neuraminidase inhibitor-resistant virus. Use of an analogous approach to target SARS-CoV-2, prophylactic administration of LNAs specific for highly conserved RNA structures in the viral genome, protected hamsters from efficient transmission of the SARS-CoV-2 USA_WA1/2020 variant. These findings highlight the potential applicability of this approach to any virus of interest via a process we term 'programmable antivirals', with implications for antiviral prophylaxis and post-exposure therapy.

    View details for DOI 10.1038/s41591-022-01908-x

    View details for PubMedID 35982307

  • Lonafarnib combination with peginterferon Lambda diminished triphasic HDV kinetic pattrn seen under Lambda monotherapy: the LIFT HDV study Duehren, S., Koh, C., Hercun, J., Rahman, F., Surana, P., Vittal, A., Lai, W., Etzion, O., Cotler, S., Glenn, J., Dahari, H., Heller, T. ELSEVIER. 2022: S70-S71
  • Mathematical modeling of HDV RNA kinetics suggests high peginterferon Lambda efficacy in blocking viral production: insights from the LIMT-1 study Cardozo-Ojeda, E., Duehren, S., Hamid, S., Lurie, Y., Gane, E. J., Nevo-Shor, A., Yardeni, D., Cotler, S., Choong, I., Glenn, J., Dahari, H., Etzion, O. ELSEVIER. 2022: S859
  • Estimating the prevalence of hepatitis delta infection among foreign-born adults with chronic hepatitis B in the United States Wong, R., Brosgart, C., Wong, S., Feld, J., Glenn, J., Hamid, S., Moraras, K., Ward, J., Wedemeyer, H., Yurdaydin, C., Gish, R. G. ELSEVIER. 2022: S274
  • First-in-human experience using RBS2418, an oral ENPP1 inhibitor within an expanded access protocol in combination with pembrolizumab in a patient with metastatic adrenal cancer. Csiki, I., Dong, A., Tuan, B. Y., John, E., O'Toole, L., Seppa, J., Huang, N., Hawley, R. C., Exon, C., Glenn, J. S., Klumpp, K. LIPPINCOTT WILLIAMS & WILKINS. 2022
  • Combination of Novel Therapies for HDV. Viruses Elazar, M., Glenn, J. S. 2022; 14 (2)

    Abstract

    Treatment options for HDV have been limited to interferon alfa-based therapies with its poor efficacy to side effects ratio. Several novel therapies have now advanced into the clinic. As they each have a different mechanism of action, there is the potential for combination therapy. Here we review how studying the HDV life cycle has led to the development of these novel therapies, the key developments leading to, and the details of, the first combination study of novel anti-HDV therapies, and suggest what additional combinations of novel therapies can be anticipated as we enter this exciting new area of HDV treatments.

    View details for DOI 10.3390/v14020268

    View details for PubMedID 35215860

  • Efficacy and Safety of a Botanical Formula Fuzheng Huayu for Hepatic Fibrosis in Patients with CHC: Results of a Phase 2 Clinical Trial. Evidence-based complementary and alternative medicine : eCAM Hassanein, T., Tai, D., Liu, C., Box, T. D., Tong, M. J., Rossaro, L., Pozza, R., Glenn, J. S., Cheung, R., Hemaidan, A., He, Y., Behling, C., Hu, X., Makhlouf, H., Fan, H., Ren, Y., Khim Chng, E. L., Liu, P., Vierling, J. M. 2022; 2022: 4494099

    Abstract

    Background: Hepatitis C virus (HCV) is a common cause of progressive hepatic fibrosis, cirrhosis, and hepatocellular carcinoma worldwide. Despite the availability of effective direct-acting antivirals, patients often have significant hepatic fibrosis at the time of diagnosis due to delay in diagnosis and comorbidities which promote fibrogenesis. Thus, antifibrotic agents represent an attractive adjunctive therapy. Fuzheng Huayu (FZHY), a traditional Chinese medicine botanical formulation, has been used as an antifibrotic agent in chronic HBV infection. Our aim was to assess FZHY in patients with HCV infection and active viremia.Method: We randomized 118 patients with active viremia from 8 liver centers in the U.S. to receive oral FZHY (n=59) or placebo (n=59) for 48 weeks. Efficacy was assessed by histopathologic changes at the end of therapy. A subset of biopsies was further analyzed using qFibrosis to detect subtle changes in fibrosis in different zones of the hepatic lobules.Results: FZHY was well tolerated and safe. Patients with baseline Ishak fibrosis stages F3 and F4 had better response rates to FZHY than patients with baseline F0-F2 (p=0.03). qFibrosis zonal analysis showed significant improvement in fibrosis in all zones in patients with regression of the fibrosis stage.Conclusions: FZHY produced antifibrotic effects in patients with baseline Ishak F3 and F4 fibrosis stages. Reduction in fibrosis severity was zonal and correlated with the severity of inflammation. Based on its tolerability, safety, and efficacy, FZHY should be further investigated as a therapy in chronic liver diseases because of its dual anti-inflammatory and antiibrotic properties. Lay Summary. This is the first US-based, multicenter and placebo-controlled clinical trial that shows statistically significant reduction in fibrosis in patients with active HCV using an antifibrotic botanical formula. This has important implications as there is an immediate need for effective antifibrotic agents in treating many chronic diseases including NASH that lead to scarring of the liver. With artificial intelligence-based methodology, qFibrosis, we may provide a more reliable way to assess the FZHY as a therapy in chronic liver diseases because of its dual anti-inflammatory and antifibrotic properties.

    View details for DOI 10.1155/2022/4494099

    View details for PubMedID 35873630

  • A Phase ii dose finding study of lonafarnib and ritonavir with or without interferon alpha for chronic delta hepatitis. Hepatology (Baltimore, Md.) Yurdaydin, C., Keskin, O., Yurdcu, E., Caliskan, A., Onem, S., Karakaya, F., Kalkan, C., Karatayli, E., Karatayli, S., Choong, I., Apelian, D., Koh, C., Heller, T., Idilman, R., Bozdayi, M. A., Glenn, J. S. 2021

    Abstract

    BACKGROUND & AIMS: Proof-of-concept studies demonstrated lonafarnib (LNF), a first-in-class, oral prenylation inhibitor, efficacy in HDV-infected patients. The LOnafarnib With Ritonavir for HDV-2 (LOWR-2) study's aim was to identify optimal combination regimens of lonafarnib + ritonavir (RTV) ± pegylated interferon alfa (PEG-IFNalpha) with efficacy and tolerability for longer term dosing. Here we report the safety and efficacy at end of treatment (EOT) for up to 24 weeks.APPROACH & RESULTS: 55 patients with chronic HDV were consecutively enrolled in an open-label single-center phase 2 dose-finding study. There were 3 main treatment groups: high dose lonafarnib (lonafarnib ≥ 75 mg po bid + ritonavir) (N=19, 12 wks); all-oral low dose lonafarnib (lonafarnib 25 or 50 mg po bid + ritonavir) (N=24, 24 wks) and combination low dose lonafarnib with PEG-IFNalpha (lonafarnib 25 or 50 mg po bid + ritonavir + PEG-IFNalpha) (N=12, 24 wks). The primary endpoint, ≥ 2 log10 decline or < LLOQ of HDV-RNA from baseline at EOT, was reached in 46% (6/13) and 89% (8/9) of patients receiving the all-oral regimen of lonafarnib 50 mg bid + ritonavir, and combination regimens of lonafarnib (25 or 50 mg bid) + ritonavir + PEG-IFNalpha, respectively. In addition, multiple patients experienced well-tolerated transient post-treatment ALT increases resulting in HDV RNA negativity and ALT normalization. The proportions of grade 2 and 3 GI adverse events in the high vs low dose groups were 49% (37/76) and only 22% (18/81), respectively.CONCLUSIONS: Lonafarnib, boosted with low dose ritonavir, is a promising all-oral therapy, and maximal efficacy achieved with PEG-IFNalpha addition. The identified optimal regimens support the first Phase 3 (D-LIVR) study of lonafarnib for the treatment of HDV.

    View details for DOI 10.1002/hep.32259

    View details for PubMedID 34860418

  • Hepatitis Delta Virus Testing and Research. Gastroenterology & hepatology Glenn, J. S. 2021; 17 (10): 482-484

    View details for PubMedID 35462727

  • HEPATITIS D VIRUS INFECTION IN UNITED STATES WOMEN WITH OR AT RISK FOR HIV Argirion, I., Mahale, P., Kuniholm, M. H., Pfeiffer, R., Koshiol, J., Glenn, J., O'Brien, T. R. WILEY. 2021: 422A
  • Cryo-EM and antisense targeting of the 28-kDa frameshift stimulation element from the SARS-CoV-2 RNA genome. Nature structural & molecular biology Zhang, K., Zheludev, I. N., Hagey, R. J., Haslecker, R., Hou, Y. J., Kretsch, R., Pintilie, G. D., Rangan, R., Kladwang, W., Li, S., Wu, M. T., Pham, E. A., Bernardin-Souibgui, C., Baric, R. S., Sheahan, T. P., D'Souza, V., Glenn, J. S., Chiu, W., Das, R. 2021

    Abstract

    Drug discovery campaigns against COVID-19 are beginning to target the SARS-CoV-2 RNA genome. The highly conserved frameshift stimulation element (FSE), required for balanced expression of viral proteins, is a particularly attractive SARS-CoV-2 RNA target. Here we present a 6.9A resolution cryo-EM structure of the FSE (88nucleotides, ~28kDa), validated through an RNA nanostructure tagging method. The tertiary structure presents a topologically complex fold in which the 5' end is threaded through a ring formed inside a three-stem pseudoknot. Guided by this structure, we develop antisense oligonucleotides that impair FSE function in frameshifting assays and knock down SARS-CoV-2 virus replication in A549-ACE2 cells at 100nM concentration.

    View details for DOI 10.1038/s41594-021-00653-y

    View details for PubMedID 34426697

  • Addiction to Golgi-resident PI4P synthesis in chromosome 1q21.3-amplified lung adenocarcinoma cells. Proceedings of the National Academy of Sciences of the United States of America Shi, L., Tan, X., Liu, X., Yu, J., Bota-Rabassedas, N., Niu, Y., Luo, J., Xi, Y., Zong, C., Creighton, C. J., Glenn, J. S., Wang, J., Kurie, J. M. 2021; 118 (25)

    Abstract

    A chromosome 1q21.3 region that is frequently amplified in diverse cancer types encodes phosphatidylinositol (PI)-4 kinase IIIbeta (PI4KIIIbeta), a key regulator of secretory vesicle biogenesis and trafficking. Chromosome 1q21.3-amplified lung adenocarcinoma (1q-LUAD) cells rely on PI4KIIIbeta for Golgi-resident PI-4-phosphate (PI4P) synthesis, prosurvival effector protein secretion, and cell viability. Here, we show that 1q-LUAD cells subjected to prolonged PI4KIIIbeta antagonist treatment acquire tolerance by activating an miR-218-5p-dependent competing endogenous RNA network that up-regulates PI4KIIalpha, which provides an alternative source of Golgi-resident PI4P that maintains prosurvival effector protein secretion and cell viability. These findings demonstrate an addiction to Golgi-resident PI4P synthesis in a genetically defined subset of cancers.

    View details for DOI 10.1073/pnas.2023537118

    View details for PubMedID 34155143

  • Publisher Correction: A mouse-adapted model of SARS-CoV-2 to test COVID-19 countermeasures. Nature Dinnon, K. H., Leist, S. R., Schäfer, A. n., Edwards, C. E., Martinez, D. R., Montgomery, S. A., West, A. n., Yount, B. L., Hou, Y. J., Adams, L. E., Gully, K. L., Brown, A. J., Huang, E. n., Bryant, M. D., Choong, I. C., Glenn, J. S., Gralinski, L. E., Sheahan, T. P., Baric, R. S. 2021

    View details for DOI 10.1038/s41586-020-03107-5

    View details for PubMedID 33469219

  • Durable virological response and functional cure of chronic hepatitis D after long-term peginterferon therapy. Alimentary pharmacology & therapeutics Hercun, J., Kim, G. E., Da, B. L., Rotman, Y., Kleiner, D. E., Chang, R., Glenn, J. S., Hoofnagle, J. H., Koh, C., Heller, T. 2021

    Abstract

    Hepatitis delta virus (HDV) infection is the most aggressive form of chronic viral hepatitis. Response rates to therapy with 1- to 2-year courses of pegylated interferon alpha (peginterferon) treatment are suboptimal.To evaluate the long-term outcomes of patients with chronic hepatitis D after an extended course of peginterferon.Patients were followed after completion of trial NCT00023322 and classified based on virological response defined as loss of detectable serum HDV RNA at last follow-up. During extended follow-up, survival and liver-related events were recorded.All 12 patients who received more than 6 months of peginterferon in the original study were included in this analysis. The cohort was mostly white (83%) and male (92%) and ranged in age from 18 to 58 years (mean = 42.6). Most patients had advanced but compensated liver disease at baseline, a median HBV DNA level of 536 IU per mL and median HDV RNA level of 6.86 log10 genome equivalents per mL. The treatment duration averaged 6.1 years (range 0.8-14.3) with a total follow-up of 8.8 years (range 1.7-17.6). At last follow-up, seven (58%) patients had durable undetectable HDV RNA in serum, and four (33%) cleared HBsAg. Overall, one of seven (14%) responders died or had a liver-related event vs four of five (80%) non-responders.With further follow-up, an extended course of peginterferon therapy was found to result in sustained clearance of HDV RNA and favourable clinical outcomes in more than half of patients and loss of HBsAg in a third.

    View details for DOI 10.1111/apt.16408

    View details for PubMedID 34048594

  • Signatures of immune dysfunction in HIV and HCV infection share features with chronic inflammation in aging and persist after viral reduction or elimination. Proceedings of the National Academy of Sciences of the United States of America Lopez Angel, C. J., Pham, E. A., Du, H. n., Vallania, F. n., Fram, B. J., Perez, K. n., Nguyen, T. n., Rosenberg-Hasson, Y. n., Ahmed, A. n., Dekker, C. L., Grant, P. M., Khatri, P. n., Maecker, H. T., Glenn, J. S., Davis, M. M., Furman, D. n. 2021; 118 (14)

    Abstract

    Chronic inflammation is thought to be a major cause of morbidity and mortality in aging, but whether similar mechanisms underlie dysfunction in infection-associated chronic inflammation is unclear. Here, we profiled the immune proteome, and cellular composition and signaling states in a cohort of aging individuals versus a set of HIV patients on long-term antiretroviral therapy therapy or hepatitis C virus (HCV) patients before and after sofosbuvir treatment. We found shared alterations in aging-associated and infection-associated chronic inflammation including T cell memory inflation, up-regulation of intracellular signaling pathways of inflammation, and diminished sensitivity to cytokines in lymphocytes and myeloid cells. In the HIV cohort, these dysregulations were evident despite viral suppression for over 10 y. Viral clearance in the HCV cohort partially restored cellular sensitivity to interferon-α, but many immune system alterations persisted for at least 1 y posttreatment. Our findings indicate that in the HIV and HCV cohorts, a broad remodeling and degradation of the immune system can persist for a year or more, even after the removal or drastic reduction of the pathogen load and that this shares some features of chronic inflammation in aging.

    View details for DOI 10.1073/pnas.2022928118

    View details for PubMedID 33811141

  • Peginterferon lambda for the treatment of outpatients with COVID-19: a phase 2, placebo-controlled randomised trial. The Lancet. Respiratory medicine Feld, J. J., Kandel, C. n., Biondi, M. J., Kozak, R. A., Zahoor, M. A., Lemieux, C. n., Borgia, S. M., Boggild, A. K., Powis, J. n., McCready, J. n., Tan, D. H., Chan, T. n., Coburn, B. n., Kumar, D. n., Humar, A. n., Chan, A. n., O'Neil, B. n., Noureldin, S. n., Booth, J. n., Hong, R. n., Smookler, D. n., Aleyadeh, W. n., Patel, A. n., Barber, B. n., Casey, J. n., Hiebert, R. n., Mistry, H. n., Choong, I. n., Hislop, C. n., Santer, D. M., Lorne Tyrrell, D. n., Glenn, J. S., Gehring, A. J., Janssen, H. L., Hansen, B. E. 2021

    Abstract

    To date, only monoclonal antibodies have been shown to be effective for outpatients with COVID-19. Interferon lambda-1 is a type III interferon involved in innate antiviral responses with activity against respiratory pathogens. We aimed to investigate the safety and efficacy of peginterferon lambda in the treatment of outpatients with mild-to-moderate COVID-19.In this double-blind, placebo-controlled trial, outpatients with laboratory-confirmed COVID-19 were randomly assigned to a single subcutaneous injection of peginterferon lambda 180 μg or placebo within 7 days of symptom onset or first positive swab if asymptomatic. Participants were randomly assigned (1:1) using a computer-generated randomisation list created with a randomisation schedule in blocks of four. At the time of administration, study nurses received a sealed opaque envelope with the treatment allocation number. The primary endpoint was the proportion of patients who were negative for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA on day 7 after the injection, analysed by a χ2 test following an intention-to-treat principle. Prespecified analysis of the primary endpoint, adjusted for baseline viral load, using bivariate logistic regression was done. The trial is now complete. This trial is registered with ClinicalTrials.gov, NCT04354259.Between May 18, and Sept 4, 2020, we recruited 30 patients per group. The decline in SARS-CoV-2 RNA was greater in those treated with peginterferon lambda than placebo from day 3 onwards, with a difference of 2·42 log copies per mL at day 7 (p=0·0041). By day 7, 24 (80%) participants in the peginterferon lambda group had an undetectable viral load, compared with 19 (63%) in the placebo group (p=0·15). After controlling for baseline viral load, patients in the peginterferon lambda group were more likely to have undetectable virus by day 7 than were those in the placebo group (odds ratio [OR] 4·12 [95% CI 1·15-16·73; p=0·029). Of those with baseline viral load above 106 copies per mL, 15 (79%) of 19 patients in the peginterferon lambda group had undetectable virus on day 7, compared with six (38%) of 16 in the placebo group (OR 6·25 [95% CI 1·49-31·06]; p=0·012). Peginterferon lambda was well tolerated, and adverse events were similar between groups with mild and transient aminotransferase, concentration increases more frequently observed in the peginterferon lambda group. Two individuals met the threshold of grade 3 increase, one in each group, and no other grade 3 or 4 laboratory adverse events were reported.Peginterferon lambda accelerated viral decline in outpatients with COVID-19, increasing the proportion of patients with viral clearance by day 7, particularly in those with high baseline viral load. Peginterferon lambda has potential to prevent clinical deterioration and shorten duration of viral shedding.The Toronto COVID-19 Action Initiative, University of Toronto, and the Ontario First COVID-19 Rapid Research Fund, Toronto General & Western Hospital Foundation.

    View details for DOI 10.1016/S2213-2600(20)30566-X

    View details for PubMedID 33556319

  • Peginterferon Lambda-1a for treatment of outpatients with uncomplicated COVID-19: a randomized placebo-controlled trial. Nature communications Jagannathan, P. n., Andrews, J. R., Bonilla, H. n., Hedlin, H. n., Jacobson, K. B., Balasubramanian, V. n., Purington, N. n., Kamble, S. n., de Vries, C. R., Quintero, O. n., Feng, K. n., Ley, C. n., Winslow, D. n., Newberry, J. n., Edwards, K. n., Hislop, C. n., Choong, I. n., Maldonado, Y. n., Glenn, J. n., Bhatt, A. n., Blish, C. n., Wang, T. n., Khosla, C. n., Pinsky, B. A., Desai, M. n., Parsonnet, J. n., Singh, U. n. 2021; 12 (1): 1967

    Abstract

    Type III interferons have been touted as promising therapeutics in outpatients with coronavirus disease 2019 (COVID-19). We conducted a randomized, single-blind, placebo-controlled trial (NCT04331899) in 120 outpatients with mild to moderate COVID-19 to determine whether a single, 180 mcg subcutaneous dose of Peginterferon Lambda-1a (Lambda) within 72 hours of diagnosis could shorten the duration of viral shedding (primary endpoint) or symptoms (secondary endpoint). In both the 60 patients receiving Lambda and 60 receiving placebo, the median time to cessation of viral shedding was 7 days (hazard ratio [HR] = 0.81; 95% confidence interval [CI] 0.56 to 1.19). Symptoms resolved in 8 and 9 days in Lambda and placebo, respectively, and symptom duration did not differ significantly between groups (HR 0.94; 95% CI 0.64 to 1.39). Both Lambda and placebo were well-tolerated, though liver transaminase elevations were more common in the Lambda vs. placebo arm (15/60 vs 5/60; p = 0.027). In this study, a single dose of subcutaneous Peginterferon Lambda-1a neither shortened the duration of SARS-CoV-2 viral shedding nor improved symptoms in outpatients with uncomplicated COVID-19.

    View details for DOI 10.1038/s41467-021-22177-1

    View details for PubMedID 33785743

  • Progenitor identification and SARS-CoV-2 infection in human distal lung organoids. Nature Salahudeen, A. A., Choi, S. S., Rustagi, A., Zhu, J., van Unen, V., de la O, S. M., Flynn, R. A., Margalef-Catala, M., Santos, A. J., Ju, J., Batish, A., Usui, T., Zheng, G. X., Edwards, C. E., Wagar, L. E., Luca, V., Anchang, B., Nagendran, M., Nguyen, K., Hart, D. J., Terry, J. M., Belgrader, P., Ziraldo, S. B., Mikkelsen, T. S., Harbury, P. B., Glenn, J. S., Garcia, K. C., Davis, M. M., Baric, R. S., Sabatti, C., Amieva, M. R., Blish, C. A., Desai, T. J., Kuo, C. J. 2020

    Abstract

    The distal lung contains terminal bronchioles and alveoli that facilitate gas exchange. Three-dimensional in vitro human distal lung culture systems would strongly facilitate investigation of pathologies including interstitial lung disease, cancer, and SARS-CoV-2-associated COVID-19 pneumonia. We generated long-term feeder-free, chemically defined culture of distal lung progenitors as organoids derived from single adult human alveolar epithelial type II (AT2) or KRT5+ basal cells. AT2 organoids exhibited AT1 transdifferentiation potential while basal cell organoids developed lumens lined by differentiated club and ciliated cells. Single cell analysis of basal organoid KRT5+ cells revealed a distinct ITGA6+ITGB4+ mitotic population whose proliferation further segregated to a TNFRSF12Ahi subfraction comprising ~10% of KRT5+ basal cells, residing in clusters within terminal bronchioles and exhibiting enriched clonogenic organoid growth activity. Distal lung organoids were created with apical-out polarity to display ACE2 on the exposed external surface, facilitating SARS-CoV-2 infection of AT2 and basal cultures and identifying club cells as a novel target population. This long-term, feeder-free organoid culture of human distal lung, coupled with single cell analysis, identifies unsuspected basal cell functional heterogeneity and establishes a facile in vitro organoid model for human distal lung infections including COVID-19-associated pneumonia.

    View details for DOI 10.1038/s41586-020-3014-1

    View details for PubMedID 33238290

  • High Prevalence of Chronic Viral Hepatitis and Liver Fibrosis Among Mongols in Southern California. Digestive diseases and sciences Fong, T., Lee, B. T., Chang, M., Nasanbayar, K., Tsogtoo, E., Boldbaatar, D., Dashdorj, E. D., Clifford, N. E., Dashdorj, A. N., Bang, B., Chida, T., Lim, C., Sugiyama, M., Mizokami, M., Dashdorj, N. J., Liu, P., Glenn, J. S., Dashdorj, N. D., Saito, T. 2020

    Abstract

    BACKGROUND: Mongolia is a highly endemic region for chronic hepatitis B (HBV), hepatitis delta (HDV), and hepatitis C (HCV) infections. Aim of this study was to comprehensively characterize chronic viral hepatitis among Mongols living in Southern California.METHODS: Three screening events were conducted between August and November 2018, with 528 adult Mongols tested for HBV and HCV. HBsAg (+) individuals (CHB) underwent additional testing for HDV RNA and anti-HDV. Liver tests, platelet count, and FibroScan were performed on CHB and chronic HCV (CHC) individuals.RESULTS: Fifty-one out of 534 were HBsAg reactive (9.7%), and all were foreign-born. Mean age of CHB individuals was 37.8 (range 18-69) years. Forty-six out of 51 were HBeAg (-). HBV genotypes were exclusively D2 or A1. Twenty-one out of 51 (41.2%) were anti-HDV (+) and 17/51 (33.3%) were HDV RNA (+). HDV RNA (+) individuals had significantly higher ALT, fibrosis-4 score, and liver stiffness compared to HDV RNA (-) individuals. Incidence of advanced fibrosis was higher in HDV RNA (+) individuals (57% vs. 13%, p=0.013). Forty-eight (9.1%) individuals were anti-HCV (+) and 19 (3.6%) were HCV RNA (+). Mean age of CHC individuals was 40.2 (range 28-71) years. Prevalence of anti-HCV (+) was higher among those born between 1945 and 1965 versus those born after 1965 (18.8% vs. 7.9%, p=0.025). Genotype 1b was predominant. Incidence of cirrhosis was 7% among all participants.CONCLUSIONS: Mongols living in the USA are at high risk for CHB and CHC infections. One-third of CHB individuals had CHD superinfection with advanced fibrosis. Universal screening for viral hepatitis in Mongols in the USA is mandatory.

    View details for DOI 10.1007/s10620-020-06499-6

    View details for PubMedID 32770488

  • A Phase 2 Study of Peginterferon Lambda, Lonafarnib and Ritonavir for 24 Weeks: End-of-Treatment Results from the LIFT HDV Study Koh, C., Hercun, J., Rahman, F., Huang, A., Da, B., Surana, P., Kapuria, D., Rotman, Y., Vittal, A., Gilman, C., Ben Yakov, G., Lai, W., Dahari, H., Glenn, J., Heller, T. ELSEVIER. 2020: S130
  • RNA genome conservation and secondary structure in SARS-CoV-2 and SARS-related viruses: a first look. RNA (New York, N.Y.) Rangan, R., Zheludev, I. N., Das, R. 2020

    Abstract

    As the COVID-19 outbreak spreads, there is a growing need for a compilation of conserved RNA genome regions in the SARS-CoV-2 virus along with their structural propensities to guide development of antivirals and diagnostics. Here we present a first look at RNA sequence conservation and structural propensities in the SARS-CoV-2 genome. Using sequence alignments spanning a range of betacoronaviruses, we rank genomic regions by RNA sequence conservation, identifying 79 regions of length at least 15 nucleotides as exactly conserved over SARS-related complete genome sequences available near the beginning of the COVID-19 outbreak. We then confirm the conservation of the majority of these genome regions across 739 SARS-CoV-2 sequences subsequently reported from the COVID-19 outbreak, and we present a curated list of 30 'SARS-related-conserved' regions. We find that known RNA structured elements curated as Rfam families and in prior literature are enriched in these conserved genome regions, and we predict additional conserved, stable secondary structures across the viral genome. We provide 106 'SARS-CoV-2-conserved-structured' regions as potential targets for antivirals that bind to structured RNA. We further provide detailed secondary structure models for the extended 5' UTR, frame-shifting element, and 3' UTR. Last, we predict regions of the SARS-CoV-2 viral genome that have low propensity for RNA secondary structure and are conserved within SARS-CoV-2 strains. These 59 'SARS-CoV-2-conserved-unstructured' genomic regions may be most easily targeted in primer-based diagnostic and oligonucleotide-based therapeutic strategies.

    View details for DOI 10.1261/rna.076141.120

    View details for PubMedID 32398273

  • COVID-19 and emerging viral infections: The case for interferon lambda. The Journal of experimental medicine Prokunina-Olsson, L., Alphonse, N., Dickenson, R. E., Durbin, J. E., Glenn, J. S., Hartmann, R., Kotenko, S. V., Lazear, H. M., O'Brien, T. R., Odendall, C., Onabajo, O. O., Piontkivska, H., Santer, D. M., Reich, N. C., Wack, A., Zanoni, I. 2020; 217 (5)

    Abstract

    With the first reports on coronavirus disease 2019 (COVID-19), which is caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the scientific community working in the field of type III IFNs (IFN-lambda) realized that this class of IFNs could play an important role in this and other emerging viral infections. In this Viewpoint, we present our opinion on the benefits and potential limitations of using IFN-lambda to prevent, limit, and treat these dangerous viral infections.

    View details for DOI 10.1084/jem.20200653

    View details for PubMedID 32289152

  • Extracellular cGAMP is a cancer-cell-produced immunotransmitter involved in radiation-induced anticancer immunity NATURE CANCER Carozza, J. A., Bohnert, V., Nguyen, K. C., Skariah, G., Shaw, K. E., Brown, J. A., Rafat, M., von Eyben, R., Graves, E. E., Glenn, J. S., Smith, M., Li, L. 2020; 1 (2): 184-+
  • Extracellular cGAMP is a cancer cell-produced immunotransmitter involved in radiation-induced anti-cancer immunity. Nature cancer Carozza, J. A., Böhnert, V., Nguyen, K. C., Skariah, G., Shaw, K. E., Brown, J. A., Rafat, M., von Eyben, R., Graves, E. E., Glenn, J. S., Smith, M., Li, L. 2020; 1 (2): 184-196

    Abstract

    2'3'-cyclic GMP-AMP (cGAMP) is an intracellular second messenger that is synthesized in response to cytosolic double-stranded DNA and activates the innate immune STING pathway. Our previous discovery of its extracellular hydrolase ENPP1 hinted at the existence of extracellular cGAMP. Here, we detected that cGAMP is continuously exported but then efficiently cleared by ENPP1, explaining why it has previously escaped detection. By developing potent, specific, and cell impermeable ENPP1 inhibitors, we found that cancer cells continuously export cGAMP in culture at steady state and at higher levels when treated with ionizing radiation (IR). In mouse tumors, depletion of extracellular cGAMP decreased tumor-associated immune cell infiltration and abolished the curative effect of IR. Boosting extracellular cGAMP with ENPP1 inhibitors synergized with IR to delay tumor growth. In conclusion, extracellular cGAMP is an anti-cancer immunotransmitter that could be harnessed to treat cancers with low immunogenicity.

    View details for DOI 10.1038/s43018-020-0028-4

    View details for PubMedID 33768207

    View details for PubMedCentralID PMC7990037

  • Upregulation of CD47 Is a Host Checkpoint Response to Pathogen Recognition. mBio Tal, M. C., Torrez Dulgeroff, L. B., Myers, L. n., Cham, L. B., Mayer-Barber, K. D., Bohrer, A. C., Castro, E. n., Yiu, Y. Y., Lopez Angel, C. n., Pham, E. n., Carmody, A. B., Messer, R. J., Gars, E. n., Kortmann, J. n., Markovic, M. n., Hasenkrug, M. n., Peterson, K. E., Winkler, C. W., Woods, T. A., Hansen, P. n., Galloway, S. n., Wagh, D. n., Fram, B. J., Nguyen, T. n., Corey, D. n., Kalluru, R. S., Banaei, N. n., Rajadas, J. n., Monack, D. M., Ahmed, A. n., Sahoo, D. n., Davis, M. M., Glenn, J. S., Adomati, T. n., Lang, K. S., Weissman, I. L., Hasenkrug, K. J. 2020; 11 (3)

    Abstract

    It is well understood that the adaptive immune response to infectious agents includes a modulating suppressive component as well as an activating component. We now show that the very early innate response also has an immunosuppressive component. Infected cells upregulate the CD47 "don't eat me" signal, which slows the phagocytic uptake of dying and viable cells as well as downstream antigen-presenting cell (APC) functions. A CD47 mimic that acts as an essential virulence factor is encoded by all poxviruses, but CD47 expression on infected cells was found to be upregulated even by pathogens, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), that encode no mimic. CD47 upregulation was revealed to be a host response induced by the stimulation of both endosomal and cytosolic pathogen recognition receptors (PRRs). Furthermore, proinflammatory cytokines, including those found in the plasma of hepatitis C patients, upregulated CD47 on uninfected dendritic cells, thereby linking innate modulation with downstream adaptive immune responses. Indeed, results from antibody-mediated CD47 blockade experiments as well as CD47 knockout mice revealed an immunosuppressive role for CD47 during infections with lymphocytic choriomeningitis virus and Mycobacterium tuberculosis Since CD47 blockade operates at the level of pattern recognition receptors rather than at a pathogen or antigen-specific level, these findings identify CD47 as a novel potential immunotherapeutic target for the enhancement of immune responses to a broad range of infectious agents.IMPORTANCE Immune responses to infectious agents are initiated when a pathogen or its components bind to pattern recognition receptors (PRRs). PRR binding sets off a cascade of events that activates immune responses. We now show that, in addition to activating immune responses, PRR signaling also initiates an immunosuppressive response, probably to limit inflammation. The importance of the current findings is that blockade of immunomodulatory signaling, which is mediated by the upregulation of the CD47 molecule, can lead to enhanced immune responses to any pathogen that triggers PRR signaling. Since most or all pathogens trigger PRRs, CD47 blockade could be used to speed up and strengthen both innate and adaptive immune responses when medically indicated. Such immunotherapy could be done without a requirement for knowing the HLA type of the individual, the specific antigens of the pathogen, or, in the case of bacterial infections, the antimicrobial resistance profile.

    View details for DOI 10.1128/mBio.01293-20

    View details for PubMedID 32576678

  • Materials science approaches in the development of broad-spectrum antiviral therapies. Nature materials Cho, N. J., Glenn, J. S. 2020

    View details for DOI 10.1038/s41563-020-0698-4

    View details for PubMedID 32427958

  • PI4KIIIβ is a therapeutic target in chromosome 1q-amplified lung adenocarcinoma. Science translational medicine Tan, X. n., Banerjee, P. n., Pham, E. A., Rutaganira, F. U., Basu, K. n., Bota-Rabassedas, N. n., Guo, H. F., Grzeskowiak, C. L., Liu, X. n., Yu, J. n., Shi, L. n., Peng, D. H., Rodriguez, B. L., Zhang, J. n., Zheng, V. n., Duose, D. Y., Solis, L. M., Mino, B. n., Raso, M. G., Behrens, C. n., Wistuba, I. I., Scott, K. L., Smith, M. n., Nguyen, K. n., Lam, G. n., Choong, I. n., Mazumdar, A. n., Hill, J. L., Gibbons, D. L., Brown, P. H., Russell, W. K., Shokat, K. n., Creighton, C. J., Glenn, J. S., Kurie, J. M. 2020; 12 (527)

    Abstract

    Heightened secretion of protumorigenic effector proteins is a feature of malignant cells. Yet, the molecular underpinnings and therapeutic implications of this feature remain unclear. Here, we identify a chromosome 1q region that is frequently amplified in diverse cancer types and encodes multiple regulators of secretory vesicle biogenesis and trafficking, including the Golgi-dedicated enzyme phosphatidylinositol (PI)-4-kinase IIIβ (PI4KIIIβ). Molecular, biochemical, and cell biological studies show that PI4KIIIβ-derived PI-4-phosphate (PI4P) synthesis enhances secretion and accelerates lung adenocarcinoma progression by activating Golgi phosphoprotein 3 (GOLPH3)-dependent vesicular release from the Golgi. PI4KIIIβ-dependent secreted factors maintain 1q-amplified cancer cell survival and influence prometastatic processes in the tumor microenvironment. Disruption of this functional circuitry in 1q-amplified cancer cells with selective PI4KIIIβ antagonists induces apoptosis and suppresses tumor growth and metastasis. These results support a model in which chromosome 1q amplifications create a dependency on PI4KIIIβ-dependent secretion for cancer cell survival and tumor progression.

    View details for DOI 10.1126/scitranslmed.aax3772

    View details for PubMedID 31969487

  • A mouse-adapted model of SARS-CoV-2 to test COVID-19 countermeasures. Nature Dinnon, K. H., Leist, S. R., Schäfer, A. n., Edwards, C. E., Martinez, D. R., Montgomery, S. A., West, A. n., Yount, B. L., Hou, Y. J., Adams, L. E., Gully, K. L., Brown, A. J., Huang, E. n., Bryant, M. D., Choong, I. C., Glenn, J. S., Gralinski, L. E., Sheahan, T. P., Baric, R. S. 2020

    Abstract

    Coronaviruses are prone to emergence into new host species most recently evidenced by SARS-CoV-2, the causative agent of the COVID-19 pandemic1. Small animal models that recapitulate SARS-CoV-2 disease are desperately needed to rapidly evaluate medical countermeasures (MCMs)2,3. SARS-CoV-2 cannot infect wildtype laboratory mice due to inefficient interactions between the viral spike (S) protein and the murine ortholog of the human receptor, ACE24. We used reverse genetics5 to remodel the interaction between S and mACE2 resulting in a recombinant virus (SARS-CoV-2 MA) that could utilize mACE2 for entry. SARS-CoV-2 MA replicated in both the upper and lower airways of both young adult and aged BALB/c mice. Importantly, disease was more severe in aged mice, and showed more clinically relevant phenotypes than those seen in HFH4-hACE2 transgenic mice. We then demonstrated the utility of this model through vaccine challenge studies in immune competent mice with native expression of mACE2. Lastly, we show that clinical candidate interferon (IFN) lambda-1a can potently inhibit SARS-CoV-2 replication in primary human airway epithelial cells in vitro, and both prophylactic and therapeutic administration diminished replication in mice. Our mouse-adapted SARS-CoV-2 model demonstrates age-related disease pathogenesis and supports the clinical use of pegylated IFN lambda-1a treatment in human COVID-19 infections6.

    View details for DOI 10.1038/s41586-020-2708-8

    View details for PubMedID 32854108

  • Structure of the neurotensin receptor 1 in complex with β-arrestin 1. Nature Huang, W. n., Masureel, M. n., Qianhui, Q. n., Janetzko, J. n., Inoue, A. n., Kato, H. E., Robertson, M. J., Nguyen, K. C., Glenn, J. S., Skiniotis, G. n., Kobilka, B. K. 2020

    Abstract

    Arrestin proteins bind to active, phosphorylated G-protein-coupled receptors (GPCRs), thereby preventing G-protein coupling, triggering receptor internalization, and affecting various downstream signalling pathways1,2. Although there is a wealth of structural information delineating the interactions between GPCRs and G proteins, less is known about how arrestins engage GPCRs. Here we report a cryo-EM structure of full-length human neurotensin receptor 1 (NTSR1) in complex with truncated human β-arrestin 1 (βarr1ΔCT). We found that phosphorylation of NTSR1 was critical for obtaining a stable complex with βarr1ΔCT, and identified phosphorylated sites in both the third intracellular loop and the C terminus that may promote this interaction. In addition, we observed a phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2) molecule forming a bridge between the membrane side of NTSR1 transmembrane segments 1 and 4 and the C-lobe of arrestin. Compared to a structure of rhodopsin-arrestin-1, our structure displays an approximately 85° rotation of arrestin relative to the receptor. These findings highlight both conserved aspects but also the plasticity of arrestin-receptor interactions.

    View details for DOI 10.1038/s41586-020-1953-1

    View details for PubMedID 31945771

  • CHARACTERIZATION OF HDV, HBsAg AND ALT KINETICS UNDER PEGINTERFERON-LAMBDA MONOTHERAPY: THE PHASE 2 LIMT STUDY Etzion, O., Duehren, S., Hamid, S. S., Lurie, Y., Gane, E. J., Yardeni, D., Nevo-Shor, A., Channa, S., Parkash, O., Uprichard, S. L., Gish, R. G., Cotler, S. J., Glenn, J., Apelian, D., Dahari, H., Mawani, M. WILEY. 2019: 1496A–1497A
  • A PHASE 2 STUDY OF LONAFARNIB, RITONAVIR AND PEGINTERFERON LAMBDA FOR 24 WEEKS: INTERIM END-OF-TREATMENT RESULTS FROM THE LIFT HDV STUDY. Koh, C., Da, B. L., Surana, P., Huang, A., Kapuria, D., Rotman, Y., Vittal, A., Gilman, C., Ben-Yakov, G., Lai, C., Kleiner, D., Rahman, F., Hercun, J., Dahari, H., Glenn, J., Heller, T. WILEY. 2019: 1483A
  • 2 ' 3 '-cGAMP is an immunotransmitter produced by cancer cells and regulated by ENPP1 Carozza, J., Bohnert, V., Shaw, K., Khanh Nyugen, Skariah, G., Brown, J., Rafat, M., von Eyben, R., Graves, E., Glenn, J., Smith, M., Li, L. AMER CHEMICAL SOC. 2019
  • HBV/HDV Coinfection: A Challenge for Therapeutics. Clinics in liver disease Koh, C., Da, B. L., Glenn, J. S. 2019; 23 (3): 557–72

    Abstract

    Chronic hepatitis D (CHD) results from an infection with the hepatitis B virus and hepatitis D virus (HDV). CHD is the most severe form of human viral hepatitis. Current treatment options consist of interferon alfa, which is effective only in a minority of patients. Study of HDV molecular virology has resulted in new approaches entering clinical trials, with phase-3 studies the most advanced. These include the entry inhibitor bulevirtide, the nucleic acid polymer REP 2139-Ca, the farnesyltransferase inhibitor lonafarnib, and pegylated interferon lambda. This article summarizes the available data on these emerging therapeutics.

    View details for DOI 10.1016/j.cld.2019.04.005

    View details for PubMedID 31266627

  • Hepatitis D virus infection, cirrhosis and hepatocellular carcinoma in The Gambia JOURNAL OF VIRAL HEPATITIS Mahale, P., Aka, P., Chen, X., Pfeiffer, R. M., Liu, P., Groover, S., Mendy, M., Njie, R., Goedert, J. J., Kirk, G. D., Glenn, J. S., O'Brien, T. R. 2019; 26 (6): 738–49

    View details for DOI 10.1111/jvh.13065

    View details for Web of Science ID 000469027000013

  • Treating chronic hepatitis delta: The need for surrogate markers of treatment efficacy JOURNAL OF HEPATOLOGY Yurdaydin, C., Abbas, Z., Buti, M., Cornberg, M., Esteban, R., Etzion, O., Ganes, E. J., Gish, R. G., Glenn, J. S., Hamids, S., Heller, T., Koh, C., Lampertico, P., Lurie, Y., Manns, M., Parana, R., Rizzetto, M., Urban, S., Wedemeyer, H., Wranke, A., Borzacov, P. M., Lobato, C., Hamid, S., Ceausu, E., Dalekos, G. N., Turcanu, A., Niro, G. A., Lubna, F., Abbas, M., Ingiliz, P., Ferenci, P., Vanwolleghem, T., Hayden, T., Dashdorj, N., Motoc, A., Hardtke, S., Hepatitis Delta Int Network HDIN 2019; 70 (5): 1008–15
  • Treating chronic hepatitis delta: The need for surrogate markers of treatment efficacy. Journal of hepatology Yurdaydin, C., Abbas, Z., Buti, M., Cornberg, M., Esteban, R., Etzion, O., Gane, E. J., Gish, R. G., Glenn, J. S., Hamid, S., Heller, T., Koh, C., Lampertico, P., Lurie, Y., Manns, M., Parana, R., Rizzetto, M., Urban, S., Wedemeyer, H., Hepatitis Delta International Network (HDIN), Wranke, A., Pinheiro Borzacov, L. M., Lobato, C., Hamid, S., Ceausu, E., Dalekos, G. N., Turcanu, A., Niro, G. A., Lubna, F., Abbas, M., Ingiliz, P., Ferenci, P., Vanwolleghem, T., Hayden, T., Dashdorj, N., Motoc, A., Hardtke, S. 2019; 70 (5): 1008–15

    Abstract

    Chronic hepatitis delta represents the most severe form of chronic viral hepatitis. The current treatment of hepatitis delta virus (HDV) infection consists of the use of interferons and is largely unsatisfactory. Several new compounds are currently in development for the treatment of HDV infection. However, surrogate markers that can be used to develop clinical endpoints in HDV infection are not well defined. In the current manuscript, we aimed to evaluate the existing data on treatment of HDV infection and to suggest treatment goals (possible "trial endpoints") that could be used across different clinical trials.

    View details for PubMedID 30982526

  • End of study results from LIMT HDV study: 36% durable virologic response at 24 weeks post-treatment with pegylated interferon lambda monotherapy in patients with chronic hepatitis delta virus infection Etzion, O., Hamid, S., Lurie, Y., Gane, E., Bader, N., Yardeni, D., Nevo-Shor, A., Channa, S. M., Mawani, M., Parkash, O., Yang, K., Longo, D., Gish, R. G., Glenn, J., Apelian, D. ELSEVIER SCIENCE BV. 2019: E32
  • The balance of type 1 and type 2 immune responses in the contexts of hepatitis B infection and hepatitis D infection JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY Townsend, E. C., Zhang, G. Y., Ali, R., Firke, M., Moon, M., Han, M., Fram, B., Glenn, J. S., Kleiner, D. E., Koh, C., Heller, T. 2019; 34 (4): 764–75

    View details for DOI 10.1111/jgh.14617

    View details for Web of Science ID 000462611300026

  • Early Multiphasic HBV Infection Initiation Kinetics Is Not Clone-Specific and Is Not Affected by Hepatitis D Virus (HDV) Infection VIRUSES-BASEL Tsuge, M., Uchida, T., Walsh, K., Ishida, Y., Tateno, C., Kumar, U., Glenn, J. S., Koh, C., Heller, T., Uprichard, S. L., Dahari, H., Chayama, K. 2019; 11 (3)

    View details for DOI 10.3390/v11030263

    View details for Web of Science ID 000464393800001

  • Early Multiphasic HBV Infection Initiation Kinetics Is Not Clone-Specific and Is Not Affected by Hepatitis D Virus (HDV) Infection. Viruses Tsuge, M., Uchida, T., Walsh, K., Ishida, Y., Tateno, C., Kumar, U., Glenn, J. S., Koh, C., Heller, T., Uprichard, S. L., Dahari, H., Chayama, K. 2019; 11 (3)

    Abstract

    Backgrounds and Aims: We previously demonstrated that serum hepatitis B virus (HBV) DNA in HBV infected humanized mice exhibited a highly dynamic multiphasic kinetic pattern from infection initiation to steady-state. Here, we investigated whether this pattern is consistent across different HBV clones or in the presence of hepatitis D virus (HDV) co-infection. Methods: We analyzed early serum viral kinetics using 26 HBV genotype C (GtC) mono-infected mice [clones: PXB, Hiroshima GtC CL4 (CL4) and Hiroshima GtC CL5 (CL5)] and four HBV CL4/HDV genotype one co-infected mice. Results: The HBV kinetics observed with clones CL4 and CL5 were similar to that previously defined in HBV PXB infected mice. Additionally, no significant differences in HBV DNA levels were observed between HBV mono-infected and HBV/HDV co-infected mice through 4 weeks post-inoculation (p.i.). However, HBV DNA levels at 6 weeks p.i. in HBV/HDV co-infected mice were significantly lower than those in HBV mono-infected mice (P = 0.002), consistent with HDV suppression of chronic HBV. Conclusions: HBV infection initiation is multiphasic across multiple viral clones and is not altered by HDV co-infection. The latter suggests that higher HDV titers (>8 log IU/mL) and/or longer duration of HDV infection might be needed to trigger HDV-induced suppression on HBV.

    View details for PubMedID 30875937

  • Changing Trends in Etiology-Based and Ethnicity-Based Annual Mortality Rates of Cirrhosis and Hepatocellular Carcinoma in the United States HEPATOLOGY Kim, D., Li, A. A., Perumpail, B. J., Gadiparthi, C., Kim, W., Cholankeril, G., Glenn, J. S., Harrison, S. A., Younossi, Z. M., Ahmed, A. 2019; 69 (3): 1064–74

    View details for DOI 10.1002/hep.30161

    View details for Web of Science ID 000459816500013

  • A functional subset of CD8+ T cells during chronic exhaustion is defined by SIRPalpha expression. Nature communications Myers, L. M., Tal, M. C., Torrez Dulgeroff, L. B., Carmody, A. B., Messer, R. J., Gulati, G., Yiu, Y. Y., Staron, M. M., Angel, C. L., Sinha, R., Markovic, M., Pham, E. A., Fram, B., Ahmed, A., Newman, A. M., Glenn, J. S., Davis, M. M., Kaech, S. M., Weissman, I. L., Hasenkrug, K. J. 2019; 10 (1): 794

    Abstract

    Prolonged exposure of CD8+ T cells to antigenic stimulation, as in chronic viral infections, leads to a state of diminished function termed exhaustion. We now demonstrate that even during exhaustion there is a subset of functional CD8+ T cells defined by surface expression of SIRPalpha, a protein not previously reported on lymphocytes. On SIRPalpha+ CD8+ T cells, expression of co-inhibitory receptors is counterbalanced by expression of co-stimulatory receptors and it is only SIRPalpha+ cells that actively proliferate, transcribe IFNgamma and show cytolytic activity. Furthermore, target cells that express the ligand for SIRPalpha, CD47, are more susceptible to CD8+ T cell-killing in vivo. SIRPalpha+ CD8+ T cells are evident in mice infected with Friend retrovirus, LCMV Clone 13, and in patients with chronic HCV infections. Furthermore, therapeutic blockade of PD-L1 to reinvigorate CD8+ T cells during chronic infection expands the cytotoxic subset of SIRPalpha+ CD8+ T cells.

    View details for PubMedID 30770827

  • A functional subset of CD8(+) T cells during chronic exhaustion is defined by SIRP alpha expression NATURE COMMUNICATIONS Myers, L. M., Tal, M., Dulgeroff, L., Carmody, A. B., Messer, R. J., Gulati, G., Yiu, Y., Staron, M. M., Angel, C., Sinha, R., Markovic, M., Pham, E. A., Fram, B., Ahmed, A., Newman, A. M., Glenn, J. S., Davis, M. M., Kaech, S. M., Weissman, I. L., Hasenkrug, K. J. 2019; 10
  • The balance of type 1 and type 2 immune responses in the contexts of hepatitis B infection and hepatitis D infection. Journal of gastroenterology and hepatology Townsend, E. C., Zhang, G. Y., Ali, R., Firke, M., Moon, M. S., Han, M. A., Fram, B., Glenn, J. S., Kleiner, D. E., Koh, C., Heller, T. 2019

    Abstract

    BACKGROUND: Hepatitis delta virus (HDV) infection is the most rapidly progressive chronic viral hepatitis. Little is understood about the immune responses to HDV.AIM: To characterize the systemic immune environments of hepatitis B virus (HBV) and HDV patients at various disease stages.METHODS: Evaluated 129 subjects; 53 HBV, 43 HDV, 33 healthy controls. HBV and HDV subjects were categorized by aspartate aminotransferase to platelet ratio index (APRI) into mild (APRI < 0.5), moderate, and severe (APRI > 1.0). Serum cytokines and immune markers were assessed at a single treatment-naive time-point.RESULTS: Type 1 cytokines are elevated in both HBV and HDV. Both groups show higher TNFa, IL-12p40, and CXCL9 when compared to controls (all p< 0.05). However, only HBV group displayed elevated IFNg compared to controls. Type 2 cytokines are elevated in HBV. HBV group shows higher IL-4, IL-13, and CCL26 compared to healthy controls and HDV. Chemokine CCL2 and CCL13 are lower in HDV. When assessing ratios, HDV displays higher IFNg/IL-4, TNFa/IL-4, and TNFa/IL-13 ratios than HBV and controls.CONCLUSION: HBV and HDV subjects show similarly elevated type 1 cytokines. HDV subjects display relatively lower type 2 cytokines. These differences in the systemic immune environments, particularly the predominance of type 1 responses, may contribute to the comparatively rapid progression of HDV disease.TRANSLATIONAL IMPACT: Characterization of the imbalance in type 1 and type 2 immunity unique HDV has the potential to provide immunological insights for designing therapeutic targets in HDV associated disease progression.

    View details for PubMedID 30695096

  • Hepatitis D virus infection, cirrhosis, and hepatocellular carcinoma in The Gambia. Journal of viral hepatitis Mahale, P., Aka, P., Chen, X., Pfeiffer, R. M., Liu, P., Groover, S., Mendy, M., Njie, R., Goedert, J. J., Kirk, G. D., Glenn, J. S., O'Brien, T. R. 2019

    Abstract

    Hepatocellular carcinoma (HCC) incidence is high in The Gambia and hepatitis B virus (HBV) infection is the main cause. People co-infected with HBV and hepatitis D virus (HDV) have an even greater risk of HCC and cirrhosis. Using a new HDV quantitative microarray antibody capture (Q-MAC) assay, we evaluated the association between HDV infection and HCC or cirrhosis among participants in The Gambia Liver Cancer Study. In this case-control study, cases had HCC (n=312) or cirrhosis (n=119). Controls (n=470) had no clinical evidence of liver disease and normal serum alfa-fetoprotein. Participants were previously tested for hepatitis B surface antigen (HBsAg); we tested HBsAg+ specimens by HDV Q-MAC, western blot, and RNA assays. We evaluated separate cutoffs of the Q-MAC assay for predicting anti-HDV and RNA positivity. Q-MAC correctly identified 29/29 subjects who were western blot positive (sensitivity=100%, specificity=99.4%) and 16/17 who were RNA positive (sensitivity=94.1%, specificity=100%). Compared to controls, cases more often had HBV monoinfection (HBsAg+/HDV RNA-; 54.1% vs. 17.0%; odds ratio [OR]= 6.28; p<0.001) or HBV-HDV coinfection (HBsAg+/HDV RNA+; 3.9% vs 0%; OR=; p<0.001). Risk estimates (for HCC or cirrhosis) based on HDV antibody status and adjusted for covariates (demographics, alcohol, smoking, body mass index, anti-HCV, and aflatoxin B1 exposure) yielded consistent results for both HBV monoinfection (adjusted OR=8.29; 95% confidence interval=5.74-11.98) and HBV-HDV coinfection (adjusted OR=30.66; 95% confidence interval=6.97-134.95). In this Gambian population, HDV Q-MAC had high sensitivity and specificity for both anti-HDV and HDV RNA. HDV infection contributed to the high risk of HCC in The Gambia. This article is protected by copyright. All rights reserved.

    View details for PubMedID 30661282

  • Trends in Mortality From Extrahepatic Complications in Patients With Chronic Liver Disease, From 2007 Through 2017. Gastroenterology Kim, D. n., Adejumo, A. C., Yoo, E. R., Iqbal, U. n., Li, A. A., Pham, E. A., Cholankeril, G. n., Glenn, J. S., Ahmed, A. n. 2019

    Abstract

    Trends of mortality associated with extrahepatic complications of chronic liver disease might be changing. We studied trends in mortality from extrahepatic complications of viral hepatitis, alcoholic liver disease (ALD), and nonalcoholic fatty liver disease in the United States (US).We performed a population-based study using US Census and the National Center for Health Statistics mortality records, from 2007 through 2017. We identified trends in age-standardized mortality using joinpoint trend analysis with estimates of annual percentage change.The liver-related mortality among patients with hepatitis C virus (HCV) infection increased from 2007 through 2013 and then decreased once patients began receiving treatment with direct-acting antiviral (DAA) agents, from 2014 through 2017. Among patients with HCV infection, the age-standardized mortality for extrahepatic cancers was 2.6%, for cardiovascular disease was 1.9%, and for diabetes was 3.3%. Among individuals with hepatitis B virus infection, liver-related mortality decreased steadily from 2007 through 2017. During the study age-standardized mortality from hepatitis B virus-related extrahepatic complications increased with an average annual percentage of 2.0%. Although liver-related mortality from ALD continued to increase, mortality from extrahepatic complications of ALD did not change significantly during the 11-year study. Among patients with nonalcoholic fatty liver disease, the cause of death was most frequently cardiovascular disease, which increased gradually over the study period, whereas liver-related mortality increased rapidly.In an analysis of US Census and the National Center for Health Statistics mortality records, we found that after widespread use of DAA agents for treatment of viral hepatitis, cause-specific mortality from extrahepatic cancers increased, whereas mortality from cardiovascular disease or diabetes increased only among patients with HCV infection. These findings indicate the need to reassess risk and risk factors for extrahepatic cancer, cardiovascular disease, and diabetes in individuals successfully treated for HCV infection with DAA agents.

    View details for DOI 10.1053/j.gastro.2019.06.026

    View details for PubMedID 31251928

  • Fuzhenghuayu Decoction ameliorates hepatic fibrosis by attenuating experimental sinusoidal capillarization and liver angiogenesis. Scientific reports Liu, H. L., Lv, J. n., Zhao, Z. M., Xiong, A. M., Tan, Y. n., Glenn, J. S., Tao, Y. Y., Weng, H. L., Liu, C. H. 2019; 9 (1): 18719

    Abstract

    Fuzhenghuayu (FZHY) is a compound extracted from natural plants. Its anti-fibrotic effect has been confirmed in experimental and clinical studies. However, precise effects and underlying mechanisms of FZHY in liver angiogenesis largely remain understood. In this study, we investigated the effects of FZHY on sinusoidal capillarization and angiogenesis with mice challenged for Carbon tetrachloride (CCl4) and dimethylnitrosamine (DMN), in vitro human hepatic sinusoidal endothelial cells (HHSEC) and Human Umbilical Vein Endothelial Cell (HUVEC) 3D fibrin gel model. Besides its anti-fibrotic effect, FZHY ameliorated CCl4 and DMN-induced sinusoidal capillarization, angiogenesis and expression of angiogenesis-associated factors, i.e. CD31, VEGF, VEGF receptor II, phosphor-ERK and HIF-1α. Consistent with the findings based on animal models, inhibitory effects of FZHY on capillarization and angiogenesis were further confirmed in HHSEC and the HUVEC 3D fibrin gel model, respectively. These data suggest that FZHY ameliorates not only liver fibrosis but also vessel remodeling in experimental models. Therefore, FZHY might be a potentially useful drug to treat liver cirrhosis in clinical practice.

    View details for DOI 10.1038/s41598-019-54663-4

    View details for PubMedID 31822697

  • Pathogenesis of and New Therapies for Hepatitis D GASTROENTEROLOGY Koh, C., Heller, T., Glenn, J. S. 2019; 156 (2): 461-+
  • Human iPS derived progenitors bioengineered into liver organoids using an inverted colloidal crystal poly (ethylene glycol) scaffold BIOMATERIALS Ng, S., Saeb-Parsy, K., Blackford, S. I., Segal, J. M., Serra, M., Horcas-Lopez, M., No, D., Mastoridis, S., Jassem, W., Frank, C. W., Cho, N., Nakauchi, H., Glenn, J. S., Rashid, S. 2018; 182: 299–311
  • Pathogenesis of and New Therapies for Hepatitis D. Gastroenterology Koh, C., Heller, T., Glenn, J. S. 2018

    Abstract

    Hepatitis delta virus (HDV) infection of humans was first reported in 1977, and now it is now estimated that 15-20 million people are infected worldwide. Infection with HDV can be an acute or chronic process that occurs only in patients with an HBV infection. Chronic HDV infection commonly results in the most rapidly progressive form of viral hepatitis; it is the chronic viral infection that is most likely to lead to cirrhosis, and it is associated with an increased risk of hepatocellular carcinoma. HDV infection is the only chronic human hepatitis virus infection without a therapy approved by the Food and Drug Administration. Peginterferon alpha is the only recommended therapy, but it produces unsatisfactory results. We review therapeutic agents in development, designed to disrupt the HDV life cycle, that might benefit patients with this devastating disease.

    View details for PubMedID 30342879

  • A new dual-targeting real-time RT-PCR assay for hepatitis D virus RNA detection DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE Wang, Y., Glenn, J. S., Winters, M. A., Shen, L., Choong, I., Shi, Y., Bi, S., Ma, L., Zeng, H., Zhang, F. 2018; 92 (2): 112–17

    Abstract

    In this study, a real-time reverse transcription-polymerase chain reaction (real time RT-PCR) assay targeting 2 genetic segments was established to detect HDV RNA. Utilizing the World Health Organization International Standard for Hepatitis D Virus RNA, the lower limit of detection was 575 IU/mL, and the linearity of quantification ranged from 575,000 IU/mL to 575 IU/mL. 384 HBsAg-positive samples collected from China were tested by this method and HDV antibody detection. Eleven samples were positive for anti-HDV IgG which may persist after HDV resolution, 6 samples were HDV RNA positive, and 5 samples were positive for anti-HDV IgM. This assay showed more sensitivity than the detection of anti-HDV IgM. These data demonstrate that the real-time RT-PCR assay for HDV RNA could be implemented in the clinical detection of HDV infection in chronic HBV-infected patients in China.

    View details for PubMedID 29941366

  • Changing Trends in Etiology-Based Annual Mortality From Chronic Liver Disease, From 2007 Through 2016 GASTROENTEROLOGY Kim, D., Li, A. A., Gadiparthi, C., Khan, M., Cholankeril, G., Glenn, J. S., Ahmed, A. 2018; 155 (4): 1154-+
  • A Challenging Case of Triple Hepatitis Virus Infection: An Accurate Diagnostic Test for Hepatitis D Virus Is Urgently Needed Choudhry, S. A., Reyes, K., Ishtiaq, R., Aslam, A., Glenn, J. S., Lau, D. Y. NATURE PUBLISHING GROUP. 2018: S1245
  • Impact of Drug Overdose Deaths on Solid Organ Transplantation in the United States JOURNAL OF GENERAL INTERNAL MEDICINE Cholankeril, G., Li, A. A., Cholankeril, R., Toll, A. E., Glenn, J. S., Ahmed, A. 2018; 33 (9): 1423–25
  • Human iPS derived progenitors bioengineered into liver organoids using an inverted colloidal crystal poly (ethylene glycol) scaffold. Biomaterials Ng, S. S., Saeb-Parsy, K., Blackford, S. J., Segal, J. M., Serra, M. P., Horcas-Lopez, M., No, D. Y., Mastoridis, S., Jassem, W., Frank, C. W., Cho, N. J., Nakauchi, H., Glenn, J. S., Rashid, S. T. 2018; 182: 299–311

    Abstract

    Generation of human organoids from induced pluripotent stem cells (iPSCs) offers exciting possibilities for developmental biology, disease modelling and cell therapy. Significant advances towards those goals have been hampered by dependence on animal derived matrices (e.g. Matrigel), immortalized cell lines and resultant structures that are difficult to control or scale. To address these challenges, we aimed to develop a fully defined liver organoid platform using inverted colloid crystal (ICC) whose 3-dimensional mechanical properties could be engineered to recapitulate the extracellular niche sensed by hepatic progenitors during human development. iPSC derived hepatic progenitors (IH) formed organoids most optimally in ICC scaffolds constructed with 140 mum diameter pores coated with type I collagen in a two-step process mimicking liver bud formation. The resultant organoids were closer to adult tissue, compared to 2D and 3D controls, with respect to morphology, gene expression, protein secretion, drug metabolism and viral infection and could integrate, vascularise and function following implantation into livers of immune-deficient mice. Preliminary interrogation of the underpinning mechanisms highlighted the importance of TGFbeta and hedgehog signalling pathways. The combination of functional relevance with tuneable mechanical properties leads us to propose this bioengineered platform to be ideally suited for a range of future mechanistic and clinical organoid related applications.

    View details for PubMedID 30149262

  • Changing Trends in Etiology- and Ethnicity-Based Annual Mortality Rates of Cirrhosis and Hepatocellular Carcinoma in the United States. Hepatology (Baltimore, Md.) Kim, D., Li, A. A., Perumpail, B. J., Gadiparthi, C., Kim, W., Cholankeril, G., Glenn, J. S., Harrison, S. A., Younossi, Z. M., Ahmed, A. 2018

    Abstract

    With recent improvements in the treatment of end-stage liver disease (ESLD), a better understanding of the burden of cirrhosis and hepatocellular carcinoma (HCC) is needed in the United States (US). A population-based study using the US Census and national mortality database was performed. We identified the age-standardized etiology-specific mortality rates for cirrhosis and HCC among US adults aged ≥ 20 years from 2007 to 2016. We determined temporal mortality rate patterns by joinpoint analysis with estimates of annual percentage change (APC). Age-standardized cirrhosis-related mortality rates increased from 19.77/100,000 persons in 2007 to 23.67 in 2016 with an annual increase of 2.3% (95% CI 2.0-2.7). The APC in mortality rates for hepatitis C virus (HCV)-cirrhosis shifted from a 2.9% increase per year during 2007-2014 to a 6.5% decline per year during 2014-2016. Meanwhile, mortality for cirrhosis from alcoholic liver disease (ALD, APC 4.5%) and nonalcoholic fatty liver disease (NAFLD, APC 15.4%) increased over the same period, while mortality for hepatitis B virus (HBV)-cirrhosis decreased with an average APC of -1.1%. HCC-related mortality increased from 3.48/100,000 persons in 2007 to 4.41 in 2016 at an annual rate of 2.0% (95% CI 1.3-2.6). Etiology-specific mortality rates of HCC were largely consistent with cirrhosis-related mortality. Minority populations had a higher burden of HCC-related mortality.CONCLUSION: Cirrhosis- and HCC-related mortality rates increased between 2007 and 2016 in the US. However, mortality rates in HCV-cirrhosis demonstrated a significant decline from 2014-2016, during the direct-acting antiviral era. Mortality rates for ALD/NAFLD-cirrhosis and HCC have continued to increase, while HBV-cirrhosis-related mortality declined during the 10-year period. Importantly, minorities had a disproportionately higher burden of ESLD-related mortality. This article is protected by copyright. All rights reserved.

    View details for PubMedID 30014489

  • Changing Trends in Etiology-based Annual Mortality From Chronic Liver Disease, From 2007 Through 2016. Gastroenterology Kim, D., Li, A. A., Gadiparthi, C., Khan, M. A., Cholankeril, G., Glenn, J. S., Ahmed, A. 2018

    Abstract

    BACKGROUND & AIMS: Although treatment of hepatitis C virus (HCV) infection has improved, the prevalence of alcoholic liver disease (ALD) has been increasing, so we need an updated estimate of the burden and etiology-specific mortality of chronic liver diseases. We studied the trends in age-standardized mortality of chronic liver diseases among adults 20 years or older in the United States (US), from 2007 through 2016.METHODS: We collected data from the US Census and National Center for Health Statistics mortality records, identifying individuals with HCV infection, ALD, nonalcoholic fatty liver disease (NAFLD), or hepatitis B virus (HBV) infection using ICD-10 codes. We obtained temporal mortality rate patterns using joinpoint trend analysis with estimates of annual percentage change (APC).RESULTS: Age-standardized HCV-related mortality increased from 7.17/100,000 persons in 2007 to 8.14/100,000 persons in 2013, followed by a marked decrease in the time period at which patients began receiving treatment with direct-acting antiviral agents (from 8.09/100,000 persons in 2014 to 7.15/100,000 persons in 2016). The APC in HCV mortality increased 2.0%/year from 2007 through 2014, but decreased 6.4%/year from 2014 through 2016. In contrast, age-standardized mortality increased for ALD (APC of 2.3% from 2007 through 2013 and APC of 5.5% from 2013 through 2016) and NAFLD (APC of 6.1% from 2007 through 2013 and APC of 11.3% from 2013 through 2016). HBV-related mortality decreased steadily from 2007 through 2016, with an average APC of -2.1% (95% CI, -3.0 to -1.2). Etiology-based mortality in minority populations were higher. HCV-related mortality (per 100,000 persons) was highest among non-Hispanic blacks (10.28) and whites (6.92), followed by Hispanics (5.94), and lowest among non-Hispanic Asians (2.33). Non-Hispanic Asians had higher mortality for HBV infection (2.82 per 100,000 vs 1.02 for non-Hispanic blacks, and 0.47 for non-Hispanic whites).CONCLUSION: In our population-based analysis of chronic liver disease mortality in the US, the decline in HCV-related mortality coincided with the introduction of direct-acting antiviral therapies, while the mortality from ALD and NAFLD increased during the same period. Minorities in the US have disproportionately higher chronic liver disease-related mortality.

    View details for PubMedID 30009816

  • Hepatitis D Viremia Among Injection Drug Users in San Francisco JOURNAL OF INFECTIOUS DISEASES Mahale, P., Aka, P., Chen, X., Liu, P., Fram, B. J., Wang, A. S., Simenel, S., Tseng, F., Chen, S., Edlin, B. R., Glenn, J. S., O'Brien, T. R. 2018; 217 (12): 1902–6

    Abstract

    People who inject drugs (PWID) are commonly exposed to hepatitis B virus (HBV) and hepatitis D virus (HDV). We evaluated the prevalence of HDV viremia among hepatitis B surface antigen (HBsAg)-positive PWID (n = 73) using a new quantitative microarray antibody capture (Q-MAC) assay, HDV western blot, and HDV RNA. HDV Q-MAC performed well in this cohort: anti-HDV, 100% sensitivity and specificity; HDV viremia, 61.5% sensitivity and 100% specificity. Hepatitis D viremia was present in 35.6% of HBsAg-positive participants and was more common in those with resolved compared to chronic hepatitis C (5.1% vs 0.6%; adjusted odds ratio, 9.80; P < .0001).

    View details for PubMedID 29800369

    View details for PubMedCentralID PMC5972608

  • Impact of Drug Overdose Deaths on Solid Organ Transplantation in the United States. Journal of general internal medicine Cholankeril, G., Li, A. A., Cholankeril, R., Toll, A. E., Glenn, J. S., Ahmed, A. 2018

    View details for PubMedID 29766381

  • Optimizing lonafarnib treatment for the management of chronic delta hepatitis: The LOWR HDV-1 study HEPATOLOGY Yurdaydin, C., Keskin, O., Kalkan, C., Karakaya, F., Caliskan, A., Karatayli, E., Karatayli, S., Bozdayi, A., Koh, C., Heller, T., Idilman, R., Glenn, J. S. 2018; 67 (4): 1224–36

    Abstract

    In a proof-of-concept (POC) study, the oral prenylation inhibitor, lonafarnib (LNF), decreased hepatitis D virus (HDV) RNA during 4 weeks of treatment. Here, we explored optimal LNF regimens. Fifteen patients (five groups; 3 per group) completed dosing as follows: (1) LNF 200 mg twice-daily (BID; 12 weeks); (2) LNF 300 mg BID (12 weeks); (3) LNF 100 mg thrice-daily (5 weeks); (4) LNF 100 mg BID + pegylated interferon alfa (PEG-IFNα) 180 μg once-weekly (QW; 8 weeks); and (5) LNF 100 mg BID + ritonavir (RTV) 100 mg once-daily (QD; 8 weeks). Tolerability and efficacy were assessed. Higher LNF monotherapy doses had greater decreases in HDV viral load than achieved in the original POC study. However, this was associated with increased gastrointestinal adverse events. Addition of RTV 100 mg QD to a LNF 100 mg BID regimen yielded better antiviral responses than LNF 300 mg BID monotherapy and with less side effects. A similar improvement was observed with LNF 100 mg BID + PEG-IFNα 180 μg QW. Two of 6 patients who received 12 weeks of LNF experienced transient posttreatment alanine aminotransferase (ALT) increases resulting in HDV-RNA negativity and ALT normalization.The cytochrome P450 3A4 inhibitor, RTV, allows a lower LNF dose to be used while achieving higher levels of postabsorption LNF, yielding better antiviral responses and tolerability. In addition, combining LNF with PEG-IFNα achieved more substantial and rapid HDV-RNA reduction, compared to historical responses with PEG-IFNα alone. Twelve weeks of LNF can result in posttreatment HDV-RNA negativity in some patients, which we speculate results from restoring favorable immune responses. These results support further development of LNF with RTV boosting and exploration of the combination of LNF with PEG-IFN. (Hepatology 2018;67:1224-1236).

    View details for DOI 10.1002/hep.29658

    View details for Web of Science ID 000428332600010

    View details for PubMedID 29152762

  • A research agenda for curing chronic hepatitis B virus infection HEPATOLOGY Alter, H., Block, T., Brown, N., Brownstein, A., Brosgart, C., Chang, K., Chen, P., Chisari, F. V., Cohen, C., El-Serag, H., Feld, J., Gish, R., Glenn, J., Greten, T., Guo, H., Guo, J., Hoshida, Y., Hu, J., Kowdley, K. V., Li, W., Liang, J., Locarnini, S., Lok, A. S., Mason, W., McMahon, B., Mehta, A., Perrillo, R., Revill, P., Rice, C. M., Rinaudo, J., Schinazi, R., Seeger, C., Shetty, K., Tavis, J., Zoulim, F. 2018; 67 (3): 1127–31

    View details for PubMedID 28877549

  • Research priorities for the discovery of a cure for chronic hepatitis B: Report of a workshop ANTIVIRAL RESEARCH Block, T. M., Alter, H., Brown, N., Brownstein, A., Brosgart, C., Chang, K., Chen, P., Cohen, C., El-Serag, H., Feld, J., Gish, R., Glenn, J., Greten, T. F., Guo, J., Hoshida, Y., Kowdley, K. V., Li, W., Lok, A. S., McMahon, B., Mehta, A., Perrillo, R., Rice, C. M., Rinaudo, J., Schinazi, R. F., Shetty, K. 2018; 150: 93–100

    Abstract

    In early 2017, the Hepatitis B Foundation invited 30 experts in the fields of hepatitis B and liver cancer research to identify projects they deemed important to the goal of finding a cure for chronic hepatitis B and D and the diseases with which these viral infections are associated. They were also asked to identify general categories of research and to prioritize sub-project topics within those areas. The experts generally agreed on broadly defined areas of research, but there was usually little difference between the highest and lowest scoring projects; for the most part, all programs described in this document were considered valuable and necessary. An executive summary of this discussion was recently published (Alter et al., Hepatology 2017). The present manuscript reports the areas of research identified by the workshop participants, provides a brief rationale for their selection, and attempts to express differences among the priorities assigned to each area of research, when such distinctions were expressed.

    View details for PubMedID 29248746

  • Letter to the Editor regarding article "Emerging concepts for the treatment of hepatitis delta" [Menashe Elazar and Jeffrey S Glenn, Curr Opin Virol 24 (2017) 55-59] Reply CURRENT OPINION IN VIROLOGY Elazar, M., Glenn, J. 2018; 28: 169

    View details for PubMedID 29456115

  • Emerging Therapeutic Targets for Hepatitis Delta Virus Infection. Gastroenterology & hepatology Glenn, J. S. 2018; 14 (1): 47–49

    View details for PubMedID 29491762

  • Structure of an engineered IFN-lambda/IFN-lambda R1/IL-10R beta complex provides insight into the functional dichotomy of type III versus type I IFNs Mendoza, J., Schneider, W. M., Hoffman, H., Vercauteren, K., Jude, K. M., Xiong, A., Moraga, I., Horton, T. M., Glenn, J. S., de Jonga, Y. P., Garcia, K. ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD. 2017: 46
  • Hepatitis B Virus Reactivation Associated With Direct-Acting Antiviral Therapy for Chronic Hepatitis C Virus ANNALS OF INTERNAL MEDICINE Mahale, P., Glenn, J. S., O'Brien, T. R. 2017; 167 (10): 759–60

    View details for PubMedID 29159388

  • Prevalence of hepatitis D viremia among injection drug users in San Francisco Mahale, P., Aka, P. V., Chen, X., Liu, P., Wang, A. S., Tseng, F., Chen, S., Edlin, B. R., Glenn, J. S., O'Brien, T. WILEY. 2017: 100A
  • Modeling hepatitis delta virus dynamics during ritonavir boosted lonafarnib treatment-the LOWR HDV-3 study Dubey, P., Koh, C., Surana, P., Uprichard, S. L., Han, M., Fryzek, N., Kapuria, D., Etzion, O., Takyar, V. K., Rotman, Y., Yurdaydin, C., Glenn, J. S., Cotler, S., Heller, T., Dahari, H. WILEY. 2017: 21A
  • Quantitative Evaluation of Viral Protein Binding to Phosphoinositide Receptors and Pharmacological Inhibition ANALYTICAL CHEMISTRY Kim, S., Jackman, J. A., Elazar, M., Cho, S., Glenn, J. S., Cho, N. 2017; 89 (18): 9742–50

    Abstract

    There is significant interest in developing analytical methods to characterize molecular recognition events between proteins and phosphoinositides, which are a medically important class of carbohydrate-functionalized lipids. Within this scope, one area of high priority involves quantitatively evaluating drug candidates that pharmacologically inhibit protein-phosphoinositide interactions. As full-length proteins are often difficult to produce, establishing methods to study these interactions with shorter, bioactive peptides would be advantageous. Herein, we report an atomic force microscopy (AFM)-based force spectroscopic approach to detect the specific interaction between an amphipathic, α-helical (AH) peptide derived from the hepatitis C virus NS5A protein and its biological target, the phosphatidylinositol (4,5)-bisphosphate [PI(4,5)P2] phosphoinositide receptor. After optimization of the peptide tethering strategy and measurement parameters, the binding specificity of AH peptide for PI(4,5)P2 receptors was comparatively evaluated across a panel of phosphoinositides and the influence of ionic strength on AH-PI(4,5)P2 binding strength was tested. Importantly, these capabilities were translated into the development of a novel experimental methodology to determine the inhibitory activity of a small-molecule drug candidate acting against the AH-PI(4,5)P2 interaction, and extracted kinetic parameters agree well with literature values obtained by conventional biochemical methods. Taken together, our findings provide a nanomechanical basis for explaining the high binding specificity of the NS5A AH to PI(4,5)P2 receptors, in turn establishing an analytical framework to study phosphoinositide-binding viral peptides and proteins as well as a broadly applicable approach to evaluate candidate inhibitors of protein-phosphoinositide interactions.

    View details for PubMedID 28809547

  • Long-term culture of human liver tissue with advanced hepatic functions. JCI insight Ng, S. S., Xiong, A., Nguyen, K., Masek, M., No, D. Y., Elazar, M., Shteyer, E., Winters, M. A., Voedisch, A., Shaw, K., Rashid, S. T., Frank, C. W., Cho, N. J., Glenn, J. S. 2017; 2 (11)

    Abstract

    A major challenge for studying authentic liver cell function and cell replacement therapies is that primary human hepatocytes rapidly lose their advanced function in conventional, 2-dimensional culture platforms. Here, we describe the fabrication of 3-dimensional hexagonally arrayed lobular human liver tissues inspired by the liver's natural architecture. The engineered liver tissues exhibit key features of advanced differentiation, such as human-specific cytochrome P450-mediated drug metabolism and the ability to support efficient infection with patient-derived inoculums of hepatitis C virus. The tissues permit the assessment of antiviral agents and maintain their advanced functions for over 5 months in culture. This extended functionality enabled the prediction of a fatal human-specific hepatotoxicity caused by fialuridine (FIAU), which had escaped detection by preclinical models and short-term clinical studies. The results obtained with the engineered human liver tissue in this study provide proof-of-concept determination of human-specific drug metabolism, demonstrate the ability to support infection with human hepatitis virus derived from an infected patient and subsequent antiviral drug testing against said infection, and facilitate detection of human-specific drug hepatotoxicity associated with late-onset liver failure. Looking forward, the scalability and biocompatibility of the scaffold are also ideal for future cell replacement therapeutic strategies.

    View details for DOI 10.1172/jci.insight.90853

    View details for PubMedID 28570275

  • Hepatitis delta infection - Current and new treatment options BEST PRACTICE & RESEARCH CLINICAL GASTROENTEROLOGY Elazar, M., Koh, C., Glenn, J. S. 2017; 31 (3): 321–27

    Abstract

    In humans, hepatitis D virus (HDV) infection only occurs in the presence of a concomitant hepatitis B virus (HBV) infection, and induces the most severe form of human viral hepatitis. Even though HDV is spread worldwide and is endemic in some regions, screening and treatment has been often neglected in part due to the lack of an effective therapy. Moreover, HDV prevalence rates are increasing in many countries driven by immigration from areas of high endemicity. Currently, no FDA-approved anti-HDV therapy is available, although interferon (IFN) alpha therapy has demonstrated benefit in a minority of patients. In this review, we present a current view of our understanding of the epidemiology, molecular virology and management of HDV infection. We additionally discuss new treatment approaches in development and describe the most promising results of recent and ongoing clinical trials of these new potential agents.

    View details for PubMedID 28774414

  • Pharmacokinetics and Pharmacodynamics Modeling of Lonafarnib in Patients With Chronic Hepatitis Delta Virus Infection HEPATOLOGY COMMUNICATIONS Canini, L., Koh, C., Cotler, S. J., Uprichard, S. L., Winters, M. A., Han, M., Kleiner, D. E., Idilman, R., Yurdaydin, C., Glenn, J. S., Heller, T., Dahari, H. 2017; 1 (4): 288–92

    Abstract

    The prenylation inhibitor lonafarnib (LNF) is a potent antiviral agent providing a breakthrough for the treatment of hepatitis delta virus (HDV). The current study used a maximum likelihood approach to model LNF pharmacokinetic (PK) and pharmacodynamic (PD) parameters and predict the dose needed to achieve 99% efficacy using data from 12 patients chronically infected with HDV and treated with LNF 100 mg twice daily (bid) (group 1) or 200 mg bid (group 2) for 28 days. The LNF-PK model predicted average steady-state LNF concentrations of 860 ng/mL and 1,734 ng/mL in groups 1 and 2, respectively, with an LNF absorption rate ka = 0.43/hour and elimination rate ke = 0.045/hour. The PK/PD model identified an average delay of 0.56 hours and an LNF concentration that decreases HDV production by 50%, EC50 = 227 ng/mL, with a Hill factor h = 1.48. The HDV half-life in blood was 1.87 days, and the average steady-state LNF efficacy in blocking HDV production was ɛ = 87.7% for group 1 and ɛ = 95.2% for group 2. A biphasic HDV decline with an average phase 1 decline (0.9 log10 IU/mL and 1.32 log10 IU/mL) was observed in groups 1 and 2, respectively. Phase 2 was not significantly (P = 0.94) different between the two groups, with an average slope of -0.06 log IU/mL/day. The model suggests an LNF dose of ∼610 mg bid would achieve ɛ = 99%. Conclusion: The first PK/PD modeling study in patients with chronic HDV indicates that a ∼3-fold increase in LNF dose (∼610 mg bid) would achieve 99% antiviral efficacy. A ritonavir-boosted LNF combination may provide a means to increase LNF efficacy with minimal side effects. The modeling findings provide an important advance in understanding HDV dynamics and the basis to optimize LNF therapy for hepatitis D. (Hepatology Communications 2017;1:288-292).

    View details for DOI 10.1002/hep4.1043

    View details for Web of Science ID 000453170600002

    View details for PubMedID 29404459

    View details for PubMedCentralID PMC5721397

  • Emerging concepts for the treatment of hepatitis delta. Current opinion in virology Elazar, M., Glenn, J. S. 2017; 24: 55-59

    Abstract

    Hepatitis delta virus (HDV) causes the most severe form of human viral hepatitis and is associated with a higher risk of cirrhosis, liver decompensation and liver cancer. Interferon alpha is the only agent that has demonstrated efficacy to date, although response rates are low and it is associated with significant side effects. A better understanding of the relevant molecular virology has resulted in the identification of new candidate targets. Future therapeutic options are rapidly evolving as several new agents have entered clinical development, including the entry inhibitor myrcludex-B, the nucleic acid polymer REP2139-Ca inhibiting HBV surface antigen secretion, the farnesyltransferase inhibitor lonafarnib that targets virus assembly, and a better tolerated interferon-interferon lambda.

    View details for DOI 10.1016/j.coviro.2017.04.004

    View details for PubMedID 28475945

  • The IFN-?-IFN-?R1-IL-10Rß Complex Reveals Structural Features Underlying Type III IFN Functional Plasticity. Immunity Mendoza, J. L., Schneider, W. M., Hoffmann, H., Vercauteren, K., Jude, K. M., Xiong, A., Moraga, I., Horton, T. M., Glenn, J. S., de Jong, Y. P., Rice, C. M., Garcia, K. C. 2017; 46 (3): 379-392

    Abstract

    Type III interferons (IFN-λs) signal through a heterodimeric receptor complex composed of the IFN-λR1 subunit, specific for IFN-λs, and interleukin-10Rβ (IL-10Rβ), which is shared by multiple cytokines in the IL-10 superfamily. Low affinity of IL-10Rβ for cytokines has impeded efforts aimed at crystallizing cytokine-receptor complexes. We used yeast surface display to engineer a higher-affinity IFN-λ variant, H11, which enabled crystallization of the ternary complex. The structure revealed that IL-10Rβ uses a network of tyrosine residues as hydrophobic anchor points to engage IL-10 family cytokines that present complementary hydrophobic binding patches, explaining its role as both a cross-reactive but cytokine-specific receptor. H11 elicited increased anti-proliferative and antiviral activities in vitro and in vivo. In contrast, engineered higher-affinity type I IFNs did not increase antiviral potency over wild-type type I IFNs. Our findings provide insight into cytokine recognition by the IL-10R family and highlight the plasticity of type III interferon signaling and its therapeutic potential.

    View details for DOI 10.1016/j.immuni.2017.02.017

    View details for PubMedID 28329704

  • The IFN-lambda-IFN-lambda R1-IL-10R beta Complex Reveals Structural Features Underlying Type III IFN Functional Plasticity IMMUNITY Mendoza, J. L., Schneider, W. M., Hoffmann, H., Vercauteren, K., Jude, K. M., Xiong, A., Moraga, I., Horton, T. M., Glenn, J. S., de Jong, Y. P., Rice, C. M., Garcia, K. C. 2017; 46 (3): 379-392

    Abstract

    Type III interferons (IFN-λs) signal through a heterodimeric receptor complex composed of the IFN-λR1 subunit, specific for IFN-λs, and interleukin-10Rβ (IL-10Rβ), which is shared by multiple cytokines in the IL-10 superfamily. Low affinity of IL-10Rβ for cytokines has impeded efforts aimed at crystallizing cytokine-receptor complexes. We used yeast surface display to engineer a higher-affinity IFN-λ variant, H11, which enabled crystallization of the ternary complex. The structure revealed that IL-10Rβ uses a network of tyrosine residues as hydrophobic anchor points to engage IL-10 family cytokines that present complementary hydrophobic binding patches, explaining its role as both a cross-reactive but cytokine-specific receptor. H11 elicited increased anti-proliferative and antiviral activities in vitro and in vivo. In contrast, engineered higher-affinity type I IFNs did not increase antiviral potency over wild-type type I IFNs. Our findings provide insight into cytokine recognition by the IL-10R family and highlight the plasticity of type III interferon signaling and its therapeutic potential.

    View details for DOI 10.1016/j.immuni.2017.02.017

    View details for Web of Science ID 000396818100011

  • Extracellular Matrix Functionalization and Huh-7.5 Cell Coculture Promote the Hepatic Differentiation of Human Adipose-Derived Mesenchymal Stem Cells in a 3D ICC Hydrogel Scaffold. ACS biomaterials science & engineering Wang, Y., Lee, J. H., Shirahama, H., Seo, J., Glenn, J. S., Cho, N. J. 2016; 2 (12): 2255-2265

    Abstract

    In this study, we constructed a microporous hydrogel scaffold with hexagonally packed interconnected cavities and extracellular matrix (ECM)-functionalized interior surface, and systematically investigated the hepatic differentiation of human adipose-derived mesenchymal stem cells (hAD-MSCs) under the influence of three key factors: three-dimensional (3D) geometry, ECM presence, and coculture with hepatocyte-derived cell line. Results confirmed that (i) hepatic differentiation of hAD-MSC is more efficient in a 3D microporous scaffold than in 2D monolayer culture; (ii) the presence of both ECM components (fibronectin and collagen-I) in the scaffold is superior to collagen-I only, highlighting the importance of fibronectin; and (iii) coculture with Huh-7.5 hepatocyte-derived cells promoted liver-specific functions of the hAD-MSC-derived hepatocytes. The optimized differentiation process only took 21 days to complete, a time length that is shorter or at least comparable to previous reports, and more importantly, yielded an albumin production more than 10-fold higher than conventional 2D culture. Our approach of optimizing hAD-MSC hepatic differentiation could provide a potential solution to the challenges such as hepatocyte transplantation or the establishment of human physiologically relevant liver models in vitro.

    View details for DOI 10.1021/acsbiomaterials.6b00487

    View details for PubMedID 33465898

  • Extracellular Matrix Functionalization and Huh-7.5 Cell Coculture Promote the Hepatic Differentiation of Human Adipose-Derived Mesenchymal Stem Cells in a 3D ICC Hydrogel Scaffold ACS BIOMATERIALS SCIENCE & ENGINEERING Wang, Y., Lee, J., Shirahama, H., Seo, J., Glenn, J. S., Cho, N. 2016; 2 (12): 2255-2265
  • ECM proteins in a microporous scaffold influence hepatocyte morphology, function, and gene expression SCIENTIFIC REPORTS Wang, Y., Kim, M. H., Shirahama, H., Lee, J. H., Ng, S. S., Glenn, J. S., Cho, N. 2016; 6

    Abstract

    It is well known that a three-dimensional (3D) culture environment and the presence of extracellular matrix (ECM) proteins facilitate hepatocyte viability and maintenance of the liver-specific phenotype in vitro. However, it is not clear whether specific ECM components such as collagen or fibronectin differentially regulate such processes, especially in 3D scaffolds. In this study, a series of ECM-functionalized inverted colloidal crystal (ICC) microporous scaffolds were fabricated and their influence on Huh-7.5 cell proliferation, morphology, hepatic-specific functions, and patterns of gene expression were compared. Both collagen and fibronectin promoted albumin production and liver-specific gene expression of Huh-7.5 cells, compared with the bare ICC scaffold. Interestingly, cells in the fibronectin-functionalized scaffold exhibited different aggregation patterns to those in the collagen-functionalized scaffold, a variation that could be related to the distinct mRNA expression levels of cell adhesion-related genes. Based on these results, we can conclude that different ECM proteins, such as fibronectin and collagen, indeed play distinct roles in the phenotypic regulation of cells cultured in a 3D environment.

    View details for DOI 10.1038/srep37427

    View details for PubMedID 27897167

  • A novel quantitative microarray antibody capture (Q-MAC) assay identifies an extremely high HDV prevalence amongst HBV infected Mongolians. Hepatology Chen, X., Oidovsambuu, O., Liu, P., Grosely, R., Elazar, M., Winn, V. D., Fram, B., Boa, Z., Dai, H., Dashtseren, B., Yagaanbuyant, D., Genden, Z., Dashdorj, N., Bungert, A., Dashdorj, N., Glenn, J. S. 2016

    Abstract

    Hepatitis delta virus (HDV) causes the most severe form of human viral hepatitis. HDV requires a hepatitis B virus (HBV) co-infection to provide HDV with HBV surface antigen envelope proteins. The net effect of HDV is to make the underlying HBV disease worse, including higher rates of hepatocellular carcinoma (HCC). Accurate assessments of current HDV prevalence have been hampered by the lack of readily available and reliable quantitative assays, combined with the absence of an FDA-approved therapy. We sought to develop a convenient assay for accurately screening populations and to use this assay to determine HDV prevalence in a population with abnormally high rates of HCC. We developed a high throughput quantitative microarray antibody capture (Q-MAC) assay for anti-HDV IgG wherein recombinant HDV delta antigen is printed by microarray on slides coated with a noncontinuous, nanostructured plasmonic gold film, enabling quantitative fluorescent detection of anti-HDV antibody in small aliquots of patient serum. This assay was then used to screen all HBV-infected patients identified in a large randomly selected cohort designed to represent the Mongolian population. We identified two quantitative thresholds of captured antibody that were 100% predictive of the sample either being positive on standard western blot, or harboring HDV RNA detectable by qPCR, respectively. Subsequent screening of the HBV-positive cohort revealed that a remarkable 57% were RNA positive and an additional 4% were positive on western blot alone.The Q-MAC assay's unique performance characteristics make it ideal for population screening. Its application to the Mongolian HBsAg+ population reveals an apparent ∼60% prevalence of HDV co-infection amongst these HBV-infected Mongolian subjects, which may help explain the extraordinarily high rate of HCC in Mongolia. This article is protected by copyright. All rights reserved.

    View details for DOI 10.1002/hep.28957

    View details for PubMedID 27880976

  • Exploring Optimal Dosing Of Lonafarnib With Ritonavir For The Treatment Of Chronic Delta Hepatitis-Interim Results From The Lowr Hdv-2 Study Yurdaydin, C., Idilman, R., Kalkan, C., Karakaya, F., Kartal, A., Keskin, O., Karatayli, E., Karatayli, S. C., Bozdayi, A., Koh, C., Heller, T., Glenn, J. WILEY. 2016: 910A–911A
  • The Prenylation Inhibitor Lonafarnib Can Induce Post-Treatment Alt Flares With Viral Clearance In Patients With Chronic Delta Hepatitis Yurdaydin, C., Idilman, R., Kalkan, C., Karakaya, F., Kartal, A., Keskin, O., Karatayli, E., Karatayli, S. C., Bozdayi, A., Koh, C., Heller, T., Glenn, J. WILEY. 2016: 927A
  • Characterization of HDV and HBV kinetics during acute co-infection and interferon-alpha treatment in uPA/SCID chimeric mice with humanized livers Uchida, T., Walsh, K., Hiraga, N., Imamura, M., Koh, C., Glenn, J., Uprichard, S. L., Heller, T., Dahari, H., Chayama, K. WILEY. 2016: 9A–10A
  • A Phase 2 Study Of Titrating-Dose Lonafarnib Plus Ritonavir In Patients With Chronic Hepatitis D: Interim Results From The Lonafarnib With Ritonavir In HDV-4 (LOWR HDV-4) Study Wedemeyer, H., Port, K., Deterding, K., Wranke, A., Kirschner, J., Martins, E. B., Glenn, J., Cornberg, M., Manns, M. P. WILEY. 2016: 121A–122A
  • Reconstitution and Functional Analysis of a Full-Length Hepatitis C Virus NS5B Polymerase on a Supported Lipid Bilayer. ACS central science Cho, N., Pham, E. A., Hagey, R. J., Lévêque, V. J., Ma, H., Klumpp, K., Glenn, J. S. 2016; 2 (7): 456-466

    Abstract

    Therapeutic targeting of membrane-associated viral proteins is complicated by the challenge of investigating their enzymatic activities in the native membrane-bound state. To permit functional characterization of these proteins, we hypothesized that the supported lipid bilayer (SLB) can support in situ reconstitution of membrane-associated viral protein complexes. As proof-of-principle, we selected the hepatitis C virus (HCV) NS5B polymerase which is essential for HCV genome replication, and determined that the SLB platform enables functional reconstitution of membrane protein activity. Quartz crystal microbalance with dissipation (QCM-D) monitoring enabled label-free detection of full-length NS5B membrane association, its interaction with replicase subunits NS3, NS5A, and template RNA, and most importantly its RNA synthesis activity. This latter activity could be inhibited by the addition of candidate small molecule drugs. Collectively, our results demonstrate that the SLB platform can support functional studies of membrane-associated viral proteins engaged in critical biological activities.

    View details for DOI 10.1021/acscentsci.6b00112

    View details for PubMedID 27504492

  • Multistep Compositional Remodeling of Supported Lipid Membranes by Interfacially Active Phosphatidylinositol Kinases ANALYTICAL CHEMISTRY Tabaei, S. R., Guo, F., Rutaganira, F. U., Vafaei, S., Choong, I., Shokat, K. M., Glenn, J. S., Cho, N. 2016; 88 (10): 5042-5045

    Abstract

    The multienzyme catalytic phosphorylation of phosphatidylinositol (PI) in a supported lipid membrane platform is demonstrated for the first time. One-step treatment with PI 4-kinase IIIβ (PI4Kβ) yielded PI 4-phosphate (PI4P), while a multistep enzymatic cascade of PI4Kβ followed by PIP 5-kinase produced PI-4,5-bisphosphate (PI(4,5)P2 or PIP2). By employing quartz crystal microbalance with dissipation monitoring, we were able to track membrane association of kinase enzymes for the first time as well as detect PI4P and PI(4,5)P2 generation based on subsequent antibody binding to the supported lipid bilayers. Pharmacologic inhibition of PI4Kβ by a small molecule inhibitor was also quantitatively assessed, yielding an EC50 value that agrees well with conventional biochemical readout. Taken together, the development of a PI-containing supported membrane platform coupled with surface-sensitive measurement techniques for kinase studies opens the door to exploring the rich biochemistry and pharmacological targeting of membrane-associated phosphoinositides.

    View details for DOI 10.1021/acs.analchem.6b01293

    View details for PubMedID 27118725

  • Profiling system-wide immune signaling in chronic viral infection and its response to viral clearance Angel, C., Furman, D., Pham, E., Fram, B., Thai Nguyen, Ahmed, A., Grant, P., Maecker, H. T., Glenn, J., Davis, M. M. AMER ASSOC IMMUNOLOGISTS. 2016
  • Parallel shRNA and CRISPR-Cas9 screens enable antiviral drug target identification NATURE CHEMICAL BIOLOGY Deans, R. M., Morgens, D. W., Okesli, A., Pillay, S., Horlbeck, M. A., Kampmann, M., Gilbert, L. A., Li, A., Mateo, R., Smith, M., Glenn, J. S., Carette, J. E., Khosla, C., Bassik, M. C. 2016; 12 (5): 361-?

    Abstract

    Broad-spectrum antiviral drugs targeting host processes could potentially treat a wide range of viruses while reducing the likelihood of emergent resistance. Despite great promise as therapeutics, such drugs remain largely elusive. Here we used parallel genome-wide high-coverage short hairpin RNA (shRNA) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 screens to identify the cellular target and mechanism of action of GSK983, a potent broad-spectrum antiviral with unexplained cytotoxicity. We found that GSK983 blocked cell proliferation and dengue virus replication by inhibiting the pyrimidine biosynthesis enzyme dihydroorotate dehydrogenase (DHODH). Guided by mechanistic insights from both genomic screens, we found that exogenous deoxycytidine markedly reduced GSK983 cytotoxicity but not antiviral activity, providing an attractive new approach to improve the therapeutic window of DHODH inhibitors against RNA viruses. Our results highlight the distinct advantages and limitations of each screening method for identifying drug targets, and demonstrate the utility of parallel knockdown and knockout screens for comprehensive probing of drug activity.

    View details for DOI 10.1038/NCHEMBIO.2050

    View details for PubMedID 27018887

  • Design and Structural Characterization of Potent and Selective Inhibitors of Phosphatidylinositol 4 Kinase III beta JOURNAL OF MEDICINAL CHEMISTRY Rutaganira, F. U., Fowler, M. L., McPhail, J. A., Gelman, M. A., Nguyen, K., Xiong, A., Doman, G. L., Tayshanjian, B., Glenn, J. S., Shokat, K. M., Burke, J. E. 2016; 59 (5): 1830-1839

    Abstract

    Type III phosphatidylinositol 4-kinase (PI4KIIIβ) is an essential enzyme in mediating membrane trafficking and is implicated in a variety of pathogenic processes. It is a key host factor mediating replication of RNA viruses. The design of potent and specific inhibitors of this enzyme will be essential to define its cellular roles and may lead to novel antiviral therapeutics. We previously reported the PI4K inhibitor PIK93, and this compound has defined key functions of PI4KIIIβ. However, this compound showed high cross reactivity with class I and III PI3Ks. Using structure-based drug design, we have designed novel potent and selective (>1000-fold over class I and class III PI3Ks) PI4KIIIβ inhibitors. These compounds showed antiviral activity against hepatitis C virus. The co-crystal structure of PI4KIIIβ bound to one of the most potent compounds reveals the molecular basis of specificity. This work will be vital in the design of novel PI4KIIIβ inhibitors, which may play significant roles as antiviral therapeutics.

    View details for DOI 10.1021/acs.jmedchem.5b01311

    View details for Web of Science ID 000372043400012

  • Design and Structural Characterization of Potent and Selective Inhibitors of Phosphatidylinositol 4 Kinase IIIß. Journal of medicinal chemistry Rutaganira, F. U., Fowler, M. L., McPhail, J. A., Gelman, M. A., Nguyen, K., Xiong, A., Dornan, G. L., Tavshanjian, B., Glenn, J. S., Shokat, K. M., Burke, J. E. 2016; 59 (5): 1830-1839

    Abstract

    Type III phosphatidylinositol 4-kinase (PI4KIIIβ) is an essential enzyme in mediating membrane trafficking and is implicated in a variety of pathogenic processes. It is a key host factor mediating replication of RNA viruses. The design of potent and specific inhibitors of this enzyme will be essential to define its cellular roles and may lead to novel antiviral therapeutics. We previously reported the PI4K inhibitor PIK93, and this compound has defined key functions of PI4KIIIβ. However, this compound showed high cross reactivity with class I and III PI3Ks. Using structure-based drug design, we have designed novel potent and selective (>1000-fold over class I and class III PI3Ks) PI4KIIIβ inhibitors. These compounds showed antiviral activity against hepatitis C virus. The co-crystal structure of PI4KIIIβ bound to one of the most potent compounds reveals the molecular basis of specificity. This work will be vital in the design of novel PI4KIIIβ inhibitors, which may play significant roles as antiviral therapeutics.

    View details for DOI 10.1021/acs.jmedchem.5b01311

    View details for PubMedID 26885694

  • Future Therapy for Hepatitis B Virus: Role of Immunomodulators. Current hepatology reports Pham, E. A., Perumpail, R. B., Fram, B. J., Glenn, J. S., Ahmed, A., Gish, R. G. 2016; 15 (4): 237-244

    Abstract

    Although currently available therapies for chronic hepatitis B virus infection can suppress viremia and provide long-term benefits for patients, they do not lead to a functional cure for most patients. Advances in our understanding of the virus-host interaction and the recent remarkable success of immunotherapy in cancer offer new and promising strategies for developing immune modulators that may become important components of a total therapeutic approach to hepatitis B, some of which are now in clinical development. Among the immunomodulatory agents currently being investigated to combat chronic HBV are toll-like receptor agonists, immune checkpoint inhibitors, therapeutic vaccines, and engineered T cells. The efficacy of some immune modulatory therapies is compromised by high viral antigen levels. Cutting edge strategies, including RNA interference and CRISPR/Cas9, are now being studied that may ultimately be shown to have the capacity to lower viral antigen levels sufficiently to substantially increase the efficacy of these agents. The current advances in therapies for chronic hepatitis B are leading us toward the possibility of a functional cure.

    View details for PubMedID 27917363

  • Decline in Hepatitis C Virus-related Liver Transplantation Waitlist Registrations among Patients without Hepatocellular Carcinoma: Early Effect of Direct-Acting Antivirals? Perumpail, R. B., Wong, R. J., Jayasekera, C. R., Gonzalez, S. A., Glenn, J. S., Younossi, Z. M., Ahmed, A. WILEY-BLACKWELL. 2015: 1397A