Innate Immune Factors in CF and COPD Patients; Development of Gene Therapy for Viral Infection
To better understand how common respiratory viruses infect the human lung and how cell-type tropism and innate immune factors influence the pathophysiological consequences of viral infection especially in individuals with underlying lung diseases such as cystic fibrosis (CF) and Chronic Obstructive Pulmonary Disease (COPD).The Pickles laboratory is located in the Cystic Fibrosis/Pulmonary Research and Treatment Center in Marsico Hall at UNC. We study how human respiratory virus pathogens infect the human lung with focus on how viruses interact with lung cells and how virus infection alters the normal functions of the lung. Our research has three major goals related to CF lung disease.
- To exploit the efficient mechanisms that respiratory viruses have evolved to infect respiratory epithelium for the development of strategies to deliver therapeutic transgenes to human airways. This is particularly relevant to the genetic delivery of CFTR to the airways of CF patients.
- To understand the role of ciliated airway epithelial cells in respiratory virus infection. These cells are predominant in the airways of humans and are often the target of common respiratory virus infection. We study the innate defense mechanisms provided by ciliated cells both at the level of mechanical innate defense (Mucociliary transport) and chemical innate defense (inflammatory mediators). Understanding the role of ciliated cells in these innate mechanisms is important for virus pathogenesis, gene therapy strategies and vaccine design.
Currently, we focus on three common respiratory viral pathogens: respiratory syncytial virus (RSV); the parainfluenza viruses (PIV) and influenza viruses (IFV) since these viruses are amongst the most common cause of viral infections in infants and children. These viruses are also a concern in individuals with compromised lungs, e.g., CF patients, and those who have undergone lung transplantation. However, we have enjoyed many successful collaborations with other groups who study coronaviruses (e.g., SARS-CoV) and rhinoviruses.
Given the high complexity of the airway epithelium it is clear that characteristics of virus-host cell interactions determined with common laboratory cell-lines need to be interpreted with caution. In addition, most human respiratory viruses have limited species tropism with human being predominant (sometimes only) host. Therefore, human airway epithelium models derived from human primary airway epithelial cells obtained from CF or non-CF patients have provided novel culture models that allow new insights into the infection characteristics of human respiratory viruses. Isolated airway epithelial cells grown over a period of 1-2 months at an air-liquid interface result in the generation of a pseudostratified, mucociliary airway epithelium that displays similar morphologic and phenotypic characteristics of the in vivo human cartilaginous airway epithelium. Recent studies have revealed that this model system recapitulates the phenotypic differences that occur between CF and non-CF airway epithelium and important characteristics of respiratory virus-host cell interactions.
- Chen A, Wessler T, Daftari K, Hinton K, Boucher RC, Pickles R, Freeman R, Lai SK, Forest MG. Modeling insights into SARS-CoV-2 respiratory tract infections prior to immune protection. Biophys J. 2022 Apr 1:S0006-3495(22)00277-6. doi: 10.1016/j.bpj.2022.04.003. Epub ahead of print. PMID: 35378080; PMCID: PMC8975607.
- Shi Y, Zeida A, Edwards CE, Mallory ML, Sastre S, Machado MR, Pickles RJ, Fu L, Liu K, Yang J, Baric RS, Boucher RC, Radi R, Carroll KS. Thiol-based chemical probes exhibit antiviral activity against SARS-CoV-2 via allosteric disulfide disruption in the spike glycoprotein. Proc Natl Acad Sci U S A. 2022 Feb 8;119(6):e2120419119. doi: 10.1073/pnas.2120419119. PMID: 35074895; PMCID: PMC8833197.
- Esther CR Jr, Kimura KS, Mikami Y, Edwards CE, Das SR, Freeman MH, Strickland BA, Brown HM, Wessinger BC, Gupta VC, Von Wahlde K, Sheng Q, Huang LC, Bacon DR, Kimple AJ, Ceppe AS, Kato T, Pickles RJ, Randell SH, Baric RS, Turner JH, Boucher RC. Pharmacokinetic-based failure of a detergent virucidal for severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) nasal infections: A preclinical study and randomized controlled trial. Int Forum Allergy Rhinol. 2022 Jan 18. doi: 10.1002/alr.22975. Epub ahead of print. PMID: 35040594. PMCID: PMC9011886
- Wahl A, Gralinski LE, Johnson CE, Yao W, Kovarova M, Dinnon KH 3rd, Liu H, Madden VJ, Krzystek HM, De C, White KK, Gully K, Schäfer A, Zaman T, Leist SR, Grant PO, Bluemling GR, Kolykhalov AA, Natchus MG, Askin FB, Painter G, Browne EP, Jones CD, Pickles RJ, Baric RS, Garcia JV. SARS-CoV-2 infection is effectively treated and prevented by EIDD-2801. Nature. 2021 Mar;591(7850):451-457. doi: 10.1038/s41586-021-03312-w. PMID: 33561864. PMCID: PMC7979515.
- Lai SK, McSweeney MD, Pickles RJ. Learning from past failures: Challenges with monoclonal antibody therapies for COVID-19. J Control Release. 2021 Jan 10;329:87-95. doi: 10.1016/j.jconrel.2020.11.057. PMID: 33276017; PMCID: PMC7836766.
- Wahl A, De C, Abad Fernandez M, Lenarcic EM, Xu Y, Cockrell AS, Cleary RA, Johnson CE, Schramm NJ, Rank LM, Newsome IG, Vincent HA, Sanders W, Aguilera-Sandoval CR, Boone A, Hildebrand WH, Dayton PA, Baric RS, Pickles RJ, Braunstein M, Moorman NJ, Goonetilleke N, Victor Garcia J. Precision mouse models with expanded tropism for human pathogens. Nat Biotechnol. 2019 Oct;37(10):1163-1173. doi: 10.1038/s41587-019-0225-9.PMID: 31451733. PMCID: PMC6776695.
- Stobart CC, Rostad CA, Ke Z, Dillard RS, Hampton CM, Strauss JD, Yi H, Hotard AL, Meng J, Pickles RJ, Sakamoto K, Lee S, Currier MG, Moin SM, Graham BS, Boukhvalova MS, Gilbert BE, Blanco JC, Piedra PA, Wright ER, Moore ML. A live RSV vaccine with engineered thermostability is immunogenic in cotton rats despite high attenuation. Nat Commun. 2016 Dec 21;7:13916. doi: 10.1038/ncomms13916. PMID: 28000669. PMCID: PMC5187593.
- Rostad CA, Stobart CC, Gilbert BE, Pickles RJ, Hotard AL, Meng J, Blanco JCG, Moin SM, Graham BS, Piedra PA, Moore ML. A recombinant respiratory syncytial virus vaccine candidate attenuated by a low-fusion F protein is immunogenic and protective against challenge in cotton rats. J Virol. 2016 Jul 27;90(16):7508-7518. doi: 10.1128/JVI.00012-16. PMID: 27279612. PMCID: PMC4984630.
- Menachery VD, Yount BL Jr, Sims AC, Debbink K, Agnihothram SS, Gralinski LE, Graham RL, Scobey T, Plante JA, Royal SR, Swanstrom J, Sheahan TP, Pickles RJ, Corti D, Randell SH, Lanzavecchia A, Marasco WA, Baric RS. SARS-like WIV1-CoV poised for human emergence. Proc Natl Acad Sci U S A. 2016 Mar 15;113(11):3048-53. doi: 10.1073/pnas.1517719113. PMID: 26976607. PMCID: PMC4801244.
- Pickles RJ, DeVincenzo JP. Respiratory syncytial virus (RSV) and its propensity for causing bronchiolitis. J Pathol. 2015 Jan;235(2):266-76. doi: 10.1002/path.4462. PMID: 25302625. PMCID: PMC5638117.
- Liesman RM, Buchholz UJ, Luongo CL, Yang L, Proia AD, DeVincenzo JP, Collins PL, Pickles RJ. RSV-encoded NS2 promotes epithelial cell shedding and distal airway obstruction. J Clin Invest. 2014 May;124(5):2219-33. doi: 10.1172/JCI72948. PMID: 24713657. PMCID: PMC4001550.
- Brockman-Schneider RA, Pickles RJ, Gern JE. Effects of vitamin D on airway epithelial cell morphology and rhinovirus replication. PLoS One. 2014 Jan 24;9(1):e86755. doi: 10.1371/journal.pone.0086755. PMID: 24475177. PMCID: PMC3901706.
- Pickles RJ. Human airway epithelial cell cultures for modeling respiratory syncytial virus infection. Curr Top Microbiol Immunol. 2013;372:371-87. doi: 10.1007/978-3-642-38919-1_19. PMID: 24362700.
- Kesimer M, Ehre C, Burns KA, Davis CW, Sheehan JK, Pickles RJ. Molecular organization of the mucins and glycocalyx underlying mucus transport over mucosal surfaces of the airways. Mucosal Immunol. 2013 Mar;6(2):379-92. doi: 10.1038/mi.2012.81. PMID: 22929560; PMCID: PMC3637662.
- Ehre C, Worthington EN, Liesman RM, Grubb BR, Barbier D, O’Neal WK, Sallenave JM, Pickles RJ, Boucher RC. Overexpressing mouse model demonstrates the protective role of Muc5ac in the lungs. Proc Natl Acad Sci U S A. 2012 Oct 9;109(41):16528-33. doi: 10.1073/pnas.1206552109. Erratum in: Proc Natl Acad Sci U S A. 2014 Apr 15;111(15):5753. PMID: 23012413; PMCID: PMC3478656.
- Nguyen Y, Procario MC, Ashley SL, O’Neal WK, Pickles RJ, Weinberg JB. Limited effects of Muc1 deficiency on mouse adenovirus type 1 respiratory infection. Virus Res. 2011 Sep;160(1-2):351-9. doi: 10.1016/j.virusres.2011.07.012. PMID: 21816184; PMCID: PMC3163747.
- Okada SF, Zhang L, Kreda SM, Abdullah LH, Davis CW, Pickles RJ, Lazarowski ER, Boucher RC. Coupled nucleotide and mucin hypersecretion from goblet-cell metaplastic human airway epithelium. Am J Respir Cell Mol Biol. 2011 Aug;45(2):253-60. doi: 10.1165/rcmb.2010-0253OC. PMID: 20935191; PMCID: PMC3175555.
- Johnson JS, Gentzsch M, Zhang L, Ribeiro CM, Kantor B, Kafri T, Pickles RJ, Samulski RJ. AAV exploits subcellular stress associated with inflammation, endoplasmic reticulum expansion, and misfolded proteins in models of cystic fibrosis. PLoS Pathog. 2011 May;7(5):e1002053. doi: 10.1371/journal.ppat.1002053. PMID: 21625534; PMCID: PMC3098238.