Studies have shown that transfer of the human cystic fibrosis transmembrane conductance regulator (CFTR) cDNA into cystic fibrosis (CF) epithelial corrects the defective cAMP-mediated chloride (CI-) transport that characterizes CF. However, no long term approach to somatic cell gene transfer has been developed. Gene transfer with integrating vectors such as retrovirus offers the exciting potential to provide long term correction. However, studies to date suggest that gene transfer with Moloney murine leukemia virus (MMLV) based vectors is inefficient in differentiated airway epithelia, in part because of the low rates of proliferation. Our recent demonstrate that we can overcome the limitation of low rates of cell division by stimulating cells with growth factors that cause differentiated epithelia to divide. Another limitation is the apparent lack of accessible receptors on the apical surface. We found that cells stimulated to divide with growth factors can be readily infected by applying the vector to the basolateral surface or by apply vector to the apical surface when tight junctions are transiently opened by Va/2+ chelation. An exciting recent development is the hybrid lentivirus-based vectors that can infect non- dividing cells. Our preliminary studies show these vectors share the same problem as MMLV with access to receptors from the apical surface. It is not yet clear if lentiviral vectors will be useful for gene transfer to airway epithelia. Now with this preliminary data and these insights into airway epithelial cell biology, we have the tools and reagents to address several important questions.
In specific aims we will answer 3 questions: 1) Does infection of dividing cells with integrating vectors produce persistent expression and correction of the CF defect?, 2) Can an integrating vector target non-dividing cells and produce persistent expression and correction of the CF defect? 3) Can integrating vectors correct the CF defect in differentiated epithelia in vivo? The results from these studies are relevant to future work with any integrating vector.
| Meyerholz, David K; Sieren, Jessica C; Beck, Amanda P et al. (2018) Approaches to Evaluate Lung Inflammation in Translational Research. Vet Pathol 55:42-52 |
| Rosen, Bradley H; Evans, T Idil Apak; Moll, Shashanna R et al. (2018) Infection Is Not Required for Mucoinflammatory Lung Disease in CFTR-Knockout Ferrets. Am J Respir Crit Care Med 197:1308-1318 |
| Mao, Suifang; Shah, Alok S; Moninger, Thomas O et al. (2018) Motile cilia of human airway epithelia contain hedgehog signaling components that mediate noncanonical hedgehog signaling. Proc Natl Acad Sci U S A 115:1370-1375 |
| Montoro, Daniel T; Haber, Adam L; Biton, Moshe et al. (2018) A revised airway epithelial hierarchy includes CFTR-expressing ionocytes. Nature 560:319-324 |
| Lynch, Thomas J; Anderson, Preston J; Rotti, Pavana G et al. (2018) Submucosal Gland Myoepithelial Cells Are Reserve Stem Cells That Can Regenerate Mouse Tracheal Epithelium. Cell Stem Cell 22:653-667.e5 |
| Meyerholz, David K; Stoltz, David A; Gansemer, Nick D et al. (2018) Lack of cystic fibrosis transmembrane conductance regulator disrupts fetal airway development in pigs. Lab Invest 98:825-838 |
| Gray, Robert D; Hardisty, Gareth; Regan, Kate H et al. (2018) Delayed neutrophil apoptosis enhances NET formation in cystic fibrosis. Thorax 73:134-144 |
| Thornell, Ian M; Li, Xiaopeng; Tang, Xiao Xiao et al. (2018) Nominal carbonic anhydrase activity minimizes airway-surface liquid pH changes during breathing. Physiol Rep 6: |
| Reznikov, Leah R; Meyerholz, David K; Abou Alaiwa, Mahmoud et al. (2018) The vagal ganglia transcriptome identifies candidate therapeutics for airway hyperreactivity. Am J Physiol Lung Cell Mol Physiol 315:L133-L148 |
| Meyerholz, David K; Beck, Amanda P; Goeken, J Adam et al. (2018) Glycogen depletion can increase the specificity of mucin detection in airway tissues. BMC Res Notes 11:763 |
Showing the most recent 10 out of 184 publications
