@article{gilger_crabtree_song_llanga_cullen_blanchard_salmon_patel_zarnitsyn_hirsch_2020, title={A Fixed-Depth Microneedle Enhances Reproducibility and Safety for Corneal Gene Therapy}, volume={39}, ISSN={["1536-4798"]}, DOI={10.1097/ICO.0000000000002182}, abstractNote={
            Purpose:
            Drug delivery directly to the corneal stroma currently relies on microscopic injections that demonstrate low reproducibility and clinician-dependent variability. With use of biological drugs such as adeno-associated viral (AAV) vectors, precise and consistent drug deposition is critical to reduce concerns related to off-target transduction and the host's immune response to the viral capsid and/or transgene-derived product. Therefore, a precise corneal injection (PCI) microneedle was designed to allow accurate depth-specific injections into the corneal stroma in a macroscopic setting.
          
          
            Methods:
            High-frequency ultrasound and confocal microscopy demonstrated the consistent ability to predetermine the precise injection depth using PCI needles of varying sizes. Next, a comparison between a standard 31-G needle and PCI needles was performed in vivo using AAV vector gene delivery.
          
          
            Results:
            Intrastromal corneal injections using the PCI microneedle resulted in less vector leakage at the site of injection and fewer anterior chamber penetrations compared with a standard 31-G needle. Although reporter gene expression appeared similar when the vector was administered with either needle type, a trend toward increased vector genomes was noted in the PCI-injected corneas at the experimental conclusion. As hypothesized, corneal perforation resulted in increased detection of AAV vector genomes in nontarget tissues, highlighting the importance of consistency for biological drug applications in the cornea.
          
          
            Conclusions:
            Further development of the PCI microneedle is warranted especially for AAV corneal gene therapy and offers the potential to enhance transduction while significantly reducing safety concerns and intraclinician and interclinician injection variability.
          }, number={3}, journal={CORNEA}, author={Gilger, Brian C. and Crabtree, Elizabeth and Song, Liujiang and Llanga, Telmo and Cullen, Megan and Blanchard, Allison and Salmon, Jacklyn and Patel, Samirkumar and Zarnitsyn, Vladimir and Hirsch, Matthew}, year={2020}, month={Mar}, pages={362–369} }
 @article{crabtree_song_llanga_bower_cullen_salmon_hirsch_gilger_2019, title={AAV-mediated expression of HLA-G1/5 reduces severity of experimental autoimmune uveitis}, volume={9}, ISSN={2045-2322}, url={http://dx.doi.org/10.1038/s41598-019-56462-3}, DOI={10.1038/s41598-019-56462-3}, abstractNote={AbstractNon-infectious uveitis (NIU) is an intractable, recurrent, and painful disease that is a common cause of vision loss. Available treatments of NIU, such as the use of topical corticosteroids, are non-specific and have serious side effects which limits them to short-term use; however, NIU requires long-term treatment to prevent vision loss. Therefore, a single dose therapeutic that mediates long-term immunosuppression with minimal side effects is desirable. In order to develop an effective long-term therapy for NIU, an adeno-associated virus (AAV) gene therapy approach was used to exploit a natural immune tolerance mechanism induced by the human leukocyte antigen G (HLA-G). To mimic the prevention of NIU, naïve Lewis rats received a single intravitreal injection of AAV particles harboring codon-optimized cDNAs encoding HLA-G1 and HLA-G5 isoforms one week prior to the induction of experimental autoimmune uveitis (EAU). AAV-mediated expression of the HLA-G-1 and -5 transgenes in the targeted ocular tissues following a single intravitreal injection of AAV-HLA-G1/5 significantly decreased clinical and histopathological inflammation scores compared to untreated EAU eyes (p < 0.04). Thus, localized ocular gene delivery of AAV-HLA-G1/5 may reduce the off-target risks and establish a long-term immunosuppressive effect that would serve as an effective and novel therapeutic strategy for NIU, with the potential for applications to additional ocular immune-mediated diseases.}, number={1}, journal={Scientific Reports}, publisher={Springer Science and Business Media LLC}, author={Crabtree, E. and Song, L. and Llanga, T. and Bower, J.J. and Cullen, M. and Salmon, J.H. and Hirsch, M.L. and Gilger, B.C.}, year={2019}, month={Dec}, pages={19864} }
 @article{cullen_jacob_cornish_vanderschel_cotter_cubeta_carbone_gilger_2019, title={Multi-locus DNA sequence analysis, antifungal agent susceptibility, and fungal keratitis outcome in horses from Southeastern United States}, volume={14}, ISSN={1932-6203}, url={http://dx.doi.org/10.1371/journal.pone.0214214}, DOI={10.1371/journal.pone.0214214}, abstractNote={Morphological characterization and multi-locus DNA sequence analysis of fungal isolates obtained from 32 clinical cases of equine fungal keratitis (FK) was performed to identify species and determine associations with antifungal susceptibility, response to therapy and clinical outcome. Two species of Aspergillus (A. flavus and A. fumigatus) and three species of Fusarium (F. falciforme, F. keratoplasticum, and F. proliferatum) were the most common fungi isolated and identified from FK horses. Most (91%) equine FK Fusarium nested within the Fusarium solani species complex (FSSC) with nine genetically diverse strains/lineages, while 83% of equine FK Aspergillus nested within the A. flavus clade with three genetically diverse lineages. Fungal species and evolutionary lineage were not associated with clinical outcome. However, species of equine FK Fusarium were more likely (p = 0.045) to be associated with stromal keratitis. Species of Aspergillus were more susceptible to voriconazole and terbinafine than species of Fusarium, while species of Fusarium were more susceptible to thiabendazole than species of Aspergillus. At the species level, A. fumigatus and A. flavus were more susceptible to voriconazole and terbinafine than F. falciforme. Natamycin susceptibility was higher for F. falciforme and A. fumigatus compared to A. flavus. Furthermore, F. falciforme was more susceptible to thiabendazole than A. flavus and A. fumigatus. These observed associations of antifungal sensitivity to natamycin, terbinafine, and thiabendazole demonstrate the importance of fungal identification to the species rather than genus level. The results of this study suggest that treatment of equine FK with antifungal agents requires accurate fungal species identification.}, number={3}, journal={PLOS ONE}, publisher={Public Library of Science (PLoS)}, author={Cullen, Megan and Jacob, Megan E. and Cornish, Vicki and VanderSchel, Ian Q. and Cotter, Henry Van T. and Cubeta, Marc A. and Carbone, Ignazio and Gilger, Brian C.}, editor={Kniemeyer, OlafEditor}, year={2019}, month={Mar}, pages={e0214214} }