Outcomes of human leukocyte antigen-matched allogeneic cultivated limbal epithelial transplantation in aniridia-associated keratopathy - a single-center retrospective analysis

To assess the efficacy and safety of human leukocyte antigen-matched allogeneic cultivated limbal epithelial stem cell grafts in the treatment of aniridia-associated keratopathy (AAK). Methods: Six eyes of 6 patients with severe AAK received an allogeneic stem cell graft between January 2010 and March 2017. Anatomical and functional results were assessed at 6 months, 1 year, 2 years, and the final follow-up visit available. Safety analysis was performed by considering all perioperative and postoperative adverse events and additional surgeries required during the follow-up period.Results:The mean follow-up was 53.6 months (range 24-104 months). In most patients (80%), there was an early improvement of the keratopathy postoperatively, which slowly regressed during longer follow-up. At the final follow-up, 4 of the eyes were graded as failure and 1 eye was graded as partial success. Grading the sixth eye was not possible because of an adverse event. None of the patients maintained a total anatomical success in the long-term. Only 1 patient maintained a modest improvement in best-corrected visual acuity from hand motion to counting fingers. Four serious adverse events were recorded in 2 patients.Conclusions:Severe AAK remains a challenging condition to manage. Transplantation of allogenic ex vivo cultivated limbal stem cells may provide a temporary improvement in ocular surface stability, but anatomical and functional results are poor in the long-term. The eyes are prone to adverse events, and any surgical treatment should take this into consideration

Matrix metalloproteinase-3 (MMP-3)-mediated gene therapy for glaucoma.

Approximately 80 million people globally are affected by glaucoma, with a projected increase to over 110 million by 2040. Substantial issues surrounding patient compliance remain with topical eye drops, and up to 10% of patients become treatment resistant, putting them at risk of permanent vision loss. The major risk factor for glaucoma is elevated intraocular pressure, which is regulated by the balance between the secretion of aqueous humor and the resistance to its flow across the conventional outflow pathway. Here, we show that adeno-associated virus 9 (AAV9)-mediated expression of matrix metalloproteinase-3 (MMP-3) can increase outflow in two murine models of glaucoma and in nonhuman primates. We show that long-term AAV9 transduction of the corneal endothelium in the nonhuman primate is safe and well tolerated. Last, MMP-3 increases outflow in donor human eyes. Collectively, our data suggest that glaucoma can be readily treated with gene therapy-based methods, paving the way for deployment in clinical trials

Total Evidence: A Test of Principle Using the Malacostraca

The phylogeny of Eumalacostraca (Crustacea) has remained elusive, despite over a century of morphological and more recently molecular studies. Prior to this, no large scale combined evidence analyses of all eumalacostracan orders has been carried out. Evidence from four nuclear ribosomal and mitochondrial loci (18S rRNA, 28S rRNA, 16S rRNA, and cytochrome c oxidase subunit I) were combined with a newly synthesized morphological dataset to examine the utility of available data. With an aim to resolve conflict in the morphological dataset, fossils were added to the morphological dataset and their effects examined. The stratigraphic congruence of crustaceans was examined to determine the quality of existing trees. A modified protocol was developed to generate new molecular markers for the investigation of eumalacostracan phylogeny. Finally the effects of the new data on existing datasets were examined in a combined evidence approach. Significant conflict was detected between data partitions, especially between morphology and molecules. The addition of fossil data revealed the morphological dataset to be very sensitive to taxon sampling. Stratigraphic congruence was found to be poor for the five crustacean groups examined. Histone 3 and alanyl-tRNA synthetase were shown to be capable of successfully recovering relationships at the level of genus and above. Glutamyl-prolyl-tRNA synthetase failed to recover monophyletic groupings when analysed alone. Combining all molecular data produced well-supported phylogenies, but significant conflict between data partitions was identified and trees were very sensitive to taxon sampling. Rate heterogeneity and conflict between data partitions mean that the total sum of molecular and morphological evidence as presented here is currently unable to resolve a wellsupported eumalacostracan phylogeny. We recommend additional taxon sampling and further sampling of novel markers using the modified protocol developed here as well as the addition of fossil data and the exploitation of next generation sequencing technologies.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Genetic and environmental risk factors in age-related macular degeneration in vitro, in vivo and population studies

THESIS 10017The aims of this project are to show the potential sites of interaction between underlying genetic susceptibilities and environmental influences (cigarette smoke) in neovascular AMD. Our current hypothesis proposes that cigarette smoke exerts its toxic effects by altering the expression of vasoactive proteins at the level of the RPE, ultimately promoting angiogenesis. To test this we used in vitro and in vivo menthods. We also consider the possibility that known genetic risk factors in regions coding for vasoactive proteins may contribute to this process of angiogenesis in Irish patients with AMD. [exerpt Page 69

Outcomes of Human Leukocyte Antigen–Matched Allogeneic Cultivated Limbal Epithelial Transplantation in Aniridia-Associated Keratopathy—A Single-Center Retrospective Analysis

To assess the efficacy and safety of human leukocyte antigen-matched allogeneic cultivated limbal epithelial stem cell grafts in the treatment of aniridia-associated keratopathy (AAK). Methods: Six eyes of 6 patients with severe AAK received an allogeneic stem cell graft between January 2010 and March 2017. Anatomical and functional results were assessed at 6 months, 1 year, 2 years, and the final follow-up visit available. Safety analysis was performed by considering all perioperative and postoperative adverse events and additional surgeries required during the follow-up period.Results:The mean follow-up was 53.6 months (range 24-104 months). In most patients (80%), there was an early improvement of the keratopathy postoperatively, which slowly regressed during longer follow-up. At the final follow-up, 4 of the eyes were graded as failure and 1 eye was graded as partial success. Grading the sixth eye was not possible because of an adverse event. None of the patients maintained a total anatomical success in the long-term. Only 1 patient maintained a modest improvement in best-corrected visual acuity from hand motion to counting fingers. Four serious adverse events were recorded in 2 patients.Conclusions:Severe AAK remains a challenging condition to manage. Transplantation of allogenic ex vivo cultivated limbal stem cells may provide a temporary improvement in ocular surface stability, but anatomical and functional results are poor in the long-term. The eyes are prone to adverse events, and any surgical treatment should take this into consideration