Femtosecond Laser Nanoparticle-Mediated Gene Therapy for Corneal Endothelial Treatment in Animal Models
Bibliographic record
Abstract
RÉSUMÉ: La motivation de cette recherche réside dans le nombre élevé de personnes affectées par les maladies endothéliales cornéennes. Plus de 2,2 milliards de personnes souffrent de troubles de la vision, et pour au moins 1 milliard d’entre elles, ces troubles auraient pu être évités ou restent non pris en charge. Les maladies endothéliales cornéennes affectent près de 250 millions de personnes dans le monde. Parmi toutes les affections endothéliales, la dystro-phie cornéenne endothéliale de Fuchs (FEDC) se distingue comme étant la plus répandue et constitue la principale cause de transplantations cornéennes à l’échelle mondiale. Pour répondre aux limites associées à la transplantation, notre proposition introduit une approche de thérapie génique utilisant un laser femtoseconde (fs) cliniquement approuvé et des nanoparticules d’or (AuNPs) pour délivrer des gènes dans les cellules endothéliales cornéennes (CECs). L’hypothèse principale est qu’en utilisant un laser femtoseconde fortement fo-calisé (actuellement utilisé à des fins ophtalmiques) pour irradier les CECs dont la membrane sera décorée avec des AuNPs (matériau biocompatible), des pores transitoires (régis par la présence des AuNPs) peuvent être ouverts dans la mem-brane cellulaire avec une grande précision, facilitant ainsi la livraison de molécules exogènes comme l’ARN messager qui permet de réaliser la thérapie génique. Dans le cadre de ce travail, des expériences in vitro et ex vivo ont été réalisées pour atteindre quatre sous-objectifs: (i) établir une preuve de concept en utilisant des cellules oculaires in vitro pour une application ultérieure chez les souris; (ii) développer un protocole plus sécuritaire pour l’injection intracamérulaire chez les modèles murins afin d’améliorer la délivrance des AuNPs dans la chambre antérieure de l’oeil; (iii) obtenir une transfection significative d’ARNm dans le modèle murin avec une haute viabilité cellulaire ex vivo; (iv) développer une technique adaptable à différentes espèces animales pour garantir une application future sécuritaire pour les cornées humaines. ABSTRACT: Worldwide, over 2.2 billion people suffer from near or distance vision impairment, and for at least 1 billion of them, this impairment could have been prevented or remains unaddressed. Within the various causes of vision impairment, corneal endothelial diseases are affecting life quality of close to 250 million people worldwide. The only alternative nowadays, is corneal transplantation, a highly invasive method with undesired side effects such immune response leading to tissue rejection, enfeebling of endothelial cells and transplant failure. Among all the endothelial ailing, Fuchs endothelial corneal dystrophy (FEDC) is the most common and the number one reason for corneal transplantation worldwide. An extra concern is the shortage of healthy tissue for transplantation. Virus-based therapy and electropora-tion are under investigation in ophthalmology, nevertheless, high immunogenicity and low specificity are still a concern. In terms of annual costs, the health care system projects a rise in cost of 30 billion dollars by 2032 only in Canada. In order to minimize some of the drawbacks of current procedures, we propose a gene therapy treatment technique using a clinically approved femtosecond (fs) laser and gold nanoparticles (AuNPs) for the delivery of genes in corneal endothelial cells (CECs). The main contribution that our technique offers is the in situ treatment, avoiding transplantation, having a high efficacy and virus-free technique with double specificity, given by the focalized laser and the implementation of functionalized AuNPs situated at the membrane of targeted cells. The main hypothesis is that by using a tightly focused femtosecond laser (cur-rently used for ophthalmic purposes) to irradiate CECs whose membrane will be decorated with AuNPs (biocompatible material), transient pores (governed by the presence of the AuNPs) can be opened in the cellular membrane with high precision facilitating the delivery of exogenous molecules such as mRNA to perform gene therapy.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.001 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.002 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".