TS-002356 — Education coordinator, Adrianna Matos-Nieves, PhD, at Nationwide Children’s Hospital created an educational board game called Viruses and Vaccines. The board game teaches the benefits of vaccines, the danger of contracting diseases, and the impact of future health decisions. The gamification of vaccine learning makes it the perfect educational tool for parents and children. The game illustrates a long healthy lifespan by going up a ladder and diseases with no prior vaccination or vaccine push the player down a ladder with each space having corresponding card of information about the vaccine and disease. Viruses and Vaccines encourages players to make informed decisions about their health to promote healthy living.

TS-002317 — FOXG1 syndrome is a rare neurological and developmental disorder that usually affects children at infancy. Individuals with this disorder experience seizures, delayed development, intellectual disability, and mobility issues. Currently, there is no cure. Researchers at Nationwide Children’s Hospital designed a conventional gene therapy approach for FOXG1 by establishing vitro models and comparing data for FOXG1 cell lines and Rhett Syndrome cell lines. (need info. about the results of the 2021 pilot study)

TS-002304 — Researchers at Nationwide Children’s Hospital have created a new, improved method of mRNA exon replacement for any disorder through the combination of a transplicing molecule (PTM) and U7 small nuclear RNA constructs exon skipping tool in an AAV vector. To establish proof of concept, the team tested for mutations in exon 1 and 1 of the SCN2A gene. Two constructs will mediate both exclusions of the endogenous exons 1-2, where one bears a disease-causing mutation, and swaps them with a wild type or enhanced exon 1-2 transplicing molecule. A self-complementary (sc) and/or single stranded adeno-associated virus (AAV) serotypes that target the central nervous system and muscles will deliver the combined approach. These first two products force replacement of mutated SCN2A exon 1-2 for a subpopulation of patients affected by SCN2A disorders. This combination shows immense potential application in its replacement of any mutated mRNA piece.

TS-002303 — WHO: Scott Harper WHAT: RNAi therapy WHY: DNM1 Developmental and Epileptic Encephalopathy (DEE) HOW: (from IDF) The DNM1 gene encodes dynamin-1, a large GTPase involved in clathrin-mediated endocytosis of synaptic vesicles in neurons and in related processes. Dynamin monomers assemble into multimers that interact with each other and with various other proteins to form ring structures for GTPase-catalyzed membrane scission. Dominant mutations in one allele of DNM1 cause neurological disease in humans and mice. Children with DNM1 mutations suffer from intractable conditions manifesting as early-onset seizures, global developmental delay, profound intellectual disability, lack of speech, muscular hypotonia, dystonia, and spasticity. As is the case with many severe DEEs, affected individuals do not respond well to antiepileptic drugs, leaving >80 % of patients with intractable seizures and little to no improvement of the severely debilitating other neurological features. At least 20 de novo pathogenic DNM1 variants have been identified, and all are presumed to operate with dominant effect, most likely by interfering with the assembly or function of normal dynamin-1 monomers. Importantly, knockout studies in mice support that at least 50% normal DNM1 levels are required, as homozogous DNM1 null animals die by postnatal day 8. We propose that a successful gene therapy approach must eliminate or reduce the expression or translation of the mutant DNM1 variant mRNA or protein, while still enabling expression of the remaining wild-type allele. We are pursuing two different approaches to accomplish this: (1) isoform-specific silencing of mutant DNM1 transcripts; (2) a knockdown-and-replace strategy; expressing exogenous wildtype Dnm1 via AAV while eliminating endogenous Dnm1 mRNA entirely. We already filed an IDF and provisional patent application on approach 1. This IDF covers the knockdown-and-replace strategy. In brief, we will develop artificial microRNAs that non-selectively knockdown both mutant and wild-type DNM1 alleles using RNAi, while at the same time adding back an RNAi-resistant, wild-type DNM1 cDNA. We will test this in mouse models of DNM1-related DEE in the Frankel lab at Columbia University.

TS-002282 — Rare variants in genes GRIN2A, GRIN2B and GRIN2D are associated with severe childhood-onset neurological diseases like epileptic encephalopathies and autism. There are no treatments for these chronic disorders that cause developmental delays or developmental skill loss. Researcher and principal investigator, Scott Harper, at Nationwide Children’s Hospital developed a genetic therapy for GRIN2 disease through a knockdown-and-replace strategy by using artificial microRNAs that non-selectively knockdown both mutant and wildtype GRIN2A alleles using RNAi and adding back an RNAi-resistant, wildtype GRIN2A cDNA. The therapy will express functional GRIN2 exogenously while eliminating endogenous GRIN2 mRNA entirely through AAV delivery.

TS-002280 — **based on incomplete invention disclosure What: RNAi Therapy Why: Developmental and Epileptic Encephalopathy --> currently no treatment for DNM1- related How: Decrease expression of pathogenic variants in the dynamin-1 gene (DNM1 gene) which causes a particularly severe form of dominant developmental and epileptic encephalopathy (DEE)

TS-002279 — What: gene therapy for pediatric epileptic encephalopathy resultant from mutations in SLC6A1 gene Six new gene replacement products New shRNAs for gene knockdown New delivery route and dosing ranges Why: pediatric epileptic encephalopathy Current treatment is limited to symptomatic treatment, primarily by use of antiepileptic drugs How: single stranded AAV serotype 9 that will provide a wildtype copy of SLC6A1 to address haploinsufficiency resultant from mutations in one copy of the SLC6A1 gene Delivered through cerebrospinal fluid (CSF) via injection

TS-002231 — About 1 in 50,000 people in the United States are affected by Friedreich’s Ataxia, an inherited disorder caused by a gene which damages the nervous system causing difficulties walking, slow speech, fatigue and sensation changes. Researchers at Nationwide Children’s Hospital developed an innovative way to modulate promoter activity to alter Frataxin gene expression using U7 small nuclear RNA. The U7 snRNA has mammalian origin, can have sustained and stable expression as it acts as a promoter and is safe for human patients. Additionally, the method suggests using U& snRNA with a tail to attract endogenous activator proteins which bind to DNA. The method can be combined with other gene replacement strategies to add additional U7-activators and using promoter regulation to activate promoters to upregulate a gene expression of a certain gene like Friedreich’s Ataxia.

TS-002230 — Currently, targeting the retina is difficult due to the thickness of vitreous preventing easy diffusing to the retina and restricted access due to the inner-limiting membrane (ILM). These challenges are especially difficult in the context of retinal AAV therapies. Researchers at Nationwide Children’s Hospital propose using Omnipaque to increase the density of injection solutions containing AAVs to allow for increased delivery to the retina combined with supine positioning of the patient. This addition would eliminate the need for risky, complicated injection methods.

TS-002229 — Adeno-associated virus vectors (AAV) are used to deliver normal copies of genes or therapies to a targeted tissue or organ as treatment for many genetic disorders. Researchers at Nationwide Children’s Hospital created a process that will improve the production process for AAV viral vectors in flatware and bioreactors for suspension and adherent cells. They performed placid transfection prior to seeding into the tissue culture flasks which decreased the incubation time for cells in the bioreactor and yielded a higher crude AAV viral vector yield by 8-10-fold. Therefore, this process will reduce production time and costs of production.