Research Updates

Statement of Progress (Haldar lab). In 2021-2022 we have advanced to show that our gene therapy (GT) for P protein mutations, reduces glycine levels in NKH mice. This occurs even when the therapy is administered to mice that are 23-36 days in age (equivalent in humans would be late teenager). When administered to very young mice (within a few days after birth) GT also prevents post-natal mouse pup death, suggesting it benefits both early, severe as well as late neurometabolic disease. We show GT acts by raising the level of P protein in the liver and are investigating expression in the brain. In 2022-2023 we will complete studies on brain and plasma analyses as well as seizure and cognition and start the process of transferring our gene therapy to pharma companies to develop clinical trials.

To date NKH Crusaders has donated over 700,000 to NKH Research at the University of Colorado and the University of Notre Dame. Research has moved forward so much in the last few years due to the hard work of the researchers. We are now working hard on creating a NKH Crusaders Patient Registry and hope to launch it in October 2022.

Dr. Johan Van Hove, We are excited to be able to donate 150,000 towards NKH research. This funding is separate from the commitment that we made to you back in November for a 100,000 budget from November 1,2021 to October 31,2022. Leading to total funding of $250,000 between November 1, 2021 and February 2023. Enclosed please find a check for $50,000 towards the new 150,000 donation. We are grateful for all the time and dedication that you put into NKH research and supporting our families. We are equally committed to providing the funds needed to keep the research moving and you supported. Together we are moving research forward and towards better treatment and a cure. Thank you for your commitment to our families. Kristin Archibald and the Leadership Committee

Summary: ND-NKH Gene Therapy. Prepared by Dr. Kasturi Haldar, Director Center for Rare and Neglected Diseases, University of Notre Dame. Submitted Oct 4th 2019. Non-ketotic hyperglycinemia (NKH) is a neuro-metabolic disorder characterized by intractable seizures, failure to thrive, lack of developmental milestones, and premature death. Loss of function of glycine decarboxylase (GLDC; also known as P protein) is a leading cause of NKH. Mutations and deletions in P protein are responsible for greater than 80% of NKH cases. Dr. Haldar and the Notre Dame team have developed pre-clinical mouse models of NKH with genetically engineered mutations in P protein. These mice show the full spectrum (severe to mild) of neuro-metabolic disease with measureable increase in glycine levels and defects in brain, liver and kidney function. Our goal for 2019-2020 is to develop gene therapy for NKH in pre-clinical mouse models to evaluate their potential for human clinical trials. The work will be led by Dr. Haldar and Notre Dame and partner with Dr. James Wilson and team at the Penn Vector Core at the University of Pennsylvania (who will be prepare gene therapy vectors) Budget Period 12 months 10_15_2019 to 10_14_2020. 1.University of Notre Dame. 1.1.Direct Costs Charged to Project. Animals: $ 34,177 Expansion of NKH mutants. Requires breeding and overall husbandry associated with feeding, drinking water changing bedding and cages, all done by the ND animal facility (charges are based on their rates). Organ Analyses Brain, Liver and blood analyses $ 20,000 Immunological reagents to determine state/health/inflammation of organs/neuroinflammation. Metabolomics and genomic marker analyses of brain, liver and blood in untreated animals and response to treatment. Personnel Junior Tech 100% (salary $35,000; benefits $9730) $ 44,730 Molecular genotyping, track litters to obtain requisite numbers of animals with desired timing, track weight, survival, undertake neurocognitive testing of live animals, seizure assays, conduct organ harvests. Personnel Travel from ND to U Penn for Training brain injection protocol (airfare, 1 week stay) $ 2,000 Subtotal: $100, 907 1.2. UND Cost share : Direct costs not charged to project. Dr. Alam. Research Asst. Professor 75% (salary + benefits S71,884). $ 0 Dr. Alam will be directly responsible for delivery of gene therapy to the brain and other organs. He will undertake molecular, biochemical and tissue analyses of brain, liver and kidney. Dr. Alam will also intensively train the technician in procedures of genotyping, tracking litters, neurocognitive assessment, seizure assays conducting organ harvests. He will also supervise the technician on a daily basis and assist Dr. Haldar with overall project design and management. Joseph Harris. Graduate Student 50% (salary and benefits $19,000) $ 0 Joe Harris will be directly responsible for computational synthesis of metabolomics and genomic data needed to prioritize predictive lead blood and tissue markers that will be tested by Dr. Alam. Dr. Kasturi Haldar. Director, (1.5 month summer salary or 12.5% annual effort). $ 0 Dr. Haldar will be responsible with overall strategy for project design and management. She will be assisted by Dr. Alam in assessing quantitative power analyses of pre-clinical findings. She will conduct weekly meetings with Dr. Alam, Joe Farris and the technician and communicate findings to the NKH community. Dr. Haldar will also be the primary liason with the University of Pennsylvania, Wilson team and Penn Vector core, hold inter-institutional video conferences needed so that Material Transfer Agreements and invoices are completed in order that the work can proceed in a timely way. Subtotal $ 0 1.3 Indirect Costs (IDC). UND’s IDC rate of 54% of direct costs are waived. $ 0 Subtotal: $ 0 _____________________________ 2. ND_Subcontract Account with Penn Vector Core University of Pennsylvania. 2.1. Direct costs for AAV9 vectors estimated. $ 45,000* This covers the production of small scale and large scale preparation and packaging of plasmids and AAV9 vectors needed. First delivery of vectors expected in December 2019 to initiate gene therapy studies in mice. * This is the lowest cost projected. It could increase to maximum of $52,000, if we need to refine vector design. 2.2 UPenn Cost Share. UPenn Consultation costs and indirect costs are waived. ____________________________________________________________________________ Total Direct Costs for Components 1& 2: $145,907