One IV HEDGES DNA vector administration encoding hGLA or hGH produces durable hGLA and hGH serum lvels in immunocompetent mice

Abstract

The great majority of human monogenic, single protein deficiency disease patients, who comprise ~ 0.5% of the population, are incurable. When available, Enzyme Replacement Therapy (ERT) is current state-of-the art therapy for the vast majority of the subset of these diseases caused by enzyme deficiencies. For example, Fabry disease, caused by hemizygous- or heterozygous-pathogenic variants in GLA encoding human-galactocersbrosidase-α (hGLA), is a rare, single protein-deficiency disease. Fabry patients require intravenous-administration of hGLA Enzyme Replacement Therapy (ERT) every two-weeks for-life. ERT costs ~ $300,000 per-year and can cause frequent infusion reactions, which can be life-threatening. The very-high yearly GLA ERT costs, as well as the recurrent, life-threatening hGLA IV infusion-reactions experienced by some patients, can cause them to permanently-discontinue ERT. This can accelerate Fabry-disease progression, leading to premature-death. Therefore, new, more effective-, safe-, durable-, cost-effective, single deficient-protein replacement platforms are urgently-needed to more-effectively treat a wide-spectrum of these rare, monogenic single protein deficiency diseases. Here we demonstrate that one intravenous-administration of our 1 st -generation HEDGES DNA-vector encoding wildtype-hGLA (T 1 / 2 < 20-minutes) produced hGLA serum-protein levels in the normal human 1,000-10,000 pg/ml range for only < 14 days. We then-created our 2 nd -generation HEDGES hGLA DNA-vector. One intravenous-administration of this DNA-vector produced durable (>550 days) serum hGLA serum levels in the 1–10 ng/ml, thus increasing the duration of hGLA serum-protein levels produced by > 38,100 fold versus administering bioreactor-produced, wildtype hGLA-protein. We also showed one intravenous-administration of our 1 st -generation HEDGES DNA-vector encoding the wildtype human growth hormone (hGH) protein, (T 1 / 2 < 20-minutes), produced serum hGH levels in the 1–10 ng/ml for > 330 days, thus increasing the duration of hGLA serum-protein levels by > 22,860 fold versus administering wildtype hGH-protein. Last, one intravenous-administration of our 2 nd -generation HEDGES hGLA DNA-vector produced serum hGLA levels in the normal human 1–10 ng/ml range for > 160 days in GLA knockout-mice, a 2,800-fold increase versus wildtype hGLA-protein. hGLA-ERT produces major therapeutic-responses in GLA knockout-mice. These substantial ERT-responses in GLA knockout-mice have been shown to be accurately-recapitulated in Fabry patients. Thus, Fabry disease appears a promising-target for subsequent phase-1 HEDGES-based human clinical trials.