Document text
Principal Investigator: James Dahlman
Organization: GEORGIA INSTITUTE OF TECHNOLOGY
Fiscal Year: 2019
Award: $710,671
Funding agency: National Center for Advancing Translational Sciences
Summary. Hemoglobinopathies such as β-thalassemia and sickle cell disease are genetic disorders caused
by mutations in the HBB gene that codes for the β-globin component of hemoglobin. Currently, the only gene
therapy available for these prevalent hereditary diseases is based on transplantation of genetically corrected
hematopoietic stem cells (HSPCs) from fully matched donors. However, the efficacy of this approach is
limited by multiple factors. Gene editing is a promising alternative approach for curing hemoglobinopathies.
Using this approach, synthetic mRNA-based drugs encoding nucleases that target the HBB gene can be
utilized to permanently correct the patient’s DNA. Combining nanoparticle-based drug delivery with zinc-
finger nucleases (ZFNs) has the potential to facilitate targeted gene-editing in HSPCs. However, the reliance
on in vitro screening of nanoparticles impedes the discovery of safe and efficient in vivo delivery vehicles.
Furthermore, current ZFN-mRNA based drugs targeting the HBB gene in HSPCs exhibit immunogenicity and
are expressed in off-target cells. The PIs have recently been shown that DNA barcoded nanoparticles can
‘evolve’ nanoparticles to target endothelial cells more efficiently than hepatocytes directly in vivo. The team
has also demonstrated that it is possible to (i) design low immune stimulating mRNA via nucleotide
modification and HPLC purification, and that (ii) mRNAs can be designed to completely preclude translation
in hepatocytes using rationally designed ‘on’ and ‘off’ switches. Based on these supporting data, it is posited
that nanoparticles can be evolved to specifically target HSPCs while avoiding hepatocytes, and that ZFN-
mRNA based drugs can be rationally optimized to generate safe gene editing therapeutics targeting
HSPCs. Thus, the team proposes to create an mRNA-based drug that safely and specifically edits HSPCs in
non-human primes in two phases. The development (UG3) phase will address 2 aims: (1) to iteratively evolve
nanoparticles that target HSPCs and avoid hepatocytes in vivo, and (2) to reduce mRNA immunogenicity
and improve cell type specific delivery to HSPCs. The demonstration (UH3) phase will address the aim (3) to
analyze functional gene editing in non-human primates (Rhesus macaques). These will be achieved using a
cutting edge multidisciplinary approaches recently developed. Specifically, the team will combine a DNA
barcoded nanoparticle technology to screen 4,500 nanoparticles in vivo, synthesize mRNA-based drugs with
low immunogenicity and cell type-specific expression, and utilize customized bioinformatics pipeline that
facilitates ‘big data’ experiments with a statistical power new to nanomedicine. By creating an mRNA-based
drug that safely edits HSPCs, the project is poised to advance gene editing as a viable therapeutic
approach for curing genetic blood disorders and pave the way for clinical trials.
Terms: <Address><Affect><Assay><B-globin><B-thalassemia><Big Data><BigData><Bio-Informatics><Bioassay><Biodistribution><Bioinformatics><Biologic Assays><Biological Assay><Blood Diseases><Blood Precursor Cell><Cell Body><Cell Culture Techniques><Cells><Chemicals><Clinical><Clinical Trials><Code><Coding System><Codon><Codon Nucleotides><Custom><DNA><DNA Therapy><Data><Deoxyribonucleic Acid><Development><Dose><Drug Delivery><Drug Delivery Systems><Drug Targeting><Drugs><Effectiveness><Endothelial Cells><Exhibits><Gene Expression><Gene Transfer><Gene Transfer Clinical><Genes><Genetic><Genetic Alteration><Genetic Change><Genetic Diseases><Genetic Intervention><Genetic defect><Genomics><Goals><HPLC><Hb SS disease><HbSS disease><Hematologic Diseases><Hematological Disease><Hematological Disorder><Hematopoietic Progenitor Cells><Hematopoietic stem cells><Hemoglobin><Hemoglobin S Disease><Hemoglobin sickle cell disease><Hemoglobin sickle cell disorder><Hemoglobinopathies><Hepatic Cells><Hepatic Parenchymal Cell><Hepatocyte><Hereditary Disease><High Performance Liquid Chromatography><High Pressure Liquid Chromatography><High Speed Liquid Chromatography><Immune><Immunes><In Vitro><Inborn Genetic Diseases><Inherited disorder><Liver Cells><Macaca mulatta><Medication><Messenger RNA><Methods><Mice><Mice Mammals><Modification><Murine><Mus><Mutation><Names><Non-Polyadenylated RNA><Nucleotides><Patients><Pharmaceutic Preparations><Pharmaceutical Preparations><Phase><Position><Positioning Attribute><Progenitor Cells><RNA><RNA Gene Products><Regimen><Rhesus><Rhesus Macaque><Rhesus Monkey><Ribonucleic Acid><Safety><Sickle Cell Anemia><Site><Stem cells><System><Technology><Testing><Therapeutic><Time><Toxic effect><Toxicities><Toxicity due to chemotherapy><Translations><Transplant Recipients><Transplantation><ZNFs><Zn-finger nuclease><base><beta Globin><beta Thalassemia><blood disorder><blood stem cell><cell culture><cell type><chemical property><chemotherapy toxicity><clinical translation><conditioning><design><designing><developmental><drug/agent><experiment><experimental research><experimental study><gene therapy><gene-based therapy><genetic condition><genetic disorder><genetic therapy><genome mutation><genomic therapy><hematopoietic progenitor><hematopoietic stem progenitor cell><hemopoietic progenitor><hemopoietic stem cell><hereditary disorder><immunogenicity><improved><in vivo><inborn error><inherited diseases><inherited genetic disease><inherited genetic disorder><interdisciplinary approach><lipid nanoparticle><mRNA><multidisciplinary approach><nano medicinal><nano medicine><nano particle><nano-sized particle><nanomedicinal><nanomedicine><nanoparticle><nanosized particle><non-human primate><nonhuman primate><novel><p-Thalassemia><physical property><protein expression><response><screening><sickle cell disease><sickle disease><sicklemia><targeted endonucleases><targeted nucleases><therapeutic target><tool><transplant><transplant patient><zinc finger nuclease><zinc finger nucleases><β-globin><β-thalassemia>