This proposal is a 5 year training program for the development of an academic career in gene therapy. The principal investigator has separately completed Ph.D. and M.D. programs. She is board certified in Pediatrics and Medicine, as well as board eligible in Pediatric/Hematology Oncology. The K01 grant will help to expand the applicant's skills, in gene therapy using multiple resources at the Powell Gene Therapy Center at the University of Florida. This program will promote the command of gene therapy as it applies to hematopoietic stem cells. The applicant's mentor is a recognized leader in the field of gene therapy. The mentor is Chief of Division of Cellular and Molecular Therapy, and also serves as Assistant Director of the UF General Clinical Research Center. The applicant and mentor have established an internal advisory committee composed of accomplished scientist and clinicians affiliated with UF, who will provide guidance and training in globin gene regulation, bone marrow environment, and development of clinical models. The research component of the K01 will focus on AAV capsid engineering to facilitate expression of therapeutic levels of beta-globin in hematopoietic stem cells. Novel AAV capsids will be generated by two methods: (1) systematic mutation of tyrosine to phenylalanine and (2) DNA shuffling, which involves randomization of capsid amino acids of different AAV serotypes. The transduction efficiency of the novel AAV capsids will be tested using reporter genes expressing enhanced green fluorescence protein (EGFP) and luciferase, and the optimum vector selected to design an antisickling 2-globin gene therapy construct. There are two specific aims: 1. To create an AAV vector with an engineered capsid that facilitates high level gene expression in both human and mouse hematopoietic stem cells. 2. To determine whether an optimized AAV vector expressing anti-sickling 2-globin can correct sickle cell disease in transgenic mice. If successful this will be the first therapeutic anti-sickling AAV gene therapy vector. The Powell Gene Therapy Department at the UF provides an ideal setting for this K01 training in gene therapy, by incorporating expertise from multiple resources into a tailored program. The environment at the UF will maximize the applicant's potential to establish a career as an independent translational investigator.

Public Health Relevance

The main aim of this proposal is to determine whether AAV capsid can be engineered to facilitate therapeutic gene delivery in hematopoietic stem cells. Recombinant AAV vectors are currently being used in a number of Phase I/II clinical trials for a wide variety of human diseases. Our proposed studies are expected to yield new and useful information that will further development of these vectors for the treatment of blood diseases.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Scientist Development Award - Research & Training (K01)
Project #
Application #
Study Section
Special Emphasis Panel (ZHL1-CSR-H (M1))
Program Officer
Werner, Ellen
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Florida
Schools of Medicine
United States
Zip Code
Jagadeeswaran, Ramasamy; Vazquez, Benjamin A; Thiruppathi, Muthusamy et al. (2017) Pharmacological inhibition of LSD1 and mTOR reduces mitochondrial retention and associated ROS levels in the red blood cells of sickle cell disease. Exp Hematol 50:46-52
Rivers, Angela; Vaitkus, Kestis; Ruiz, Maria Armila et al. (2015) RN-1, a potent and selective lysine-specific demethylase 1 inhibitor, increases ?-globin expression, F reticulocytes, and F cells in a sickle cell disease mouse model. Exp Hematol 43:546-53.e1-3
Han, Jin; Kemiki, Opeyemi; Hsu, Lewis L et al. (2015) Adverse Reactions to Pneumococcal Vaccine in Pediatric and Adolescent Patients with Sickle Cell Disease. Pharmacotherapy 35:696-700
Choe, Se-woon; Terman, David S; Rivers, Angela E et al. (2013) Drug-loaded sickle cells programmed ex vivo for delayed hemolysis target hypoxic tumor microvessels and augment tumor drug delivery. J Control Release 171:184-92
Aslanidi, George V; Rivers, Angela E; Ortiz, Luis et al. (2013) Optimization of the capsid of recombinant adeno-associated virus 2 (AAV2) vectors: the final threshold? PLoS One 8:e59142
Song, Liujiang; Kauss, M Ariel; Kopin, Etana et al. (2013) Optimizing the transduction efficiency of capsid-modified AAV6 serotype vectors in primary human hematopoietic stem cells in vitro and in a xenograft mouse model in vivo. Cytotherapy 15:986-98
Aslanidi, George V; Rivers, Angela E; Ortiz, Luis et al. (2012) High-efficiency transduction of human monocyte-derived dendritic cells by capsid-modified recombinant AAV2 vectors. Vaccine 30:3908-17