Home WakeSpace Scholarship › Electronic Theses and Dissertations

EVALUATING THERAPEUTIC APPROACHES FOR PRENATAL CELL-GENE THERAPY FOR HEMOPHILIA A

Electronic Theses and Dissertations

Item Files

Item Details

abstract
Hemophilia A (HA) is an X-linked inheritable monogenic disorder occurring in 1 in 5000 male births. Mutations in the gene for clotting factor VIII (FVIII) cause defective or absent coagulation activity, which in patients with a severe phenotype can lead to spontaneous bleeds, debilitating hemarthrosis, and life-threatening intracranial hemorrhages. The standard of care is prophylactic infusions of recombinant FVIII which is an expensive lifelong therapy ($20 million lifetime estimate) and not accessible to 75% of patients with HA worldwide. Moreover, 30% of patients who receive the therapy develop inhibitors against the protein. The inaccessibility and complications of current therapy and the severe psychological and economic burden it causes on HA patients, their families and the healthcare system calls attention to the compelling need to develop therapies that would provide a clinical cure for the disease or at the least improve the quality of life of the patients. Currently, there is strong interest in the development of cell-based platforms for FVIII delivery, especially for prenatal therapies as it would allow for safety testing prior to administration. There are two components of the therapy that are critical for a successful outcome: the cell type that can efficiently engraft and produce FVIII and the gene delivery machinery that can effectively insert FVIII transgene in the cells without deleterious effects. The studies in this thesis were performed with the goal of identifying effective and safe gene delivery tools, to develop an optimal cellular platform, and to study the impact of HLA matching on levels of cell engraftment in fetal recipients. First, we evaluated CRISPR/Cas9 as a gene delivery tool that could insert FVIII transgene at a genomic safe harbor with less off-target effects. Second, we compared the engraftment of bone marrow–derived mesenchymal stromal progenitors derived from the mother (haploidentical) and an allogeneic source post in utero transplantation to assess whether allogeneic “off-the-shelf” source can achieve the same efficiency in engraftment as cells which are a better match to the fetal recipient. It was hoped that the completion of these studies would provide us insights that could help us develop optimal cellular platforms to cure HA.
subject
CRISPR/Cas9
Hemophilia A
HLA/MHC Matching
lcoET3
Lentivirus Gene Delivery
Prenatal Cell-Gene Therapy
contributor
Mahesh Ramamurthy, Ritu (author)
Almeida-Porada, Graca (committee chair)
Soker, Shay (committee member)
Walker, Stephen J (committee member)
date
2022-01-15T09:35:31Z (accessioned)
2022 (issued)
degree
Biomedical Engineering (discipline)
2027-01-14 (liftdate)
embargo
2027-01-14 (terms)
identifier
http://hdl.handle.net/10339/99390 (uri)
language
en (iso)
publisher
Wake Forest University
title
EVALUATING THERAPEUTIC APPROACHES FOR PRENATAL CELL-GENE THERAPY FOR HEMOPHILIA A
type
Thesis

Usage Statistics