Item Details

title

DEFINING THE SAFETY AND EFFICACY OF AAV GENE THERAPY USING HUMAN ORGAN TISSUE EQUIVALENTS

author

Mahesh Ramamurthy, Ritu

abstract

Adeno-associated virus (AAV)-based gene therapies have emerged as a promising therapeutic modality to treat several monogenic disorders, including hemophilia A. However, current preclinical animal models often fail to predict clinical outcomes accurately due to significant biological differences between species, creating a major roadblock to AAV gene therapy and a demand for more human-specific models. Human-derived 3D organoids or organ tissue equivalents (OTEs) can serve as potential tools in overcoming the knowledge gap by more accurately predicting therapeutic response. These OTEs are fabricated using major cell types derived from their respective native tissue and can more accurately recapitulate their biology and function. Human liver tissue equivalents were employed to study the two clinically-relevant serotypes, AAV5 and AAV3b. We performed detailed analysis of transduction efficiency, cell tropism, inflammation/ hepatotoxicity/fibrosis, liver function biomarker alterations, and genotoxicity. Having established the utility of human liver tissue equivalents (hLTEs) in predicting response to liver-directed AAV gene therapy, we applied the model to study an AAV5-based gene therapy targeting the liver for the treatment of hemophilia A, involving a bioengineered FVIII transgene “lcoET3.” We performed analysis of vector copy number, transgene mRNA and protein expression, and, more importantly, FVIII transgene-specific response in the liver. Finally, given the lack of definitive data on true-human tropism, liver, BBB, lung, and testicular OTEs were utilized to define the tropism of AAV5 and AAV3b. Overall, the findings of this dissertation demonstrate the potential of OTEs to serve as critical tools in bridging the translational gap, accelerating preclinical studies, and paving the way for more efficient and safer AAV gene therapies.

subject

AAV

Gene Therapy

Hemophilia A

Human Organoid Models

Liver Organoid

Tropism

contributor

Almeida-Porada, Graca (advisor)

Hall, Adam R (committee member)

Seeds, Michael (committee member)

Murphy, Sean V (committee member)

Soker, Shay (committee member)

date

2024-09-13T08:36:35Z (accessioned)

2024 (issued)

degree

Biomedical Engineering (discipline)

embargo

2029-09-05 (terms)

2029-09-05 (liftdate)

identifier

http://hdl.handle.net/10339/109852 (uri)

language

en (iso)

publisher

Wake Forest University

type

Dissertation