Therapeutic genome editing for hereditary connective tissue disorders Lauren Testa

Format:

Book

Thesis/Dissertation

Author/Creator:

Testa, Lauren, author.

Contributor:

University of Pennsylvania. Cell and Molecular Biology., degree granting institution.

Language:

English

Subjects (All):

Genetics.

Therapy.

Molecular biology.

0369.

0212.

0307.

Local Subjects:

Genetics.

Therapy.

Molecular biology.

0369.

0212.

0307.

Genre:

Academic theses

Physical Description:

1 online resource (134 pages)

Contained In:

Dissertations Abstracts International 87-12B

Place of Publication:

Ann Arbor : ProQuest Dissertations and Theses, 2026

Language Note:

English

Summary:

Pseudoxanthoma elasticum (PXE) and Marfan syndrome (MFS) are two prototypical hereditary connective tissue disorders caused by pathogenic variants in ABCC6 and FBN1, respectively. While both conditions exhibit distinct clinical manifestations and mechanisms of disease, both are characterized by deficiency of elastic fibers within the extracellular matrix of multiple organ systems. Elastic fiber fragmentation of MFS and elastic fiber calcification of PXE can lead to multisystemic pathology, including arterial disease, a main contributor of morbidity and mortality in both disorders. There are no targeted therapies for these conditions. Genetic therapies (base, prime, and epigenome editing) may offer durable, one-and-done protection against one or many of their clinical manifestations. Here, we used three different CRISPR-based strategies to advance therapeutic development for Marfan syndrome and PXE. We used adenine base editing with a DNA/RNA hybrid guide RNA to correct the R1164X pathogenic variant in ABCC6 in the liver with reduced bystander and off-target editing compared to a traditional guide RNA, which prevented ectopic calcification and restored a blood biomarker in variant-humanized PXE mice. We applied this approach toward the ABCC6 R1141X variant, the most prevalent variant in PXE patients. We used epigenome editing to methylate the promoter and silence expression of the AGT gene, a gene that contributes to aortic aneurysm progression in Marfan syndrome by crosstalk with TGFβ signaling. While CpG dinucleotide methylation within promoters is generally stable across cell divisions, we showed that CpG methylation of the AGT promoter is reversed over time, which provided valuable insight into AGT promoter methylation dynamics. We developed and characterized a novel murine model of Marfan syndrome harboring a frameshift variant in Fbn1 and used prime editing delivered via an AAV with a capsid engineered to target vascular smooth muscle cells to correct the frameshift variant. These three approaches each had strengths and limitations, but all contributed toward a deeper understanding of the biology of hereditary connective tissue disorders and how best to develop therapies for these debilitating conditions

Notes:

Source: Dissertations Abstracts International, Volume: 87-12, Section: B.

Advisors: Prosser, Benjamin Committee members: Ahrens-Nicklas, Rebecca; Bhoj, Elizabeth; Volk, Susan

Ph.D. University of Pennsylvania 2026

Vendor supplied data

Local Notes:

School code: 0175

ISBN:

9798247972624

Access Restriction:

Restricted for use by site license