Human Gene Therapy - April 2023 - 303

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Ex Vivo Gene Delivery to Porcine Cardiac Allografts Using
a Myocardial-Enhanced Adeno-Associated Viral Vector
Michelle Mendiola Pla,1,{ Yuting Chiang,2,{ Antonio Roki,3 Chunbo Wang,3 Franklin H. Lee,3 Matthew F. Smith,3
Ryan T. Gross,3 Jun-Neng Roan,4 Muath Bishawi,1 Amy Evans,5 Lynden E. Gault,6 Sam Ho,6 Carolyn Glass,7
Jacob N. Schroder,1 Paul Lezberg,8 Carmelo A. Milano,1 and Dawn E. Bowles3,*
1Division of Cardiothoracic Surgery, Department of Surgery, 3Division of Surgical Sciences, Department of Surgery, 5Perfusion Services, and 7Department of
Pathology, Duke University Medical Center, Durham, North Carolina, USA.
2Division of Cardiothoracic Surgery, Department of Surgery, Columbia University Medical Center, New York City, New York, USA.
4Division of Cardiovascular Surgery, Department of Surgery, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan,
Taiwan.
6Gift of Hope Organ and Tissue Donor Network, Itasca, Illinois, USA.
8TransMedics, Inc., Andover, Massachusetts, USA.
{Co-first authors.
Presented at the Ninety-fifth American Heart Association Scientific Sessions, Chicago, IL, November 6, 2022.
Transplantation, the gold standard intervention for organ failure, is a clinical field that is ripe for applications of gene
therapy. One of the major challenges in applying gene therapy to this field is the need for a method that achieves
consistent and robust gene delivery to allografts. Normothermic ex vivo perfusion is a growing organ preservation
method and a device for cardiac preservation was recently approved by the Food and Drug Administration (FDA) (Organ
Care System, OCS; TransMedics, Inc., Andover, MA); this device maintains donor hearts in a near physiologic state
while they are transported from the donor to the recipient. This study describes the administration of recombinant adenoassociated
viral vectors (rAAVs) during ex vivo normothermic perfusion for the delivery of transgenes to porcine cardiac
allografts. We utilized a myocardial-enhanced AAV3b variant, SASTG, assessing its transduction efficiency in the OCS
perfusate relative to other AAV serotypes. We describe the use of normothermic ex vivo perfusion to deliver SASTG
carrying the Firefly Luciferase transgene to porcine donor hearts in four heterotopic transplant procedures. Durable and
dose-dependent transgene expression was achieved in the allografts in 30 days, with no evidence of off-target transgene
expression. This study demonstrates the feasibility and efficiency of delivering genes to a large animal allograft utilizing
AAV vectors during ex vivo perfusion. These findings support the idea of gene therapy interventions to enhance
transplantation outcomes.
Keywords: gene therapy, gene delivery, viral vectors, adeno-associated virus, transplantation, ex vivo machine perfusion
INTRODUCTION
INCREASED CLINICAL USE of ex vivo machine perfusion
(EVMP) devices has improved transplant outcomes by
reducing ischemic and hypothermic injury, but this technology
may also introduce a way to effectively deliver
therapeutics to allografts, including gene and cell
therapy.1-4 The recent success of the first porcine-tohuman
cardiac xenotransplantation using a gene-edited
pig highlights the important role that genetically based
interventions may play in the future of transplantation.5
Cardiac transplantation remains the gold standard therapy
for end-stage heart failure; however, transplantation is
*Correspondence: Dr. Dawn E. Bowles, Division of Surgical Sciences, Department of Surgery, Duke University Medical Center, Box 2642, Research Drive, Durham, NC
27710, USA. E-mail: dawn.bowles@duke.edu
HUMAN GENE THERAPY, VOLUME 34, NUMBERS 7 and 8
ª 2023 by Mary Ann Liebert, Inc.
DOI: 10.1089/hum.2022.241 j 303

Human Gene Therapy - April 2023

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