AAV9-mediated Gene Delivery to Liver Grafts During Static Cold Storage in a Rat Liver Transplant Model

Overview

Journal Front Transplant

Specialty General Surgery

Date 2024 Jul 12

PMID 38993865

Authors

Qimeng Gao

Samuel J Kesseli

Trevor Gonzalez

Min Zhang

Riley Kahan

Madison Krischak

Samantha E Halpern

Mingqing Song

Hongzhi Xu

Nader Abraham

Imran J Anwar

Isaac Alderete

Aravind Asokan

Matthew G Hartwig

Andrew S Barbas

Affiliations

Soon will be listed here.

Abstract

Introduction: Recombinant adeno-associated virus (rAAV) is a novel strategy used clinically for gene delivery, but has not been characterized in the context of organ transplantation. We sought to determine the efficacy of rAAV-mediated gene delivery during static cold storage (SCS) prior to liver transplantation.

Methods: A triple-plasmid transfection protocol was used to produce rAAV subtype-9 vectors containing firefly luciferase genomes in HEK293 cells. Lewis rat liver grafts were flushed and stored in cold HTK solution. Three experimental groups received rAAV at different doses, administered via the portal vein as a bolus during SCS. A control group did not receive rAAV ( = 2). Recipients then underwent syngeneic liver transplantation. Bioluminescence imaging to quantify luciferase expression was performed on post-operative days 7, 14, 28, and 56.

Results: Control animals demonstrated no bioluminescent activity, while animals receiving rAAV-treated livers had increasing bioluminescence, peaking at four weeks but sustained to the eight-week endpoint. This result was confirmed by experimental endpoint tissue luciferase activity assay.

Discussion: rAAV mediates gene transduction in liver grafts when administered during SCS and has potential for gene therapy applications in solid organ transplantation.

References

Svensson E, Marshall D, Woodard K, Lin H, Jiang F, Chu L . Efficient and stable transduction of cardiomyocytes after intramyocardial injection or intracoronary perfusion with recombinant adeno-associated virus vectors. Circulation. 1999; 99(2):201-5. DOI: 10.1161/01.cir.99.2.201. View

Ertl H . Immunogenicity and toxicity of AAV gene therapy. Front Immunol. 2022; 13:975803. PMC: 9411526. DOI: 10.3389/fimmu.2022.975803. View

Wang D, Tai P, Gao G . Adeno-associated virus vector as a platform for gene therapy delivery. Nat Rev Drug Discov. 2019; 18(5):358-378. PMC: 6927556. DOI: 10.1038/s41573-019-0012-9. View

Benigni A, Tomasoni S, Remuzzi G . Impediments to successful gene transfer to the kidney in the context of transplantation and how to overcome them. Kidney Int. 2002; 61(1 Suppl):S115-9. DOI: 10.1046/j.1523-1755.2002.0610s1115.x. View

Kesseli S, Krischak M, Gao Q, Gonzalez T, Zhang M, Halpern S . Adeno-associated virus mediates gene transduction after static cold storage treatment in rodent lung transplantation. J Thorac Cardiovasc Surg. 2024; 166(1):e38-e49. DOI: 10.1016/j.jtcvs.2022.08.050. View

Csete M, Drazan K, Van Bree M, McIntee D, McBride W, Bett A . Adenovirus-mediated gene transfer in the transplant setting. I. Conditions for expression of transferred genes in cold-preserved hepatocytes. Transplantation. 1994; 57(10):1502-7. View

Yang Z, Wu X, Tsui T, Hou Y, Luk J, Fan S . Recombinant adeno-associated virus vector: Is it ideal for gene delivery in liver transplantation?. Liver Transpl. 2003; 9(4):411-20. DOI: 10.1053/jlts.2003.50058. View

Shaked A, Csete M, Drazan K, Bullington D, Wu L, Busuttil R . Adenovirus-mediated gene transfer in the transplant setting. II. Successful expression of transferred cDNA in syngeneic liver grafts. Transplantation. 1994; 57(10):1508-11. View

Nathwani A, Reiss U, Tuddenham E, Rosales C, Chowdary P, McIntosh J . Long-term safety and efficacy of factor IX gene therapy in hemophilia B. N Engl J Med. 2014; 371(21):1994-2004. PMC: 4278802. DOI: 10.1056/NEJMoa1407309. View

10.

Abraham N, Zhang M, Cray P, Gao Q, Samy K, Neill R . Two Compartment Evaluation of Liver Grafts During Acellular Room Temperature Machine Perfusion (acRTMP) in a Rat Liver Transplant Model. Front Med (Lausanne). 2022; 9:804834. PMC: 8907827. DOI: 10.3389/fmed.2022.804834. View

11.

Bonaccorsi-Riani E, Gillooly A, Bruggenwirth I, Martins P . Delivery of genetic load during ex situ liver machine perfusion with potential for CRISPR-Cas9 gene editing: An innovative strategy for graft treatment. Hepatobiliary Pancreat Dis Int. 2021; 20(5):503-505. DOI: 10.1016/j.hbpd.2021.04.006. View

12.

Raper S, Chirmule N, Lee F, Wivel N, Bagg A, Gao G . Fatal systemic inflammatory response syndrome in a ornithine transcarbamylase deficient patient following adenoviral gene transfer. Mol Genet Metab. 2003; 80(1-2):148-58. DOI: 10.1016/j.ymgme.2003.08.016. View

13.

. High-dose AAV gene therapy deaths. Nat Biotechnol. 2020; 38(8):910. DOI: 10.1038/s41587-020-0642-9. View

14.

Hinderer C, Katz N, Buza E, Dyer C, Goode T, Bell P . Severe Toxicity in Nonhuman Primates and Piglets Following High-Dose Intravenous Administration of an Adeno-Associated Virus Vector Expressing Human SMN. Hum Gene Ther. 2018; 29(3):285-298. PMC: 5865262. DOI: 10.1089/hum.2018.015. View

15.

Yeung J, Wagnetz D, Cypel M, Rubacha M, Koike T, Chun Y . Ex vivo adenoviral vector gene delivery results in decreased vector-associated inflammation pre- and post-lung transplantation in the pig. Mol Ther. 2012; 20(6):1204-11. PMC: 3369301. DOI: 10.1038/mt.2012.57. View

16.

Hori T, Nguyen J, Zhao X, Ogura Y, Hata T, Yagi S . Comprehensive and innovative techniques for liver transplantation in rats: a surgical guide. World J Gastroenterol. 2010; 16(25):3120-32. PMC: 2896749. DOI: 10.3748/wjg.v16.i25.3120. View

17.

George L, Sullivan S, Giermasz A, Rasko J, Samelson-Jones B, Ducore J . Hemophilia B Gene Therapy with a High-Specific-Activity Factor IX Variant. N Engl J Med. 2017; 377(23):2215-2227. PMC: 6029626. DOI: 10.1056/NEJMoa1708538. View

18.

Olthoff K, Judge T, Gelman A, da Shen X, Hancock W, Turka L . Adenovirus-mediated gene transfer into cold-preserved liver allografts: survival pattern and unresponsiveness following transduction with CTLA4Ig. Nat Med. 1998; 4(2):194-200. DOI: 10.1038/nm0298-194. View

19.

Cypel M, Liu M, Rubacha M, Yeung J, Hirayama S, Anraku M . Functional repair of human donor lungs by IL-10 gene therapy. Sci Transl Med. 2010; 1(4):4ra9. DOI: 10.1126/scitranslmed.3000266. View