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El-Akabawy N, Rodriguez M, Ramamurthy R, et al. Defining the Optimal FVIII Transgene for Placental Cell-Based Gene Therapy to Treat Hemophilia A. Mol Ther Methods Clin Dev. 2020;17:465-477doi: 10.1016/j.omtm.2020.03.001.
El-Akabawy, N., Rodriguez, M., Ramamurthy, R., Rabah, A., Trevisan, B., Morsi, A., George, S., Shields, J., Meares, D., Farland, A., Atala, A., Doering, C. B., Spencer, H. T., Porada, C. D., & Almeida-Porada, G. (2020). Defining the Optimal FVIII Transgene for Placental Cell-Based Gene Therapy to Treat Hemophilia A. Molecular therapy. Methods & clinical development, 17465-477. https://doi.org/10.1016/j.omtm.2020.03.001
El-Akabawy, Nadia, et al. "Defining the Optimal FVIII Transgene for Placental Cell-Based Gene Therapy to Treat Hemophilia A." Molecular therapy. Methods & clinical development vol. 17 (2020): 465-477. doi: https://doi.org/10.1016/j.omtm.2020.03.001
El-Akabawy N, Rodriguez M, Ramamurthy R, Rabah A, Trevisan B, Morsi A, George S, Shields J, Meares D, Farland A, Atala A, Doering CB, Spencer HT, Porada CD, Almeida-Porada G. Defining the Optimal FVIII Transgene for Placental Cell-Based Gene Therapy to Treat Hemophilia A. Mol Ther Methods Clin Dev. 2020 Mar 14;17:465-477. doi: 10.1016/j.omtm.2020.03.001. eCollection 2020 Jun 12. PMID: 32258210; PMCID: PMC7109377.
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Mol Ther Methods Clin Dev. 2020 Mar 14;17:465-477. doi: 10.1016/j.omtm.2020.03.001. eCollection 2020 Jun 12.

Defining the Optimal FVIII Transgene for Placental Cell-Based Gene Therapy to Treat Hemophilia A.

Molecular therapy. Methods & clinical development

Nadia El-Akabawy, Martin Rodriguez, Ritu Ramamurthy, Andrew Rabah, Brady Trevisan, Alshaimaa Morsi, Sunil George, Jordan Shields, Diane Meares, Andrew Farland, Anthony Atala, Christopher B Doering, H Trent Spencer, Christopher D Porada, Graça Almeida-Porada

Affiliations

  1. Wake Forest Institute for Regenerative Medicine, Fetal Research and Therapy Program, Wake Forest School of Medicine, Winston-Salem, NC 27101, USA.
  2. Zagazig University Faculty of Medicine, Zagazig, Egypt.
  3. Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Atlanta, GA, USA.
  4. Department of Pediatrics, Emory University, Atlanta, GA, USA.
  5. Special Hematology Laboratory, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA.

PMID: 32258210 PMCID: PMC7109377 DOI: 10.1016/j.omtm.2020.03.001

Abstract

The delivery of factor VIII (FVIII) through gene and/or cellular platforms has emerged as a promising hemophilia A treatment. Herein, we investigated the suitability of human placental cells (PLCs) as delivery vehicles for FVIII and determined an optimal FVIII transgene to produce/secrete therapeutic FVIII levels from these cells. Using three PLC cell banks we demonstrated that PLCs constitutively secreted low levels of FVIII, suggesting their suitability as a transgenic FVIII production platform. Furthermore, PLCs significantly increased FVIII secretion after transduction with a lentiviral vector (LV) encoding a myeloid codon-optimized bioengineered FVIII containing high-expression elements from porcine FVIII. Importantly, transduced PLCs did not upregulate cellular stress or innate immunity molecules, demonstrating that after transduction and FVIII production/secretion, PLCs retained low immunogenicity and cell stress. When LV encoding five different bioengineered FVIII transgenes were compared for transduction efficiency, FVIII production, and secretion, data showed that PLCs transduced with LV encoding hybrid human/porcine FVIII transgenes secreted substantially higher levels of FVIII than did LV encoding B domain-deleted human FVIII. In addition, data showed that in PLCs, myeloid codon optimization is needed to increase FVIII secretion to therapeutic levels. These studies have identified an optimal combination of FVIII transgene and cell source to achieve clinically meaningful levels of secreted FVIII.

© 2020 The Author(s).

Keywords: ET3; FVIII; HSQ; cell therapy; codon-optimization; gene therapy; hemophilia A; placental cells

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