Prenatal AAV9-GFP administration in fetal lambs results in transduction of female germ cells and maternal exposure to virus

Beltran Borges; Antonia Varthaliti; Marisa Schwab; Maria T. Clarke; Christopher Pivetti; Nalin Gupta; Cathryn R. Cadwell; Ghiabe Guibinga; Shirley Phillips; Tony Del Rio; Fatih Ozsolak; Denise Imai-Leonard; Lingling Kong; Diana J. Laird; Akos Herzeg; Charlotte J. Sumner; Tippi C. MacKenzie

Molecular Therapy: Methods & Clinical Development (Jun 2024)

Prenatal AAV9-GFP administration in fetal lambs results in transduction of female germ cells and maternal exposure to virus

Beltran Borges,
Antonia Varthaliti,
Marisa Schwab,
Maria T. Clarke,
Christopher Pivetti,
Nalin Gupta,
Cathryn R. Cadwell,
Ghiabe Guibinga,
Shirley Phillips,
Tony Del Rio,
Fatih Ozsolak,
Denise Imai-Leonard,
Lingling Kong,
Diana J. Laird,
Akos Herzeg,
Charlotte J. Sumner,
Tippi C. MacKenzie

Affiliations

Beltran Borges: Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; UCSF Center for Maternal-Fetal Precision Medicine, San Francisco, CA 94158, USA; The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA
Antonia Varthaliti: Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA
Marisa Schwab: Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA
Maria T. Clarke: Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; UCSF Center for Maternal-Fetal Precision Medicine, San Francisco, CA 94158, USA; The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA
Christopher Pivetti: Department of Surgery, University of California, Davis, Davis, CA 95817, USA
Nalin Gupta: Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Pediatrics and Benioff Children’s Hospital, University of California, San Francisco, San Francisco, CA 94158, USA
Cathryn R. Cadwell: Department of Neurological Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Pathology, University of California, San Francisco, San Francisco, CA 94143, USA; Weill Neurohub, University of California, San Francisco, San Francisco, CA 94158, USA
Ghiabe Guibinga: Novartis Institutes for BioMedical Research Biologics Center, San Diego, CA 92121, USA
Shirley Phillips: Novartis Institutes for BioMedical Research Biologics Center, San Diego, CA 92121, USA
Tony Del Rio: Novartis Institutes for BioMedical Research Biologics Center, San Diego, CA 92121, USA
Fatih Ozsolak: Novartis Institutes for BioMedical Research Biologics Center, San Diego, CA 92121, USA
Denise Imai-Leonard: Comparative Pathology Laboratory, University of California, Davis, Davis, CA 95616, USA
Lingling Kong: Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21218, USA
Diana J. Laird: The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Obstetrics and Gynecology, University of California, San Francisco, San Francisco, CA 94158, USA
Akos Herzeg: Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; UCSF Center for Maternal-Fetal Precision Medicine, San Francisco, CA 94158, USA; The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA
Charlotte J. Sumner: Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21218, USA; Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21218, USA; Department of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21218, USA
Tippi C. MacKenzie: Department of Surgery, University of California, San Francisco, San Francisco, CA 94143, USA; UCSF Center for Maternal-Fetal Precision Medicine, San Francisco, CA 94158, USA; The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Pediatrics and Benioff Children’s Hospital, University of California, San Francisco, San Francisco, CA 94158, USA; Department of Obstetrics and Gynecology, University of California, San Francisco, San Francisco, CA 94158, USA; Corresponding author: Tippi C. MacKenzie, MD UCSF School of Medicine Regeneration Medicine, Building 35 Medical Center Way RMB 900E, Campus Box 0665, San Francisco, CA, USA.

Journal volume & issue: Vol. 32, no. 2
p. 101263

Abstract

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Prenatal somatic cell gene therapy (PSCGT) could potentially treat severe, early-onset genetic disorders such as spinal muscular atrophy (SMA) or muscular dystrophy. Given the approval of adeno-associated virus serotype 9 (AAV9) vectors in infants with SMA by the U.S. Food and Drug Administration, we tested the safety and biodistribution of AAV9-GFP (clinical-grade and dose) in fetal lambs to understand safety and efficacy after umbilical vein or intracranial injection on embryonic day 75 (E75) . Umbilical vein injection led to widespread biodistribution of vector genomes in all examined lamb tissues and in maternal uteruses at harvest (E96 or E140; term = E150). There was robust GFP expression in brain, spinal cord, dorsal root ganglia (DRGs), without DRG toxicity and excellent transduction of diaphragm and quadriceps muscles. However, we found evidence of systemic toxicity (fetal growth restriction) and maternal exposure to the viral vector (transient elevation of total bilirubin and a trend toward elevation in anti-AAV9 antibodies). There were no antibodies against GFP in ewes or lambs. Analysis of fetal gonads demonstrated GFP expression in female (but not male) germ cells, with low levels of integration-specific reads, without integration in select proto-oncogenes. These results suggest potential therapeutic benefit of AAV9 PSCGT for neuromuscular disorders, but warrant caution for exposure of female germ cells.

Published in Molecular Therapy: Methods & Clinical Development

ISSN: 2329-0501 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Biology (General): Genetics; Science: Biology (General): Cytology
Website: http://www.cell.com/molecular-therapy-family/methods/latest-content

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