Go to JCI Insight
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
  • Clinical Research and Public Health
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Gastroenterology
    • Immunology
    • Metabolism
    • Nephrology
    • Neuroscience
    • Oncology
    • Pulmonology
    • Vascular biology
    • All ...
  • Videos
    • Conversations with Giants in Medicine
    • Video Abstracts
  • Reviews
    • View all reviews ...
    • Complement Biology and Therapeutics (May 2025)
    • Evolving insights into MASLD and MASH pathogenesis and treatment (Apr 2025)
    • Microbiome in Health and Disease (Feb 2025)
    • Substance Use Disorders (Oct 2024)
    • Clonal Hematopoiesis (Oct 2024)
    • Sex Differences in Medicine (Sep 2024)
    • Vascular Malformations (Apr 2024)
    • View all review series ...
  • Viewpoint
  • Collections
    • In-Press Preview
    • Clinical Research and Public Health
    • Research Letters
    • Letters to the Editor
    • Editorials
    • Commentaries
    • Editor's notes
    • Reviews
    • Viewpoints
    • 100th anniversary
    • Top read articles

  • Current issue
  • Past issues
  • Specialties
  • Reviews
  • Review series
  • Conversations with Giants in Medicine
  • Video Abstracts
  • In-Press Preview
  • Clinical Research and Public Health
  • Research Letters
  • Letters to the Editor
  • Editorials
  • Commentaries
  • Editor's notes
  • Reviews
  • Viewpoints
  • 100th anniversary
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
Preexisting maternal immunity to AAV but not Cas9 impairs in utero gene editing in mice
John S. Riley, … , Philip W. Zoltick, William H. Peranteau
John S. Riley, … , Philip W. Zoltick, William H. Peranteau
Published June 17, 2024
Citation Information: J Clin Invest. 2024;134(12):e179848. https://doi.org/10.1172/JCI179848.
View: Text | PDF
Research Article Immunology Article has an altmetric score of 6

Preexisting maternal immunity to AAV but not Cas9 impairs in utero gene editing in mice

  • Text
  • PDF
Abstract

In utero gene editing (IUGE) is a potential treatment for inherited diseases that cause pathology before or soon after birth. Preexisting immunity to adeno-associated virus (AAV) vectors and Cas9 endonuclease may limit postnatal gene editing. The tolerogenic fetal immune system minimizes a fetal immune barrier to IUGE. However, the ability of maternal immunity to limit fetal gene editing remains a question. We investigated whether preexisting maternal immunity to AAV or Cas9 impairs IUGE. Using a combination of fluorescent reporter mice and a murine model of a metabolic liver disease, we demonstrated that maternal anti-AAV IgG antibodies were efficiently transferred from dam to fetus and impaired IUGE in a maternal titer–dependent fashion. By contrast, maternal cellular immunity was inefficiently transferred to the fetus, and neither maternal cellular nor humoral immunity to Cas9 impaired IUGE. Using human umbilical cord and maternal blood samples collected from mid- to late-gestation pregnancies, we demonstrated that maternal-fetal transmission of anti-AAV IgG was inefficient in midgestation compared with term, suggesting that the maternal immune barrier to clinical IUGE would be less relevant at midgestation. These findings support immunologic advantages for IUGE and inform maternal preprocedural testing protocols and exclusion criteria for future clinical trials.

Authors

John S. Riley, Valerie L. Luks, Cara L. Berkowitz, Ana Maria Dumitru, Nicole J. Kus, Apeksha Dave, Pallavi Menon, Monique E. De Paepe, Rajan Jain, Li Li, Lorraine Dugoff, Christina Paidas Teefey, Mohamad-Gabriel Alameh, Philip W. Zoltick, William H. Peranteau

×

Figure 1

Preexisting maternal humoral immunity to AAV impairs fetal gene editing.

Options: View larger image (or click on image) Download as PowerPoint
Preexisting maternal humoral immunity to AAV impairs fetal gene editing....
(A) Experimental design. (B) Anti-AAV9 IgG binding antibody (BAb). BAb concentration was assessed by ELISA and compared among the groups using 1-way ANOVA. (C) Maternal-fetal transmission of anti-AAV9 antibodies. At delivery, serum was collected from dams and newborn fetuses, and the concentration of anti-AAV9 IgG and IgM was compared to generate a fetal/maternal ratio. (D) Comparison of maternal and fetal anti-AAV9 BAb and neutralizing antibody (NAb) titer. Notably, neonatal blood collection is prone to hemolysis, which interferes with luciferase-based assays such as NAb titration. (E) Fetal antibody concentration by group. Anti-AAV9 IgG BAb concentrations were assessed in the serum of uninjected offspring at birth and compared among the groups using 1-way ANOVA. (F) Anti-AAV9 IFN-γ enzyme-linked immunosorbent spot (ELISPOT). Spot-forming units (SFU) per million cells were compared for each AAV9 peptide library by 2-way ANOVA. (G) Ultraviolet stereomicroscopy and immunofluorescence microscopy. Livers of unsensitized offspring and AAV8 directly sensitized offspring showed numerous bright green (mG+) cells by ultraviolet stereomicroscopy and immunofluorescence microscopy, confirming successful gene editing. By contrast, livers of AA9 directly and indirectly sensitized offspring showed no mG+ cells, indicating impaired gene editing. (H) Quantification of fetal liver gene editing by flow cytometry. Shown is percent mG+mT–/(mG+mT– + mG–mT+) among live CD45–TER119–CD31–EpCAM–E-cadherin+ offspring hepatocytes. Mean liver editing was statistically equal among unsensitized offspring, AAV8 directly sensitized offspring, and AAV9 indirectly sensitized (serum diluted 1:100) offspring by 1-way ANOVA. Low-level editing was observed among AAV9 indirectly sensitized (1:10) offspring, and no editing was observed among AAV9 directly and indirectly sensitized (undiluted serum) offspring. (I) Fetal liver editing correlated with maternal anti-AAV9 IgG titer. Fetal gene editing was absent when maternal titer was greater than 1:25 and restored when it was less than 1:25. **P < 0.01, ****P < 0.0001.

Copyright © 2025 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

Sign up for email alerts

Posted by 10 X users
Referenced by 1 Bluesky users
9 readers on Mendeley
See more details