TY - JOUR T1 - Investigation of Cas9 antibodies in the human eye. JF - Nat Commun Y1 - 2022 A1 - Toral, Marcus A A1 - Charlesworth, Carsten T A1 - Ng, Benjamin A1 - Chemudupati, Teja A1 - Homma, Shota A1 - Nakauchi, Hiromitsu A1 - Bassuk, Alexander G A1 - Porteus, Matthew H A1 - Mahajan, Vinit B KW - Animals KW - Antibodies KW - CRISPR-Associated Protein 9 KW - CRISPR-Cas Systems KW - Humans KW - Mice KW - Streptococcus pyogenes KW - T-Lymphocytes AB -

Preexisting immunity against Cas9 proteins in humans represents a safety risk for CRISPR-Cas9 technologies. However, it is unclear to what extent preexisting Cas9 immunity is relevant to the eye as it is targeted for early in vivo CRISPR-Cas9 clinical trials. While the eye lacks T-cells, it contains antibodies, cytokines, and resident immune cells. Although precise mechanisms are unclear, intraocular inflammation remains a major cause of vision loss. Here, we used immunoglobulin isotyping and ELISA platforms to profile antibodies in serum and vitreous fluid biopsies from human adult subjects and Cas9-immunized mice. We observed high prevalence of preexisting Cas9-reactive antibodies in serum but not in the eye. However, we detected intraocular antibodies reactive to S. pyogenes-derived Cas9 after S. pyogenes intraocular infection. Our data suggest that serum antibody concentration may determine whether specific intraocular antibodies develop, but preexisting immunity to Cas9 may represent a lower risk in human eyes than systemically.

VL - 13 IS - 1 ER -