Rescuing the cytolytic function of APDS1 patient T cells via TALEN-mediated PIK3CD gene correction

Lucie Poggi; Loïc Chentout; Sabrina Lizot; Alex Boyne; Alexandre Juillerat; Arianna Moiani; Marine Luka; Francesco Carbone; Mickael Ménager; Marina Cavazzana; Philippe Duchateau; Julien Valton; Sven Kracker

doi:10.1016/j.omtm.2023.101133

Rescuing the cytolytic function of APDS1 patient T cells via TALEN-mediated PIK3CD gene correction

Mol Ther Methods Clin Dev. 2023 Oct 10:31:101133. doi: 10.1016/j.omtm.2023.101133. eCollection 2023 Dec 14.

Authors

Lucie Poggi^{1

2}, Loïc Chentout^{1

2}, Sabrina Lizot³, Alex Boyne⁴, Alexandre Juillerat⁴, Arianna Moiani³, Marine Luka^{5

6}, Francesco Carbone^{5

6}, Mickael Ménager^{5

6}, Marina Cavazzana^{1

7}, Philippe Duchateau⁴, Julien Valton³, Sven Kracker^{1

2}

Affiliations

¹ Université de Paris Cité, Imagine Institute, Paris, France.
² Laboratory of Human Lymphohematopoiesis, INSERM UMR 1163, Paris, France.
³ Cellectis, 8 rue de la Croix Jarry, 75013 Paris, France.
⁴ Cellectis, Inc., 430 East 29th Street, New York, NY 10016, USA.
⁵ Université de Paris Cité, Imagine Institute, Laboratory of Inflammatory Responses and Transcriptomic Networks in Diseases, Atip-Avenir Team, INSERM UMR 1163, 75015 Paris, France.
⁶ Labtech Single-Cell@Imagine, Imagine Institute, INSERM UMR 1163, 75015 Paris, France.
⁷ Biotherapy Clinical Investigation Center, Groupe Hospitalier Universitaire Ouest, Assistance Publique-Hôpitaux de Paris, INSERM, Paris, France.

Abstract

Gain-of-function mutations in the PIK3CD gene result in activated phosphoinositide 3-kinase δ syndrome type 1 (APDS1). This syndrome is a life-threatening combined immunodeficiency and today there are neither optimal nor long-term therapeutic solutions for APDS1 patients. Thus, new alternative treatments are highly needed. The aim of the present study is to explore one therapeutic avenue that consists of the correction of the PIK3CD gene through gene editing. Our proof-of-concept shows that TALEN-mediated gene correction of the mutated PIK3CD gene in APDS1 T cells results in normalized phospho-AKT levels in basal and activated conditions. Normalization of PI3K signaling was correlated to restored cytotoxic functions of edited CD8+ T cells. At the transcriptomic level, single-cell RNA sequencing revealed corrected signatures of CD8+ effector memory and CD8+ proliferating T cells. This proof-of-concept study paves the way for the future development of a gene therapy candidate to cure activated phosphoinositide 3-kinase δ syndrome type 1.

Keywords: APDS1; PI3K signaling; TALEN; gene editing; serial killing assay; single-cell RNA sequencing.