Rescue of STAT3 Function in Hyper-IgE Syndrome Using Adenine Base Editing

Abstract

STAT3-hyper IgE syndrome (STAT3-HIES) is a primary immunodeficiency presenting with destructive lung disease along with other symptoms. CRISPR-Cas9-mediated adenine base editors (ABEs) have the potential to correct one of the most common STAT3-HIES causing heterozygous STAT3 mutations (c.1144C>T/p.R382W). As a proof-of-concept, we successfully applied ABEs to correct STAT3 p.R382W in patient fibroblasts and induced pluripotent stem cells (iPSCs). Treated primary STAT3-HIES patient fibroblasts showed a correction efficiency of 29% -7% without detectable off-target effects evaluated through whole-genome and high-throughput sequencing. Compared with untreated patient fibroblasts, corrected single-cell clones showed functional rescue of STAT3 signaling with significantly increased STAT3 DNA-binding activity and target gene expression of CCL2 and SOCS3. Patient-derived iPSCs were corrected with an efficiency of 30% -6% and differentiated to alveolar organoids showing preserved plasticity in treated cells. In conclusion, our results are supportive for ABE-based gene correction as a potential causative treatment of STAT3-HIES.

European Respiratory Society International Congress 2021: highlights from best-abstract awardees

“…As stated at this Congress, their synergistic interaction increases this risk even more and deserves to be more widely studied in the future [18]. For the future in paediatric lung diseases, CRISPR-Cas9 gene editing may represent a therapeutic tool for genetic disease such as cystic fibrosis [19], STAT3 hyper-IgE syndrome [20] and sickle cell disease [21].…”

A novel human monogenic obesity trait: severe early-onset childhood obesity caused by aberrant expression of agouti-signaling protein (ASIP): a case report

“…The generation of induced pluripotent cells SVF cells isolated from adipose tissue of the patient and two control patients, 1 and 10, was performed by the iPSC Core Facility, Institute of Stem Cell Research, Helmholtz Zentrum München, Neuherberg, Germany and pluripotency was con rmed by staining of OCT4, SOX2, LIN28 and NANOG 47 . The iPSCs were cultivated on Geltrex (Thermo Fisher Scienti c) coated cell culture plates in mTeSR TM 1 medium (Stem Cell TM Technologies) at 37 °C and 5% CO 2 .…”

“…Characterization of the pluripotency of patient-derived induced pluripotent stem cells (iPSCs) was performed as monolayers by directed differentiation into the ectodermal 48 , mesodermal 49 and endodermal 50 germ layers for a period of 5 days and subsequent qRT-PCR analyses of marker genes for each of the germ layers 47 . Relative expression levels were calculated using the Delta-Delta Ct method normalized to β-actin.…”

Gene Editing in Pluripotent Stem Cells and Their Derived Organoids

“…Ethical issues have always been unavoidable in the context of gene editing [ 102 ]. CRISPR/Cas9-mediated adenine base editors can correct STAT3 p.R382W in patient-derived iPSCs, providing a potential treatment for STAT3-hyper IgE syndrome; however, for clinical translation, safety and ethical implications still need to be resolved [ 103 ]. For human medical development, ethics should be a priority and appliance stringently monitored, but also not be a stumbling block.…”