Gene Correction for SCID-X1 in Long-term Hematopoietic Stem Cells

Overview

Journal Nat Commun

Specialty Biology

Date 2019 Apr 11

PMID 30967552

Citations 112

Authors

Mara Pavel-Dinu

Volker Wiebking

Beruh T Dejene

Waracharee Srifa

Sruthi Mantri

Carmencita E Nicolas

Ciaran Lee

Gang Bao

Eric J Kildebeck

Niraj Punjya

Camille Sindhu

Matthew A Inlay

Nivedita Saxena

Suk See DeRavin

Harry Malech

Maria Grazia Roncarolo

Kenneth I Weinberg

Matthew H Porteus

Affiliations

Soon will be listed here.

Abstract

Gene correction in human long-term hematopoietic stem cells (LT-HSCs) could be an effective therapy for monogenic diseases of the blood and immune system. Here we describe an approach for X-linked sSevere cCombined iImmunodeficiency (SCID-X1) using targeted integration of a cDNA into the endogenous start codon to functionally correct disease-causing mutations throughout the gene. Using a CRISPR-Cas9/AAV6 based strategy, we achieve up to 20% targeted integration frequencies in LT-HSCs. As measures of the lack of toxicity we observe no evidence of abnormal hematopoiesis following transplantation and no evidence of off-target mutations using a high-fidelity Cas9 as a ribonucleoprotein complex. We achieve high levels of targeting frequencies (median 45%) in CD34 HSPCs from six SCID-X1 patients and demonstrate rescue of lymphopoietic defect in a patient derived HSPC population in vitro and in vivo. In sum, our study provides specificity, toxicity and efficacy data supportive of clinical development of genome editing to treat SCID-Xl.

Citing Articles

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