Our Science: Publications
Graphite Bio’s scientific approach is supported by extensive peer-reviewed preclinical research. Explore select publications featuring our novel targeted integration platform below.
Sickle Cell Disease (SCD)
This study details the successful correction of the mutation that causes sickle cell disease (SCD) in patient-derived hematopoietic stem and progenitor cells using the CRISPR/Cas9 system with AAV6-mediated donor delivery. These findings provide early support for gene correction-based therapies and its potential to cure SCD by targeting the causative mutation.
This study describes the development of a streamlined protocol that optimizes hematopoietic stem cell editing at any gene of interest, enabling precise single-nucleotide genome changes or longer gene cassette introduction. Selective gene editing via this protocol may be utilized in hematopoietic stem cell transplantation-based therapies to correct underlying gene mutations that are the cause of a broad range of genetic diseases.
This manuscript describes the identification of a HiFi Cas9 variant that induces robust AAV6-mediated correction of the HBB gene in SCD patient-derived hematopoietic stem and progenitor cells, while significantly reducing off-target editing compared to wild-type Cas9. The HiFi Cas9 variant identified in this study has broad potential applicability in CRISPR/Cas9 genome editing approaches including our HBB gene editing program to significantly reduce off-target editing.
This study demonstrates proof of concept for precise, CRISPR/Cas9 mediated gene correction in a murine model of sickle cell disease (SCD). The authors demonstrate that engraftment of gene-corrected HSCs resulted in improvement of underlying SCD pathophysiology in this preclinical study. These findings show that CRISPR/Cas9-AAV6 gene correction therapies have the potential to provide a curative treatment for SCD and support their further clinical investigation.
This Science Translational Medicine publication presents the preclinical research and key scale-up processes enabling the clinical translation of a first-in-human gene correction strategy for SCD. Study findings demonstrate the safety, efficacy, and reproducibility of a CRISPR/Cas9-AAV6 based gene correction strategy to support initiation of a Phase 1/2 clinical trial for SCD patients.
This paper highlights the ability of the CRISPR/Cas9-AAV6 system to enable the correction of an entire gene, while preserving endogenous regulatory control. In this proof-of-concept study, researchers focused on x-linked severe combined immunodeficiency (XSCID), an inherited disorder that can result from one of a number of previously identified loss-of-function mutations throughout the IL2RG gene. By demonstrating the efficiency of the CRISPR/Cas9-AAV6 platform to integrate a complete IL2RG cDNA into patient-derived hematopoietic stem and progenitor cells, findings of this preclinical study provide early efficacy data for the use of this novel genome-editing approach to treat XSCID patients.
This study showcases the potential for using the CRISPR/Cas9-AAV6 system to introduce gene expression cassettes into hematopoietic stem and progenitor cells, resulting in lineage-specific expression of therapeutic proteins. The authors utilized this system to overexpress macrophage-specific glucocerebrosidase (GCase) within the CCR5 gene, a validated safe harbor locus, providing in vitro and in vivo proof-of-concept for a possible treatment for Gaucher disease. Gaucher disease is a lysosomal storage disorder that is caused by GCase deficiency and accumulation of glycolipids in specific cell types, especially macrophages.
This study demonstrates the successful correction of the loss-of-function mutations throughout the HBB gene that cause β-thalassemia by using the CRISPR/Cas9-AAV6 system to replace the entire coding region of the HBA1 gene with an HBB transgene at high frequencies in patient-derived hematopoietic stem and progenitor cells. These data demonstrate that a single Cas9-induced double-stranded break can mediate replacement of an entire endogenous gene with an exogenous transgene at high frequencies, and that this technology may be utilized to correct the loss-of-function mutations that are the cause of a broad range of genetic diseases.