CRISPR-based rapid molecular diagnostic tests for fusion-driven leukemias

• Published in: Blood Vol. 144; no. 12; pp. 1290 - 1299
• Main Authors: Vedula, Rahul S., Karp, Hannah Q., Koob, Jeremy, Lim, Felicia, Garcia, Jacqueline S., Winer, Eric S., Luskin, Marlise R., Ghiaur, Gabriel, Kim, Annette S., Beppu, Lan W., Sala-Torra, Olga, Radich, Jerald, Gootenberg, Jonathan, Abudayyeh, Omar, Zhang, Feng, Lindsley, R. Coleman
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 19.09.2024
• Subjects: Clustered Regularly Interspaced Short Palindromic Repeats; CRISPR-Cas Systems; Humans; Leukemia - diagnosis; Leukemia - genetics; Leukemia - therapy; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - diagnosis; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - genetics; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - therapy; Leukemia, Promyelocytic, Acute - diagnosis; Leukemia, Promyelocytic, Acute - genetics; Leukemia, Promyelocytic, Acute - therapy; Molecular Diagnostic Techniques - methods; Oncogene Proteins, Fusion - genetics
• ISSN: 0006-4971; 1528-0020; 1528-0020
• DOI: 10.1182/blood.2023022908

•SHERLOCK is 100% sensitive and specific for the diagnosis of APL and CML in patient samples.•SHERLOCK has the potential to improve access to life-saving therapies for clinically actionable fusion-driven leukemias. [Display omitted] Fusion oncogenes can be cancer-defining molecular alterations that are essential for diagnosis and therapy selection.1,2 Rapid and accessible molecular diagnostics for fusion-driven leukemias such as acute promyelocytic leukemia (APL), Philadelphia chromosome–positive acute lymphoblastic leukemia, and chronic myeloid leukemia (CML) are unavailable, creating a barrier to timely diagnosis and effective targeted therapy in many health care settings, including community hospitals and low-resource environments. We developed CRISPR-based RNA-fusion transcript detection assays using SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) for the diagnosis of fusion-driven leukemias. We validated these assays using diagnostic samples from patients with APL and CML from academic centers and dried blood spots from low-resource environments, demonstrating 100% sensitivity and specificity. We identified assay optimizations to enable the use of these tests outside of tertiary cancer centers and clinical laboratories, enhancing the potential impact of this technology. Rapid point-of-care diagnostics can improve outcomes for patients with cancer by expanding access to therapies for highly treatable diseases that would otherwise lead to serious adverse outcomes due to delayed or missed diagnoses.