High-Throughput CRISPR Screening in Hematological Neoplasms

Abstract

CRISPR is becoming an indispensable tool in biological research, revolutionizing diverse fields of medical research and biotechnology. In the last few years, several CRISPR-based genome-targeting tools have been translated for the study of hematological neoplasms. However, there is a lack of reviews focused on the wide uses of this technology in hematology. Therefore, in this review, we summarize the main CRISPR-based approaches of high throughput screenings applied to this field. Here we explain several libraries and algorithms for analysis of CRISPR screens used in hematology, accompanied by the most relevant databases. Moreover, we focus on (1) the identification of novel modulator genes of drug resistance and efficacy, which could anticipate relapses in patients and (2) new therapeutic targets and synthetic lethal interactions. We also discuss the approaches to uncover novel biomarkers of malignant transformations and immune evasion mechanisms. We explain the current literature in the most common lymphoid and myeloid neoplasms using this tool. Then, we conclude with future directions, highlighting the importance of further gene candidate validation and the integration and harmonization of the data from CRISPR screening approaches.

Preclinical Anticipation of On- and Off-Target Resistance Mechanisms to Anti-Cancer Drugs: A Systematic Review

“…In addition, coupling the CRISPR technology with cutting-edge bioinformatic tools has dramatically improved our knowledge on cancer initiation and progression, and also on therapy resistance, by elucidating the role of individual genes, enabling the identification of multiple interacting pathways, determining the part played by non-coding RNAs, and identifying numerous regulatory elements of oncogenes and tumor suppressor genes. The first examples of pathways and targets identified via these approaches and leading to clinical evaluation of novel drugs have now been reported, mainly in hematological tumors [211,217]. This provides hope for major advancements in the understanding and treatment of hematological and solid tumors in the near future.…”

“…There are now a growing number of examples where a CRISPR/Cas-based screen allowed to identify pathways responsible for de novo or acquired drug resistance pathways, including activation of alternative pathways, chromatin reprogramming, and cell lineage switches [126,129,130]. Recently, CRISPR screenings have been performed in patient samples to guide subsequent treatments, mostly in hematological tumors where tumor samples are more easily accessible, but subsequent clinical translation has proven difficult [211].…”

Integrated drug profiling and CRISPR screening identify BCR::ABL1-independent vulnerabilities in chronic myeloid leukemia

“…CRISPR-Cas9 screens have been increasingly used to identify druggable hits to overcome therapy resistance in cancer, 42 including hematological malignancies. 43 They have, however, seldom been applied to study BCR::ABL1-independent TKI resistance in CML. One previous study employed the genome-scale GeCKO v.2 library to investigate mechanisms of imatinib resistance in K562 cells.…”

Relapse after CAR-T cell therapy in B-cell malignancies: challenges and future approaches

“…Moreover, specific knockout of transcription factors, such as transducin-like enhancer of split 4 (TLE4) and IKAROS family zinc finger 2 (IKZF2) based on genome-wide CRISPR screening, promoted CAR-T cell effector function and inhibited CAR-T cell exhaustion (Wang DR et al, 2021). Equally, novel modulator genes and biomarkers of malignant transformation in tumor cells could be identified and used as new therapeutic targets in combination with CAR-T cell therapy to prevent tumor relapse (Ancos-Pintado et al, 2022).…”