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CRISPR-Cas9 system: a novel and promising era of genotherapy for beta-hemoglobinopathies, hematological malignancy, and hemophilia

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Abstract

Gene therapy represents a significant potential to revolutionize the field of hematology with applications in correcting genetic mutations, generating cell lines and animal models, and improving the feasibility and efficacy of cancer immunotherapy. Compared to different genetic engineering tools, clustered regularly interspaced short palindromic repeats (CRISPR) CRISPR-associated protein 9 (Cas9) emerged as an effective and versatile genetic editor with the ability to precisely modify the genome. The applications of genetic engineering in various hematological disorders have shown encouraging results. Monogenic hematological disorders can conceivably be corrected with single gene modification. Through the use of CRISPR-CAS9, restoration of functional red blood cells and hemostasis factors were successfully attained in sickle cell anemia, beta-thalassemia, and hemophilia disorders. Our understanding of hemato-oncology has been advanced via CRIPSR-CAS9 technology. CRISPR-CAS9 aided to build a platform of mutated genes responsible for cell survival and proliferation in leukemia. Therapeutic application of CRISPR-CAS9 when combined with chimeric antigen receptor (CAR) T cell therapy in multiple myeloma and acute lymphoblastic leukemia was feasible with attenuation of CAR T cell therapy pitfalls. Our review outlines the latest literature on the utilization of CRISPR-Cas9 in the treatment of beta-hemoglobinopathies and hemophilia disorders. We present the strategies that were employed and the findings of preclinical and clinical trials. Also, the review will discuss gene engineering in the field of hemato-oncology as a proper tool to facilitate and overcome the drawbacks of chimeric antigen receptor T cell therapy (CAR-T).

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Data sharing is not applicable to this article as no datasets were generated or analyzed during the current study.

Abbreviations

DNA :

deoxyribonucleic acid

ZFN :

zinc finger nuclease

TALEN :

transcription activator-like effector nuclease

CRISPR :

clustered regulatory interspaced short palindromic repeats

Cas9 :

CRISPR-associated protein 9

DSB :

DNA double-strand break

NHEJ :

non-homologous end joining

HDR :

homology-directed repair

INDELs :

insertion and deletion mutations

RNA :

ribonucleic acid

SCD :

sickle cell disease

HSCT :

hematopoietic stem cell transplantation

Cr-RNA :

CRISPR RNA

Tracr-RNA :

transactivating crRNA

PAM :

protospacer adjacent motif

sgRNA :

single-strand small-guide RNA

CAR :

chimeric antigen receptor

dCas9 :

deactivated Cas9

RT :

reverse transcriptase

pegRNA :

prime-editing single guide RNA

SCA :

sickle cell anemia

TM :

thalassemia major

HbS :

hemoglobin S

HbF :

fetal hemoglobin

HbA :

adult hemoglobin

HPFH :

hereditary persistence of fetal hemoglobin

IPSCs :

induced pluripotent stem cells

UTR :

untranslated region

FDA :

Food and Drug Administration

HLA :

human leukocyte antigens

TRAC :

T-cell alpha receptor

PD-1 :

programmed cell death protein 1

GvH :

graft-versus-host

TCR-B :

T cell receptor beta

ASXL1 :

additional sex combs-like 1

AML :

acute myeloid leukemia

CML :

chronic myeloid leukemia

PRC2 :

polycomb repressive complex 2

VRPEB1 :

V-set pre-B-cell surrogate light chain 1

MM :

multiple myeloma

BDDF8 :

B domain-deleted factor 8

EF1α :

human elongation factor 1 alpha

ssODN :

single-stranded-oligodeoxynucleotide

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Medicine, College of Medicine, Taibah University, Madinah, Saudi Arabia

    Abdulfatah M. Alayoubi

  2. College of Medicine, Taibah University, Madinah, Saudi Arabia

    Zakaria Y. Khawaji & Mohammed A. Mohammed

  3. Divisions of Experimental Medicine & Hematology, Department of Medicine, Faculty of Medicine, McGill University, Montreal, Quebec, Canada

    François E. Mercier

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The review topic was suggested by ZK and MM. The manuscript idea was reviewed and expanded by AA. Manuscript outlines were laid out by AA. Literature review was done by ZK, MM, AA, and FM. First draft was created by ZK and MM. Subsequent version was produced by AA. Revision and enhancement was done by FM.

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Correspondence to Zakaria Y. Khawaji.

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Alayoubi, A.M., Khawaji, Z.Y., Mohammed, M.A. et al. CRISPR-Cas9 system: a novel and promising era of genotherapy for beta-hemoglobinopathies, hematological malignancy, and hemophilia. Ann Hematol 103, 1805–1817 (2024). https://doi.org/10.1007/s00277-023-05457-2

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