Genetic engineering of hematopoiesis: current stage of clinical translation and future perspectives

Naldini, Luigi

doi:10.15252/emmm.201809958

Cited by 91 publications

(86 citation statements)

References 108 publications

(182 reference statements)

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“…The application of genome editing to SCID and other PIDs is obviously an attractive option, because the end product would (in most cases) result from expression of the right gene in the right place, i.e., in its physiological environment (Porteus, 2019; Naldini, 2019). This is important for both safety and efficacy.…”

Section: Conclusion and The Futurementioning

confidence: 99%

Gene therapy for severe combined immunodeficiencies and beyond

Fischer

Hacein‐Bey‐Abina

2019

Journal of Experimental Medicine

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Ex vivo retrovirally mediated gene therapy has been shown within the last 20 yr to correct the T cell immunodeficiency caused by γc-deficiency (SCID X1) and adenosine deaminase (ADA) deficiency. The rationale was brought up by the observation of the revertant of SCIDX1 and ADA deficiency as a kind of natural gene therapy. Nevertheless, the first attempts of gene therapy for SCID X1 were associated with insertional mutagenesis causing leukemia, because the viral enhancer induced transactivation of oncogenes. Removal of this element and use of a promoter instead led to safer but still efficacious gene therapy. It was observed that a fully diversified T cell repertoire could be generated by a limited set (<1,000) of progenitor cells. Further advances in gene transfer technology, including the use of lentiviral vectors, has led to success in the treatment of Wiskott–Aldrich syndrome, while further applications are pending. Genome editing of the mutated gene may be envisaged as an alternative strategy to treat SCID diseases.

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Section: Conclusion and The Futurementioning

confidence: 99%

Gene therapy for severe combined immunodeficiencies and beyond

Fischer

Hacein‐Bey‐Abina

2019

Journal of Experimental Medicine

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“…New precision techniques based on gene editing are being developed preclinically and have been recently applied in vitro and in vivo through the use of specific endonucleases . Although very promising, many aspects related to the safety and effectiveness of gene editing have yet to be verified.…”

Section: Introductionmentioning

confidence: 99%

“…With gene editing, genes can be targeted and corrected in situ by enzymes that catalyze the breaking of the double helix of DNA. This can be achieved through meganucleases, Zinc‐finger nucleases (ZFN), transcriptor activator‐like effector nucleases (TALENs) and clustered regulatory interspaced short palindromic repeats CRISPR‐associated (CRISPR/Cas) based RNA‐guided DNA endonucleases …”

Section: Introductionmentioning

confidence: 99%

“…An advantage of gene editing is the gene‐targeted correction maintaining the gene expression by endogenous, specific, and physiological regulatory elements. This should avoid the vectors’ potential risk of insertional mutagenesis; however, surveillance on potential risks of cutting the DNA with nucleases with potential recombination outside the target locus will need to be verified over time, before this strategy could be applicable in clinics …”

Section: Introductionmentioning

confidence: 99%

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New perspectives in gene therapy for inherited disorders

Cicalese

Aiuti

2020

Pediatric Allergy Immunology

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Gene therapy has become promising in many fields of medicine, as a single treatment could allow long-lasting and curative benefits. New medicines based on cell gene correction are expected to occur in upcoming years and will be hopefully part of the therapeutic armamentarium for inherited and acquired diseases. Issues related to the costs of these new therapies and access to care for all patients, and procedures and expertise needed to facilitate their application worldwide require to be addressed, together with long-term safety and efficacy monitoring. K E Y W O R D S cancer, gene therapy, immunodeficiency, inherited disordersHow to cite this article: Cicalese MP, Aiuti A. New perspectives in gene therapy for inherited disorders. Pediatr Allergy Immunol. 2020;31(Suppl 24):5-7. https ://doi.

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“…Because infused hCD34+ HSPC dose correlates with time to engraftment and this is limited in UCB, efforts to enhance HSPCs in UCB units have included appropriate donor UCB selection, the use of a second UCB or haploidentical hematopoietic stem cell (HSC) donation for HSCTs, ex vivo UCB HSPC expansion before transplant and up regulating homing and/or bone marrow retention cell surface molecules on the UCB HSPCs . In the latter two instances, thawed UCB with or without hCD34+ or hCD133+ HSPC enrichment are cultured for relatively short periods of time (typically 12 h to 21 days) in serum‐free, cytokine and/or small molecule–supplemented media . In the majority of cases, isolation of hCD133+ or hCD34+ HSPCs from fresh UCB for preclinical studies has achieved high purities, viabilities, and recoveries .…”

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confidence: 99%

A modified CD34+ hematopoietic stem and progenitor cell isolation strategy from cryopreserved human umbilical cord blood

et al. 2019

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BACKGROUND: Umbilical cord blood (UCB) is a source of hematopoietic stem cells for transplantation, offering an alternative for patients unable to find a matched adult donor. UCB is also a versatile source of hematopoietic stem and progenitor cells (hCD34 + HSPCs) for research into hematologic diseases, in vitro expansion, ex vivo gene therapy, and adoptive immunotherapy. For these studies, there is a need to isolate hCD34 + HSPCs from cryopreserved units, and protocols developed for isolation from fresh cord blood are unsuitable. STUDY DESIGN:This study describes a modified method for isolating hCD34 + HSPCs from cryopreserved UCB. It uses the Plasmatherm system for thawing, followed by CD34 microbead magneticactivated cell sorting isolation with a cell separation kit (Whole Blood Columns, Miltenyi Biotec). hCD34 + HSPC phenotypes and functionality were assessed in vitro and hematologic reconstitution determined in vivo in immunodeficient mice. RESULTS:Total nucleated cell recovery after thawing and washing was 44.7 AE 11.7%. Recovery of hCD34 + HSPCs after application of thawed cells to Whole Blood Columns was 77.5 AE 22.6%. When assessed in two independent laboratories, the hCD34+ cell purities were 71.7 AE 10.7% and 87.8 AE 2.4%. Transplantation of the enriched hCD34 + HSPCs into NSG mice revealed the presence of repopulating hematopoietic stem cells (estimated frequency of 0.07%) and multilineage engraftment.CONCLUSION: This provides a simplified protocol for isolating high-purity human CD34 + HSPCs from banked UCB adaptable to current Good Manufacturing Practice. This protocol reduces the number of steps and associated risks and thus total production costs. Importantly, the isolated CD34 + HSPCs possess in vivo repopulating activity in immunodeficient mice, making them a suitable starting population for ex vivo culture and gene editing. ABBREVIATIONS: BMRs = batch manufacturing records; CFUs = colony-forming units; cGMP = current good manufacturing practice; HSC = hematopoietic stem cell; HSCT = hematopoietic stem cell transplantation; HSPCs = hematopoietic stem and progenitor cells; LMPP = lymphoid-primed multipotent; MPP = Multipotent progenitors; SOPs = standard operating procedures; SRCs = severe combined immunodeficiency repopulating cells; TNCs = total nucleated cells; UCB = umbilical cord blood.

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Genetic engineering of hematopoiesis: current stage of clinical translation and future perspectives

Abstract: Here I review the scientific background, current stage of development and future perspectives that I foresee in the field of genetic manipulation of hematopoietic stem cells with a special emphasis on clinical applications.

Cited by 91 publications

References 108 publications

Gene therapy for severe combined immunodeficiencies and beyond

Gene therapy for severe combined immunodeficiencies and beyond

New perspectives in gene therapy for inherited disorders

A modified CD34+ hematopoietic stem and progenitor cell isolation strategy from cryopreserved human umbilical cord blood

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