In Vivo Tracking of Human Hematopoiesis Reveals Patterns of Clonal Dynamics during Early and Steady-State Reconstitution Phases

Biasco, Luca; Pellin, Danilo; Scala, Serena; Dionisio, Francesca; Basso-Ricci, Luca; Leonardelli, Lorena; Scaramuzza, Samantha; Baricordi, Cristina; Ferrua, Francesca; Cicalese, Maria Pia; Giannelli, Stefania; Neduva, Victor; Dow, David; Schmidt, Manfred; Kalle, Christof von; Roncarolo, Maria Grazia; Ciceri, Fabio; Vicard, Paola; Wit, Ernst; Serio, Clelia Di; Naldini, Luigi; Aiuti, Alessandro

doi:10.1016/j.stem.2016.04.016

Cited by 200 publications

(225 citation statements)

References 51 publications

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“…In non-human primate and human studies with ex vivo lentivirus vector-transduced HSCs, it was estimated that one in 1 million transplanted HSCs were capable of long-term repopulation. 32,33 In the human trials, this would correspond to 200 to 300 gene-corrected, long-term engrafting cells per 1.5 3 10 12 total MNCs in the human BM. 4 The fact that our approach allows for the transduction of primitive HSPCs and that the percentage of these cells increases over time ( Figure 3B), suggests that transduced HSPCs self-renew and give rise to GFP 1 progeny cells that slowly overtake the BM.…”

Section: Discussionmentioning

confidence: 99%

In vivo transduction of primitive mobilized hematopoietic stem cells after intravenous injection of integrating adenovirus vectors

Richter

Saydaminova

Yumul

et al. 2016

Blood

145

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• Mobilized hematopoietic stem cells transduced with IV injected HD-Ad5/35 11 vectors home to the BM persist long term.• Our approach allows for the stable genetic modification of primitive, long-term persisting HSPCs.Current protocols for hematopoietic stem/progenitor cell (HSPC) gene therapy, involving the transplantation of ex vivo genetically modified HSPCs are complex and not without risk for the patient. We developed a new approach for in vivo HSPC transduction that does not require myeloablation and transplantation. It involves subcutaneous injections of granulocyte-colony-stimulating factor/AMD3100 to mobilize HSPCs from the bone marrow (BM) into the peripheral blood stream and the IV injection of an integrating, helper-dependent adenovirus (HD-Ad5/35 11 ) vector system. These vectors target CD46, a receptor that is uniformly expressed on HSPCs. We demonstrated in human CD46 transgenic mice and immunodeficient mice with engrafted human CD34 1 cells that HSPCs transduced in the periphery home back to the BM where they stably express the transgene. In hCD46 transgenic mice, we showed that our in vivo HSPC transduction approach allows for the stable transduction of primitive HSPCs. Twenty weeks after in vivo transduction, green fluorescent protein (GFP) marking in BM HSPCs (Lin 2 Sca1 1 Kit 2 cells) in most of the mice was in the range of 5% to 10%. The percentage of GFP-expressing primitive HSPCs capable of forming multilineage progenitor colonies (colony-forming units [CFUs]) increased from 4% of all CFUs at week 4 to 16% at week 12, indicating transduction and expansion of long-term surviving HSPCs. Our approach was well tolerated, did not result in significant transduction of nonhematopoietic tissues, and was not associated with genotoxicty. The ability to stably genetically modify HSPCs without the need of myeloablative conditioning is relevant for a broader clinical application of gene therapy. (Blood. 2016;128(18):2206-2217

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Section: Discussionmentioning

confidence: 99%

In vivo transduction of primitive mobilized hematopoietic stem cells after intravenous injection of integrating adenovirus vectors

Richter

Saydaminova

Yumul

et al. 2016

Blood

145

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“…Using treated WAS patients, it was therefore possible to define cellular contributions to early and late phases of reconstitution, and to show that HSC/Ps manipulated in vitro for gene transfer retained the ability to restore hematopoiesis in vivo in a way that mirrors normal HSC activity after transplantation. 91 …”

Section: Gene Therapy For Chronic Granulomatous Disease: No Selectivementioning

confidence: 99%

Evolving Gene Therapy in Primary Immunodeficiency

Thrasher

Williams

2017

Molecular Therapy

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“…3,4 As direct observation is not implementable for all systems, labeling of cells has been used to study a wide variety of questions in an assortment of experimental systems and organisms, first by dyes 5,6 and radioactive tracers 7 and later by fluorescent markers, 8 genetic recombination, 9–11 and static genetic barcoding. 12–17 …”

mentioning

confidence: 99%

Quantitative Analysis of Synthetic Cell Lineage Tracing Using Nuclease Barcoding

et al. 2017

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Lineage tracing by the determination and mapping of progeny arising from single cells is an important approach enabling the elucidation of mechanisms underlying diverse biological processes ranging from development to disease. We developed a dynamic sequence-based barcode system for synthetic lineage tracing and have demonstrated its performance in C. elegans, a model organism whose lineage tree is well established. The strategy we use creates lineage trees based upon the introduction of synthetically controlled mutations into cells and the propagation of these mutations to daughter cells at each cell division. We analyzed this experimental proof of concept along with a corresponding simulation and analytical model to gain a deeper understanding of the coding capacity of the system. Our results provide specific bounds on the fidelity of lineage tracing using such approaches.

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In Vivo Tracking of Human Hematopoiesis Reveals Patterns of Clonal Dynamics during Early and Steady-State Reconstitution Phases

Cited by 200 publications

References 51 publications

In vivo transduction of primitive mobilized hematopoietic stem cells after intravenous injection of integrating adenovirus vectors

In vivo transduction of primitive mobilized hematopoietic stem cells after intravenous injection of integrating adenovirus vectors

Evolving Gene Therapy in Primary Immunodeficiency

Quantitative Analysis of Synthetic Cell Lineage Tracing Using Nuclease Barcoding

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