Characterization and quantification of clonal heterogeneity among hematopoietic stem cells: a model-based approach

Roeder, Ingo; Horn, Katrin; Sieburg, Hans-Bernd; Cho, Rebecca; Müller‐Sieburg, Christa E.; Loeffler, Markus

doi:10.1182/blood-2008-05-155374

Cited by 55 publications

(29 citation statements)

References 53 publications

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“…Initially, such studies were performed using the unique marks that virus insertion form when they integrate into the host genome (20,21). From these studies, several models for behavior of the hematopoietic system were postulated and mathematically modeled (22). DNA barcoding technology allowed a much more refined insight into the composition of the blood compartments and allowed the study of aging HSC (13) and distribution of HSC clones through the different locations in the body (14), in addition to the clonal dynamics of repopulation of the BM subset hierarchy after transplantation (23,24).…”

Section: Discussionmentioning

confidence: 99%

Development of a diverse human T-cell repertoire despite stringent restriction of hematopoietic clonality in the thymus

Brugman

Wiekmeijer

Eggermond

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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The fate and numbers of hematopoietic stem cells (HSC) and their progeny that seed the thymus constitute a fundamental question with important clinical implications. HSC transplantation is often complicated by limited T-cell reconstitution, especially when HSC from umbilical cord blood are used. Attempts to improve immune reconstitution have until now been unsuccessful, underscoring the need for better insight into thymic reconstitution. Here we made use of the NOD-SCID-IL-2Rγ−/− xenograft model and lentiviral cellular barcoding of human HSCs to study T-cell development in the thymus at a clonal level. Barcoded HSCs showed robust (>80% human chimerism) and reproducible myeloid and lymphoid engraftment, with T cells arising 12 wk after transplantation. A very limited number of HSC clones (<10) repopulated the xenografted thymus, with further restriction of the number of clones during subsequent development. Nevertheless, T-cell receptor rearrangements were polyclonal and showed a diverse repertoire, demonstrating that a multitude of T-lymphocyte clones can develop from a single HSC clone. Our data imply that intrathymic clonal fitness is important during T-cell development. As a consequence, immune incompetence after HSC transplantation is not related to the transplantation of limited numbers of HSC but to intrathymic events.H ematopoietic stem cell transplantation (HSCT) has become common clinical practice in the treatment of leukemia, lymphoma, and certain inherited immune and metabolic disorders. After transplantation there is an immediate need for de novo production of granulocytes, erythrocytes, and platelets, referred to as "hematopoietic reconstitution," later followed by recovery of lymphocyte numbers, termed "immune reconstitution." Although often successful, HSCT is associated with a number of complications arising from poor immune reconstitution, which presents one of the most important causes of morbidity after HSCT (1, 2). Poor myeloid reconstitution is directly linked to low numbers of HSCs in the transplant, but the reasons for poor immune and, in particular, T-lymphocyte reconstitution are much less clear.A study on the application of antithymocyte globulin in cord blood transplantation showed that antithymocyte globulin administration in pretransplantation conditioning results in decreased T-cell reconstitution and survival (3), indicating the need for a better understanding of de novo T-cell development after HSCT. As T cells develop in the thymus, in contrast to all other blood lineages that develop in the bone marrow (BM), HSCderived progenitors need to seed the thymus. Earlier work in mice has indicated that relatively few progenitors seed the thymus (4, 5), yet their numbers and the subsequent fate of the progeny derived from an HSC clone has remained elusive. The nature of the thymusseeding cell has been subject of much debate, certainly in humans. CD34+ cells (6, 7), CD34 (8), and others have been proposed as thymus-seeding cells in the human situation. In contrast to mice, the earliest hu...

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Section: Discussionmentioning

confidence: 99%

Development of a diverse human T-cell repertoire despite stringent restriction of hematopoietic clonality in the thymus

Brugman

Wiekmeijer

Eggermond

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…As well, such data would be useful to corroborate or further inform long-standing theoretical concepts and quantitative models on the clonal dynamics of HSCs. 26,27 Collectively, these applications of the cellular barcoding method have the potential to greatly increase our understanding of clonal dynamics in a BM transplantation setting.…”

Section: Discussionmentioning

confidence: 99%

Cellular barcoding tool for clonal analysis in the hematopoietic system

Gerrits

Dykstra

Kalmykowa

et al. 2010

Blood

227

188

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Clonal analysis is important for many areas of hematopoietic stem cell research, including in vitro cell expansion, gene therapy, and cancer progression and treatment. A common approach to measure clonality of retrovirally transduced cells is to perform integration site analysis using Southern blotting or polymerase chain reaction-based methods. Although these methods are useful in principle, they generally provide a lowresolution, biased, and incomplete assessment of clonality. To overcome those limitations, we labeled retroviral vectors with random sequence tags or "barcodes." On integration, each vector introduces a unique, identifiable, and heritable mark into the host cell genome, allowing the clonal progeny of each cell to be tracked over time. By coupling the barcoding method to a sequencing-based detection system, we could identify major and minor clones in 2 distinct cell culture systems in vitro and in a long-term transplantation setting. In addition, we demonstrate how clonal analysis can be complemented with transgene expression and integration site analysis. This cellular barcoding tool permits a simple, sensitive assessment of clonality and holds great promise for future gene therapy protocols in humans, and any other applications when clonal tracking is important. (Blood. 2010;115(13):2610-2618)

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“…Rather, current findings suggest the involvement of highly complex and possibly alternative molecular networks controlling LTRC self-renewal that may require systems approaches to elucidate. 53,54 Indeed, one might advance the somewhat heretical idea that intracellular states responsible for different HSC self-renewal potentialities may not be well-matched to cellular phenotypes that display different transplant-derived endpoints of repopulating activity.…”

Section: Hsc Purificationmentioning

confidence: 99%

Hematopoietic stem cells: concepts, definitions, and the new reality

Eaves

2015

Blood

449

315

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Hematopoietic stem cell (HSC) research took hold in the 1950s with the demonstration that intravenously injected bone marrow cells can rescue irradiated mice from lethality by reestablishing blood cell production. Attempts to quantify the cells responsible led to the discovery of serially transplantable, donor-derived, macroscopic, multilineage colonies detectable on the spleen surface 1 to 2 weeks posttransplant. The concept of self-renewing multipotent HSCs was born, but accompanied by perplexing evidence of great variability in the outcomes of HSC self-renewal divisions. The next 60 years saw an explosion in the development and use of more refined tools for assessing the behavior of prospectively purified subsets of hematopoietic cells with blood cell–producing capacity. These developments have led to the formulation of increasingly complex hierarchical models of hematopoiesis and a growing list of intrinsic and extrinsic elements that regulate HSC cycling status, viability, self-renewal, and lineage outputs. More recent examination of these properties in individual, highly purified HSCs and analyses of their perpetuation in clonally generated progeny HSCs have now provided definitive evidence of linearly transmitted heterogeneity in HSC states. These results anticipate the need and use of emerging new technologies to establish models that will accommodate such pluralistic features of HSCs and their control mechanisms.

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Characterization and quantification of clonal heterogeneity among hematopoietic stem cells: a model-based approach

Cited by 55 publications

References 53 publications

Development of a diverse human T-cell repertoire despite stringent restriction of hematopoietic clonality in the thymus

Development of a diverse human T-cell repertoire despite stringent restriction of hematopoietic clonality in the thymus

Cellular barcoding tool for clonal analysis in the hematopoietic system

Hematopoietic stem cells: concepts, definitions, and the new reality

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