Decreased homing of retrovirally transduced human bone marrow CD34+ cells in the NOD/SCID mouse model

Hall, Kristin M.; Horváth, Tamara; Abonour, Rafat; Cornetta, Kenneth; Srour, Edward F.

doi:10.1016/j.exphem.2005.12.014

Cited by 21 publications

(22 citation statements)

References 43 publications

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“…Further expansion of a CXCR4-overexpressing system may serve as an effective tool to study regulation and functionality of this unique receptor as well as to improve the compromised homing and engraftment following gene therapy protocols. 7,8 A significant clinical breakthrough in gene therapy was made in patients with human SCID-X1 resulting in full correction of disease phenotype, 60,61 proving that gene therapy can work in practice. However, the risk of malignant transformation is prevalent in some patients.…”

Section: Discussionmentioning

confidence: 99%

“…[1][2][3] One of the disadvantages of BM transplantation is the long-lasting reduced levels of immature progenitors, including long-term culture-initiating cells (LTCICs; 1 log reduction), in the BM of patients who have received transplants compared with healthy individuals. [4][5][6] Furthermore, emerging evidence exists for impaired homing 7 and low engraftment 8 of retrovirally transduced human CD34 ϩ cells. Enhanced efficacy of HSC engraftment could improve the outcome of clinical transplantations as well as gene therapy protocols and might be achieved by modulating the ability of stem cells to home to and repopulate the recipient BM.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Overexpression of CXCR4 on human CD34+ progenitors increases their proliferation, migration, and NOD/SCID repopulation

Kahn

Byk

Jansson‐Sjöstrand

et al. 2004

Blood

211

158

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A major limitation to clinical stem cellmediated gene therapy protocols is the low levels of engraftment by transduced progenitors. We report that CXCR4 overexpression on human CD34 ؉ progenitors using a lentiviral gene transfer technique helped navigate these cells to the murine bone marrow and spleen in response to stromal-derived factor 1 (SDF-1) signaling. Cells overexpressing CXCR4 exhibited significant increases in SDF-1-mediated chemotaxis and actin polymerization compared with control cells. A major advantage of CXCR4 overexpression was demonstrated by the ability of transduced CD34 ؉ cells to respond to lower, physiologic levels of SDF-1 when compared to control cells, leading to improved SDF-1-induced migration and proliferation/survival, and finally resulting in significantly higher levels of in vivo repopulation of nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice including primitive CD34 ؉ /CD38 ؊/low cells. Importantly, no cellular transformation was observed following transduction with the CXCR4 vector. Unexpectedly, we documented lack of receptor internalization in response to high levels of SDF-1, which can also contribute to increased migration and proliferation by the transduced CD34 ؉ cells. Our results suggest CXCR4 overexpression for improved definitive human stem cell motility, retention, and multilineage repopulation, which could be beneficial for in vivo navigation and expansion of hematopoietic progenitors. ( IntroductionGene transfer into human hematopoietic stem cells (HSCs) may be a promising tool in the correction of a wide variety of hematopoietic and genetic disorders. HSC transplantation can be used to durably deliver these genetically modified cells to the bone marrow (BM), which in turn will release mature cells with the corrected gene into the circulation throughout life. Clinical and experimental HSC transplantation procedures mimic the physiologic process of HSC migration from the circulation into the BM occurring during late embryonic development and steady-state hematopoiesis in adults throughout life. [1][2][3] One of the disadvantages of BM transplantation is the long-lasting reduced levels of immature progenitors, including long-term culture-initiating cells (LTCICs; 1 log reduction), in the BM of patients who have received transplants compared with healthy individuals. [4][5][6] Furthermore, emerging evidence exists for impaired homing 7 and low engraftment 8 of retrovirally transduced human CD34 ϩ cells. Enhanced efficacy of HSC engraftment could improve the outcome of clinical transplantations as well as gene therapy protocols and might be achieved by modulating the ability of stem cells to home to and repopulate the recipient BM.Interactions between the chemokine stromal-derived factor 1 (SDF-1), also referred to as CXCL12, and its receptor CXCR4 play an essential role in stem cell seeding of the BM during murine embryonic development. 9,10 Moreover, we have previously demonstrated in a functional preclinical model, using nonobese diabetic/ severe ...

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Overexpression of CXCR4 on human CD34+ progenitors increases their proliferation, migration, and NOD/SCID repopulation

Kahn

Byk

Jansson‐Sjöstrand

et al. 2004

Blood

211

158

View full text Add to dashboard Cite

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“…For example, gene transfer into human hematopoietic stem cells is a promising tool for the correction of a wide variety of hematopoietic and genetic disorders [35]. Since ex vivotransduced hematopoietic stem cells are reported to exhibit reduced homing ability [36], IBMT is an ideal transplantation method for transferring engineered hematopoietic stem cells to bone marrow compartments. Nonetheless, our data reveal that consideration and manipulation of the migration property are important for IBMT of ex vivo-treated hematopoietic stem cells in future clinical settings.…”

Section: Cd34mentioning

confidence: 99%

Ex Vivo Culture of Human Cord Blood Hematopoietic Stem/Progenitor Cells Adversely Influences Their Distribution to Other Bone Marrow Compartments After Intra-Bone Marrow Transplantation

et al. 2007

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Intra-bone marrow injection is a novel strategy for hematopoietic stem cell transplantation. Here, we investigated whether ex vivo culture of cord blood hematopoietic stem/ progenitor cells influences their reconstitution in bone marrow after intra-bone marrow transplantation. Freshly isolated AC133؉ cells or cells derived from AC133 ؉ cells cultured with cytokines (stem cell factor, flt-3 ligand, and thrombopoietin) for 5 days were injected into the bone marrow of the left tibia in irradiated NOD/SCID mice. In the bone marrow of the injected left tibia, the engraftment levels of human CD45 ؉ cells at 6 weeks after transplantation did not differ considerably between transplantation of noncultured and cytokine-cultured cells. However, the migration and distribution of transplanted cells to the bone marrow of other, noninjected bones were extremely reduced for cytokine-treated cells compared with noncultured cells. Similar findings were observed for engraftment of CD34 ؉ cells. Administration of granulocyte colony-stimulating factor to mice after transplantation induced the migration of cytokine-cultured cells to the bone marrow of previously aspirated bone but not to other intact bones. These data suggest that ex vivo manipulation of hematopoietic progenitor/stem cells significantly affects their migration properties to other bone marrow compartments after intra-bone marrow transplantation. Our data raise a caution for future clinical applications of the intra-bone marrow transplantation method using ex vivo-manipulated hematopoietic stem cells. STEM CELLS 2008;26:543-549 Disclosure of potential conflicts of interest is found at the end of this article.

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“…This could be owing to loss of HSC content with the enrichment and culture manipulations or to a homing and engraftment defect in the manipulated cell populations, as has been previously reported. 20,21 To further assess the long-term repopulating capacity of the transduced SLAM-HSCs, we performed a secondary transplantation experiment. Primary donor CD45.2 CD150 + /CD48 À cells were transduced using the described abbreviated protocol.…”

Section: Slam-enriched Hematopoietic Stem Cells P Laje Et Almentioning

confidence: 99%

“…7,8 Protocols for lentiviral transduction of HSCs have evolved over the years with a recent trend toward abbreviated culture times and minimal cytokine exposure, to avoid differentiation, and loss of long-term repopulating capacity after transplantation. [9][10][11][12] In addition to loss of repopulating capacity, transduction of HSCs has been shown to result in loss of homing capacity 13 and may have other detrimental effects. Thus, the process of HSC enrichment and subsequent gene transfer may result in significant reduction of HSC repopulating capacity by a variety of mechanisms.…”

Section: Introductionmentioning

confidence: 99%

SLAM-enriched hematopoietic stem cells maintain long-term repopulating capacity after lentiviral transduction using an abbreviated protocol

2009

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Gene transfer to long-term repopulating hematopoietic stem cells (HSCs) using integrating viral vectors is an important goal in gene therapy. The SLAM (signaling lymphocyte activation molecule)-family receptors have recently been used for the isolation of highly enriched murine HSCs. This HSC enrichment protocol is relatively simple, and results in an HSC population with comparable repopulating capacity to c-kit + lin À Sca-1 + (KSL) HSCs. The capacity to withstand genetic manipulation and, most importantly, to maintain long-term repopulating capacity of SLAM-enriched HSC populations has not been reported. In this study, SLAMenriched HSCs were assessed for transduction efficiency and in vivo long-term repopulating capacity after lentiviral transduction using an abbreviated transduction protocol and KSL-enriched HSCs as a reference population. SLAM-and KSL-enriched HSCs were efficiently transduced by lentiviral vector using a simple protocol that involves minimal in vitro manipulation and no pre-stimulation. SLAM-HSCs are at least equal to KSL-HSCs with respect to efficiency of transduction and maintenance of long-term repopulating capacity. Although there was a reduction in repopulating capacity related to enrichment and culture manipulations relative to freshly isolated bone marrow (BM) cells, no detrimental effects were identified on long-term competitive capacity related to transduction, as transduced cells maintained stable levels of chimerism in competition with nontransduced cells and freshly isolated BM cells. These results support the SLAM-HSC enrichment protocol as a simple and efficient method for HSC enrichment for gene transfer studies.

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Decreased homing of retrovirally transduced human bone marrow CD34+ cells in the NOD/SCID mouse model

Cited by 21 publications

References 43 publications

Overexpression of CXCR4 on human CD34+ progenitors increases their proliferation, migration, and NOD/SCID repopulation

Overexpression of CXCR4 on human CD34+ progenitors increases their proliferation, migration, and NOD/SCID repopulation

Ex Vivo Culture of Human Cord Blood Hematopoietic Stem/Progenitor Cells Adversely Influences Their Distribution to Other Bone Marrow Compartments After Intra-Bone Marrow Transplantation

SLAM-enriched hematopoietic stem cells maintain long-term repopulating capacity after lentiviral transduction using an abbreviated protocol

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