High incidence of leukemia in large animals after stem cell gene therapy with a HOXB4-expressing retroviral vector

Zhang, Xiao-Bing; Beard, Brian C.; Trobridge, Grant D.; Wood, Brent L.; Sale, George E.; Sud, Reeteka; Humphries, R. Keith; Kiem, Hans Peter

doi:10.1172/jci34371

Cited by 104 publications

(121 citation statements)

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“…However, this system requires adaptation for hESCs and application of effective human promoters rather than the CMV promoter [48,49]. Also, new nonviral transposon-mediated gene transfer systems may be a safer option [50], also promising high efficiency and inducibility in embryonic stem cells [51,52]. Generally, HoxB4 regulation could avoid possible side effects caused by expression above the therapeutic level.…”

Section: Discussionmentioning

confidence: 99%

Lentiviral-Mediated HoxB4 Expression in Human Embryonic Stem Cells Initiates Early Hematopoiesis in a Dose-Dependent Manner but Does Not Promote Myeloid Differentiation

et al. 2008

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The variation of HoxB4 expression levels might be a key regulatory mechanism in the differentiation of human embryonic stem cell (hESC)-derived hematopoietic stem cells (HSCs). In this study, hESCs ectopically expressing high and low levels of HoxB4 were obtained using lentiviral gene transfer. Quantification throughout differentiation revealed a steady increase in transcription levels from our constructs. The effects of the two expression levels of HoxB4 were compared regarding the differentiation potential into HSCs. High levels of HoxB4 expression correlated to an improved yield of cells expressing CD34, CD38, the stem cell leukemia gene, and vascular epitheliumcadherin. However, no improvement in myeloid cell maturation was observed, as determined by colony formation assays. In contrast, hESCs with low HoxB4 levels did not show any elevated hematopoietic development. In addition, we found that the total population of HoxB4-expressing cells, on both levels, decreased in developing embryoid bodies. Notably, a high HoxB4 expression in hESCs also seemed to interfere with the formation of germ layers after xenografting into immunodeficient mice. These data suggest that HoxB4-induced effects on hESC-derived HSCs are concentrationdependent during in vitro development and reduce proliferation of other cell types in vitro and in vivo. The application of the transcription factor HoxB4 during early hematopoiesis from hESCs might provide new means for regenerative medicine, allowing efficient differentiation and engraftment of genetically modified hESC clones. Our study highlights the importance of HoxB4 dosage and points to the need for experimental systems allowing controlled gene expression.

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Section: Discussionmentioning

confidence: 99%

Lentiviral-Mediated HoxB4 Expression in Human Embryonic Stem Cells Initiates Early Hematopoiesis in a Dose-Dependent Manner but Does Not Promote Myeloid Differentiation

et al. 2008

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“…Cell Populations Studied Gene Symbols* 7 In order to improve in vitro HSC expansion, and to circumvent potential drawbacks such as those observed by Zhang et al, 4 we transduced the HOXC4 protein into human HSC using the same method as for HOXB4. 7 We provide evidence that, like HOXB4, the human HOXC4 protein behaves as a growth factor able to enhance the number of hematopoietic progenitors and stem cells.…”

Section: Referencementioning

confidence: 99%

“…[1][2][3] Although that gene was first described as non-leukemogenic, a recent study revealed that transduction of HoxB4 into the HSC of large animals could lead to the late emergence of acute myeloid leukemias. 4 Thus, retrovirus-mediated genetic alterations of HSC along with constitutive expression of human HOXB4 can be hazardous for therapeutic applications.…”

Section: Introductionmentioning

confidence: 99%

HOXC4 homeoprotein efficiently expands human hematopoietic stem cells and triggers similar molecular alterations as HOXB4

Auvray¹,

Delahaye²,

Pflumio³

et al. 2012

Haematologica

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The online version of this article has a Supplementary Appendix. BackgroundExpansion of hematopoietic stem cells represents an important objective for improving cell and gene therapy protocols. Retroviral transduction of the HoxB4 homeogene in mouse and human hematopoietic stem cells and hematopoietic progenitors is known to promote the cells' expansion. A safer approach consists in transferring homeobox proteins into hematopoietic stem cells taking advantage of the natural ability of homeoproteins to cross cell membranes. Thus, HOXB4 protein transfer is operative for expanding human hematopoietic cells, but such expansion needs to be improved. Design and MethodsTo that aim, we evaluated the effects of HOXC4, a protein encoded by a HOXB4 paralog gene, by co-culturing HOXC4-producing stromal cells with human CD34 + hematopoietic cells. Numbers of progenitors and stem cells were assessed by in vitro cloning assays and injection into immuno-deficient mice, respectively. We also looked for activation or inhibition of target downstream gene expression. ResultsWe show that the HOXC4 homeoprotein expands human hematopoietic immature cells by 3 to 6 times ex vivo and significantly improves the level of in vivo engraftment. Comparative transcriptome analysis of CD34 + cells subjected or not to HOXB4 or HOXC4 demonstrated that both homeoproteins regulate the same set of genes, some of which encode key hematopoietic factors and signaling molecules. Certain molecules identified herein are factors reported to be involved in stem cell fate or expansion in other models, such as MEF2C, EZH2, DBF4, DHX9, YPEL5 and Pumilio. ConclusionsThe present study may help to identify new HOX downstream key factors potentially involved in hematopoietic stem cell expansion or in leukemogenesis.

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“…In addition to our study, HOXB4 has been suggested as a cancer-related gene in the various types of cancers, such as leukemia, breast cancer, osteosarcoma and lung cancer. [34][35][36][37] In leukemia, leukemic cells had dysregulated expression of oncogenes, a block in myeloid differentiation, and overexpression of HOXB4, whereas HOXB4 knockdown restored differentiation in leukemic cells. 34 Bodey et al [35][36][37] confirmed the expression of HOXB3, B4, and C6 genes in breast cancer, osteosarcoma, and lung cancer by immunocytochemical analysis and suggested their association with the development of these cancers.…”

mentioning

confidence: 99%

“…[34][35][36][37] In leukemia, leukemic cells had dysregulated expression of oncogenes, a block in myeloid differentiation, and overexpression of HOXB4, whereas HOXB4 knockdown restored differentiation in leukemic cells. 34 Bodey et al [35][36][37] confirmed the expression of HOXB3, B4, and C6 genes in breast cancer, osteosarcoma, and lung cancer by immunocytochemical analysis and suggested their association with the development of these cancers. In contrast to our results, Naora et al 17 reported that ovarian carcinoma was found to express HOXB7 at markedly higher levels than normal ovarian surface epithelium.…”

mentioning

confidence: 99%

Expression pattern of the class I homeobox genes in ovarian carcinoma

et al. 2010

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Objective: Although some sporadic reports reveal the link between the homeobox (HOX) genes and ovarian carcinoma, there is no comprehensive analysis of the expression pattern of the class I homeobox genes in ovarian carcinoma that determines the candidate genes involved in ovarian carcinogenesis. Methods: The different patterns of expression of 36 HOX genes were analyzed, including 4 ovarian cancer cell lines and 4 normal ovarian tissues. Using a reverse transcription-polymerase chain reaction (RT-PCR) and quantification analysis, the specific gene that showed a significantly higher expression in ovarian cancer cell lines than in normal ovaries was selected, and western blot analysis was performed adding 7 ovarian cancer tissue specimens. Finally, immunohistochemical and immunocytochemical analyses were performed to compare the pattern of expression of the specific HOX gene between ovarian cancer tissue and normal ovaries. Results: Among 36 genes, 11 genes had a different level of mRNA expression between the cancer cell lines and the normal ovarian tissues. Of the 11 genes, only HOXB4 had a significantly higher level of expression in ovarian cancer cell lines than in normal ovaries (p=0.029). Based on western blot, immunohistochemical, and immunocytochemical analyses, HOXB4 was expressed exclusively in the ovarian cancer cell lines or cancer tissue specimens, but not in the normal ovaries. Conclusion:We suggest HOXB4 may be a novel candidate gene involved in ovarian carcinogenesis.

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High incidence of leukemia in large animals after stem cell gene therapy with a HOXB4-expressing retroviral vector

Cited by 104 publications

References 41 publications

Lentiviral-Mediated HoxB4 Expression in Human Embryonic Stem Cells Initiates Early Hematopoiesis in a Dose-Dependent Manner but Does Not Promote Myeloid Differentiation

Lentiviral-Mediated HoxB4 Expression in Human Embryonic Stem Cells Initiates Early Hematopoiesis in a Dose-Dependent Manner but Does Not Promote Myeloid Differentiation

HOXC4 homeoprotein efficiently expands human hematopoietic stem cells and triggers similar molecular alterations as HOXB4

Expression pattern of the class I homeobox genes in ovarian carcinoma

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