Evidence from a leukaemia model for maintenance of vascular endothelium by bone-marrow-derived endothelial cells

doi:10.1016/S0140-6736(00)02241-8

The Lancet

Volume 355, Issue 9216, 13 May 2000, Pages 1688-1691

https://doi.org/10.1016/S0140-6736(00)02241-8 Get rights and content

Summary

Background

Vascular endothelial cells lost from the bloodvessel endothelium through necrosis or apoptosis must be replaced. We investigated in a leukaemia model whether bone-marrow-derived endothelial cells contribute to this maintenance angiogenesis.

Methods

We studied six patients with chronic myelogenous leukaemia (CML) carrying the BCR/ABL fusion gene in their bone-marrow-derived cells. We screened endothelial cells generated in vitro from bone-marrow-derived progenitor cells and vascular endothelium in myocardial tissue for the BCR/ABL fusion gene by in-situ hybridisation. For detection of donor-type endothelial cells after transplantation of haemopoietic stem cells, recipient tissue was stained with monoclonal antibodies against donor-type HLA antigens.

Findings

We identified the BCR/ABL fusion gene in variable proportions (0·56%) of endothelial cells generated in vitro. Endothelial cells expressing the fusion gene were found in the vascular endothelium of a patient. In a recipient of an allogeneic stem-cell transplant, normal donor-type endothelial cells were detected in the vascular endothelium.

Interpretation

These findings suggest that CML is not solely a haematological disease but originates from a bone-marrow-derived haemangioblastic precursor cell that can give rise to both blood cells and endothelial cells. Moreover, normal bone-marrow-derived endothelial cells can contribute to the maintenance of the blood vascular endothelium. The integration of bone-marrow-derived endothelial cells into the vascular endothelium provides a rationale for developing vascular targeting strategies in vasculopathies, inflammatory diseases, and cancer.

Introduction

Angiogenesis is the formation of new blood vessels from pre-existing vessels.¹ In adults, angiogenesis takes place during placental development, ovulation, and wound healing, and in disease states such as growth of malignant tumours.² Given that vascular endothelial cells have a limited lifespan,³ there must be a mechanism by which those undergoing necrosis or apoptosis are replaced. Two possible mechanisms for this maintenance angiogenesis are the migration and proliferation of existing endothelial cells within the vascular endothelium⁴ and the recruitment and homing of circulating endothelial cells or their progenitors released from the bone marrow.

Postulated a century ago,⁵ a progenitor cell that can give rise to blood cells of many lineages as well as endothelial cells has been characterised during embryonic development.⁶ Cells from peripheral blood, bone marrow, and fetal liver expressing CD34 antigen (which is found on both haemopoietic cells and endothelial cells in the developing embryo and adult) are progenitors of sustained multilineage haemopoiesis⁷ and can be driven to produce endothelial progenitor cells in vitro.8, 9 Thus, we hypothesised that precursor cells with haemangioblastic features are present also in the adult bone marrow and bring about the maintenance angiogenesis of the blood vascular endothelium.

This hypothesis was tested in patients with chronic myelogenous leukaemia (CML). Emerging after the malignant transformation of a progenitor cell with haemopoietic multilineage differentiation capacity,¹⁰ CML is characterised by a unique chromosomal translocation, t(9;22),11, 12 resulting in the BCR/ABL fusion gene¹³ (which is present in bone-marrow-derived immature and mature myelomonocytic cells including dendritic cells,¹⁴ and in T lymphocytes¹¹). Multipotent haemopoietic progenitor cells, committed blood precursors, and endothelial cells generated in vitro from peripheral blood or bone marrow of patients with CML were screened for this BCR/ABL fusion gene. Furthermore, a BCR/ABL-specific gene probe was applied to endothelial cells from the intimal layer of blood vessels of a patient with CML.

Section snippets

Methods

Blood and bone-marrow samples were collected from six patients with CML (all white men, aged 22–61 years). All had Philadelphia-chromosome-positive CML in chronic phase as judged by bone-marrow histology and cytogenetic analysis (with >80% Philadelphia-chromosome-positive cells in bone marrow before mobilisation treatment). All were pretreated with hydroxyurea, and no interferon had been given. All patients received mobilisation treatment within 6 months of diagnosis. After patients had given

Results

In the six patients with CML, endothelial cells were generated in vitro from blood or bone-marrow-derived mononuclear cells. Mononuclear cells became adherent within 2 h of culture, and after 5 days clusters containing cells with cytological features of endothelial cells such as spindle shape and granularity were present (figure, A, B). These cells were identified as endothelial cells by their antigen expression repertoire (CD34 positive, CD31 positive, VE-cadherin negative, E-selectin

Discussion

We have shown that cells of the endothelial lineage are part of the malignant cell clone in CML. Since FLK-T^−/− mouse embryos (knockouts for the gene encoding the receptor for vascular endothelial growth factor) do not show vasculogenesis or develop blood islands,²⁰ a common progenitor of endothelial cells and haemopoietic progenitor cells has been postulated. Our observations support the existence of this precursor cell with haemangioblastic features in human adults, because both endothelial

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Early ReportEvidence from a leukaemia model for maintenance of vascular endothelium by bone-marrow-derived endothelial cells

Summary

Background

Methods

Findings

Interpretation

Introduction

Section snippets

Methods

Results

Discussion

Cell

Blood

Am J Med

Cell

Blood

Vasculogenesis and angiogenesis in embryonic-stem-cell-derived embryoid bodies

Development

Angiogenesis in cancer, vascular, rheumatoid and other disease

Nat Med

Vessel cooption, regression, and growth in tumors mediated by angiopoietins and VEGF

Science

Lecithoblast und Angioblast der Wirbelthiere

Abhandl KS Ges Wiss Math Phys

A common precursor for hematopoietic and endothelial cells

Development

Early Report
Evidence from a leukaemia model for maintenance of vascular endothelium by bone-marrow-derived endothelial cells