Characterizing how the microenvironment, or niche, regulates stem cell activity is central to understanding stem cell biology and to developing strategies for therapeutic manipulation of stem cells1. Low oxygen tension (hypoxia) is commonly thought to be a shared niche characteristic in maintaining quiescence in multiple stem cell types2–4. However, support for the existence of a hypoxic niche has largely come from indirect evidence such as proteomic analysis5, expression of HIF-1 and related genes6, and staining with surrogate hypoxic markers (e.g. pimonidazole)6–8. Here we perform direct in vivo measurements of local oxygen tension (pO2) in the bone marrow (BM) of live mice. Using two-photon phosphorescence lifetime microscopy (2PLM), we determined the absolute pO2 of the BM to be quite low (<32 mmHg) despite very high vascular density. We further uncovered heterogeneities in local pO2, with the lowest pO2 (~9.9 mmHg, or 1.3%) found in deeper peri-sinusoidal regions. The endosteal region, by contrast, is less hypoxic as it is perfused with small arteries that are often positive for the marker nestin. These pO2 values change dramatically after radiation and chemotherapy, pointing to the role of stress in altering the stem cell metabolic microenvironment.

The organization of cellular niches has been shown to play a key role in regulating normal stem cell differentiation and regeneration, yet relatively little is known about the architecture of microenvironments that support malignant metastasis. 1,2 Using dynamic in vivo confocal imaging, we show that the murine bone marrow (BM) contains unique anatomic regions defined by specialized endothelium. This vasculature expresses the adhesion molecule E-selectin and the chemoattractant SDF-1 in discrete, discontinuous areas that localize the homing of a variety of tumor cell lines. Disruption of SDF-1/CXCR4 interactions inhibits Nalm-6 cell (acute lymphoblastic leukaemia) homing to these vessels. Further studies revealed that circulating leukemic cells engraft surrounding these vessels, suggesting that this molecularly distinct vasculature denotes a microenvironment for early metastatic tumor spread in BM. Finally, purified hematopoietic stem/progenitor cells and lymphocytes also localize to the same microdomains, indicating that this vasculature may function in benign states to demarcate specific portals for entry of cells into the marrow space. Specialized vascular structures therefore appear to delineate a microenvironment with unique physiology that is exploited by circulating malignant cells.It has been thought that tumor cells derive their ability to transit to specific organs by co-opting the same tissue-homing mechanisms used by benign leukocytes. 3 Substantial in vitro and more limited in vivo data provide evidence that tumors depend on selectin-, integrin-, and chemokinemediated vascular cell adhesion events in order to identify and bind to vascular beds at sites of tissue entry. 4,5 These molecular mechanisms are thought to enable the efficient spread of malignancies to target organs. Differential expression of these endothelial signals among tissues is known to control the destination of cellular traffic, but the contributions of the vascular molecular framework to the regulation of complex cellular microenvironments remain to be fully elucidated. Competing interests statementThe authors declare that they have no competing financial interests. The bone marrow (BM) is a frequent site for solid tumor spread. It can also be considered the most ubiquitous site for leukemic cell metastasis, as disease is seen to migrate from the initial birthplace of the leukemic clone to marrow spaces in distant sites throughout the body. These observations suggest that BM provides an avid environment for circulating tumor lodgement and growth. Moreover, the BM is commonly the source of latent or "minimal residual disease" following treatment, raising the possibility that specific anti-apoptotic "niches" for metastatic growth may exist. Understanding the biologic architecture of this host microenvironment therefore has significant implications for our approach to tumor treatment. NIH Public AccessWhile a variety of in vitro and in vivo techniques exist to study cell transit through BM, these are limited in their ability...

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