The CXCR4-SDF-1 axis plays a central role in the trafficking and retention of normal and malignant stem cells in the bone marrow (BM) microenvironment. Here, we used a mouse model of acute promyelocytic leukemia (APL) and a small molecule competitive antagonist of CXCR4, AMD3100, to examine the interaction of mouse APL cells with the BM microenvironment. APL cells from a murine cathepsin G-PML-RAR␣ knockin mouse were genetically modified with firefly luciferase (APL luc ) to allow tracking by bioluminescence imaging. Coculture of APL luc cells with M2-10B4 stromal cells protected the leukemia cells from chemotherapy-induced apoptosis in vitro. Upon injection into syngeneic recipients, APL luc cells rapidly migrated to the BM followed by egress to the spleen then to the peripheral blood with death due to leukostasis by day 15. Administration of AMD3100 to leukemic mice induced a 1.6-fold increase in total leukocytes and a 9-fold increase of circulating APL blast counts, which peak at 3 hours and return to baseline by 12 hours. Treatment of leukemic mice with chemotherapy plus AMD3100 resulted in decreased tumor burden and improved overall survival compared with mice treated with chemotherapy alone. These studies provide a proof-of-principle for directing therapy to the critical tethers that promote AMLniche interactions. (Blood. 2009;113: 6206-6214) IntroductionHematopoietic stem cells (HSCs) reside in the bone marrow (BM) and interact with a highly organized microenvironment composed of a diverse population of stromal cells and an extracellular matrix rich in fibronectin, collagens, and various proteoglycans. The interaction between HSCs and the BM microenvironment is critical in regulating HSC processes such as trafficking, self-renewal, proliferation, and differentiation.Egress (mobilization) of stem cells from the BM to the peripheral blood can be induced by cytokines (G-CSF and GM-CSF), chemokines (Gro-␤ and IL-8), or by small molecule inhibitors of both CXCR4 and VLA-4. 1 Interaction between the chemokine, SDF-1, and its cognate receptor CXCR4 functions as a key regulator of stem cell mobilization and trafficking. 2,3 Constitutive secretion of SDF-1 by marrow stromal cells creates a gradient by which HSCs expressing the receptor CXCR4 home to and interact with its marrow niche. 4 In response to factors such as G-CSF, SDF-1 production is down-regulated by stromal cells that release HSCs into the peripheral circulation. 5 AMD3100 is a bicyclam molecule that potently, selectively, and reversibly antagonizes the binding of SDF-1 to CXCR4. 6 In multiple clinical studies, AMD3100 rapidly and effectively mobilizes HSCs into the peripheral circulation and is currently under development as a stem cell mobilization agent prior to high-dose chemotherapy for multiple myeloma, non-Hodgkin lymphoma, and other hematologic malignancies. [7][8][9] In acute myeloid leukemia (AML), the bone marrow microenvironment provides the primary site of minimal residual disease after chemotherapy. [10][11][12] Similar to normal HSCs, AM...

Here we show that interruption of the VCAM-1/VLA-4 axis with a small molecule inhibitor of VLA-4, BIO5192, results in a 30-fold increase in mobilization of murine hematopoietic stem and progenitors (HSPCs) over basal levels. An additive affect on HSPC mobilization (3-fold) was observed when plerixafor (AMD3100), a small molecule inhibitor of the CXCR-4/SDF-1 axis, was combined with BIO5192. Furthermore, the combination of granulocyte colony-stimulating factor (G-CSF), BIO5192, and plerixafor enhanced mobilization by 17-fold compared with G-CSF alone. HSPCs mobilized by BIO5192 or the combination of BIO5192 and plerixafor mobilized long-term repopulating cells, which successfully engraft and expand in a multilineage fashion in secondary transplantation recipients.

Double umbilical cord blood transplantation (dUCBT), developed as a strategy to treat larger patients with hematologic malignancies, frequently leads to the long-term establishment of a new hematopoietic system maintained by cells derived from a single UCB unit. However, predicting which unit will predominate has remained elusive. This retrospective study examined risk factor associated with unit predominance in 262 patients with hematologic malignancies who underwent dUCBT with subsequent hematopoietic recovery and complete chimerism between 2001–2009. Dual chimerism was detected at day 21–28, with subsequent single chimerism in 97% of cases by day +100 and beyond. Risk factors included nucleated cell dose, CD34+ and CD3+ cell dose, colony forming units-granulocyte macrophage dose, donor-recipient human leukocyte antigen (HLA) match, sex and ABO match, order of infusion, and cell viability. In the myeloablative setting, CD3+ cell dose was the only factor associated with unit predominance (OR 4.4, 95%CI, 1.8–10.6; p<0.01), but in the non-myeloablative setting, CD3+ cell dose (OR 2.1, 95%CI, 1.0–4.2; p=0.05) and HLA match (OR 3.4, 95%CI, 1.0–11.4; p=0.05) were independent factor associated with unit predominance. Taken together, these findings suggest that immune reactivity plays a role in unit predominance and should be considered during graft selection and graft manipulation.

Hematopoietic stem cell transplantation is the accepted therapy of choice for a variety of malignant and non-malignant diseases in children and adults. Initially developed as rescue therapy for a patient with cancer after high doses of chemotherapy and radiation as well as the correction of severe defi ciencies in the hematopoietic system, it has evolved into an adoptive immune therapy for malignancies and autoimmune disorders. The procedure has helped to obtain key information about the bone marrow environment, the biology of hematopoietic stem cells and histocompatibility. The development of this new discipline has allowed numerous groups working around the world to cure patients of diseases previously considered lethal. Together with the ever growing list of volunteer donors and umbilical cord blood banks, this has resulted in life saving therapy for thousands of patients yearly. We present an overview of the procedure from its cradle to the most novel applications, as well as the results of the HSC transplant program developed at our institution since 1989.

Delayed platelet recovery (DPR) is common after allo-SCT. Insufficient data on risk factors and association with OS and TRM are available. We conducted a retrospective analysis of all allografts at the University of Minnesota between 2000 and 2005 to characterize the frequency of DPR (platelets o50 000/lL by day 60), risk factors and related complications. A total of 850 patients with hematological malignancies and benign disorders were included. Myeloablative (MA) conditioning was used in 65% of the patients and 45% received umbilical cord blood (UCB) grafts. The 60-day cumulative incidence of platelet recovery was 40% in UCB, 57% in unrelated donor (URD) and 74% in sibling donor. Multivariate analysis confirmed that the variables associated with DPR were MA (versus reduced intensity) conditioning, graft source other than sibling donor, ABO major mismatch, recipient CMV-positive serostatus, the presence of grade II-IV acute GVHD and slower neutrophil recovery. These data demonstrate that DPR is frequent after allogeneic hematopoietic cell transplantation, especially after UCB. DPR is a significant independent risk factor for increased TRM and poorer OS along with HLA-mismatched URD, but not UCB, grade II-IV acute GVHD, old age and advanced disease stage.