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Conditioning for hematopoietic transplantation activates the complement cascade and induces a proteolytic environment in bone marrow: a novel role for bioactive lipids and soluble C5b-C9 as homing factors.

Kim CH, Wu W, Wysoczynski M, Abdel-Latif A, Sunkara M, Morris A, Kucia M, Ratajczak J, Ratajczak MZ - Leukemia (2011)

Bottom Line: As a result, BM is enriched for proteolytic enzymes and the soluble form of the terminal product of CC activation, the membrane attack complex C5b-C9 (MAC).Next, we observed that C5-deficient mice that do not generate MAC show impaired engraftment of HSPCs.We conclude that an increase in BM levels of proteolytic enzyme-resistant S1P and C1P and activation of CC, which leads to the generation of MAC, has an important and previously underappreciated role in the homing of transplanted HSPCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Stem Cell Institute at the James Graham Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA.

ABSTRACT
We have observed that conditioning for hematopoietic transplantation by lethal irradiation induces a proteolytic microenvironment in the bone marrow (BM) that activates the complement cascade (CC). As a result, BM is enriched for proteolytic enzymes and the soluble form of the terminal product of CC activation, the membrane attack complex C5b-C9 (MAC). At the same time, proteolytic enzymes induced in irradiated BM impair the chemotactic activity of α-chemokine stromal-derived factor-1 (SDF-1). As SDF-1 is considered a crucial BM chemoattractant for transplanted hematopoietic stem/progenitor cells (HSPCs), we sought to determine whether other factors that are resistant to proteolytic enzymes have a role in this process, focusing on proteolysis-resistant bioactive lipids. We found that the concentrations of sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P) increase in the BM after conditioning for transplantation and that both S1P and, as we show here for the first time, C1P are potent chemoattractants for HSPCs. Next, we observed that C5-deficient mice that do not generate MAC show impaired engraftment of HSPCs. In support of a role for MAC in homing and engraftment, we found that soluble MAC enhances in a CR3 (CD11b/CD18)-dependent manner the adhesion of HSPCs to BM stromal cells and increases the secretion of SDF-1 by BM stroma. We conclude that an increase in BM levels of proteolytic enzyme-resistant S1P and C1P and activation of CC, which leads to the generation of MAC, has an important and previously underappreciated role in the homing of transplanted HSPCs.

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Bioactive lipids show strong chemotactic activity against HSPCsPanel A: The chemotatic dose response of S1P, C1P, and SDF-1 against murine clonogenic progenitors is shown. Note that the physiological concentration of SDF-1 in biological fluids (e.g., serum) is approximately 2–5ng/ml, and at this concentration SDF-1 is not effective as a chemoattractant. In contrast, S1P is already a strong chemoattractant for murine BM-derived hematopoietic progenitors at biologically relevant concentrations. Panel B: Both C1P and S1P, like SDF-1, increase adhesion of murine clonogenic progenitors (Sca-1+ cells) to BM-derived stromal cells. The data shown represent the combined results from three independent experiments carried out in triplicate per group. Panel C: In blocking studies, Sca-1+ cells were incubated with 10μg/ml anti-VLA4 integrin mAb for 30 minutes prior to VCAM-1 adhesion assay. The data shown represent the combined results from two independent experiments carried out in quadruplicate per group. * p<0.05.
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Figure 4: Bioactive lipids show strong chemotactic activity against HSPCsPanel A: The chemotatic dose response of S1P, C1P, and SDF-1 against murine clonogenic progenitors is shown. Note that the physiological concentration of SDF-1 in biological fluids (e.g., serum) is approximately 2–5ng/ml, and at this concentration SDF-1 is not effective as a chemoattractant. In contrast, S1P is already a strong chemoattractant for murine BM-derived hematopoietic progenitors at biologically relevant concentrations. Panel B: Both C1P and S1P, like SDF-1, increase adhesion of murine clonogenic progenitors (Sca-1+ cells) to BM-derived stromal cells. The data shown represent the combined results from three independent experiments carried out in triplicate per group. Panel C: In blocking studies, Sca-1+ cells were incubated with 10μg/ml anti-VLA4 integrin mAb for 30 minutes prior to VCAM-1 adhesion assay. The data shown represent the combined results from two independent experiments carried out in quadruplicate per group. * p<0.05.

Mentions: S1P has already been reported to be a strong chemoattractant for HSPCs 15. To our surprise, we found that another bioactive lipid, C1P, in a similar way as S1P, induces several signaling pathways in murine Sca-1+ cells purified by immunomagnetic beads (Figure 3), as well as strongly chemoattacts murine HSPCs (Figure 4panel A andSupplementary Figure 1). However, both these bioactive lipids and soluble MAC (sMAC), as shown in Supplementary Figure 2 and 3 respectively, do not affect the clonogenicity of murine progenitors for all major hematopoietic lineages. The same result was found for their unphosphorylated forms (data not shown).


Conditioning for hematopoietic transplantation activates the complement cascade and induces a proteolytic environment in bone marrow: a novel role for bioactive lipids and soluble C5b-C9 as homing factors.

Kim CH, Wu W, Wysoczynski M, Abdel-Latif A, Sunkara M, Morris A, Kucia M, Ratajczak J, Ratajczak MZ - Leukemia (2011)

Bioactive lipids show strong chemotactic activity against HSPCsPanel A: The chemotatic dose response of S1P, C1P, and SDF-1 against murine clonogenic progenitors is shown. Note that the physiological concentration of SDF-1 in biological fluids (e.g., serum) is approximately 2–5ng/ml, and at this concentration SDF-1 is not effective as a chemoattractant. In contrast, S1P is already a strong chemoattractant for murine BM-derived hematopoietic progenitors at biologically relevant concentrations. Panel B: Both C1P and S1P, like SDF-1, increase adhesion of murine clonogenic progenitors (Sca-1+ cells) to BM-derived stromal cells. The data shown represent the combined results from three independent experiments carried out in triplicate per group. Panel C: In blocking studies, Sca-1+ cells were incubated with 10μg/ml anti-VLA4 integrin mAb for 30 minutes prior to VCAM-1 adhesion assay. The data shown represent the combined results from two independent experiments carried out in quadruplicate per group. * p<0.05.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3197954&req=5

Figure 4: Bioactive lipids show strong chemotactic activity against HSPCsPanel A: The chemotatic dose response of S1P, C1P, and SDF-1 against murine clonogenic progenitors is shown. Note that the physiological concentration of SDF-1 in biological fluids (e.g., serum) is approximately 2–5ng/ml, and at this concentration SDF-1 is not effective as a chemoattractant. In contrast, S1P is already a strong chemoattractant for murine BM-derived hematopoietic progenitors at biologically relevant concentrations. Panel B: Both C1P and S1P, like SDF-1, increase adhesion of murine clonogenic progenitors (Sca-1+ cells) to BM-derived stromal cells. The data shown represent the combined results from three independent experiments carried out in triplicate per group. Panel C: In blocking studies, Sca-1+ cells were incubated with 10μg/ml anti-VLA4 integrin mAb for 30 minutes prior to VCAM-1 adhesion assay. The data shown represent the combined results from two independent experiments carried out in quadruplicate per group. * p<0.05.
Mentions: S1P has already been reported to be a strong chemoattractant for HSPCs 15. To our surprise, we found that another bioactive lipid, C1P, in a similar way as S1P, induces several signaling pathways in murine Sca-1+ cells purified by immunomagnetic beads (Figure 3), as well as strongly chemoattacts murine HSPCs (Figure 4panel A andSupplementary Figure 1). However, both these bioactive lipids and soluble MAC (sMAC), as shown in Supplementary Figure 2 and 3 respectively, do not affect the clonogenicity of murine progenitors for all major hematopoietic lineages. The same result was found for their unphosphorylated forms (data not shown).

Bottom Line: As a result, BM is enriched for proteolytic enzymes and the soluble form of the terminal product of CC activation, the membrane attack complex C5b-C9 (MAC).Next, we observed that C5-deficient mice that do not generate MAC show impaired engraftment of HSPCs.We conclude that an increase in BM levels of proteolytic enzyme-resistant S1P and C1P and activation of CC, which leads to the generation of MAC, has an important and previously underappreciated role in the homing of transplanted HSPCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Stem Cell Institute at the James Graham Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA.

ABSTRACT
We have observed that conditioning for hematopoietic transplantation by lethal irradiation induces a proteolytic microenvironment in the bone marrow (BM) that activates the complement cascade (CC). As a result, BM is enriched for proteolytic enzymes and the soluble form of the terminal product of CC activation, the membrane attack complex C5b-C9 (MAC). At the same time, proteolytic enzymes induced in irradiated BM impair the chemotactic activity of α-chemokine stromal-derived factor-1 (SDF-1). As SDF-1 is considered a crucial BM chemoattractant for transplanted hematopoietic stem/progenitor cells (HSPCs), we sought to determine whether other factors that are resistant to proteolytic enzymes have a role in this process, focusing on proteolysis-resistant bioactive lipids. We found that the concentrations of sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P) increase in the BM after conditioning for transplantation and that both S1P and, as we show here for the first time, C1P are potent chemoattractants for HSPCs. Next, we observed that C5-deficient mice that do not generate MAC show impaired engraftment of HSPCs. In support of a role for MAC in homing and engraftment, we found that soluble MAC enhances in a CR3 (CD11b/CD18)-dependent manner the adhesion of HSPCs to BM stromal cells and increases the secretion of SDF-1 by BM stroma. We conclude that an increase in BM levels of proteolytic enzyme-resistant S1P and C1P and activation of CC, which leads to the generation of MAC, has an important and previously underappreciated role in the homing of transplanted HSPCs.

Show MeSH
Related in: MedlinePlus