Both pharmacological mobilization of hematopoietic stem progenitor cells (HSPCs) and myeloablative conditioning for transplantation activate the complement cascade (CC) in bone marrow (BM), and in the previous funding period we focused on the role of the third complement component (C3) in mobilization and homing of HSPCs. In this competing renewal we will focus on the fifth complement component (C5), which is located downstream from C3. C5 cleavage leads to generation of the potent anaphylatoxins C5a and desArgC5a and generation of non-lytic and lytic C5b-C9, known as the membrane attack complex (MAC). Our recently published data support a crucial role for C5a/C5b-C9 in trafficking of HSPCs. In particular, C5-deficient mice are poor mobilizers in response to granulocyte colony stimulating factor (G-CSF) and exhibit delayed hematopoietic reconstitution after transplantation with wild type (WT) BM cells. Based on these findings, the central hypothesis of this competing renewal is that the activation of distal steps of the complement cascade and release of C5a and C5b-C9 (MAC) are important regulators of mobilization and homing of HSPCs. To address this hypothesis, we propose:
Specific Aim 1. The molecular basis of the mobilization defect in C5-deficient mice. Based on our observation that C5-deficient mice are poor G-CSF mobilizers, we will address whether C5 cleavage is i) required for circadian HSPC mobilization, ii) mobilization of HSPC induced by other agents and iii) whether C5 activation and S1P plasma level correlates with mobilization efficacy in patients. We will also focus on the role of erythrocyte-released S1P in mobilization by performing mobilization studies in CD55/CD59-deficient mice that are highly sensitive to C5b-C9 (MAC) lysis. Finally, we will focus on the role of C5b-C9-induced release of S1P in mobilization observed in patients with hemolytic syndromes.
Specific Aim 2. The molecular basis of the homing defect in C5-deficient mice. We found that the CC is activated in lethally irradiated mice and that C5-deficient mice engraft poorly with WT BM cells. To shed more light on the role of the distal part of the CC (C5a/C5b-C9) in homing/engraftment, we will study whether conditioning for transplantation by chemotherapy also activates the CC in BM and study the effect of C5a/C5b-C9 on expression of i) HSPC chemoattractants in BM, ii) adhesion/tethering molecules for HSPCs on BM endothelium, and ii) on secretion of factors facilitating homing.
Specific Aim 3. The C5a/C5b-C9-induced BM stroma-derived cathelicidin (LL-37) as a potent homing sensitizing factor. We found that LL-37, like C3a, is a very strong priming factor for SDF-1 that becomes upregulated in BM stroma after conditioning for transplantation in a C5a/C5b-C9-dependent manner. In an immunodeficient mouse model, we will test whether short exposure of HSPCs to LL-37 before transplantation increases homing of human BM- and umbilical cord blood (UCB)-derived HSPCs. We will also address whether LL-37 also primes the responsiveness of HSPCs to other homing factors and focus on the molecular mechanism of these phenomena.

Public Health Relevance

Section In this competing renewal, we will focus on the role of a major component of innate immunity, namely, activation of the complement cascade in trafficking of hematopoietic stem/progenitor cells. This application will allow us to better understand this process, as well as lead to the development of novel therapeutic strategies to treat patients with leukemias and inborn defects of hematopoiesis by transplantation of hematopoietic stem cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK074720-07
Application #
8628110
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Bishop, Terry Rogers
Project Start
2006-04-01
Project End
2017-02-28
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
7
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Louisville
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
City
Louisville
State
KY
Country
United States
Zip Code
40202
Adamiak, Mateusz; Abdel-Latif, Ahmed; Ratajczak, Mariusz Z (2017) Mannan binding lectin triggers mobilization of hematopoietic stem cells. Oncotarget 8:73368-73369
Ratajczak, Mariusz Z; Ratajczak, Janina (2017) Extracellular Microvesicles as Game Changers in Better Understanding the Complexity of Cellular Interactions-From Bench to Clinical Applications. Am J Med Sci 354:449-452
Adamiak, Mateusz; Abdelbaset-Ismail, Ahmed; Moore 4th, Joseph B et al. (2017) Inducible Nitric Oxide Synthase (iNOS) Is a Novel Negative Regulator of Hematopoietic Stem/Progenitor Cell Trafficking. Stem Cell Rev 13:92-103
Abdelbaset-Ismail, Ahmed; Pedziwiatr, Daniel; Schneider, Gabriela et al. (2017) Pituitary sex hormones enhance the pro?metastatic potential of human lung cancer cells by downregulating the intracellular expression of heme oxygenase?1. Int J Oncol 50:317-328
Abdelbaset-Ismail, A; Borkowska-Rzeszotek, S; Kubis, E et al. (2017) Activation of the complement cascade enhances motility of leukemic cells by downregulating expression of HO-1. Leukemia 31:446-458
Ratajczak, Mariusz Z; Ratajczak, Janina; Suszynska, Malwina et al. (2017) A Novel View of the Adult Stem Cell Compartment From the Perspective of a Quiescent Population of Very Small Embryonic-Like Stem Cells. Circ Res 120:166-178
Ratajczak, Mariusz Z; Bartke, Andrzej; Darzynkiewicz, Zbigniew (2017) Prolonged Growth Hormone/Insulin/Insulin-like Growth Factor Nutrient Response Signaling Pathway as a Silent Killer of Stem Cells and a Culprit in Aging. Stem Cell Rev 13:443-453
Bujko, Kamila; Rzeszotek, Sylwia; Hoehlig, Kai et al. (2017) Signaling of the Complement Cleavage Product Anaphylatoxin C5a Through C5aR (CD88) Contributes to Pharmacological Hematopoietic Stem Cell Mobilization. Stem Cell Rev 13:793-800
Sielatycka, Katarzyna; Poniewierska-Baran, Agata; Nurek, Karolina et al. (2017) Novel View on Umbilical Cord Blood and Maternal Peripheral Blood-an Evidence for an Increase in the Number of Circulating Stem Cells on Both Sides of the Fetal-Maternal Circulation Barrier. Stem Cell Rev 13:774-780
Ratajczak, M Z (2017) Why are hematopoietic stem cells so 'sexy'? on a search for developmental explanation. Leukemia 31:1671-1677

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