Pharmacological inhibition of caspase and calpain proteases: a novel strategy to enhance the homing responses of cord blood HSPCs during expansion
| Autor: | Darshana Kadekar, Vaijayanti Kale, V M Sangeetha, Lalita Limaye |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
rho GTP-Binding Proteins
Proteases Receptors CXCR4 Anatomy and Physiology Integrin alpha4 lcsh:Medicine Integrin alpha5 Amino Acid Chloromethyl Ketones Mice Cell Movement medicine Cell Adhesion Animals Humans Protease Inhibitors Cell adhesion lcsh:Science Protein Structure Quaternary Biology Caspase Cell Size Multidisciplinary biology Calpain Stem Cells lcsh:R Cell Polarity Hematology Fetal Blood Hematopoietic Stem Cells Caspase Inhibitors Actins Chemokine CXCL12 Cell biology Transplantation Haematopoiesis medicine.anatomical_structure Gene Expression Regulation biology.protein Medicine lcsh:Q Female Bone marrow Protein Multimerization Cell Adhesion Molecules Homing (hematopoietic) Research Article Developmental Biology |
| Zdroj: | PLoS ONE PLoS ONE, Vol 7, Iss 1, p e29383 (2012) |
| ISSN: | 1932-6203 |
| Popis: | Background Expansion of hematopoietic stem/progenitor cells (HSPCs) is a well-known strategy employed to facilitate the transplantation outcome. We have previously shown that the prevention of apoptosis by the inhibition of cysteine proteases, caspase and calpain played an important role in the expansion and engraftment of cord blood (CB) derived HSPCs. We hypothesize that these protease inhibitors might have maneuvered the adhesive and migratory properties of the cells rendering them to be retained in the bone marrow for sustained engraftment. The current study was aimed to investigate the mechanism of the homing responses of CB cells during expansion. Methodology/Principal Findings CB derived CD34+ cells were expanded using a combination of growth factors with and without Caspase inhibitor -zVADfmk or Calpain 1 inhibitor- zLLYfmk. The cells were analyzed for the expression of homing-related molecules. In vitro adhesive/migratory interactions and actin polymerization dynamics of HSPCs were assessed. In vivo homing assays were carried out in NOD/SCID mice to corroborate these observations. We observed that the presence of zVADfmk or zLLYfmk (inhibitors) caused the functional up regulation of CXCR4, integrins, and adhesion molecules, reflecting in a higher migration and adhesive interactions in vitro. The enhanced actin polymerization and the RhoGTPase protein expression complemented these observations. Furthermore, in vivo experiments showed a significantly enhanced homing to the bone marrow of NOD/SCID mice. Conclusion/Significance Our present study reveals another novel aspect of the regulation of caspase and calpain proteases in the biology of HSPCs. The priming of the homing responses of the inhibitor-cultured HSPCs compared to the cytokine-graft suggests that the modulation of these proteases may help in overcoming the major homing defects prevalent in the expansion cultures thereby facilitating the manipulation of cells for transplant procedures. |
| Databáze: | OpenAIRE |
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