CD133+ adult bone marrow stem cells to promote liver regeneration - the role of platelets for hepatic homing and analyses of released factors for differentiation as well as proliferation subsequent to clinical hepatic resection
Final Report Abstract
In up to 45 % of patients with primary or secondary liver tumors extended hepatectomy is necessary to achieve tumor negative resection margins. Patients with an anticipated future liver remnant volume (FLRV) below 25% of total liver volume have an increased incidence of post-operative morbidity and mortality. Bone marrow stem cells (BMSC) have been shown to participate in liver regeneration subsequent to hepatic damage. Our group was the first to report clinical experience with therapeutic application of BMSC to promote liver proliferation subsequent to partial hepatic ischemia but the molecular mechanisms of BMSC mobilization and homing to the liver after partial hepatectomy (PH) remain largely unexplored. Correlation of peripheral CD133+ BMSC mobilization with the extent of resected liver volume and its regain and with paracrine factors revealed significantly higher levels of peripheral BMSC which were positively correlated with the extent of resected liver volume and its regain after 3 weeks and increased serum levels of HGF, SDF-1 and IGF-1 in the first 6h and of AFP beyond 24h in patients with extended liver resections. The evaluation of peripheral human AFP expression demonstrated pronounced sternness following increased CD133+ BMSC in the course of liver regeneration following PH. Migration assays of human BMSC in vitro showed a specific target-demonstrated migration towards recombinant HGF and SDF-1 gradients in a concentration and specific receptor (c-Met and CXCR4) dependent manner. These results show that HGF and SDF-1 are required for effective HSC mobilization and homing to the liver after hepatic resection. These findings have significant implications for potential therapeutic strategies targeting chemotactant modulation and stem cell mobilization for liver protection and regeneration. Analyzing the potential of platelets to promote CD133+ BMSC participation in liver regeneration we showed that CD133+ BMSC inhibit ADP-dependent activation of platelets in a contact dependent manner. Our data suggest this effect is CD39- dependent and is preferentially associated with BM derived cells, rather than peripheral blood apheresis samples. We hypothesize CD133+ BMSC might interact with platelets and endothelium of regenerating organs and tissue. Further studies will investigate the relevance of CD39 expression by BMSC and dissect how these interactions might impact liver regeneration in vivo. To further investigate the promoting potential of platelets for the homing of CD133+ BMSC along endothelium we established co-culture models of CD133+ BMSC with human and mouse endothelial cells (HMEC-1/mDMEC) and mouse hepatic sinusoidal endothelial cells (mLSEC) in capillaries of a live cell imaging system. Our data indicate platelets to trigger adhesion of CD133+ BMSC to human and murine micro vasculature in a stimulation independent manner. Further we demonstrate here for the first time that platelets bear the capacity to promote BMSC-adhesion to hepatic vasculature. These data may add to the understanding of mechanisms by which platelets support hepatic generation and offer novel strategies to increase the efficacy of therapeutic BMSC-application in clinical liver disease. Further data suggest SDF-1 and P-selectin with its ligand PSGL-1 to possess an independent promoting effect for homing of CD133+ BMSC along vasculature in a nonsynergistically mode, which may have relevance for BMSC homing along hepatic sinusoids during liver regeneration. To gain insight In faith and mechanisms of CD133+ BMSC applied to improve physiological liver regeneration processes we evaluated early aspects of BMSC-homing along (hepatic) vasculature following liver damage and subsequent regeneration processes utilising experimental models of isolated rat liver perfusion (IPRL). We were able to demonstrate hepatic homing and extravasation of CD133+BMSC infused to an IPRL subsequent to warm ischemia. Platelets demonstrated to have a positive effect on BMSC homing to the extrasinusoidal space of the liver. Future investigations will deeper characterize the interaction of CD133+BMSC and platelets with the hepatic microenvironment.
Publications
- (2012) Infusion of CD133+ bone marrow derived stem cells after selective portal embolization enhances functional hepatic reserves after extended right hepatectomy: A retrospective single center study. Ann Surg. 255(1): 79-85
J. Schulte am Esch, M. Schmelzle, G. Fürst, S. C. Robson, A. Krieg, C. Duhme, R. Y. Tustas, A. Alexander. H. M. Klein, S. A. Topp, J. G. Bode, D. Häussinger, C. F. Eisenberger, W. T. Knoefel
(See online at https://doi.org/10.1097/SLA.0b013e31823d7d08) - (2013) CD39 modulates hematopoietic stem cell recruitment and promotes liver regeneration in mice and humans after partial hepatectomy. Ann Surg. 257(4): 693-701
M. Schmelzle, C. Duhme, W. Junger, S.D. Salhanick, Y. Chen, Y. Wu, V. Toxavidis, E. Csizmadia, L. Han, S. Bian, G. Fürst, M. Nowak, S. J. Karp, W. T. Knoefel, J. Schulte Esch, S. C. Robson
(See online at https://doi.org/10.1097/SLA.0b013e31826c3ec2) - (2013) Increased plasma levels of microparticles expressing CD39 and CD133 in acute liver injury. Transplantation 15; 95(1): 63-9
Schmelzle M, Splith K, Andersen LW, Kornek M, Schuppen D, Jones- Bamman C, Nowak M, Toxavidis V, Salhanick SD, Han L, Schulte am Esch J, Jonas S, Donnino MW, Robson SC
(See online at https://doi.org/10.1097/TP.0b013e318278d3cd) - (2013) Stern cell-induced liver regeneration. Zentralbl Chir. 138(2): 166-72
Knoefel WT, Alexander A, Tustas RY, Schmelzle M, Klein HM, Krieg A, Topp SA, Eisenberger CF, Fuerst G, Schulte am Esch J
- (2014) HGF and SDF-1-mediated mobilization of CD133+ BMSC for hepatic regeneration following extensive liver resection. Liver Int. 34(1): 89-101
N. Lehwald, C. Duhme, M. Wildner, S. Kuhn, G. Fürst, S. J. Forbes, S. Jonas, S. C. Robson, W. T. Knoefel, M. Schmelzle, J. Schulte Am Esch
(See online at https://doi.org/10.1111/liv.12195) - (2016) Reply to Letter: Infusion of CD 7 33+ Bone Marrow-derived Stem Cell after Selective Portal Vein Embolization Enhances Functional Hepatic Reserves after Extended Right Hepatectomy”. Ann Surg. 263(3): e44-5
J. Schulte am Esch, M. Schmelzle, C. Duhme, G. Fürst, S. C. Robson, J. G. Bode, A. Krieg, S. A. Topp, D. Häussinger, W. T. Knoefel
(See online at https://dx.doi.org/10.1097/SLA.0000000000000849)
