PATTERNS OF INTERACTION BETWEEN MULTIPOTENT MESENCHYMAL STROMA CELLS (MSC) AND ACUTE MYELOID LEUKEMIA (AML) CELLS IN VITRO AND IN VIVO
(Abstract release date: 05/19/16)
EHA Library. Schelker R. 06/09/16; 132456; E907
Disclosure(s): There is no need to disclose actual financial value
Dr. Roland Schelker
Contributions
Contributions
Abstract
Abstract: E907
Type: Eposter Presentation
Background
MSC are a key factor within the bone marrow (BM) niche by regulating hematopoietic stem cells (HSC) within the endosteal and perivascular region. Moreover, MSC are suspected to support the aberrant hematopoiesis in a similar manner.
Aims
In our study we sought to investigate the effect of MSC on AML cells in vitro and in vivo.
Methods
MSC from AML patients (LD) and donors without BM disorder (BD) were obtained from BM aspirates. Phenotypic and functional criteria of MSC were tested subsequently. Liquid culture assays were performed from LD and BD samples to compare expansion capacity. Co-cultures of AML cells (LC) on MSC were initiated and compared to those on mouse fibroblasts (MS5). Additionally, the effect of blocking CXCR4 and TGF-b1 on proliferation of LC was tested in these co-cultures with and without cytarabine supplementation. Ultrastructural intercellular communication of MSC and LC was investigated by electron microscopy (ECM). Gene expression profiles of cultured LD-MSC vs. BD-MSC were established by using Affimetrix gene chip. Finally, homing and engraftment of MSC, LC and combinations thereof in NSG mice were investigated.
Results
MSC were successfully harvested from BM aspirates and showed a typical differentiation pattern and surface marker profile. LD-MSC showed significant decreased expansion capacity compared to BD-MSC. Co-culture of LC on MSC resulted in a higher proliferation than on MS5. Blocking TGF-b1 but not CXCR4 increased proliferation of LC and enhanced chemo-sensitivity towards cytarabine. ECM analysis showed no evidence for junctional bridging between LC and MSC. Gene analysis revealed a marked overexpression of stanniocalcin-1 (STC-1) in LD-MSC compared to BD-MSC. mRNA and western blot analysis as well as ELISA and immunohistological findings confirmed this observation. In vivo experiments emphasized that HSC and LC exhibited significant higher homing efficiency than MSC, the latter being mainly trapped within the lung. Mice treated (transplanted) with MSC showed decreased cytarabine sensibility as compared to control mice without MSC.
Conclusion
MSC derived from patients with AML show distinct differences to MSC from BD. MSC support LC in vitro and in vivo. TGF-b1, CXCR4 and STC-1 signaling may play an important role within the leukemic niche.
Session topic: E-poster
Keyword(s): CXCR4, Leukemogenesis, Stem cell niche, TGF-
Type: Eposter Presentation
Background
MSC are a key factor within the bone marrow (BM) niche by regulating hematopoietic stem cells (HSC) within the endosteal and perivascular region. Moreover, MSC are suspected to support the aberrant hematopoiesis in a similar manner.
Aims
In our study we sought to investigate the effect of MSC on AML cells in vitro and in vivo.
Methods
MSC from AML patients (LD) and donors without BM disorder (BD) were obtained from BM aspirates. Phenotypic and functional criteria of MSC were tested subsequently. Liquid culture assays were performed from LD and BD samples to compare expansion capacity. Co-cultures of AML cells (LC) on MSC were initiated and compared to those on mouse fibroblasts (MS5). Additionally, the effect of blocking CXCR4 and TGF-b1 on proliferation of LC was tested in these co-cultures with and without cytarabine supplementation. Ultrastructural intercellular communication of MSC and LC was investigated by electron microscopy (ECM). Gene expression profiles of cultured LD-MSC vs. BD-MSC were established by using Affimetrix gene chip. Finally, homing and engraftment of MSC, LC and combinations thereof in NSG mice were investigated.
Results
MSC were successfully harvested from BM aspirates and showed a typical differentiation pattern and surface marker profile. LD-MSC showed significant decreased expansion capacity compared to BD-MSC. Co-culture of LC on MSC resulted in a higher proliferation than on MS5. Blocking TGF-b1 but not CXCR4 increased proliferation of LC and enhanced chemo-sensitivity towards cytarabine. ECM analysis showed no evidence for junctional bridging between LC and MSC. Gene analysis revealed a marked overexpression of stanniocalcin-1 (STC-1) in LD-MSC compared to BD-MSC. mRNA and western blot analysis as well as ELISA and immunohistological findings confirmed this observation. In vivo experiments emphasized that HSC and LC exhibited significant higher homing efficiency than MSC, the latter being mainly trapped within the lung. Mice treated (transplanted) with MSC showed decreased cytarabine sensibility as compared to control mice without MSC.
Conclusion
MSC derived from patients with AML show distinct differences to MSC from BD. MSC support LC in vitro and in vivo. TGF-b1, CXCR4 and STC-1 signaling may play an important role within the leukemic niche.
Session topic: E-poster
Keyword(s): CXCR4, Leukemogenesis, Stem cell niche, TGF-
Abstract: E907
Type: Eposter Presentation
Background
MSC are a key factor within the bone marrow (BM) niche by regulating hematopoietic stem cells (HSC) within the endosteal and perivascular region. Moreover, MSC are suspected to support the aberrant hematopoiesis in a similar manner.
Aims
In our study we sought to investigate the effect of MSC on AML cells in vitro and in vivo.
Methods
MSC from AML patients (LD) and donors without BM disorder (BD) were obtained from BM aspirates. Phenotypic and functional criteria of MSC were tested subsequently. Liquid culture assays were performed from LD and BD samples to compare expansion capacity. Co-cultures of AML cells (LC) on MSC were initiated and compared to those on mouse fibroblasts (MS5). Additionally, the effect of blocking CXCR4 and TGF-b1 on proliferation of LC was tested in these co-cultures with and without cytarabine supplementation. Ultrastructural intercellular communication of MSC and LC was investigated by electron microscopy (ECM). Gene expression profiles of cultured LD-MSC vs. BD-MSC were established by using Affimetrix gene chip. Finally, homing and engraftment of MSC, LC and combinations thereof in NSG mice were investigated.
Results
MSC were successfully harvested from BM aspirates and showed a typical differentiation pattern and surface marker profile. LD-MSC showed significant decreased expansion capacity compared to BD-MSC. Co-culture of LC on MSC resulted in a higher proliferation than on MS5. Blocking TGF-b1 but not CXCR4 increased proliferation of LC and enhanced chemo-sensitivity towards cytarabine. ECM analysis showed no evidence for junctional bridging between LC and MSC. Gene analysis revealed a marked overexpression of stanniocalcin-1 (STC-1) in LD-MSC compared to BD-MSC. mRNA and western blot analysis as well as ELISA and immunohistological findings confirmed this observation. In vivo experiments emphasized that HSC and LC exhibited significant higher homing efficiency than MSC, the latter being mainly trapped within the lung. Mice treated (transplanted) with MSC showed decreased cytarabine sensibility as compared to control mice without MSC.
Conclusion
MSC derived from patients with AML show distinct differences to MSC from BD. MSC support LC in vitro and in vivo. TGF-b1, CXCR4 and STC-1 signaling may play an important role within the leukemic niche.
Session topic: E-poster
Keyword(s): CXCR4, Leukemogenesis, Stem cell niche, TGF-
Type: Eposter Presentation
Background
MSC are a key factor within the bone marrow (BM) niche by regulating hematopoietic stem cells (HSC) within the endosteal and perivascular region. Moreover, MSC are suspected to support the aberrant hematopoiesis in a similar manner.
Aims
In our study we sought to investigate the effect of MSC on AML cells in vitro and in vivo.
Methods
MSC from AML patients (LD) and donors without BM disorder (BD) were obtained from BM aspirates. Phenotypic and functional criteria of MSC were tested subsequently. Liquid culture assays were performed from LD and BD samples to compare expansion capacity. Co-cultures of AML cells (LC) on MSC were initiated and compared to those on mouse fibroblasts (MS5). Additionally, the effect of blocking CXCR4 and TGF-b1 on proliferation of LC was tested in these co-cultures with and without cytarabine supplementation. Ultrastructural intercellular communication of MSC and LC was investigated by electron microscopy (ECM). Gene expression profiles of cultured LD-MSC vs. BD-MSC were established by using Affimetrix gene chip. Finally, homing and engraftment of MSC, LC and combinations thereof in NSG mice were investigated.
Results
MSC were successfully harvested from BM aspirates and showed a typical differentiation pattern and surface marker profile. LD-MSC showed significant decreased expansion capacity compared to BD-MSC. Co-culture of LC on MSC resulted in a higher proliferation than on MS5. Blocking TGF-b1 but not CXCR4 increased proliferation of LC and enhanced chemo-sensitivity towards cytarabine. ECM analysis showed no evidence for junctional bridging between LC and MSC. Gene analysis revealed a marked overexpression of stanniocalcin-1 (STC-1) in LD-MSC compared to BD-MSC. mRNA and western blot analysis as well as ELISA and immunohistological findings confirmed this observation. In vivo experiments emphasized that HSC and LC exhibited significant higher homing efficiency than MSC, the latter being mainly trapped within the lung. Mice treated (transplanted) with MSC showed decreased cytarabine sensibility as compared to control mice without MSC.
Conclusion
MSC derived from patients with AML show distinct differences to MSC from BD. MSC support LC in vitro and in vivo. TGF-b1, CXCR4 and STC-1 signaling may play an important role within the leukemic niche.
Session topic: E-poster
Keyword(s): CXCR4, Leukemogenesis, Stem cell niche, TGF-
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