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Inhibitory receptors bind ANGPTLs and support blood stem cells and leukaemia development.

Zheng J, Umikawa M, Cui C, Li J, Chen X, Zhang C, Huynh H, Hyunh H, Kang X, Silvany R, Wan X, Ye J, Cantó AP, Chen SH, Wang HY, Ward ES, Zhang CC - Nature (2012)

Bottom Line: Angiopoietin-like proteins (ANGPTLs), a family of seven secreted glycoproteins, are known to support the activity of haematopoietic stem cells (HSCs) in vitro and in vivo.ANGPTLs also have important roles in lipid metabolism, angiogenesis and inflammation, but were considered 'orphan ligands' because no receptors were identified.In mouse transplantation acute myeloid leukaemia models, a deficiency in intracellular signalling of PIRB resulted in increased differentiation of leukaemia cells, revealing that PIRB supports leukaemia development.

View Article: PubMed Central - PubMed

Affiliation: Departments of Physiology and Developmental Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.

ABSTRACT
How environmental cues regulate adult stem cell and cancer cell activity through surface receptors is poorly understood. Angiopoietin-like proteins (ANGPTLs), a family of seven secreted glycoproteins, are known to support the activity of haematopoietic stem cells (HSCs) in vitro and in vivo. ANGPTLs also have important roles in lipid metabolism, angiogenesis and inflammation, but were considered 'orphan ligands' because no receptors were identified. Here we show that the immune-inhibitory receptor human leukocyte immunoglobulin-like receptor B2 (LILRB2) and its mouse orthologue paired immunoglobulin-like receptor (PIRB) are receptors for several ANGPTLs. LILRB2 and PIRB are expressed on human and mouse HSCs, respectively, and the binding of ANGPTLs to these receptors supported ex vivo expansion of HSCs. In mouse transplantation acute myeloid leukaemia models, a deficiency in intracellular signalling of PIRB resulted in increased differentiation of leukaemia cells, revealing that PIRB supports leukaemia development. Our study indicates an unexpected functional significance of classical immune-inhibitory receptors in maintenance of stemness of normal adult stem cells and in support of cancer development.

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PirB suppresses differentiation and enhances development of MLL-AF9 AMLa, PirB expression on YFP+Mac-1+Kit+ AML cells as determined by flow cytometery. b, Survival curve of mice receiving MLL-AF9-infected WT or PirBTM hematopoietic progenitors (n = 15); p < 0.05. c, Comparison of the sizes of spleen, liver, and numbers of peripheral blood cells of the mice transplanted with WT MLL-AF9 cells and those with PirBTM MLL-AF9 cells at 28 d after transplantation (n = 6). d, Representative flow cytometry plots showing that PirBTM AML mice have decreased Mac-1+Kit+ cells and increased differentiated cells relative to mice transplanted with WT cells at 28 d after transplantation. e, Comparison of colony forming activity of WT and PirBTM MLL-AF9+ BM cells. Shown is typical morphology of WT and PirBTM CFUs. f, PirBTM MLL-AF9 BM cells have dramatically decreased CFU forming ability in second replating (n = 3). g, GSEA plots evaluating changes in leukemia initiation/maintenance and myeloid differentiation gene signatures upon PirB signaling depletion in WT or PirBTM MLL-AF9 Mac-1+Kit+ AML cells. * p < 0.05. Error bars, s.e.m.
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Figure 4: PirB suppresses differentiation and enhances development of MLL-AF9 AMLa, PirB expression on YFP+Mac-1+Kit+ AML cells as determined by flow cytometery. b, Survival curve of mice receiving MLL-AF9-infected WT or PirBTM hematopoietic progenitors (n = 15); p < 0.05. c, Comparison of the sizes of spleen, liver, and numbers of peripheral blood cells of the mice transplanted with WT MLL-AF9 cells and those with PirBTM MLL-AF9 cells at 28 d after transplantation (n = 6). d, Representative flow cytometry plots showing that PirBTM AML mice have decreased Mac-1+Kit+ cells and increased differentiated cells relative to mice transplanted with WT cells at 28 d after transplantation. e, Comparison of colony forming activity of WT and PirBTM MLL-AF9+ BM cells. Shown is typical morphology of WT and PirBTM CFUs. f, PirBTM MLL-AF9 BM cells have dramatically decreased CFU forming ability in second replating (n = 3). g, GSEA plots evaluating changes in leukemia initiation/maintenance and myeloid differentiation gene signatures upon PirB signaling depletion in WT or PirBTM MLL-AF9 Mac-1+Kit+ AML cells. * p < 0.05. Error bars, s.e.m.

Mentions: Based on our in silico analysis of a pool of 9004 samples described previously 21, the level of LILRB2 mRNA is at least 4-fold higher in the human acute monoblastic and monocytic leukemia cells (M5 subtype of acute myeloid leukemia (AML)) than in other AML cells (Supplementary Fig. 15). Since human acute monoblastic and monocytic leukemia cells are often associated with rearrangement of MLL (a histone methyltransferase deemed a positive global regulator of gene transcription), we used a retroviral MLL-AF9 transplantation mouse model 22,23 to further examine the role of PirB in regulation of AML development. WT or PirBTM donor Lin− cells infected by retroviral MLL-AF9-IRES-YFP were used to induce AML as previously described 22,23. We examined PirB expression in YFP+Mac-1+Kit+ cells that may be enriched for AML initiating activity 22,23, and found that about 80% YFP+Mac-1+Kit+ cells were PirB+ (Fig. 4a). We next investigated whether PirB was required for the induction of AML by MLL-AF9. Mice transplanted with MLL-AF9-transduced WT cells developed AML and died within approximately 5 weeks, whereas those transplanted with MLL-AF9-transduced PirBTM cells were resistant to the induction of MLL-AF9 and developed AML much more slowly (Fig. 4b, Supplementary Fig. 16). The significantly delayed development of the PirBTM leukemia was correlated with about 50% lower numbers of white blood cells in circulation and a much less severe infiltration of myeloid leukemia cells into the liver and spleen (Fig. 4c–d). Consistently, PirB deficiency caused an approximately 50% reduction of YFP+Mac-1+Kit+ cells in both bone marrow and peripheral blood (Fig. 4d). There were more CD3+ or B220+ cells in mice that received MLL-AF9-transduced PirBTM donor cells than in those given WT cells (Fig. 4d). These results demonstrate that PirB mediated signaling is associated with faster AML development and greater numbers of YFP+Mac-1+Kit+ AML cells in vivo.


Inhibitory receptors bind ANGPTLs and support blood stem cells and leukaemia development.

Zheng J, Umikawa M, Cui C, Li J, Chen X, Zhang C, Huynh H, Hyunh H, Kang X, Silvany R, Wan X, Ye J, Cantó AP, Chen SH, Wang HY, Ward ES, Zhang CC - Nature (2012)

PirB suppresses differentiation and enhances development of MLL-AF9 AMLa, PirB expression on YFP+Mac-1+Kit+ AML cells as determined by flow cytometery. b, Survival curve of mice receiving MLL-AF9-infected WT or PirBTM hematopoietic progenitors (n = 15); p < 0.05. c, Comparison of the sizes of spleen, liver, and numbers of peripheral blood cells of the mice transplanted with WT MLL-AF9 cells and those with PirBTM MLL-AF9 cells at 28 d after transplantation (n = 6). d, Representative flow cytometry plots showing that PirBTM AML mice have decreased Mac-1+Kit+ cells and increased differentiated cells relative to mice transplanted with WT cells at 28 d after transplantation. e, Comparison of colony forming activity of WT and PirBTM MLL-AF9+ BM cells. Shown is typical morphology of WT and PirBTM CFUs. f, PirBTM MLL-AF9 BM cells have dramatically decreased CFU forming ability in second replating (n = 3). g, GSEA plots evaluating changes in leukemia initiation/maintenance and myeloid differentiation gene signatures upon PirB signaling depletion in WT or PirBTM MLL-AF9 Mac-1+Kit+ AML cells. * p < 0.05. Error bars, s.e.m.
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Figure 4: PirB suppresses differentiation and enhances development of MLL-AF9 AMLa, PirB expression on YFP+Mac-1+Kit+ AML cells as determined by flow cytometery. b, Survival curve of mice receiving MLL-AF9-infected WT or PirBTM hematopoietic progenitors (n = 15); p < 0.05. c, Comparison of the sizes of spleen, liver, and numbers of peripheral blood cells of the mice transplanted with WT MLL-AF9 cells and those with PirBTM MLL-AF9 cells at 28 d after transplantation (n = 6). d, Representative flow cytometry plots showing that PirBTM AML mice have decreased Mac-1+Kit+ cells and increased differentiated cells relative to mice transplanted with WT cells at 28 d after transplantation. e, Comparison of colony forming activity of WT and PirBTM MLL-AF9+ BM cells. Shown is typical morphology of WT and PirBTM CFUs. f, PirBTM MLL-AF9 BM cells have dramatically decreased CFU forming ability in second replating (n = 3). g, GSEA plots evaluating changes in leukemia initiation/maintenance and myeloid differentiation gene signatures upon PirB signaling depletion in WT or PirBTM MLL-AF9 Mac-1+Kit+ AML cells. * p < 0.05. Error bars, s.e.m.
Mentions: Based on our in silico analysis of a pool of 9004 samples described previously 21, the level of LILRB2 mRNA is at least 4-fold higher in the human acute monoblastic and monocytic leukemia cells (M5 subtype of acute myeloid leukemia (AML)) than in other AML cells (Supplementary Fig. 15). Since human acute monoblastic and monocytic leukemia cells are often associated with rearrangement of MLL (a histone methyltransferase deemed a positive global regulator of gene transcription), we used a retroviral MLL-AF9 transplantation mouse model 22,23 to further examine the role of PirB in regulation of AML development. WT or PirBTM donor Lin− cells infected by retroviral MLL-AF9-IRES-YFP were used to induce AML as previously described 22,23. We examined PirB expression in YFP+Mac-1+Kit+ cells that may be enriched for AML initiating activity 22,23, and found that about 80% YFP+Mac-1+Kit+ cells were PirB+ (Fig. 4a). We next investigated whether PirB was required for the induction of AML by MLL-AF9. Mice transplanted with MLL-AF9-transduced WT cells developed AML and died within approximately 5 weeks, whereas those transplanted with MLL-AF9-transduced PirBTM cells were resistant to the induction of MLL-AF9 and developed AML much more slowly (Fig. 4b, Supplementary Fig. 16). The significantly delayed development of the PirBTM leukemia was correlated with about 50% lower numbers of white blood cells in circulation and a much less severe infiltration of myeloid leukemia cells into the liver and spleen (Fig. 4c–d). Consistently, PirB deficiency caused an approximately 50% reduction of YFP+Mac-1+Kit+ cells in both bone marrow and peripheral blood (Fig. 4d). There were more CD3+ or B220+ cells in mice that received MLL-AF9-transduced PirBTM donor cells than in those given WT cells (Fig. 4d). These results demonstrate that PirB mediated signaling is associated with faster AML development and greater numbers of YFP+Mac-1+Kit+ AML cells in vivo.

Bottom Line: Angiopoietin-like proteins (ANGPTLs), a family of seven secreted glycoproteins, are known to support the activity of haematopoietic stem cells (HSCs) in vitro and in vivo.ANGPTLs also have important roles in lipid metabolism, angiogenesis and inflammation, but were considered 'orphan ligands' because no receptors were identified.In mouse transplantation acute myeloid leukaemia models, a deficiency in intracellular signalling of PIRB resulted in increased differentiation of leukaemia cells, revealing that PIRB supports leukaemia development.

View Article: PubMed Central - PubMed

Affiliation: Departments of Physiology and Developmental Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.

ABSTRACT
How environmental cues regulate adult stem cell and cancer cell activity through surface receptors is poorly understood. Angiopoietin-like proteins (ANGPTLs), a family of seven secreted glycoproteins, are known to support the activity of haematopoietic stem cells (HSCs) in vitro and in vivo. ANGPTLs also have important roles in lipid metabolism, angiogenesis and inflammation, but were considered 'orphan ligands' because no receptors were identified. Here we show that the immune-inhibitory receptor human leukocyte immunoglobulin-like receptor B2 (LILRB2) and its mouse orthologue paired immunoglobulin-like receptor (PIRB) are receptors for several ANGPTLs. LILRB2 and PIRB are expressed on human and mouse HSCs, respectively, and the binding of ANGPTLs to these receptors supported ex vivo expansion of HSCs. In mouse transplantation acute myeloid leukaemia models, a deficiency in intracellular signalling of PIRB resulted in increased differentiation of leukaemia cells, revealing that PIRB supports leukaemia development. Our study indicates an unexpected functional significance of classical immune-inhibitory receptors in maintenance of stemness of normal adult stem cells and in support of cancer development.

Show MeSH
Related in: MedlinePlus