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TCGA data and patient-derived orthotopic xenografts highlight pancreatic cancer-associated angiogenesis.

Gore J, Craven KE, Wilson JL, Cote GA, Cheng M, Nguyen HV, Cramer HM, Sherman S, Korc M - Oncotarget (2015)

Bottom Line: Inhibition of the type I TGF-β receptor with SB505124 does not alter endothelial activation in vitro, but decreases pro-angiogenic gene expression and suppresses angiogenesis in vivo.Conversely, STAT3 silencing or JAK1-2 inhibition with ruxolitinib blocks CM-enhanced EC proliferation.Thus, targeting JAK1-2 with ruxolitinib blocks a final pathway that is common to multiple pro-angiogenic factors, suppresses EC-mediated PCC proliferation, and may be useful in PDACs with a strong pro-angiogenic signature.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Indiana University School of Medicine, Indianapolis, IN 46202, USA.

ABSTRACT
Pancreatic ductal adenocarcinomas (PDACs) overexpress pro-angiogenic factors but are not viewed as vascular. Using data from The Cancer Genome Atlas we demonstrate that a subset of PDACs exhibits a strong pro-angiogenic signature that includes 37 genes, such as HDAC9, that are overexpressed in PDAC arising in KRC mice, which express mutated Kras and lack RB. Moreover, patient-derived orthotopic xenografts can exhibit tumor angiogenesis, whereas conditioned media (CM) from KRC-derived pancreatic cancer cells (PCCs) enhance endothelial cell (EC) growth and migration, and activate canonical TGF-β signaling and STAT3. Inhibition of the type I TGF-β receptor with SB505124 does not alter endothelial activation in vitro, but decreases pro-angiogenic gene expression and suppresses angiogenesis in vivo. Conversely, STAT3 silencing or JAK1-2 inhibition with ruxolitinib blocks CM-enhanced EC proliferation. STAT3 disruption also suppresses endothelial HDAC9 and blocks CM-induced HDAC9 expression, whereas HDAC9 re-expression restores CM-enhanced endothelial proliferation. Moreover, ruxolitinib blocks mitogenic EC/PCC cross-talk, and suppresses endothelial p-STAT3 and HDAC9, and PDAC progression and angiogenesis in vivo, while markedly prolonging survival of KRC mice. Thus, targeting JAK1-2 with ruxolitinib blocks a final pathway that is common to multiple pro-angiogenic factors, suppresses EC-mediated PCC proliferation, and may be useful in PDACs with a strong pro-angiogenic signature.

No MeSH data available.


Related in: MedlinePlus

A Subset of Human PDACs exhibit a strong angiogenic gene signature(A) Hierarchical clustering of TCGA data show that genes annotated to angiogenesis (white lines) are up-regulated in some human PDACs (left). Extraction (middle) and re-clustering (right) shows that many of these angiogenesis genes are up-regulated in a subset of PDACs (S = strong), whereas some (M = moderate) or few of these genes (W = weak) are increased in other PDACs. (B) H&E staining shows cytology of EUS-FNA samples (left) and histology of EUS-PDOX tumors in athymic mice. CD31 immunohistochemistry shows that EUS-PDOX tumors harbor ECs in the collagen-rich stroma highlighted by Masson's Trichrome staining of serial sections. (C) Human-specific CD34 (top), VE-Cadherin (middle) and CD105 (bottom) antibodies react with ECs in EUS-PDOX tumors, but fail to react with ECs in normal murine pancreata. Quantitation (right) of CD31 and Masson's Trichrome (B), or CD34 (upper), VE-Cadherin (middle) and CD105 (lower panel) (C) pixel intensity shows that stroma content and the abundance of ECs in PDOX2 are decreased compared with PDOX1 and PDOX3. Shown in (B–C) are representative images from three EUS-PDOX tumors. Insets show magnified images of boxed areas. Scale bars, 50 μm.
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Figure 1: A Subset of Human PDACs exhibit a strong angiogenic gene signature(A) Hierarchical clustering of TCGA data show that genes annotated to angiogenesis (white lines) are up-regulated in some human PDACs (left). Extraction (middle) and re-clustering (right) shows that many of these angiogenesis genes are up-regulated in a subset of PDACs (S = strong), whereas some (M = moderate) or few of these genes (W = weak) are increased in other PDACs. (B) H&E staining shows cytology of EUS-FNA samples (left) and histology of EUS-PDOX tumors in athymic mice. CD31 immunohistochemistry shows that EUS-PDOX tumors harbor ECs in the collagen-rich stroma highlighted by Masson's Trichrome staining of serial sections. (C) Human-specific CD34 (top), VE-Cadherin (middle) and CD105 (bottom) antibodies react with ECs in EUS-PDOX tumors, but fail to react with ECs in normal murine pancreata. Quantitation (right) of CD31 and Masson's Trichrome (B), or CD34 (upper), VE-Cadherin (middle) and CD105 (lower panel) (C) pixel intensity shows that stroma content and the abundance of ECs in PDOX2 are decreased compared with PDOX1 and PDOX3. Shown in (B–C) are representative images from three EUS-PDOX tumors. Insets show magnified images of boxed areas. Scale bars, 50 μm.

Mentions: To assess the angiogenic potential of human PDACs we analyzed an RNASeq dataset from 85 PDACs in The Cancer Genome Atlas (TCGA), focusing on genes annotated to angiogenesis Gene Ontology (GO) terms. Hierarchical clustering revealed that out of 384 genes annotated to angiogenesis, approximately 128 were up-regulated in some tumors (Figure 1A). We therefore extracted this gene set, and performed a second cluster analysis to determine whether any patients exhibited similar angiogenesis gene expression profiles (Figure 1A). Based on this analysis, 10/85 (~12%) PDAC patients were identified that harbored tumors in which multiple angiogenesis genes were up-regulated, suggesting that they exhibit a strong angiogenesis gene signature (Figure 1A). By contrast, 59/85 PDACs (~69%) exhibited variable expression levels of these genes, suggesting that they have a moderate angiogenesis signature, whereas 16/85 PDACs (~19%) displayed a weak signature in which few genes were elevated (Figure 1A). Differential expression analysis of tumors exhibiting a strong signature with those exhibiting a weak signature revealed that 77 angiogenesis genes were significantly up-regulated, 63 of which were pro-angiogenic (Supplementary Table 1).


TCGA data and patient-derived orthotopic xenografts highlight pancreatic cancer-associated angiogenesis.

Gore J, Craven KE, Wilson JL, Cote GA, Cheng M, Nguyen HV, Cramer HM, Sherman S, Korc M - Oncotarget (2015)

A Subset of Human PDACs exhibit a strong angiogenic gene signature(A) Hierarchical clustering of TCGA data show that genes annotated to angiogenesis (white lines) are up-regulated in some human PDACs (left). Extraction (middle) and re-clustering (right) shows that many of these angiogenesis genes are up-regulated in a subset of PDACs (S = strong), whereas some (M = moderate) or few of these genes (W = weak) are increased in other PDACs. (B) H&E staining shows cytology of EUS-FNA samples (left) and histology of EUS-PDOX tumors in athymic mice. CD31 immunohistochemistry shows that EUS-PDOX tumors harbor ECs in the collagen-rich stroma highlighted by Masson's Trichrome staining of serial sections. (C) Human-specific CD34 (top), VE-Cadherin (middle) and CD105 (bottom) antibodies react with ECs in EUS-PDOX tumors, but fail to react with ECs in normal murine pancreata. Quantitation (right) of CD31 and Masson's Trichrome (B), or CD34 (upper), VE-Cadherin (middle) and CD105 (lower panel) (C) pixel intensity shows that stroma content and the abundance of ECs in PDOX2 are decreased compared with PDOX1 and PDOX3. Shown in (B–C) are representative images from three EUS-PDOX tumors. Insets show magnified images of boxed areas. Scale bars, 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4480696&req=5

Figure 1: A Subset of Human PDACs exhibit a strong angiogenic gene signature(A) Hierarchical clustering of TCGA data show that genes annotated to angiogenesis (white lines) are up-regulated in some human PDACs (left). Extraction (middle) and re-clustering (right) shows that many of these angiogenesis genes are up-regulated in a subset of PDACs (S = strong), whereas some (M = moderate) or few of these genes (W = weak) are increased in other PDACs. (B) H&E staining shows cytology of EUS-FNA samples (left) and histology of EUS-PDOX tumors in athymic mice. CD31 immunohistochemistry shows that EUS-PDOX tumors harbor ECs in the collagen-rich stroma highlighted by Masson's Trichrome staining of serial sections. (C) Human-specific CD34 (top), VE-Cadherin (middle) and CD105 (bottom) antibodies react with ECs in EUS-PDOX tumors, but fail to react with ECs in normal murine pancreata. Quantitation (right) of CD31 and Masson's Trichrome (B), or CD34 (upper), VE-Cadherin (middle) and CD105 (lower panel) (C) pixel intensity shows that stroma content and the abundance of ECs in PDOX2 are decreased compared with PDOX1 and PDOX3. Shown in (B–C) are representative images from three EUS-PDOX tumors. Insets show magnified images of boxed areas. Scale bars, 50 μm.
Mentions: To assess the angiogenic potential of human PDACs we analyzed an RNASeq dataset from 85 PDACs in The Cancer Genome Atlas (TCGA), focusing on genes annotated to angiogenesis Gene Ontology (GO) terms. Hierarchical clustering revealed that out of 384 genes annotated to angiogenesis, approximately 128 were up-regulated in some tumors (Figure 1A). We therefore extracted this gene set, and performed a second cluster analysis to determine whether any patients exhibited similar angiogenesis gene expression profiles (Figure 1A). Based on this analysis, 10/85 (~12%) PDAC patients were identified that harbored tumors in which multiple angiogenesis genes were up-regulated, suggesting that they exhibit a strong angiogenesis gene signature (Figure 1A). By contrast, 59/85 PDACs (~69%) exhibited variable expression levels of these genes, suggesting that they have a moderate angiogenesis signature, whereas 16/85 PDACs (~19%) displayed a weak signature in which few genes were elevated (Figure 1A). Differential expression analysis of tumors exhibiting a strong signature with those exhibiting a weak signature revealed that 77 angiogenesis genes were significantly up-regulated, 63 of which were pro-angiogenic (Supplementary Table 1).

Bottom Line: Inhibition of the type I TGF-β receptor with SB505124 does not alter endothelial activation in vitro, but decreases pro-angiogenic gene expression and suppresses angiogenesis in vivo.Conversely, STAT3 silencing or JAK1-2 inhibition with ruxolitinib blocks CM-enhanced EC proliferation.Thus, targeting JAK1-2 with ruxolitinib blocks a final pathway that is common to multiple pro-angiogenic factors, suppresses EC-mediated PCC proliferation, and may be useful in PDACs with a strong pro-angiogenic signature.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Indiana University School of Medicine, Indianapolis, IN 46202, USA.

ABSTRACT
Pancreatic ductal adenocarcinomas (PDACs) overexpress pro-angiogenic factors but are not viewed as vascular. Using data from The Cancer Genome Atlas we demonstrate that a subset of PDACs exhibits a strong pro-angiogenic signature that includes 37 genes, such as HDAC9, that are overexpressed in PDAC arising in KRC mice, which express mutated Kras and lack RB. Moreover, patient-derived orthotopic xenografts can exhibit tumor angiogenesis, whereas conditioned media (CM) from KRC-derived pancreatic cancer cells (PCCs) enhance endothelial cell (EC) growth and migration, and activate canonical TGF-β signaling and STAT3. Inhibition of the type I TGF-β receptor with SB505124 does not alter endothelial activation in vitro, but decreases pro-angiogenic gene expression and suppresses angiogenesis in vivo. Conversely, STAT3 silencing or JAK1-2 inhibition with ruxolitinib blocks CM-enhanced EC proliferation. STAT3 disruption also suppresses endothelial HDAC9 and blocks CM-induced HDAC9 expression, whereas HDAC9 re-expression restores CM-enhanced endothelial proliferation. Moreover, ruxolitinib blocks mitogenic EC/PCC cross-talk, and suppresses endothelial p-STAT3 and HDAC9, and PDAC progression and angiogenesis in vivo, while markedly prolonging survival of KRC mice. Thus, targeting JAK1-2 with ruxolitinib blocks a final pathway that is common to multiple pro-angiogenic factors, suppresses EC-mediated PCC proliferation, and may be useful in PDACs with a strong pro-angiogenic signature.

No MeSH data available.


Related in: MedlinePlus