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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

Ruxolitinib suppresses mitogenic cross-talk between endothelial cells and PCCs(A) 3D co-cultures of ECs (red) and KRC PCCs (green) shows that compared with vehicle (DMSO, left), ruxolitinib ([100 nM], right) suppresses PCC growth. Shown are representative phase contrast and fluorescent images taken on day 8. Scale bars, 200 μm. (B) Fluorescence intensity quantitation shows that compared with 3D cultures in which ECs and PCCs are cultured independently (single culture), culturing ECs and PCCs together in 3D (co-culture) significantly enhances PCC growth, which is blocked by ruxolitinib (open bars). Data are mean ± SEM from three independent experiments. *P < 0.05, and **P < 0.01. (C) Schematic representation of PCC and EC cross-talk. TGF-β activates canonical Smad-dependent signaling in PCCs (top) leading to enhanced production of pro-angiogenic factors, which can be blocked by SB505124. These factors activate JAK/STAT3 signaling in ECs (bottom), which promotes EC proliferation through HDAC9, and ruxolitinib blocks these effects. ECs also produce factors (angiocrine factors) that can exert growth-stimulatory effects on PCCs through JAK/STAT3 signaling, which can also be targeted with ruxolitinib.
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Figure 6: Ruxolitinib suppresses mitogenic cross-talk between endothelial cells and PCCs(A) 3D co-cultures of ECs (red) and KRC PCCs (green) shows that compared with vehicle (DMSO, left), ruxolitinib ([100 nM], right) suppresses PCC growth. Shown are representative phase contrast and fluorescent images taken on day 8. Scale bars, 200 μm. (B) Fluorescence intensity quantitation shows that compared with 3D cultures in which ECs and PCCs are cultured independently (single culture), culturing ECs and PCCs together in 3D (co-culture) significantly enhances PCC growth, which is blocked by ruxolitinib (open bars). Data are mean ± SEM from three independent experiments. *P < 0.05, and **P < 0.01. (C) Schematic representation of PCC and EC cross-talk. TGF-β activates canonical Smad-dependent signaling in PCCs (top) leading to enhanced production of pro-angiogenic factors, which can be blocked by SB505124. These factors activate JAK/STAT3 signaling in ECs (bottom), which promotes EC proliferation through HDAC9, and ruxolitinib blocks these effects. ECs also produce factors (angiocrine factors) that can exert growth-stimulatory effects on PCCs through JAK/STAT3 signaling, which can also be targeted with ruxolitinib.

Mentions: To determine if ruxolitinib exerts tumor suppressive effects in KRC mice by targeting the endothelium, the neoplastic epithelium, or both compartments, we next co-cultured fluorescently-labeled SVEC4–10 ECs and KRC PCCs in a 3-dimensional (3D) culture system [32]. Remarkably, PCC growth was enhanced in the co-culture model compared with 3D cultures in which PCCs were cultured separately from ECs, and this enhanced growth in co-culture was completely suppressed by ruxolitinib (Figure 6A). By contrast, ruxolitinib failed to inhibit the growth of either PCCs or ECs when cultured separately (Figure 6B). Thus, ECs can enhance PCC growth through an angiocrine mechanism, which is suppressible by targeting JAK1–2 with ruxolitinib (Figure 6C).


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)

Ruxolitinib suppresses mitogenic cross-talk between endothelial cells and PCCs(A) 3D co-cultures of ECs (red) and KRC PCCs (green) shows that compared with vehicle (DMSO, left), ruxolitinib ([100 nM], right) suppresses PCC growth. Shown are representative phase contrast and fluorescent images taken on day 8. Scale bars, 200 μm. (B) Fluorescence intensity quantitation shows that compared with 3D cultures in which ECs and PCCs are cultured independently (single culture), culturing ECs and PCCs together in 3D (co-culture) significantly enhances PCC growth, which is blocked by ruxolitinib (open bars). Data are mean ± SEM from three independent experiments. *P < 0.05, and **P < 0.01. (C) Schematic representation of PCC and EC cross-talk. TGF-β activates canonical Smad-dependent signaling in PCCs (top) leading to enhanced production of pro-angiogenic factors, which can be blocked by SB505124. These factors activate JAK/STAT3 signaling in ECs (bottom), which promotes EC proliferation through HDAC9, and ruxolitinib blocks these effects. ECs also produce factors (angiocrine factors) that can exert growth-stimulatory effects on PCCs through JAK/STAT3 signaling, which can also be targeted with ruxolitinib.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 6: Ruxolitinib suppresses mitogenic cross-talk between endothelial cells and PCCs(A) 3D co-cultures of ECs (red) and KRC PCCs (green) shows that compared with vehicle (DMSO, left), ruxolitinib ([100 nM], right) suppresses PCC growth. Shown are representative phase contrast and fluorescent images taken on day 8. Scale bars, 200 μm. (B) Fluorescence intensity quantitation shows that compared with 3D cultures in which ECs and PCCs are cultured independently (single culture), culturing ECs and PCCs together in 3D (co-culture) significantly enhances PCC growth, which is blocked by ruxolitinib (open bars). Data are mean ± SEM from three independent experiments. *P < 0.05, and **P < 0.01. (C) Schematic representation of PCC and EC cross-talk. TGF-β activates canonical Smad-dependent signaling in PCCs (top) leading to enhanced production of pro-angiogenic factors, which can be blocked by SB505124. These factors activate JAK/STAT3 signaling in ECs (bottom), which promotes EC proliferation through HDAC9, and ruxolitinib blocks these effects. ECs also produce factors (angiocrine factors) that can exert growth-stimulatory effects on PCCs through JAK/STAT3 signaling, which can also be targeted with ruxolitinib.
Mentions: To determine if ruxolitinib exerts tumor suppressive effects in KRC mice by targeting the endothelium, the neoplastic epithelium, or both compartments, we next co-cultured fluorescently-labeled SVEC4–10 ECs and KRC PCCs in a 3-dimensional (3D) culture system [32]. Remarkably, PCC growth was enhanced in the co-culture model compared with 3D cultures in which PCCs were cultured separately from ECs, and this enhanced growth in co-culture was completely suppressed by ruxolitinib (Figure 6A). By contrast, ruxolitinib failed to inhibit the growth of either PCCs or ECs when cultured separately (Figure 6B). Thus, ECs can enhance PCC growth through an angiocrine mechanism, which is suppressible by targeting JAK1–2 with ruxolitinib (Figure 6C).

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